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Sample records for phytophthora infestans sporangia

  1. Infection Efficiency of Four Phytophthora infestans Clonal Lineages and DNA-Based Quantification of Sporangia

    PubMed Central

    Fall, Mamadou Lamine; Tremblay, David Mathieu; Gobeil-Richard, Mélanie; Couillard, Julie; Rocheleau, Hélène; Van der Heyden, Hervé; Lévesque, Camile André; Beaulieu, Carole; Carisse, Odile

    2015-01-01

    The presence and abundance of pathogen inoculum is with host resistance and environmental conditions a key factor in epidemic development. Therefore, several spore-sampling devices have been proposed to monitor pathogen inoculum above fields. However, to make spore sampling more reliable as a management tool and to facilitate its adoption, information on infection efficiency and molecular tools for estimating airborne sporangia concentration are needed. Experiments were thus undertaken in a growth chamber to study the infection efficiency of four clonal lineages of P. infestans (US-8, US-11, US-23, and US-24) by measuring the airborne sporangia concentration and resulting disease intensity. The relationship between the airborne sporangia concentration and the number of lesions per leaf was exponential. For the same concentration, the sporangia of US-23 caused significantly more lesions than the sporangia of the other clonal lineages did. Under optimal conditions, an airborne sporangia concentration of 10 sporangia m−3 for US-23 was sufficient to cause one lesion per leaf, whereas for the other clonal lineages, it took 15 to 25 sporangia m−3 to reach the same disease intensity. However, in terms of diseased leaf area, there was no difference between clonal lineages US-8, US-23 and US-24. Also, a sensitive quantitative real-time polymerase chain reaction (qPCR) tool was developed to quantify P. infestans airborne sporangia with detection sensitivity of one sporangium. The specificity of the qPCR assay was rigorously tested for airborne inoculum and was either similar to, or an improvement on, other published PCR assays. This assay allows rapid and reliable detection and quantification of P. infestans airborne sporangia and thereby, facilitates the implementation of spores-sampling network. PMID:26301826

  2. GK4, a G-protein-coupled receptor with a phosphatidylinositol phosphate kinase domain in Phytophthora infestans, is involved in sporangia development and virulence.

    PubMed

    Hua, Chenlei; Meijer, Harold J G; de Keijzer, Jeroen; Zhao, Wei; Wang, Yuanchao; Govers, Francine

    2013-04-01

    For dispersal and host infection plant pathogens largely depend on asexual spores. Pathogenesis and sporulation are complex processes that are governed by cellular signalling networks including G-protein and phospholipid signalling. Oomycetes possess a family of novel proteins called GPCR-PIPKs (GKs) that are composed of a seven-transmembrane spanning (7-TM) domain fused to a phosphatidylinositol phosphate kinase (PIPK) domain. Based on this domain structure GKs are anticipated to link G-protein and phospholipid signal pathways; however, their functions are currently unknown. Expression analyses of the 12 GK genes in Phytophthora infestans and their orthologues in Phytophthora sojae, revealed differential expression during asexual development. PiGK1 and PiGK4 were fused to monomeric red fluorescent protein (mRFP) and ectopically expressed in P. infestans. In growing hyphae different subcellular distribution patterns were observed indicating that these two GKs act independently during development. We focused on the functional analyses of PiGK4. Its localization suggested involvement in cell differentiation and elongation and its 7-TM domain showed a canonical GPCR membrane topology. Silencing of GK4 and overexpression of full-length and truncated constructs in P. infestans revealed that PiGK4 is not only involved in spore germination and hyphal elongation but also in sporangia cleavage and infection.

  3. Comparing necrosis of Rhododendron leaf tissue inoculated with Phytophthora ramorum sporangia or zoospores

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora species produce sporangia that either germinate directly or release zoospores depending upon environmental conditions. Phytophthora ramorum is no exception producing abundant sporangia that are disseminated by wind and rain. Past research involving inoculation trials or screening host...

  4. Characterization of Phytophthora infestans resistance to mefenoxam using FTIR spectroscopy.

    PubMed

    Pomerantz, A; Cohen, Y; Shufan, E; Ben-Naim, Y; Mordechai, S; Salman, A; Huleihel, M

    2014-12-01

    Phytophthora infestans (P. infestans) is the causal agent of late blight in potato and tomato. This pathogen devastated the potato crops in Ireland more than a century years ago and is still causing great losses worldwide. Although fungicides controlling P. infestans have been used successfully for almost 100 years, some isolates have developed resistance to most common fungicides. Identification and characterization of these resistant isolates is required for better control of the disease. Current methods that are based on microbiological and molecular techniques are both expensive and time consuming. Fourier Transform Infra-Red spectroscopy (FTIR) is an inexpensive and reagent-free technique that provides accurate results in only a few minutes. In this study the infrared absorption spectra of the sporangia of P. infestans were measured to evaluate the potential of FTIR spectroscopy in tandem with multivariate analysis in order to classify those sporangia into those that were resistant and those that were non-resistant to the phenylamide fungicide mefenoxam. Based on individual measurements, our results show that FTIR spectroscopy enables classification of P. infestans isolates into mefenoxam resistant and mefenoxam non-resistant types with specificity of 81.9% and sensitivity of 75.5%. Using average spectra per leaf, it was possible to improve the classification results to 88% sensitivity and 95% specificity.

  5. MiR1918 enhances tomato sensitivity to Phytophthora infestans infection

    PubMed Central

    Luan, Yushi; Cui, Jun; Wang, Weichen; Meng, Jun

    2016-01-01

    Late blight of tomato is caused by the oomycete pathogen Phytophthora infestans. In our previous work, we identified and characterized a miR1918 in P. infestans (pi-miR1918), and showed that its sequence is similar to the sequence of tomato miR1918 (sly-miR1918). In this study, we used Arabidopsis thaliana pre-miR159a as a backbone to synthesize pi-miR1918 via PCR and mutagenesis. The artificial pi-miR1918 was used to investigate the role of miR1918 in tomato-P. infestans interaction. Trangenic tomato plants that overexpressed the artificial pi-miR1918 displayed more serious disease symptoms than wild-type tomato plants after infection with P. infestans, as shown by increased number of necrotic cells, lesion sizes and number of sporangia per leaf. The target genes of pi-miR1918 and sly-miR1918 were also predicted for tomato and P. infestans, respectively. qPCR analysis of these targets also performed during tomato-P. infestans interaction. The expression of target gene, RING finger were negatively correlated with miR1918 in the all Lines of transgenic tomato plants. In addition, we used the 5′ RACE to determine the cleavage site of miR1918 to RING finger. These results suggested that miR1918 might be involved in the silencing of target genes, thereby enhancing the susceptibility of tomato to P. infestans infection. PMID:27779242

  6. Actin in the oomycetous fungus Phytophthora infestans is the product of several genes.

    PubMed

    Unkles, S E; Moon, R P; Hawkins, A R; Duncan, J M; Kinghorn, J R

    1991-04-01

    Actin (ACT) in Phytophthora infestans is encoded by at least two genes, in contrast to unicellular and other filamentous fungi where there is a single gene. These genes (designated actA and actB) have been isolated from a genomic library of P. infestans. The complete nucleotide sequence of both genes has been determined. Unlike the actin-encoding genes (act) of other filamentous fungi, no introns are obvious in the coding region, a feature shared with the act genes of certain protists. Northern blotting and primer extension studies of the mRNA show that actA and actB are actively transcribed in mycelium, sporangia and germinating cysts but only at a low level in the case of actB. Both genes display bias in their codon usage. This is more extreme in actA. The deduced ACTB protein is strikingly similar to that of the Phytophthora megasperma actin and is more diverged from other actins than ACTA.

  7. A novel taurocyamine kinase found in the protist Phytophthora infestans.

    PubMed

    Uda, Kouji; Hoshijima, Michihiro; Suzuki, Tomohiko

    2013-05-01

    Phosphagen kinase (PK), which is typically in the form of creatine kinase (CK; EC 2.7.3.2) in vertebrates or arginine kinase (AK; EC 2.7.3.3) in invertebrates, plays a key role in ATP buffering systems of tissues and nerves that display high and variable rates of ATP turnover. The enzyme is also found with intermittent occurrence as AK in unicellular organisms, protist and bacteria species, suggesting an ancient origin of AK. Through a database search, we identified two novel PK genes, coding 40- and 80-kDa (contiguous dimer) enzymes in the protist Phytophthora infestans. Both enzymes showed strong activity for taurocyamine and, in addition, we detected taurocyamine in cell extracts of P. infestans. Thus, the enzyme was identified to be taurocyamine kinase (TK; EC 2.7.3.4). This was the first phosphagen kinase, other than AK, to be found in unicellular organisms. Their position on the phylogenetic tree indicates that P. infestans TKs evolved uniquely at an early stage of evolution. Occurrence of TK in protists suggests that PK enzymes show flexible substrate specificity.

  8. Characterization of a Phytophthora infestans gene involved in vesicle transport.

    PubMed

    Chen, Y; Roxby, R

    1996-11-28

    Members of the Ras superfamily of monomeric GTP-binding proteins have been shown to be essential in specific steps of vesicle transport and secretion in widely divergent organisms. We report here the characterization of a gene from Phytophthora infestans encoding a deduced amino acid (aa) sequence belonging to the Ypt class of monomeric GTP-binding proteins, products shown in other organisms to be essential for vesicle transport between the endoplasmic reticulum and the cis-Golgi compartments. Analysis of genomic and cDNA sequences of this gene, Piypt1, indicates that it contains five introns, one in the 5'-untranslated region. All introns are typical in beginning with GT and ending with AG. The region of the transcription start point displays a number of features characteristic of fungi and other eukaryotes, but it does not contain TATA or CAAT motifs. A single transcript is produced from the gene, which is polyadenylated, but the gene does not contain a recognizable polyadenylation signal. Genomic DNA blots indicate that Piypt1 is a single-copy gene. Comparisons of Ypt1 aa sequences indicate that P. infestans is more closely related to algae and higher plants than to the true fungi. The protein product of the Piypt1 gene, expressed in Escherichia coli, cross-reacts with antiserum against yeast Ypt1 protein and binds GTP. Furthermore, the Piypt1 gene is able to functionally complement a mutant ypt1 gene in Saccharomyces cerevisiae. The aa sequence similarity, immunological cross-reactivity and functional attributes of Piypt1 make it likely that it is an authentic ypt1 gene which participates in vesicle transport in Phytophthora infestans.

  9. Efficacy of Fungicide Mixtures for Management of Phytophthora infestans (US-1) on Potato

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fungicide application strategies (timing, frequency, rates and mixtures) are important for control and resistance management of late blight, caused by Phytophthora infestans. The efficacy of systemic/protectant fungicide consisting of fenamidone + mancozeb, and propamocarb–HCL + mancozeb combination...

  10. Discovering the next generation of late blight resistance genes – can we battle Phytophthora infestans evolution

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Potato late blight, caused by the oomycete pathogen Phytophthora infestans, is one of the most destructive plant diseases. RB from Solanum bulbocastanum encodes a CC-NB-LRR (CNL) protein that confers partial resistance to most P. infestans isolates through its recognition of the corresponding pathog...

  11. The genome sequence of the Irish potato famine pathogen Phytophthora infestans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora infestans is the most destructive pathogen of potato and a model organism for the oomycetes, a distinct lineage of fungus-like eukaryotes that are related to photosynthetic organisms such as brown algae and diatoms. Here, we report the genome sequence of P. infestans. The ~240 Mb genome...

  12. An ephemeral sexual population of Phytophthora infestans in the northeastern United States and Canada

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora infestans, the causal agent of late blight disease, has been reported in North America since the mid-nineteenth century. In the United States the lack of or very limited sexual reproduction has resulted in largely clonal populations of P. infestans. In 2010 and 2011, but not in 2012 or ...

  13. Activation of defense against Phytophthora infestans in potato by down-regulation of syntaxin gene expression.

    PubMed

    Eschen-Lippold, Lennart; Landgraf, Ramona; Smolka, Ulrike; Schulze, Sebastian; Heilmann, Mareike; Heilmann, Ingo; Hause, Gerd; Rosahl, Sabine

    2012-03-01

    The oomycete Phytophthora infestans is the causal agent of late blight, the most devastating disease of potato. The importance of vesicle fusion processes and callose deposition for defense of potato against Phytophthora infestans was analyzed. Transgenic plants were generated, which express RNA interference constructs targeted against plasma membrane-localized SYNTAXIN-RELATED 1 (StSYR1) and SOLUBLE N-ETHYLMALEIMIDE-SENSITIVE FACTOR ADAPTOR PROTEIN 33 (StSNAP33), the potato homologs of Arabidopsis AtSYP121 and AtSNAP33, respectively. Phenotypically, transgenic plants grew normally, but showed spontaneous necrosis and chlorosis formation at later stages. In response to infection with Phytophthora infestans, increased resistance of StSYR1-RNAi plants, but not StSNAP33-RNAi plants, was observed. This increased resistance correlated with the constitutive accumulation of salicylic acid and PR1 transcripts. Aberrant callose deposition in Phytophthora infestans-infected StSYR1-RNAi plants coincided with decreased papilla formation at penetration sites. Resistance against the necrotrophic fungus Botrytis cinerea was not significantly altered. Infiltration experiments with bacterial solutions of Agrobacterium tumefaciens and Escherichia coli revealed a hypersensitive phenotype of both types of RNAi lines. The enhanced defense status and the reduced growth of Phytophthora infestans on StSYR1-RNAi plants suggest an involvement of syntaxins in secretory defense responses of potato and, in particular, in the formation of callose-containing papillae. PMID:22243492

  14. The effect of relative humidity on germination of Sporangia of Phytophthora ramorum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sporangia of three isolates of P. ramorum representing three different clonal lineages were subjected to relative humidity (RH) levels between 80 and 100% for exposure periods ranging from 1 to 24 h at 20°C in darkness. Airtight snap-lid plastic containers (21.5 x 14.5 x 5 cm) were used as humidity ...

  15. Phytophthora infestans Has a Plethora of Phospholipase D Enzymes Including a Subclass That Has Extracellular Activity

    PubMed Central

    Meijer, Harold J. G.; Hassen, Hussen Harrun; Govers, Francine

    2011-01-01

    In eukaryotes phospholipase D (PLD) is involved in many cellular processes. Currently little is known about PLDs in oomycetes. Here we report that the oomycete plant pathogen Phytophthora infestans has a large repertoire of PLDs divided over six subfamilies: PXPH-PLD, PXTM-PLD, TM-PLD, PLD-likes, and type A and B sPLD-likes. Since the latter have signal peptides we developed a method using metabolically labelled phospholipids to monitor if P. infestans secretes PLD. In extracellular medium of ten P. infestans strains PLD activity was detected as demonstrated by the production of phosphatidic acid and the PLD specific marker phosphatidylalcohol. PMID:21423760

  16. Phytophthora infestans has a plethora of phospholipase D enzymes including a subclass that has extracellular activity.

    PubMed

    Meijer, Harold J G; Hassen, Hussen Harrun; Govers, Francine

    2011-01-01

    In eukaryotes phospholipase D (PLD) is involved in many cellular processes. Currently little is known about PLDs in oomycetes. Here we report that the oomycete plant pathogen Phytophthora infestans has a large repertoire of PLDs divided over six subfamilies: PXPH-PLD, PXTM-PLD, TM-PLD, PLD-likes, and type A and B sPLD-likes. Since the latter have signal peptides we developed a method using metabolically labelled phospholipids to monitor if P. infestans secretes PLD. In extracellular medium of ten P. infestans strains PLD activity was detected as demonstrated by the production of phosphatidic acid and the PLD specific marker phosphatidylalcohol. PMID:21423760

  17. Competitive Fitness of Phytophthora infestans Isolates Under Semiarid Field Conditions.

    PubMed

    Miller, J S; Johnson, D A

    2000-03-01

    ABSTRACT Spread of US-1 and US-8 isolates of Phytophthora infestans were observed in field plots of potato (cv. Russet Burbank) grown in Pullman, WA, in 1996 and 1997. Infected greenhouse-grown potato plants with similar lesion numbers for both strains were transplanted to field plots with four replications. Spread of the pathogen was favored by sprinkler irrigation during evening hours. Diseased leaves and stems were sampled over time to determine the spread of US-1 and US-8 isolates. In 1996, late blight developed in two of the four replications (105 and 87 total isolates recovered). From those two replications, two US-1 isolates were recovered, both from the same replication. Nine isolates from one replication and six isolates from another displayed a phenotype different from the initial isolates, as determined by compatibility type, allozyme genotype, and restriction fragment length polymorphism genotype. These putative recombinant isolates may have arisen from sexual recombination between the US-1 and US-8 isolates. The remaining isolates were of the US-8 strain. In 1997, late blight developed in all four replications (123, 122, 81, and 34 total isolates recovered). One US-1 isolate was recovered (out of 123) from one replication and three (out of 122) from another, and the remaining isolates were of the US-8 strain. Isolates with phenotypes differing from the initial isolates were not recovered in 1997. In both years, oospores were not observed in the plant tissue examined. The low number of putative recombinant isolates in 1996 and their absence in 1997 suggests that sexual reproduction between US-8 and US-1 isolates in a field setting is a rare event. The predominance of US-8 isolates recovered is a measure of the increased fitness and aggressiveness of the US-8 isolates relative to the US-1 isolate used in this study. This further substantiates the increased aggressiveness of the US-8 genotype observed on excised tissues and potted plants in previous

  18. The rise and fall of the Phytophthora infestans lineage that triggered the Irish potato famine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Potato late blight, caused by the oomycete Phytophthora infestans, is a plant pathogen of historical dimension that remains one of the world’s most destructive crop diseases. Ever since its first global outbreak of the 1840s that culminated in the Great Famine in Ireland, late blight has been a majo...

  19. The Irish potato famine pathogen Phytophthora infestans originated in central Mexico rather than the Andes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora infestans is a destructive plant pathogen best known for causing the disease that triggered the Irish potato famine and continues to be the most costly potato pathogen to manage worldwide. Identification of its elusive center of origin is critical to understanding the mechanisms of repe...

  20. Genetic diversity of Phytophthora infestans in the Northern-Andean region

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora infestans (Mont.) de Bary, the causal agent of potato late blight, is responsible for tremendous crop and economic losses worldwide. Countries in the northern part of the Andes dedicate a large proportion of the highlands to the production of potato and, more recently, solanaceous fruit...

  1. Genomic Characterization of a South American Phytophthora Hybrid Mandates Reassessment of the Geographic Origins of Phytophthora infestans.

    PubMed

    Martin, Michael D; Vieira, Filipe G; Ho, Simon Y W; Wales, Nathan; Schubert, Mikkel; Seguin-Orlando, Andaine; Ristaino, Jean B; Gilbert, M Thomas P

    2016-02-01

    As the oomycete pathogen causing potato late blight disease, Phytophthora infestans triggered the famous 19th-century Irish potato famine and remains the leading cause of global commercial potato crop destruction. But the geographic origin of the genotype that caused this devastating initial outbreak remains disputed, as does the New World center of origin of the species itself. Both Mexico and South America have been proposed, generating considerable controversy. Here, we readdress the pathogen's origins using a genomic data set encompassing 71 globally sourced modern and historical samples of P. infestans and the hybrid species P. andina, a close relative known only from the Andean highlands. Previous studies have suggested that the nuclear DNA lineage behind the initial outbreaks in Europe in 1845 is now extinct. Analysis of P. andina's phased haplotypes recovered eight haploid genome sequences, four of which represent a previously unknown basal lineage of P. infestans closely related to the famine-era lineage. Our analyses further reveal that clonal lineages of both P. andina and historical P. infestans diverged earlier than modern Mexican lineages, casting doubt on recent claims of a Mexican center of origin. Finally, we use haplotype phasing to demonstrate that basal branches of the clade comprising Mexican samples are occupied by clonal isolates collected from wild Solanum hosts, suggesting that modern Mexican P. infestans diversified on Solanum tuberosum after a host jump from a wild species and that the origins of P. infestans are more complex than was previously thought.

  2. Genomic Characterization of a South American Phytophthora Hybrid Mandates Reassessment of the Geographic Origins of Phytophthora infestans.

    PubMed

    Martin, Michael D; Vieira, Filipe G; Ho, Simon Y W; Wales, Nathan; Schubert, Mikkel; Seguin-Orlando, Andaine; Ristaino, Jean B; Gilbert, M Thomas P

    2016-02-01

    As the oomycete pathogen causing potato late blight disease, Phytophthora infestans triggered the famous 19th-century Irish potato famine and remains the leading cause of global commercial potato crop destruction. But the geographic origin of the genotype that caused this devastating initial outbreak remains disputed, as does the New World center of origin of the species itself. Both Mexico and South America have been proposed, generating considerable controversy. Here, we readdress the pathogen's origins using a genomic data set encompassing 71 globally sourced modern and historical samples of P. infestans and the hybrid species P. andina, a close relative known only from the Andean highlands. Previous studies have suggested that the nuclear DNA lineage behind the initial outbreaks in Europe in 1845 is now extinct. Analysis of P. andina's phased haplotypes recovered eight haploid genome sequences, four of which represent a previously unknown basal lineage of P. infestans closely related to the famine-era lineage. Our analyses further reveal that clonal lineages of both P. andina and historical P. infestans diverged earlier than modern Mexican lineages, casting doubt on recent claims of a Mexican center of origin. Finally, we use haplotype phasing to demonstrate that basal branches of the clade comprising Mexican samples are occupied by clonal isolates collected from wild Solanum hosts, suggesting that modern Mexican P. infestans diversified on Solanum tuberosum after a host jump from a wild species and that the origins of P. infestans are more complex than was previously thought. PMID:26576850

  3. Genomic Characterization of a South American Phytophthora Hybrid Mandates Reassessment of the Geographic Origins of Phytophthora infestans

    PubMed Central

    Martin, Michael D.; Vieira, Filipe G.; Ho, Simon Y.W.; Wales, Nathan; Schubert, Mikkel; Seguin-Orlando, Andaine; Ristaino, Jean B.; Gilbert, M. Thomas P.

    2016-01-01

    As the oomycete pathogen causing potato late blight disease, Phytophthora infestans triggered the famous 19th-century Irish potato famine and remains the leading cause of global commercial potato crop destruction. But the geographic origin of the genotype that caused this devastating initial outbreak remains disputed, as does the New World center of origin of the species itself. Both Mexico and South America have been proposed, generating considerable controversy. Here, we readdress the pathogen’s origins using a genomic data set encompassing 71 globally sourced modern and historical samples of P. infestans and the hybrid species P. andina, a close relative known only from the Andean highlands. Previous studies have suggested that the nuclear DNA lineage behind the initial outbreaks in Europe in 1845 is now extinct. Analysis of P. andina’s phased haplotypes recovered eight haploid genome sequences, four of which represent a previously unknown basal lineage of P. infestans closely related to the famine-era lineage. Our analyses further reveal that clonal lineages of both P. andina and historical P. infestans diverged earlier than modern Mexican lineages, casting doubt on recent claims of a Mexican center of origin. Finally, we use haplotype phasing to demonstrate that basal branches of the clade comprising Mexican samples are occupied by clonal isolates collected from wild Solanum hosts, suggesting that modern Mexican P. infestans diversified on Solanum tuberosum after a host jump from a wild species and that the origins of P. infestans are more complex than was previously thought. PMID:26576850

  4. SNP-based differentiation of Phytophthora infestans clonal lineages using locked nucleic acid probes and high resolution melt analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora infestans, the cause of the devastating late blight disease of potato and tomato, exhibits a clonal reproductive lifestyle in North America. Phenotypes such as fungicide sensitivity and host preference are conserved among individuals within clonal lineages, while substantial phenotypic ...

  5. Application of the PCR technique to detect Phytophthora infestans in potato tubers and leaves.

    PubMed

    Niepold, F; Schöber-Butin, B

    1995-11-01

    A characterized repetitive sequence from Phytophthora infestans (P. infestans) was used to perform a PCR with the DNA from the four races 1, 3, 4, and 1-11. To obtain amplifiable DNA, all extractions had to be purified by DNA adsorbing spin columns. Only two out of four tested primers were well suited and gave an DNA amplificate of the same size for all four races. After optimization the detection limit of the PCR corresponded to 100 ng of freeze-dried mycelia per ml, specificity was established when testing a collection of the most important potato pathogenic fungi and bacteria. Using P. infestans zoospores to infect tuber slices, the detection threshold was determined to be two days post infection when 3 to 6 zoospores were applied. After infiltrating the tenfold concentration into potato leaves a visible PCR signal was obtained one day post infection. Further improvements of the sensitivity threshold in detecting P. infestans for breeding and prognosis purposes are discussed.

  6. Genome-wide Prediction and Functional Validation of Promoter Motifs Regulating Gene Expression in Spore and Infection Stages of Phytophthora infestans

    PubMed Central

    Roy, Sourav; Kagda, Meenakshi; Judelson, Howard S.

    2013-01-01

    Most eukaryotic pathogens have complex life cycles in which gene expression networks orchestrate the formation of cells specialized for dissemination or host colonization. In the oomycete Phytophthora infestans, the potato late blight pathogen, major shifts in mRNA profiles during developmental transitions were identified using microarrays. We used those data with search algorithms to discover about 100 motifs that are over-represented in promoters of genes up-regulated in hyphae, sporangia, sporangia undergoing zoosporogenesis, swimming zoospores, or germinated cysts forming appressoria (infection structures). Most of the putative stage-specific transcription factor binding sites (TFBSs) thus identified had features typical of TFBSs such as position or orientation bias, palindromy, and conservation in related species. Each of six motifs tested in P. infestans transformants using the GUS reporter gene conferred the expected stage-specific expression pattern, and several were shown to bind nuclear proteins in gel-shift assays. Motifs linked to the appressoria-forming stage, including a functionally validated TFBS, were over-represented in promoters of genes encoding effectors and other pathogenesis-related proteins. To understand how promoter and genome architecture influence expression, we also mapped transcription patterns to the P. infestans genome assembly. Adjacent genes were not typically induced in the same stage, including genes transcribed in opposite directions from small intergenic regions, but co-regulated gene pairs occurred more than expected by random chance. These data help illuminate the processes regulating development and pathogenesis, and will enable future attempts to purify the cognate transcription factors. PMID:23516354

  7. Presence of the potato late blight resistance gene RB does not promote adaptive parasitism of phytophthora infestans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The potato gene Rpi-blb1, from the wild potato species S. bulbocastanum, confers partial resistance to late blight, caused by the oomycete pathogen Phytophthora infestans. In order to determine whether a single strain of P. infestans can adapt to overcome this partial resistance source, we subjected...

  8. Presence of the potato late blight resistance gene RB does not promote adaptive parasitism of phytophthora infestans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The gene RB is derived from the wild potato species S. bulbocastanum and confers partial resistance to late blight, caused by the oomycete pathogen Phytophthora infestans. In order to investigate whether a single strain of P. infestans can adapt to overcome this partial resistance source, we subject...

  9. The Irish potato famine pathogen Phytophthora infestans originated in central Mexico rather than the Andes

    PubMed Central

    Goss, Erica M.; Tabima, Javier F.; Cooke, David E. L.; Restrepo, Silvia; Fry, William E.; Forbes, Gregory A.; Fieland, Valerie J.; Cardenas, Martha; Grünwald, Niklaus J.

    2014-01-01

    Phytophthora infestans is a destructive plant pathogen best known for causing the disease that triggered the Irish potato famine and remains the most costly potato pathogen to manage worldwide. Identification of P. infestan’s elusive center of origin is critical to understanding the mechanisms of repeated global emergence of this pathogen. There are two competing theories, placing the origin in either South America or in central Mexico, both of which are centers of diversity of Solanum host plants. To test these competing hypotheses, we conducted detailed phylogeographic and approximate Bayesian computation analyses, which are suitable approaches to unraveling complex demographic histories. Our analyses used microsatellite markers and sequences of four nuclear genes sampled from populations in the Andes, Mexico, and elsewhere. To infer the ancestral state, we included the closest known relatives Phytophthora phaseoli, Phytophthora mirabilis, and Phytophthora ipomoeae, as well as the interspecific hybrid Phytophthora andina. We did not find support for an Andean origin of P. infestans; rather, the sequence data suggest a Mexican origin. Our findings support the hypothesis that populations found in the Andes are descendants of the Mexican populations and reconcile previous findings of ancestral variation in the Andes. Although centers of origin are well documented as centers of evolution and diversity for numerous crop plants, the number of plant pathogens with a known geographic origin are limited. This work has important implications for our understanding of the coevolution of hosts and pathogens, as well as the harnessing of plant disease resistance to manage late blight. PMID:24889615

  10. Computational models in plant-pathogen interactions: the case of Phytophthora infestans

    PubMed Central

    2009-01-01

    Background Phytophthora infestans is a devastating oomycete pathogen of potato production worldwide. This review explores the use of computational models for studying the molecular interactions between P. infestans and one of its hosts, Solanum tuberosum. Modeling and conclusion Deterministic logistics models have been widely used to study pathogenicity mechanisms since the early 1950s, and have focused on processes at higher biological resolution levels. In recent years, owing to the availability of high throughput biological data and computational resources, interest in stochastic modeling of plant-pathogen interactions has grown. Stochastic models better reflect the behavior of biological systems. Most modern approaches to plant pathology modeling require molecular kinetics information. Unfortunately, this information is not available for many plant pathogens, including P. infestans. Boolean formalism has compensated for the lack of kinetics; this is especially the case where comparative genomics, protein-protein interactions and differential gene expression are the most common data resources. PMID:19909526

  11. Discovery of Phytophthora infestans Genes Expressed in Planta through Mining of cDNA Libraries

    PubMed Central

    Chaves, Diego; Pinzón, Andrés; Grajales, Alejandro; Rojas, Alejandro; Mutis, Gabriel; Cárdenas, Martha; Burbano, Daniel; Jiménez, Pedro; Bernal, Adriana; Restrepo, Silvia

    2010-01-01

    Background Phytophthora infestans (Mont.) de Bary causes late blight of potato and tomato, and has a broad host range within the Solanaceae family. Most studies of the Phytophthora – Solanum pathosystem have focused on gene expression in the host and have not analyzed pathogen gene expression in planta. Methodology/Principal Findings We describe in detail an in silico approach to mine ESTs from inoculated host plants deposited in a database in order to identify particular pathogen sequences associated with disease. We identified candidate effector genes through mining of 22,795 ESTs corresponding to P. infestans cDNA libraries in compatible and incompatible interactions with hosts from the Solanaceae family. Conclusions/Significance We annotated genes of P. infestans expressed in planta associated with late blight using different approaches and assigned putative functions to 373 out of the 501 sequences found in the P. infestans genome draft, including putative secreted proteins, domains associated with pathogenicity and poorly characterized proteins ideal for further experimental studies. Our study provides a methodology for analyzing cDNA libraries and provides an understanding of the plant – oomycete pathosystems that is independent of the host, condition, or type of sample by identifying genes of the pathogen expressed in planta. PMID:20352100

  12. Contrasting Potato Foliage and Tuber Defense Mechanisms against the Late Blight Pathogen Phytophthora infestans

    PubMed Central

    Bradeen, James M.

    2016-01-01

    The late blight pathogen Phytophthora infestans can attack both potato foliage and tubers. When inoculated with P. infestans, foliage of nontransformed ‘Russet Burbank’ (WT) develops late blight disease while that of transgenic ‘Russet Burbank’ line SP2211 (+RB) does not. We compared the foliar transcriptome responses of these two lines to P. infestans inoculation using an RNA-seq approach. A total of 515 million paired end RNA-seq reads were generated, representing the transcription of 29,970 genes. We also compared the differences and similarities of defense mechanisms against P. infestans in potato foliage and tubers. Differentially expressed genes, gene groups and ontology bins were identified to show similarities and differences in foliage and tuber defense mechanisms. Our results suggest that R gene dosage and shared biochemical pathways (such as ethylene and stress bins) contribute to RB-mediated incompatible potato-P. infestans interactions in both the foliage and tubers. Certain ontology bins such as cell wall and lipid metabolisms are potentially organ-specific. PMID:27441721

  13. Contrasting Potato Foliage and Tuber Defense Mechanisms against the Late Blight Pathogen Phytophthora infestans.

    PubMed

    Gao, Liangliang; Bradeen, James M

    2016-01-01

    The late blight pathogen Phytophthora infestans can attack both potato foliage and tubers. When inoculated with P. infestans, foliage of nontransformed 'Russet Burbank' (WT) develops late blight disease while that of transgenic 'Russet Burbank' line SP2211 (+RB) does not. We compared the foliar transcriptome responses of these two lines to P. infestans inoculation using an RNA-seq approach. A total of 515 million paired end RNA-seq reads were generated, representing the transcription of 29,970 genes. We also compared the differences and similarities of defense mechanisms against P. infestans in potato foliage and tubers. Differentially expressed genes, gene groups and ontology bins were identified to show similarities and differences in foliage and tuber defense mechanisms. Our results suggest that R gene dosage and shared biochemical pathways (such as ethylene and stress bins) contribute to RB-mediated incompatible potato-P. infestans interactions in both the foliage and tubers. Certain ontology bins such as cell wall and lipid metabolisms are potentially organ-specific. PMID:27441721

  14. Toxicity of metalaxyl, azoxystrobin, dimethomorph, cymoxanil, zoxamide and mancozeb to Phytophthora infestans isolates from Serbia.

    PubMed

    Rekanović, Emil; Potočnik, Ivana; Milijašević-Marčić, Svetlana; Stepanović, Miloš; Todorović, Biljana; Mihajlović, Milica

    2012-01-01

    A study of the in vitro sensitivity of 12 isolates of Phytophthora infestans to metalaxyl, azoxystrobin, dimethomorph, cymoxanil, zoxamide and mancozeb, was conducted. The isolates derived from infected potato leaves collected at eight different localities in Serbia during 2005-2007. The widest range of EC(50) values for mycelial growth of the isolates was recorded for metalaxyl. They varied from 0.3 to 3.9 μg mL(-1) and were higher than those expected in a susceptible population of P. infestans. The EC(50) values of the isolates were 0.16-0.30 μg mL(-1) for dimethomorph, 0.27-0.57 μg mL(-1) for cymoxanil, 0.0026-0.0049 μg mL(-1) for zoxamide and 2.9-5.0 μg mL(-1) for mancozeb. The results indicated that according to effective concentration (EC(50)) the 12 isolates of P. infestans were sensitive to azoxystrobin (0.019-0.074 μg mL(-1)), and intermediate resistant to metalaxyl, dimethomorph and cymoxanil. According to resistance factor, all P. infestans isolates were sensitive to dimethomorph, cymoxanil, mancozeb and zoxamide, 58.3% of isolates were sensitive to azoxystrobin and 50% to metalaxyl. Gout's scale indicated that 41.7% isolates were moderately sensitive to azoxystrobin and 50% to metalaxyl. PMID:22424065

  15. Plant-mediated gene silencing restricts growth of the potato late blight pathogen Phytophthora infestans

    PubMed Central

    Jahan, Sultana N.; Åsman, Anna K. M.; Corcoran, Pádraic; Fogelqvist, Johan; Vetukuri, Ramesh R.; Dixelius, Christina

    2015-01-01

    Phytophthora infestans is an oomycete that causes severe damage to potato, and is well known for its ability to evolve rapidly in order to overcome resistant potato varieties. An RNA silencing strategy was evaluated here to clarify if small interfering RNA homologous to selected genes in P. infestans could be targeted from the plant host to reduce the magnitude of the infection. As a proof-of-concept, a hairpin RNA (hp-RNA) construct using the GFP marker gene was designed and introduced in potato. At 72 hpi, a 55-fold reduction of the signal intensity of a corresponding GFP expressing P. infestans strain on leaf samples of transgenic plants, compared with wild-type potato, was detected. This suggests that an RNA interference construct in the potato host could be processed and target a transcript of the pathogen. Three genes important in the infection process of P. infestans, PiGPB1, PiCESA2, and PiPEC, together with PiGAPDH taking part in basic cell maintenance were subsequently tested using an analogous transgenic strategy. Out of these gene candidates, the hp-PiGPB1 targeting the G protein β-subunit (PiGPB1) important for pathogenicity resulted in most restricted disease progress. Further, Illumina sequencing of inoculated transgenic potato leaves revealed sRNAs of 24/25 nt size homologous to the PiGPB1 gene in the transgenic plants indicating post-transcriptional silencing of the target gene. The work demonstrates that a host-induced gene-silencing approach is functional against P. infestans but is highly dependent on target gene for a successful outcome. This finding broadens the arsenal of control strategies to this important plant disease. PMID:25788734

  16. Determination of virulence contributions from Phytophthora infestans effectors IPI-O1 and IPI-O4

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Potato late blight, caused by the oomycete pathogen Phytophthora infestans, is one of the most destructive plant diseases. Despite decades of intensive breeding efforts, it remains a threat to potato production worldwide, in part because newly evolved pathogen isolates quickly overcome major resista...

  17. Competitive Between Phytophthora Infestans Effectors Leads to Increased Aggressiveness on Plants Containing Broad-Spectrum Late Blight Resistance

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Potato late blight is a particularly destructive plant disease caused by the oomycete pathogen Phytophthora infestans (Mont.) de Bary. Late blight has remained particularly problematic despite intensive breeding efforts to integrate resistance into cultivated potato. This is due to the pathogen’s ab...

  18. Competitive Interaction Between Phytophthora Infestans Effectors Leads to Increased Aggressiveness on Plants Containing Broad-spectrum Late Blight Resistance

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The resistance (R) gene RB confers broad-spectrum resistance to potato late blight and belongs. The RB protein recognizes the presence of members of the Phytophthora infestans effector family IPI-O to elicit resistance. Most isolates of the pathogen contain IPI-O variants that are recognized by RB...

  19. Competitive Interaction Between Phytophthora Infestans Effectors Leads to Increased Aggressiveness on Plants Containing Broad-spectrum Late Blight Resistance

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The resistance (R) gene RB confers broad-spectrum resistance to potato late blight and belongs. The RB protein recognizes the presence of members of the Phytophthora infestans effector family IPI-O to elicit resistance. Most isolates of the pathogen contain IPI-O variants that are recognized by R...

  20. Effect of Flumorph on F-Actin Dynamics in the Potato Late Blight Pathogen Phytophthora infestans.

    PubMed

    Hua, Chenlei; Kots, Kiki; Ketelaar, Tijs; Govers, Francine; Meijer, Harold J G

    2015-04-01

    Oomycetes are fungal-like pathogens that cause notorious diseases. Protecting crops against oomycetes requires regular spraying with chemicals, many with an unknown mode of action. In the 1990s, flumorph was identified as a novel crop protection agent. It was shown to inhibit the growth of oomycete pathogens including Phytophthora spp., presumably by targeting actin. We recently generated transgenic Phytophthora infestans strains that express Lifeact-enhanced green fluorescent protein (eGFP), which enabled us to monitor the actin cytoskeleton during hyphal growth. For analyzing effects of oomicides on the actin cytoskeleton in vivo, the P. infestans Lifeact-eGFP strain is an excellent tool. Here, we confirm that flumorph is an oomicide with growth inhibitory activity. Microscopic analyses showed that low flumorph concentrations provoked hyphal tip swellings accompanied by accumulation of actin plaques in the apex, a feature reminiscent of tips of nongrowing hyphae. At higher concentrations, swelling was more pronounced and accompanied by an increase in hyphal bursting events. However, in hyphae that remained intact, actin filaments were indistinguishable from those in nontreated, nongrowing hyphae. In contrast, in hyphae treated with the actin depolymerizing drug latrunculin B, no hyphal bursting was observed but the actin filaments were completely disrupted. This difference demonstrates that actin is not the primary target of flumorph. PMID:25496300

  1. Methanol extract of mycelia from Phytophthora infestans-induced resistance in potato.

    PubMed

    Monjil, Mohammad Shahjahan; Nozawa, Takeshi; Shibata, Yusuke; Takemoto, Daigo; Ojika, Makoto; Kawakita, Kazuhito

    2015-03-01

    Plants recognize certain microbial compounds as elicitors in their active defence mechanisms. It has been shown that a series of defence reactions are induced in potato plant cells after treatment with water-soluble hyphal wall components prepared from Phytophthora infestans. In this study, a methanol extract from mycelia of P. infestans (MEM), which contains lipophilic compounds, was used as another elicitor for the induction of the defence reactions in potato. MEM elicitor induced reactive oxygen species (ROS), especially O2(-) and H2O2 production, and nitric oxide (NO) generation in potato leaves and suspension-cultured cells. Hypersensitive cell death was detected in potato leaves within 6-8 h after MEM elicitor treatment. The accumulation of phytoalexins was detected by MEM elicitor treatment in potato tubers. In potato suspension-cultured cells, several defence-related genes were induced by MEM elicitors, namely Strboh, Sthsr203J, StPVS3, StPR1, and StNR5, which regulate various defence-related functions. Enhanced resistance against P. infestans was found in MEM-treated potato plants. These results suggested that MEM elicitor is recognized by host and enhances defence activities to produce substances inhibitory to pathogens.

  2. An RNA symbiont enhances heat tolerance and secondary homothallism in the oomycete Phytophthora infestans.

    PubMed

    Judelson, Howard S; Ah-Fong, Audrey M V; Fabritius, Anna-Liisa

    2010-07-01

    Some strains of Phytophthora infestans, the potato late blight pathogen, harbour a small extrachromosomal RNA called PiERE1. A previous study reported that this RNA symbiont does not noticeably affect its host. Here it is revealed that PiERE1 exerts subtle effects on P. infestans, which result in greater thermotolerance during growth and an increase in secondary homothallism, i.e. oospore formation in the absence of the opposite mating type. The interaction can be considered mutualistic since these traits may increase the fitness of P. infestans in nature. Assays of biomarkers for cellular stress revealed that an Hsp70 chaperone was upregulated by PiERE1. A genome-wide search for more members of the Hsp70 family identified ten belonging to the DnaK subfamily, one in the Hsp110/SSE subfamily, and pseudogenes. Four DnaK subfamily genes encoding predicted cytoplasmic or endoplasmic reticulum proteins were upregulated in strains harbouring PiERE1. This may explain the greater thermotolerance conferred by the RNA element, and suggests that Hsp70 may be a useful biomarker for testing organisms for the cellular effects of symbiotic elements. PMID:20360179

  3. Multiple recognition of RXLR effectors is associated with nonhost resistance of pepper against Phytophthora infestans.

    PubMed

    Lee, Hyun-Ah; Kim, Shin-Young; Oh, Sang-Keun; Yeom, Seon-In; Kim, Saet-Byul; Kim, Myung-Shin; Kamoun, Sophien; Choi, Doil

    2014-08-01

    Nonhost resistance (NHR) is a plant immune response to resist most pathogens. The molecular basis of NHR is poorly understood, but recognition of pathogen effectors by immune receptors, a response known as effector-triggered immunity, has been proposed as a component of NHR. We performed transient expression of 54 Phytophthora infestansRXLR effectors in pepper (Capsicum annuum) accessions. We used optimized heterologous expression methods and analyzed the inheritance of effector-induced cell death in an F2 population derived from a cross between two pepper accessions. Pepper showed a localized cell death response upon inoculation with P. infestans, suggesting that recognition of effectors may contribute to NHR in this system. Pepper accessions recognized as many as 36 effectors. Among the effectors, PexRD8 and Avrblb2 induced cell death in a broad range of pepper accessions. Segregation of effector-induced cell death in an F2 population derived from a cross between two pepper accessions fit 15:1, 9:7 or 3:1 ratios, depending on the effector. Our genetic data suggest that a single or two independent/complementary dominant genes are involved in the recognition of RXLR effectors. Multiple loci recognizing a series of effectors may underpin NHR of pepper to P. infestans and confer resistance durability.

  4. Transcriptional dynamics of Phytophthora infestans during sequential stages of hemibiotrophic infection of tomato.

    PubMed

    Zuluaga, Andrea P; Vega-Arreguín, Julio C; Fei, Zhangjun; Ponnala, Lalit; Lee, Sang Jik; Matas, Antonio J; Patev, Sean; Fry, William E; Rose, Jocelyn K C

    2016-01-01

    Hemibiotrophic plant pathogens, such as the oomycete Phytophthora infestans, employ a biphasic infection strategy, initially behaving as biotrophs, where minimal symptoms are exhibited by the plant, and subsequently as necrotrophs, feeding on dead plant tissue. The regulation of this transition and the breadth of molecular mechanisms that modulate plant defences are not well understood, although effector proteins secreted by the pathogen are thought to play a key role. We examined the transcriptional dynamics of P. infestans in a compatible interaction with its host tomato (Solanum lycopersicum) at three infection stages: biotrophy; the transition from biotrophy to necrotrophy; and necrotrophy. The expression data suggest a tight temporal regulation of many pathways associated with the suppression of plant defence mechanisms and pathogenicity, including the induction of putative cytoplasmic and apoplastic effectors. Twelve of these were experimentally evaluated to determine their ability to suppress necrosis caused by the P. infestans necrosis-inducing protein PiNPP1.1 in Nicotiana benthamiana. Four effectors suppressed necrosis, suggesting that they might prolong the biotrophic phase. This study suggests that a complex regulation of effector expression modulates the outcome of the interaction.

  5. Metalaxyl Resistance in Phytophthora infestans: Assessing Role of RPA190 Gene and Diversity Within Clonal Lineages.

    PubMed

    Matson, Michael E H; Small, Ian M; Fry, William E; Judelson, Howard S

    2015-12-01

    Prior work has shown that the inheritance of resistance to metalaxyl, an oomycete-specific fungicide, is complex and may involve multiple genes. Recent research indicated that a single nucleotide polymorphism (SNP) in the gene encoding RPA190, the largest subunit of RNA polymerase I, confers resistance to metalaxyl (or mefenoxam) in some isolates of the potato late blight pathogen Phytophthora infestans. Using both DNA sequencing and high resolution melt assays for distinguishing RPA190 alleles, we show here that the SNP is absent from certain resistant isolates of P. infestans from North America, Europe, and Mexico. The SNP is present in some members of the US-23 and US-24 clonal lineages, but these tend to be fairly sensitive to the fungicide based on artificial media and field test data. Diversity in the level of sensitivity, RPA190 genotype, and RPA190 copy number was observed in these lineages but were uncorrelated. Controlled laboratory crosses demonstrated that RPA190 did not cosegregate with metalaxyl resistance from a Mexican and British isolate. We conclude that while metalaxyl may be used to control many contemporary strains of P. infestans, an assay based on RPA190 will not be sufficient to diagnose the sensitivity levels of isolates. PMID:26551315

  6. Metalaxyl Resistance in Phytophthora infestans: Assessing Role of RPA190 Gene and Diversity Within Clonal Lineages.

    PubMed

    Matson, Michael E H; Small, Ian M; Fry, William E; Judelson, Howard S

    2015-12-01

    Prior work has shown that the inheritance of resistance to metalaxyl, an oomycete-specific fungicide, is complex and may involve multiple genes. Recent research indicated that a single nucleotide polymorphism (SNP) in the gene encoding RPA190, the largest subunit of RNA polymerase I, confers resistance to metalaxyl (or mefenoxam) in some isolates of the potato late blight pathogen Phytophthora infestans. Using both DNA sequencing and high resolution melt assays for distinguishing RPA190 alleles, we show here that the SNP is absent from certain resistant isolates of P. infestans from North America, Europe, and Mexico. The SNP is present in some members of the US-23 and US-24 clonal lineages, but these tend to be fairly sensitive to the fungicide based on artificial media and field test data. Diversity in the level of sensitivity, RPA190 genotype, and RPA190 copy number was observed in these lineages but were uncorrelated. Controlled laboratory crosses demonstrated that RPA190 did not cosegregate with metalaxyl resistance from a Mexican and British isolate. We conclude that while metalaxyl may be used to control many contemporary strains of P. infestans, an assay based on RPA190 will not be sufficient to diagnose the sensitivity levels of isolates.

  7. Potassium phosphite primes defense responses in potato against Phytophthora infestans.

    PubMed

    Machinandiarena, Milagros Florencia; Lobato, María Candela; Feldman, Mariana Laura; Daleo, Gustavo Raúl; Andreu, Adriana Balbina

    2012-09-15

    Although phosphite is widely used to protect plants from pathogenic oomycetes on a wide range of horticultural crops, the molecular mechanisms behind phosphite induced resistance are poorly understood. The aim of this work was to assess the effects of potassium phosphite (KPhi) on potato plant defense responses to infection with Phytophtora infestans (Pi). Pathogen development was severely restricted and there was also an important decrease in lesion size in infected KPhi-treated leaves. We demonstrated that KPhi primed hydrogen peroxide and superoxide anion production in potato leaves at 12 h post-inoculation with Pi. Moreover, the KPhi-treated leaves showed an increased and earlier callose deposition as compared with water-treated plants, beginning 48 h after inoculation. In contrast, callose deposition was not detected in water-treated leaves until 72 h after inoculation. In addition, we carried out RNA gel blot analysis of genes implicated in the responses mediated by salicylic (SA) and jasmonic acid (JA). To this end, we examined the temporal expression pattern of StNPR1 and StWRKY1, two transcription factors related to SA pathway, and StPR1 and StIPII, marker genes related to SA and JA pathways, respectively. The expression of StNPR1 and StWRKY1 was enhanced in response to KPhi treatment. In contrast, StIPII was down regulated in both KPhi- and water-treated leaves, until 48 h after infection with Pi, suggesting that the regulation of this gene could be independent of the KPhi treatment. Our results indicate that KPhi primes the plant for an earlier and more intense response to infection and that SA would mediate this response.

  8. The Plant Pathogen Phytophthora andina Emerged via Hybridization of an Unknown Phytophthora Species and the Irish Potato Famine Pathogen, P. infestans

    PubMed Central

    Goss, Erica M.; Cardenas, Martha E.; Myers, Kevin; Forbes, Gregory A.; Fry, William E.; Restrepo, Silvia; Grünwald, Niklaus J.

    2011-01-01

    Emerging plant pathogens have largely been a consequence of the movement of pathogens to new geographic regions. Another documented mechanism for the emergence of plant pathogens is hybridization between individuals of different species or subspecies, which may allow rapid evolution and adaptation to new hosts or environments. Hybrid plant pathogens have traditionally been difficult to detect or confirm, but the increasing ease of cloning and sequencing PCR products now makes the identification of species that consistently have genes or alleles with phylogenetically divergent origins relatively straightforward. We investigated the genetic origin of Phytophthora andina, an increasingly common pathogen of Andean crops Solanum betaceum, S. muricatum, S. quitoense, and several wild Solanum spp. It has been hypothesized that P. andina is a hybrid between the potato late blight pathogen P. infestans and another Phytophthora species. We tested this hypothesis by cloning four nuclear loci to obtain haplotypes and using these loci to infer the phylogenetic relationships of P. andina to P. infestans and other related species. Sequencing of cloned PCR products in every case revealed two distinct haplotypes for each locus in P. andina, such that each isolate had one allele derived from a P. infestans parent and a second divergent allele derived from an unknown species that is closely related but distinct from P. infestans, P. mirabilis, and P. ipomoeae. To the best of our knowledge, the unknown parent has not yet been collected. We also observed sequence polymorphism among P. andina isolates at three of the four loci, many of which segregate between previously described P. andina clonal lineages. These results provide strong support that P. andina emerged via hybridization between P. infestans and another unknown Phytophthora species also belonging to Phytophthora clade 1c. PMID:21949727

  9. Inoculation of Transgenic Resistant Potato by Phytophthora infestans Affects Host Plant Choice of a Generalist Moth.

    PubMed

    Abreha, Kibrom B; Alexandersson, Erik; Vossen, Jack H; Anderson, Peter; Andreasson, Erik

    2015-01-01

    Pathogen attack and the plant's response to this attack affect herbivore oviposition preference and larval performance. Introduction of major resistance genes against Phytophthora infestans (Rpi-genes), the cause of the devastating late blight disease, from wild Solanum species into potato changes the plant-pathogen interaction dynamics completely, but little is known about the effects on non-target organisms. Thus, we examined the effect of P. infestans itself and introduction of an Rpi-gene into the crop on host plant preference of the generalist insect herbivore, Spodoptera littoralis (Lepidoptera: Noctuidae). In two choice bioassays, S. littoralis preferred to oviposit on P. infestans-inoculated plants of both the susceptible potato (cv. Desiree) and an isogenic resistant clone (A01-22: cv. Desiree transformed with Rpi-blb1), when compared to uninoculated plants of the same genotype. Both cv. Desiree and clone A01-22 were equally preferred for oviposition by S. littoralis when uninoculated plants were used, while cv. Desiree received more eggs compared to the resistant clone when both were inoculated with the pathogen. No significant difference in larval and pupal weight was found between S. littoralis larvae reared on leaves of the susceptible potato plants inoculated or uninoculated with P. infestans. Thus, the herbivore's host plant preference in this system was not directly associated with larval performance. The results indicate that the Rpi-blb1 based resistance in itself does not influence insect behavior, but that herbivore oviposition preference is affected by a change in the plant-microbe interaction. PMID:26053171

  10. Inoculation of Transgenic Resistant Potato by Phytophthora infestans Affects Host Plant Choice of a Generalist Moth

    PubMed Central

    Abreha, Kibrom B.; Alexandersson, Erik; Vossen, Jack H.; Anderson, Peter; Andreasson, Erik

    2015-01-01

    Pathogen attack and the plant’s response to this attack affect herbivore oviposition preference and larval performance. Introduction of major resistance genes against Phytophthora infestans (Rpi-genes), the cause of the devastating late blight disease, from wild Solanum species into potato changes the plant-pathogen interaction dynamics completely, but little is known about the effects on non-target organisms. Thus, we examined the effect of P. infestans itself and introduction of an Rpi-gene into the crop on host plant preference of the generalist insect herbivore, Spodoptera littoralis (Lepidoptera: Noctuidae). In two choice bioassays, S. littoralis preferred to oviposit on P. infestans-inoculated plants of both the susceptible potato (cv. Desiree) and an isogenic resistant clone (A01-22: cv. Desiree transformed with Rpi-blb1), when compared to uninoculated plants of the same genotype. Both cv. Desiree and clone A01-22 were equally preferred for oviposition by S. littoralis when uninoculated plants were used, while cv. Desiree received more eggs compared to the resistant clone when both were inoculated with the pathogen. No significant difference in larval and pupal weight was found between S. littoralis larvae reared on leaves of the susceptible potato plants inoculated or uninoculated with P. infestans. Thus, the herbivore’s host plant preference in this system was not directly associated with larval performance. The results indicate that the Rpi-blb1 based resistance in itself does not influence insect behavior, but that herbivore oviposition preference is affected by a change in the plant-microbe interaction. PMID:26053171

  11. Comparisons of Ribosomal Protein Gene Promoters Indicate Superiority of Heterologous Regulatory Sequences for Expressing Transgenes in Phytophthora infestans

    PubMed Central

    Khachatoorian, Careen; Judelson, Howard S.

    2015-01-01

    Molecular genetics approaches in Phytophthora research can be hampered by the limited number of known constitutive promoters for expressing transgenes and the instability of transgene activity. We have therefore characterized genes encoding the cytoplasmic ribosomal proteins of Phytophthora and studied their suitability for expressing transgenes in P. infestans. Phytophthora spp. encode a standard complement of 79 cytoplasmic ribosomal proteins. Several genes are duplicated, and two appear to be pseudogenes. Half of the genes are expressed at similar levels during all stages of asexual development, and we discovered that the majority share a novel promoter motif named the PhRiboBox. This sequence is enriched in genes associated with transcription, translation, and DNA replication, including tRNA and rRNA biogenesis. Promoters from the three P. infestans genes encoding ribosomal proteins S9, L10, and L23 and their orthologs from P. capsici were tested for their ability to drive transgenes in stable transformants of P. infestans. Five of the six promoters yielded strong expression of a GUS reporter, but the stability of expression was higher using the P. capsici promoters. With the RPS9 and RPL10 promoters of P. infestans, about half of transformants stopped making GUS over two years of culture, while their P. capsici orthologs conferred stable expression. Since cross-talk between native and transgene loci may trigger gene silencing, we encourage the use of heterologous promoters in transformation studies. PMID:26716454

  12. Comparisons of Ribosomal Protein Gene Promoters Indicate Superiority of Heterologous Regulatory Sequences for Expressing Transgenes in Phytophthora infestans.

    PubMed

    Poidevin, Laetitia; Andreeva, Kalina; Khachatoorian, Careen; Judelson, Howard S

    2015-01-01

    Molecular genetics approaches in Phytophthora research can be hampered by the limited number of known constitutive promoters for expressing transgenes and the instability of transgene activity. We have therefore characterized genes encoding the cytoplasmic ribosomal proteins of Phytophthora and studied their suitability for expressing transgenes in P. infestans. Phytophthora spp. encode a standard complement of 79 cytoplasmic ribosomal proteins. Several genes are duplicated, and two appear to be pseudogenes. Half of the genes are expressed at similar levels during all stages of asexual development, and we discovered that the majority share a novel promoter motif named the PhRiboBox. This sequence is enriched in genes associated with transcription, translation, and DNA replication, including tRNA and rRNA biogenesis. Promoters from the three P. infestans genes encoding ribosomal proteins S9, L10, and L23 and their orthologs from P. capsici were tested for their ability to drive transgenes in stable transformants of P. infestans. Five of the six promoters yielded strong expression of a GUS reporter, but the stability of expression was higher using the P. capsici promoters. With the RPS9 and RPL10 promoters of P. infestans, about half of transformants stopped making GUS over two years of culture, while their P. capsici orthologs conferred stable expression. Since cross-talk between native and transgene loci may trigger gene silencing, we encourage the use of heterologous promoters in transformation studies. PMID:26716454

  13. Comparative analysis of Phytophthora infestans induced gene expression in potato cultivars with different levels of resistance.

    PubMed

    Ros, B; Thümmler, F; Wenzel, G

    2005-11-01

    Differential gene expression was analyzed after infection with Phytophthora infestans in six potato cultivars with different levels of resistance to late blight. To verify the infection of the potato leaflets, the amount of phytopathogen mRNA within the plant material was quantified by real-time quantitative PCR. The expression of 182 genes selected from two subtracted cDNA libraries was studied with cDNA array hybridization using RNA from non-infected and infected potato leaflets. Gene up- and down-regulation were clearly detectable in all cultivars 72 h post inoculation. Gene expression patterns in susceptible cultivars differed from those in potato varieties with a higher level of resistance. In general, a stronger gene induction was observed in the susceptible cultivars compared to the moderately to highly resistant potato varieties. Five genes with the highest homology to stress and/or defence-related genes were induced specifically in the susceptible cultivars. Four genes responded to pathogen attack independently of the level of resistance of the cultivar used, and three genes were repressed in infected tissue of most cultivars. Even in the absence of P. infestans infection, six genes showed higher expression levels in the somewhat resistant cultivars Bettina and Matilda. Possible reasons for the different levels of gene expression are discussed.

  14. The rise and fall of the Phytophthora infestans lineage that triggered the Irish potato famine.

    PubMed

    Yoshida, Kentaro; Schuenemann, Verena J; Cano, Liliana M; Pais, Marina; Mishra, Bagdevi; Sharma, Rahul; Lanz, Chirsta; Martin, Frank N; Kamoun, Sophien; Krause, Johannes; Thines, Marco; Weigel, Detlef; Burbano, Hernán A

    2013-01-01

    Phytophthora infestans, the cause of potato late blight, is infamous for having triggered the Irish Great Famine in the 1840s. Until the late 1970s, P. infestans diversity outside of its Mexican center of origin was low, and one scenario held that a single strain, US-1, had dominated the global population for 150 years; this was later challenged based on DNA analysis of historical herbarium specimens. We have compared the genomes of 11 herbarium and 15 modern strains. We conclude that the 19th century epidemic was caused by a unique genotype, HERB-1, that persisted for over 50 years. HERB-1 is distinct from all examined modern strains, but it is a close relative of US-1, which replaced it outside of Mexico in the 20th century. We propose that HERB-1 and US-1 emerged from a metapopulation that was established in the early 1800s outside of the species' center of diversity. DOI:http://dx.doi.org/10.7554/eLife.00731.001. PMID:23741619

  15. The rise and fall of the Phytophthora infestans lineage that triggered the Irish potato famine

    PubMed Central

    Yoshida, Kentaro; Schuenemann, Verena J; Cano, Liliana M; Pais, Marina; Mishra, Bagdevi; Sharma, Rahul; Lanz, Chirsta; Martin, Frank N; Kamoun, Sophien; Krause, Johannes; Thines, Marco; Weigel, Detlef; Burbano, Hernán A

    2013-01-01

    Phytophthora infestans, the cause of potato late blight, is infamous for having triggered the Irish Great Famine in the 1840s. Until the late 1970s, P. infestans diversity outside of its Mexican center of origin was low, and one scenario held that a single strain, US-1, had dominated the global population for 150 years; this was later challenged based on DNA analysis of historical herbarium specimens. We have compared the genomes of 11 herbarium and 15 modern strains. We conclude that the 19th century epidemic was caused by a unique genotype, HERB-1, that persisted for over 50 years. HERB-1 is distinct from all examined modern strains, but it is a close relative of US-1, which replaced it outside of Mexico in the 20th century. We propose that HERB-1 and US-1 emerged from a metapopulation that was established in the early 1800s outside of the species' center of diversity. DOI: http://dx.doi.org/10.7554/eLife.00731.001 PMID:23741619

  16. Species Tree Estimation for the Late Blight Pathogen, Phytophthora infestans, and Close Relatives

    PubMed Central

    Blair, Jaime E.; Coffey, Michael D.; Martin, Frank N.

    2012-01-01

    To better understand the evolutionary history of a group of organisms, an accurate estimate of the species phylogeny must be known. Traditionally, gene trees have served as a proxy for the species tree, although it was acknowledged early on that these trees represented different evolutionary processes. Discordances among gene trees and between the gene trees and the species tree are also expected in closely related species that have rapidly diverged, due to processes such as the incomplete sorting of ancestral polymorphisms. Recently, methods have been developed for the explicit estimation of species trees, using information from multilocus gene trees while accommodating heterogeneity among them. Here we have used three distinct approaches to estimate the species tree for five Phytophthora pathogens, including P. infestans, the causal agent of late blight disease in potato and tomato. Our concatenation-based “supergene” approach was unable to resolve relationships even with data from both the nuclear and mitochondrial genomes, and from multiple isolates per species. Our multispecies coalescent approach using both Bayesian and maximum likelihood methods was able to estimate a moderately supported species tree showing a close relationship among P. infestans, P. andina, and P. ipomoeae. The topology of the species tree was also identical to the dominant phylogenetic history estimated in our third approach, Bayesian concordance analysis. Our results support previous suggestions that P. andina is a hybrid species, with P. infestans representing one parental lineage. The other parental lineage is not known, but represents an independent evolutionary lineage more closely related to P. ipomoeae. While all five species likely originated in the New World, further study is needed to determine when and under what conditions this hybridization event may have occurred. PMID:22615869

  17. Trade-offs and evolution of thermal adaptation in the Irish potato famine pathogen Phytophthora infestans.

    PubMed

    Yang, Li-Na; Zhu, Wen; Wu, E-Jiao; Yang, Ce; Thrall, Peter H; Burdon, Jeremy J; Jin, Li-Ping; Shang, Li-Ping; Zhan, Jiasui

    2016-08-01

    Temperature is one of the most important environmental parameters with crucial impacts on nearly all biological processes. Due to anthropogenic activity, average air temperatures are expected to increase by a few degrees in coming decades, accompanied by an increased occurrence of extreme temperature events. Such global trends are likely to have various major impacts on human society through their influence on natural ecosystems, food production and biotic interactions, including diseases. In this study, we used a combination of statistical genetics, experimental evolution and common garden experiments to investigate the evolutionary potential for thermal adaptation in the potato late blight pathogen, Phytophthora infestans, and infer its likely response to changing temperatures. We found a trade-off associated with thermal adaptation to heterogeneous environments in P. infestans, with the degree of the trade-off peaking approximately at the pathogen's optimum growth temperature. A genetic trade-off in thermal adaptation was also evidenced by the negative association between a strain's growth rate and its thermal range for growth, and warm climates selecting for a low pathogen growth rate. We also found a mirror effect of phenotypic plasticity and genetic adaptation on growth rate. At below the optimum, phenotypic plasticity enhances pathogen's growth rate but nature selects for slower growing genotypes when temperature increases. At above the optimum, phenotypic plasticity reduces pathogen's growth rate but natural selection favours for faster growing genotypes when temperature increases further. We conclude from these findings that the growth rate of P. infestans will only be marginally affected by global warming.

  18. A predicted functional gene network for the plant pathogen Phytophthora infestans as a framework for genomic biology

    PubMed Central

    2013-01-01

    Background Associations between proteins are essential to understand cell biology. While this complex interplay between proteins has been studied in model organisms, it has not yet been described for the oomycete late blight pathogen Phytophthora infestans. Results We present an integrative probabilistic functional gene network that provides associations for 37 percent of the predicted P. infestans proteome. Our method unifies available genomic, transcriptomic and comparative genomic data into a single comprehensive network using a Bayesian approach. Enrichment of proteins residing in the same or related subcellular localization validates the biological coherence of our predictions. The network serves as a framework to query existing genomic data using network-based methods, which thus far was not possible in Phytophthora. We used the network to study the set of interacting proteins that are encoded by genes co-expressed during sporulation. This identified potential novel roles for proteins in spore formation through their links to proteins known to be involved in this process such as the phosphatase Cdc14. Conclusions The functional association network represents a novel genome-wide data source for P. infestans that also acts as a framework to interrogate other system-wide data. In both capacities it will improve our understanding of the complex biology of P. infestans and related oomycete pathogens. PMID:23865555

  19. Effect of Temperature on Growth and Sporulation of US-22, US-23, and US-24 Clonal Lineages of Phytophthora infestans and Implications for Late Blight Epidemiology.

    PubMed

    Seidl Johnson, Anna C; Frost, Kenneth E; Rouse, Douglas I; Gevens, Amanda J

    2015-04-01

    Epidemics of late blight, caused by Phytophthora infestans (Mont.) de Bary, have been studied by plant pathologists and regarded with great concern by potato and tomato growers since the Irish potato famine in the 1840s. P. infestans populations have continued to evolve, with unique clonal lineages arising which differ in pathogen fitness and pathogenicity, potentially impacting epidemiology. In 2012 and 2013, the US-23 clonal lineage predominated late blight epidemics in most U.S. potato and tomato production regions, including Wisconsin. This lineage was unknown prior to 2009. For isolates of three recently identified clonal lineages of P. infestans (US-22, US-23, and US-24), sporulation rates were experimentally determined on potato and tomato foliage and the effect of temperature on lesion growth rate on tomato was investigated. The US-22 and US-23 isolates had greater lesion growth rates on tomato than US-24 isolates. Sporulation rates for all isolates were greater on potato than tomato, and the US-23 isolates had greater sporulation rates on both tomato and potato than the US-22 and US-24 isolates. Experimentally determined correlates of fitness were input to the LATEBLIGHT model and epidemics were simulated using archived Wisconsin weather data from four growing seasons (2009 to 2012) to investigate the effect of isolates of these new lineages on late blight epidemiology. The fast lesion growth rates of US-22 and US-23 isolates resulted in severe epidemics in all years tested, particularly in 2011. The greater sporulation rates of P. infestans on potato resulted in simulated epidemics that progressed faster than epidemics simulated for tomato; the high sporulation rates of US-23 isolates resulted in simulated epidemics more severe than simulated epidemics of isolates of the US-22 and US-24 isolates and EC-1 clonal lineages on potato and tomato. Additionally, US-23 isolates consistently caused severe simulated epidemics when lesion growth rate and sporulation

  20. Effect of Temperature on Growth and Sporulation of US-22, US-23, and US-24 Clonal Lineages of Phytophthora infestans and Implications for Late Blight Epidemiology.

    PubMed

    Seidl Johnson, Anna C; Frost, Kenneth E; Rouse, Douglas I; Gevens, Amanda J

    2015-04-01

    Epidemics of late blight, caused by Phytophthora infestans (Mont.) de Bary, have been studied by plant pathologists and regarded with great concern by potato and tomato growers since the Irish potato famine in the 1840s. P. infestans populations have continued to evolve, with unique clonal lineages arising which differ in pathogen fitness and pathogenicity, potentially impacting epidemiology. In 2012 and 2013, the US-23 clonal lineage predominated late blight epidemics in most U.S. potato and tomato production regions, including Wisconsin. This lineage was unknown prior to 2009. For isolates of three recently identified clonal lineages of P. infestans (US-22, US-23, and US-24), sporulation rates were experimentally determined on potato and tomato foliage and the effect of temperature on lesion growth rate on tomato was investigated. The US-22 and US-23 isolates had greater lesion growth rates on tomato than US-24 isolates. Sporulation rates for all isolates were greater on potato than tomato, and the US-23 isolates had greater sporulation rates on both tomato and potato than the US-22 and US-24 isolates. Experimentally determined correlates of fitness were input to the LATEBLIGHT model and epidemics were simulated using archived Wisconsin weather data from four growing seasons (2009 to 2012) to investigate the effect of isolates of these new lineages on late blight epidemiology. The fast lesion growth rates of US-22 and US-23 isolates resulted in severe epidemics in all years tested, particularly in 2011. The greater sporulation rates of P. infestans on potato resulted in simulated epidemics that progressed faster than epidemics simulated for tomato; the high sporulation rates of US-23 isolates resulted in simulated epidemics more severe than simulated epidemics of isolates of the US-22 and US-24 isolates and EC-1 clonal lineages on potato and tomato. Additionally, US-23 isolates consistently caused severe simulated epidemics when lesion growth rate and sporulation

  1. Pseudomonas strains naturally associated with potato plants produce volatiles with high potential for inhibition of Phytophthora infestans.

    PubMed

    Hunziker, Lukas; Bönisch, Denise; Groenhagen, Ulrike; Bailly, Aurélien; Schulz, Stefan; Weisskopf, Laure

    2015-02-01

    Bacteria emit volatile organic compounds with a wide range of effects on bacteria, fungi, plants, and animals. The antifungal potential of bacterial volatiles has been investigated with a broad span of phytopathogenic organisms, yet the reaction of oomycetes to these volatile signals is largely unknown. For instance, the response of the late blight-causing agent and most devastating oomycete pathogen worldwide, Phytophthora infestans, to bacterial volatiles has not been assessed so far. In this work, we analyzed this response and compared it to that of selected fungal and bacterial potato pathogens, using newly isolated, potato-associated bacterial strains as volatile emitters. P. infestans was highly susceptible to bacterial volatiles, while fungal and bacterial pathogens were less sensitive. Cyanogenic Pseudomonas strains were the most active, leading to complete growth inhibition, yet noncyanogenic ones also produced antioomycete volatiles. Headspace analysis of the emitted volatiles revealed 1-undecene as a compound produced by strains inducing volatile-mediated P. infestans growth inhibition. Supplying pure 1-undecene to P. infestans significantly reduced mycelial growth, sporangium formation, germination, and zoospore release in a dose-dependent manner. This work demonstrates the high sensitivity of P. infestans to bacterial volatiles and opens new perspectives for sustainable control of this devastating pathogen.

  2. Pseudomonas Strains Naturally Associated with Potato Plants Produce Volatiles with High Potential for Inhibition of Phytophthora infestans

    PubMed Central

    Hunziker, Lukas; Bönisch, Denise; Groenhagen, Ulrike; Bailly, Aurélien; Schulz, Stefan

    2014-01-01

    Bacteria emit volatile organic compounds with a wide range of effects on bacteria, fungi, plants, and animals. The antifungal potential of bacterial volatiles has been investigated with a broad span of phytopathogenic organisms, yet the reaction of oomycetes to these volatile signals is largely unknown. For instance, the response of the late blight-causing agent and most devastating oomycete pathogen worldwide, Phytophthora infestans, to bacterial volatiles has not been assessed so far. In this work, we analyzed this response and compared it to that of selected fungal and bacterial potato pathogens, using newly isolated, potato-associated bacterial strains as volatile emitters. P. infestans was highly susceptible to bacterial volatiles, while fungal and bacterial pathogens were less sensitive. Cyanogenic Pseudomonas strains were the most active, leading to complete growth inhibition, yet noncyanogenic ones also produced antioomycete volatiles. Headspace analysis of the emitted volatiles revealed 1-undecene as a compound produced by strains inducing volatile-mediated P. infestans growth inhibition. Supplying pure 1-undecene to P. infestans significantly reduced mycelial growth, sporangium formation, germination, and zoospore release in a dose-dependent manner. This work demonstrates the high sensitivity of P. infestans to bacterial volatiles and opens new perspectives for sustainable control of this devastating pathogen. PMID:25398872

  3. Population Structure of the Late Blight Pathogen Phytophthora infestans in a Potato Germplasm Nursery in Two Consecutive Years.

    PubMed

    Tian, Yuee; Yin, Junliang; Sun, Jieping; Ma, Hongmei; Ma, Yunfang; Quan, Junli; Shan, Weixing

    2015-06-01

    As the causal agent of late blight on potato, Phytophthora infestans is one of the most destructive plant pathogens worldwide and widely known as the Irish potato famine pathogen. Understanding the genetic structure of P. infestans populations is important both for breeding and deployment of resistant varieties and for development of disease control strategies. Here, we investigate the population genetic structure of P. infestans in a potato germplasm nursery in northwestern China. In total, 279 isolates were recovered from 63 potato varieties or lines in 2010 and 2011, and were genotyped by mitochondrial DNA haplotypes and a set of nine simple-sequence repeat markers. Selected isolates were further examined for virulence on a set of differential lines containing each resistance (R) gene (R1 to R11). The overall P. infestans population was characterized as having a low level of genetic diversity and resistance to metalaxyl, and containing a high percentage of individuals that virulent to all 11 R genes. Both A1 and A2 mating types as well as self-fertile P. infestans isolates were present but there was no evidence of sexual reproduction. The low level of genetic differentiation in P. infestans populations is probably due to the action of relatively high levels of migration as supported by analysis of molecular variance (P < 0.01). Migration and asexual reproduction were the predominant mechanisms influencing the P. infestans population structure in the germplasm nursery. Therefore, it is important to ensure the production of pathogen-free potato seed tubers to aid sustainable production of potato in northwestern China. PMID:25738550

  4. Molecular Determinants of Resistance Activation and Suppression by Phytophthora infestans Effector IPI-O

    PubMed Central

    Chen, Yu; Liu, Zhenyu; Halterman, Dennis A.

    2012-01-01

    Despite intensive breeding efforts, potato late blight, caused by the oomycete pathogen Phytophthora infestans, remains a threat to potato production worldwide because newly evolved pathogen strains have consistently overcome major resistance genes. The potato RB gene, derived from the wild species Solanum bulbocastanum, confers resistance to most P. infestans strains through recognition of members of the pathogen effector family IPI-O. While the majority of IPI-O proteins are recognized by RB to elicit resistance (e.g. IPI-O1, IPI-O2), some family members are able to elude detection (e.g. IPI-O4). In addition, IPI-O4 blocks recognition of IPI-O1, leading to inactivation of RB-mediated programmed cell death. Here, we report results that elucidate molecular mechanisms governing resistance elicitation or suppression of RB by IPI-O. Our data indicate self-association of the RB coiled coil (CC) domain as well as a physical interaction between this domain and the effectors IPI-O4 and IPI-O1. We identified four amino acids within IPI-O that are critical for interaction with the RB CC domain and one of these amino acids, at position 129, determines hypersensitive response (HR) elicitation in planta. IPI-O1 mutant L129P fails to induce HR in presence of RB while IPI-O4 P129L gains the ability to induce an HR. Like IPI-O4, IPI-O1 L129P is also able to suppress the HR mediated by RB, indicating a critical step in the evolution of this gene family. Our results point to a model in which IPI-O effectors can affect RB function through interaction with the RB CC domain. PMID:22438813

  5. BABA-primed defense responses to Phytophthora infestans in the next vegetative progeny of potato

    PubMed Central

    Floryszak-Wieczorek, Jolanta; Arasimowicz-Jelonek, Magdalena; Abramowski, Dariusz

    2015-01-01

    The transcript of the PR1 gene accumulation as an informative marker of systemic acquired resistance (SAR) was analyzed in β-aminobutyric acid (BABA) primed potato in the short-lasting (3 days) and long-lasting (28 days) time periods after induction and in the vegetative descendants of primed plants derived from tubers and from in vitro seedlings. BABA pretreatment resulted either in minimal or no PR1 gene expression, but sequential treatment with BABA followed by virulent Phytophthora infestans provided data on the imprint of post-stress information and its duration until fertilization, in the form of an enhanced PR1 transcript accumulation and a transient increase of basal resistance to the late blight disease. The primed state for defense of the susceptible potato cultivar was transmitted to its vegetative progeny as a potentiated PR1 mRNA accumulation following challenge inoculation. However, variation was observed between vegetative accessions of the BABA-primed potato genotype in responsiveness to disease. In contrast to plants derived from tubers, potato propagated through in vitro seedlings largely lost inducible resistance traits, although itretained primed PR1 gene expression. PMID:26528308

  6. Testing Taxonomic Predictivity of Foliar and Tuber Resistance to Phytophthora infestans in Wild Relatives of Potato.

    PubMed

    Khiutti, A; Spooner, D M; Jansky, S H; Halterman, D A

    2015-09-01

    Potato late blight, caused by the oomycete phytopathogen Phytophthora infestans, is a devastating disease found in potato-growing regions worldwide. Long-term management strategies to control late blight include the incorporation of host resistance to predominant strains. However, due to rapid genetic changes within pathogen populations, rapid and recurring identification and integration of novel host resistance traits is necessary. Wild relatives of potato offer a rich source of desirable traits, including late blight resistance, but screening methods can be time intensive. We tested the ability of taxonomy, ploidy, crossing group, breeding system, and geography to predict the presence of foliar and tuber late blight resistance in wild Solanum spp. Significant variation for resistance to both tuber and foliar late blight was found within and among species but there was no discernable predictive power based on taxonomic series, clade, ploidy, breeding system, elevation, or geographic location. We observed a moderate but significant correlation between tuber and foliar resistance within species. Although previously uncharacterized sources of both foliar and tuber resistance were identified, our study does not support an assumption that taxonomic or geographic data can be used to predict sources of late blight resistance in wild Solanum spp.

  7. A Phytophthora infestans RXLR effector targets plant PP1c isoforms that promote late blight disease

    PubMed Central

    Boevink, Petra C.; Wang, Xiaodan; McLellan, Hazel; He, Qin; Naqvi, Shaista; Armstrong, Miles R.; Zhang, Wei; Hein, Ingo; Gilroy, Eleanor M.; Tian, Zhendong; Birch, Paul R. J.

    2016-01-01

    Plant pathogens deliver effectors to alter host processes. Knowledge of how effectors target and manipulate host proteins is critical to understand crop disease. Here, we show that in planta expression of the RXLR effector Pi04314 enhances leaf colonization by Phytophthora infestans via activity in the host nucleus and attenuates induction of jasmonic and salicylic acid-responsive genes. Pi04314 interacts with three host protein phosphatase 1 catalytic (PP1c) isoforms, causing their re-localization from the nucleolus to the nucleoplasm. Re-localization of PP1c-1 also occurs during infection and is dependent on an R/KVxF motif in the effector. Silencing the PP1c isoforms or overexpression of a phosphatase-dead PP1c-1 mutant attenuates infection, demonstrating that host PP1c activity is required for disease. Moreover, expression of PP1c–1mut abolishes enhanced leaf colonization mediated by in planta Pi04314 expression. We argue that PP1c isoforms are susceptibility factors forming holoenzymes with Pi04314 to promote late blight disease. PMID:26822079

  8. A Phytophthora infestans RXLR effector targets plant PP1c isoforms that promote late blight disease.

    PubMed

    Boevink, Petra C; Wang, Xiaodan; McLellan, Hazel; He, Qin; Naqvi, Shaista; Armstrong, Miles R; Zhang, Wei; Hein, Ingo; Gilroy, Eleanor M; Tian, Zhendong; Birch, Paul R J

    2016-01-01

    Plant pathogens deliver effectors to alter host processes. Knowledge of how effectors target and manipulate host proteins is critical to understand crop disease. Here, we show that in planta expression of the RXLR effector Pi04314 enhances leaf colonization by Phytophthora infestans via activity in the host nucleus and attenuates induction of jasmonic and salicylic acid-responsive genes. Pi04314 interacts with three host protein phosphatase 1 catalytic (PP1c) isoforms, causing their re-localization from the nucleolus to the nucleoplasm. Re-localization of PP1c-1 also occurs during infection and is dependent on an R/KVxF motif in the effector. Silencing the PP1c isoforms or overexpression of a phosphatase-dead PP1c-1 mutant attenuates infection, demonstrating that host PP1c activity is required for disease. Moreover, expression of PP1c-1mut abolishes enhanced leaf colonization mediated by in planta Pi04314 expression. We argue that PP1c isoforms are susceptibility factors forming holoenzymes with Pi04314 to promote late blight disease. PMID:26822079

  9. Time-course investigation of Phytophthora infestans infection of potato leaf from three cultivars by quantitative proteomics

    PubMed Central

    Larsen, Mia Kruse Guldstrand; Jørgensen, Malene Møller; Bennike, Tue Bjerg; Stensballe, Allan

    2015-01-01

    Potato late blight is one the most important crop diseases worldwide. Even though potato has been studied for many years, the potato disease late blight still has a vast negative effect on the potato production [1], [2], [3]. Late blight is caused by the pathogen Phytophthora infestans (P. infestans), which initiates infection through leaves. However, the biological activities during different stages of infection are poorly described, and could enable novel or improved ways of defeating late blight infection [4]. Therefore, we investigated the interactions between P. infestans (mixed strain culture) and potato (Solanum tuberosum). Three commercially available field potato cultivars of different resistance to late blight infection; Kuras (moderate), Sarpo Mira (highly resistant) and Bintje (very susceptable) were grown under controlled green house conditions and inoculated with a diversity of P. infestans populations. We used label-free quantitative proteomics to investigate the infection with P. infestans in a time-course study over 258 h. Several key issues limits proteome analysis of potato leaf tissue [5], [6], [7]. Firstly, the immense complexity of the plant proteome, which is further complicated by the presence of highly abundant proteins, such as ribulose bisphosphate carboxylase/oxygenase (RuBisCO). Secondly, plant leaf and potato, in particular, contain abundant levels amounts of phenols and polyphenols, which hinder or completely prevent a successful protein extraction. Hitherto, protein profiling of potato leaf tissues have been limited to few proteome studies and only 1484 proteins have been extracted and comprehensively described [5], [8], [9]. We here present the detailed methods and raw data by optimized gel-enhanced label free quantitative approach. The methodology enabled us to detect and quantify between 3248 and 3529 unique proteins from each cultivar, and up to 758 P. infestans derived proteins. The complete dataset is available via Proteome

  10. Time-course investigation of Phytophthora infestans infection of potato leaf from three cultivars by quantitative proteomics.

    PubMed

    Larsen, Mia Kruse Guldstrand; Jørgensen, Malene Møller; Bennike, Tue Bjerg; Stensballe, Allan

    2016-03-01

    Potato late blight is one the most important crop diseases worldwide. Even though potato has been studied for many years, the potato disease late blight still has a vast negative effect on the potato production [1], [2], [3]. Late blight is caused by the pathogen Phytophthora infestans (P. infestans), which initiates infection through leaves. However, the biological activities during different stages of infection are poorly described, and could enable novel or improved ways of defeating late blight infection [4]. Therefore, we investigated the interactions between P. infestans (mixed strain culture) and potato (Solanum tuberosum). Three commercially available field potato cultivars of different resistance to late blight infection; Kuras (moderate), Sarpo Mira (highly resistant) and Bintje (very susceptable) were grown under controlled green house conditions and inoculated with a diversity of P. infestans populations. We used label-free quantitative proteomics to investigate the infection with P. infestans in a time-course study over 258 h. Several key issues limits proteome analysis of potato leaf tissue [5], [6], [7]. Firstly, the immense complexity of the plant proteome, which is further complicated by the presence of highly abundant proteins, such as ribulose bisphosphate carboxylase/oxygenase (RuBisCO). Secondly, plant leaf and potato, in particular, contain abundant levels amounts of phenols and polyphenols, which hinder or completely prevent a successful protein extraction. Hitherto, protein profiling of potato leaf tissues have been limited to few proteome studies and only 1484 proteins have been extracted and comprehensively described [5], [8], [9]. We here present the detailed methods and raw data by optimized gel-enhanced label free quantitative approach. The methodology enabled us to detect and quantify between 3248 and 3529 unique proteins from each cultivar, and up to 758 P. infestans derived proteins. The complete dataset is available via Proteome

  11. The dihydrolipoyl acyltransferase gene BCE2 participates in basal resistance against Phytophthora infestans in potato and Nicotiana benthamiana.

    PubMed

    Wang, Hongyang; Sun, Chunlian; Jiang, Rui; He, Qin; Yang, Yu; Tian, Zhejuan; Tian, Zhendong; Xie, Conghua

    2014-07-01

    Dihydrolipoyl acyltransferase (EC 2.3.1.12), a branched-chain α-ketoacid dehydrogenase E2 subunit (BCE2), catalyzes the transfer of the acyl group from the lipoyl moiety to coenzyme A. However, the role of BCE2 responding to biotic stress in plant is not clear. In this study, we cloned and characterized a BCE2 gene from potato, namely StBCE2, which was previously suggested to be involved in Phytophthora infestans-potato interaction. We found that the expression of StBCE2 was strongly induced by both P. infestans isolate HB09-14-2 and salicylic acid. Besides, when the homolog of StBCE2 in Nicotiana benthamiana named NbBCE2 was silenced, plants showed increased susceptibility to P. infestans and reduced accumulation of hydrogen peroxide (H2O2). Furthermore, we found that a marker gene NbrbohB involved in the production of reactive oxygen species, was also suppressed in NbBCE2-silenced plants. However, silencing of NbBCE2 had no significant effect on the hypersensitive responses trigged by INF1, R3a-AVR3a(KI) pair or Rpi-vnt1.1-AVR-vnt1.1 pair. Our results suggest that BCE2 is associated with the basal resistance to P. infestans by regulating H2O2 production. PMID:24913048

  12. [RNA-silencing of anionic peroxidase gene decreases the potato plant resistance to Phytophthora infestans (Mont.) de Bary].

    PubMed

    Sorokan', A V; Kuluev, B R; Burkhanova, G F; Maksimov, I V

    2014-01-01

    Transformed potato (Solanum tuberosum L.) plants expressing the antisense-fragment of M21334 gene were estimated. In transgenic plants the decrease of anionic isoperoxidase pI - 3.5 activity was detected. So, the data testify that M21334 gene encodes this isoperoxidase. Decrease of lignin accumulation and dramatic decline of resistance of transgenic potato plants to the late blight agent Phytophthora infestans emphasize an importance of isoperoxidase pI - 3.5 in defense reaction against late blight. PMID:25842867

  13. Production of Phytophthora infestans-resistant potato (Solanum tuberosum) utilising Ensifer adhaerens OV14.

    PubMed

    Wendt, Toni; Doohan, Fiona; Mullins, Ewen

    2012-06-01

    Based on the use of Agrobacterium tumefaciens-mediated transformation commodity crop improvement through genetic engineering is the fastest adopted crop technology in the world (James 2010). However, the complexity of the Agrobacterium patent landscape remains a challenge for non-patent holders who wish to generate novel varieties for a commercial purpose. The potential of non-Agrobacterium strains (Transbacter(™)) to modify a plant genome has previously been described. However, they are unlikely to be widely used without significant adjustments in transformation protocols in order to improve their gene transfer efficiencies. In this study we set out to identify alternative bacteria species that could (a) utilize vir genes for genetic transformation and (b) substitute for A. tumefaciens in existing transformation protocols, without a prerequisite for protocol modifications. To this end we isolated a collection (n=751) of plant-associated bacteria from the rhizosphere of commercially grown crops. Based on various screens, including plant transformation with the open-source vector pCAMBIA5105, we identified a strain of the bacterium Ensifer adhaerens with the capacity to transform both Arabidopsis thaliana (0.12%) and potato (mean transformation frequency 35.1%). Thereafter, Ensifer adhaerens was used to generate blight- (causative organism Phytophthora infestans) resistant potato using the Solanum bulbocastanum 'resistance to blight' (RB) gene. Resistant genotypes were confirmed by associated molecular analysis and resistant phenotypes demonstrated by the development of hypersensitive lesions on inoculated leaf tissue post-pathogen inoculation. These data confirm the potential of Ensifer-mediated transformation (EMT) as a novel platform for the high frequency generation of transgenic potato.

  14. High-Quality Draft Genome Sequence of Arthrobacter sp. OY3WO11, a Strain That Inhibits the Growth of Phytophthora infestans

    PubMed Central

    Town, Jennifer; Audy, Patrice; Boyetchko, Susan M.

    2016-01-01

    Arthrobacter sp. strain OY3WO11 inhibits the growth of the potato pathogen Phytophthora infestans in in vivo growth challenge assays. We determined the draft genome sequence of this strain, assembling it into 3 scaffolds of 4.2 Mbp total length. OY3WO11 may represent a novel species of Arthrobacter. PMID:27340067

  15. Comparative analyses of fungicide sensitivity and SSR marker variations indicate a low risk of developing azoxystrobin resistance in Phytophthora infestans.

    PubMed

    Qin, Chun-Fang; He, Meng-Han; Chen, Feng-Ping; Zhu, Wen; Yang, Li-Na; Wu, E-Jiao; Guo, Zheng-Liang; Shang, Li-Ping; Zhan, Jiasui

    2016-01-01

    Knowledge of the evolution of fungicide resistance is important in securing sustainable disease management in agricultural systems. In this study, we analyzed and compared the spatial distribution of genetic variation in azoxystrobin sensitivity and SSR markers in 140 Phytophthora infestans isolates sampled from seven geographic locations in China. Sensitivity to azoxystrobin and its genetic variation in the pathogen populations was measured by the relative growth rate (RGR) at four fungicide concentrations and determination of the effective concentration for 50% inhibition (EC50). We found that all isolates in the current study were sensitive to azoxystrobin and their EC50 was similar to that detected from a European population about 20 years ago, suggesting the risk of developing azoxystrobin resistance in P. infestans populations is low. Further analyses indicate that reduced genetic variation and high fitness cost in resistant mutations are the likely causes for the low evolutionary likelihood of developing azoxystrobin resistance in the pathogen. We also found a negative correlation between azoxystrobin tolerance in P. infestans populations and the mean annual temperature of collection sites, suggesting that global warming may increase the efficiency of using the fungicide to control the late blight. PMID:26853908

  16. Comparative analyses of fungicide sensitivity and SSR marker variations indicate a low risk of developing azoxystrobin resistance in Phytophthora infestans

    PubMed Central

    Qin, Chun-Fang; He, Meng-Han; Chen, Feng-Ping; Zhu, Wen; Yang, Li-Na; Wu, E-Jiao; Guo, Zheng-Liang; Shang, Li-Ping; Zhan, Jiasui

    2016-01-01

    Knowledge of the evolution of fungicide resistance is important in securing sustainable disease management in agricultural systems. In this study, we analyzed and compared the spatial distribution of genetic variation in azoxystrobin sensitivity and SSR markers in 140 Phytophthora infestans isolates sampled from seven geographic locations in China. Sensitivity to azoxystrobin and its genetic variation in the pathogen populations was measured by the relative growth rate (RGR) at four fungicide concentrations and determination of the effective concentration for 50% inhibition (EC50). We found that all isolates in the current study were sensitive to azoxystrobin and their EC50 was similar to that detected from a European population about 20 years ago, suggesting the risk of developing azoxystrobin resistance in P. infestans populations is low. Further analyses indicate that reduced genetic variation and high fitness cost in resistant mutations are the likely causes for the low evolutionary likelihood of developing azoxystrobin resistance in the pathogen. We also found a negative correlation between azoxystrobin tolerance in P. infestans populations and the mean annual temperature of collection sites, suggesting that global warming may increase the efficiency of using the fungicide to control the late blight. PMID:26853908

  17. Diverse mechanisms shape the evolution of virulence factors in the potato late blight pathogen Phytophthora infestans sampled from China

    PubMed Central

    Wu, E-Jiao; Yang, Li-Na; Zhu, Wen; Chen, Xiao-Mei; Shang, Li-Ping; Zhan, Jiasui

    2016-01-01

    Evolution of virulence in plant pathogens is still poorly understood but the knowledge is important for the effective use of plant resistance and sustainable disease management. Spatial population dynamics of virulence, race and SSR markers in 140 genotypes sampled from seven geographic locations in China were compared to infer the mechanisms driving the evolution of virulence in Phytophthora infestans (P. infestans). All virulence types and a full spectrum of race complexity, ranging from the race able to infect the universally susceptible cultivar only to all differentials, were detected. Eight and two virulence factors were under diversifying and constraining selection respectively while no natural selection was detected in one of the virulence types. Further analyses revealed excesses in simple and complex races but deficiency in intermediate race and negative associations of annual mean temperature at the site from which pathogen isolates were collected with frequency of virulence to differentials and race complexity in the pathogen populations. These results suggest that host selection may interact with other factors such as climatic conditions in determining the evolutionary trajectory of virulence and race structure in P. infestans and global warming may slow down the emergence of new virulence in the pathogen. PMID:27193142

  18. Comparative analyses of fungicide sensitivity and SSR marker variations indicate a low risk of developing azoxystrobin resistance in Phytophthora infestans.

    PubMed

    Qin, Chun-Fang; He, Meng-Han; Chen, Feng-Ping; Zhu, Wen; Yang, Li-Na; Wu, E-Jiao; Guo, Zheng-Liang; Shang, Li-Ping; Zhan, Jiasui

    2016-01-01

    Knowledge of the evolution of fungicide resistance is important in securing sustainable disease management in agricultural systems. In this study, we analyzed and compared the spatial distribution of genetic variation in azoxystrobin sensitivity and SSR markers in 140 Phytophthora infestans isolates sampled from seven geographic locations in China. Sensitivity to azoxystrobin and its genetic variation in the pathogen populations was measured by the relative growth rate (RGR) at four fungicide concentrations and determination of the effective concentration for 50% inhibition (EC50). We found that all isolates in the current study were sensitive to azoxystrobin and their EC50 was similar to that detected from a European population about 20 years ago, suggesting the risk of developing azoxystrobin resistance in P. infestans populations is low. Further analyses indicate that reduced genetic variation and high fitness cost in resistant mutations are the likely causes for the low evolutionary likelihood of developing azoxystrobin resistance in the pathogen. We also found a negative correlation between azoxystrobin tolerance in P. infestans populations and the mean annual temperature of collection sites, suggesting that global warming may increase the efficiency of using the fungicide to control the late blight.

  19. Genetic structure of Phytophthora infestans populations in China indicates multiple migration events.

    PubMed

    Guo, Liyun; Zhu, Xiao-Qiong; Hu, Chia-Hui; Ristaino, Jean Beagle

    2010-10-01

    One hundred isolates of Phytophthora infestans collected from 10 provinces in China between 1998 and 2004 were analyzed for mating type, metalaxyl resistance, mitochondrial DNA (mtDNA) haplotype, allozyme genotype, and restriction fragment length polymorphism (RFLP) with the RG-57 probe. In addition, herbarium samples collected in China, Russia, Australia, and other Asian countries were also typed for mtDNA haplotype. The Ia haplotype was found during the first outbreaks of the disease in China (1938 and 1940), Japan (1901, 1930, and 1931), India (1913), Peninsular Malaysia (1950), Nepal (1954), The Philippines (1910), Australia (1917), Russia (1917), and Latvia (1935). In contrast, the Ib haplotype was found after 1950 in China on both potato and tomato (1952, 1954, 1956, and 1982) and in India (1968 and 1974). Another migration of a genotype found in Siberia called SIB-1 (Glucose-6-phosphate isomerase [Gpi] 100/100, Peptidase [Pep] 100/100, IIa mtDNA haplotype) was identified using RFLP fingerprints among 72% of the isolates and was widely distributed in the north and south of China and has also been reported in Japan. A new genotype named CN-11 (Gpi 100/111, Pep 100/100, IIb mtDNA haplotype), found only in the south of China, and two additional genotypes (Gpi 100/100, Pep 100/100, Ia mtDNA haplotype) named CN-9 and CN-10 were identified. There were more diverse genotypes among isolates from Yunnan province than elsewhere. The SIB-1 (IIa) genotype is identical to those from Siberia, suggesting later migration of this genotype from either Russia or Japan into China. The widespread predominance of SIB-1 suggests that this genotype has enhanced fitness compared with other genotypes found. Movement of the pathogen into China via infected seed from several sources most likely accounts for the distribution of pathogen genotypes observed. MtDNA haplotype evidence and RFLP data suggest multiple migrations of the pathogen into China after the initial introduction of the

  20. Functionally Redundant RXLR Effectors from Phytophthora infestans Act at Different Steps to Suppress Early flg22-Triggered Immunity

    PubMed Central

    Fraiture, Malou; Liu, Xiaoyu; Boevink, Petra C.; Gilroy, Eleanor M.; Chen, Ying; Kandel, Kabindra; Sessa, Guido; Birch, Paul R. J.; Brunner, Frédéric

    2014-01-01

    Genome sequences of several economically important phytopathogenic oomycetes have revealed the presence of large families of so-called RXLR effectors. Functional screens have identified RXLR effector repertoires that either compromise or induce plant defense responses. However, limited information is available about the molecular mechanisms underlying the modes of action of these effectors in planta. The perception of highly conserved pathogen- or microbe-associated molecular patterns (PAMPs/MAMPs), such as flg22, triggers converging signaling pathways recruiting MAP kinase cascades and inducing transcriptional re-programming, yielding a generic anti-microbial response. We used a highly synchronizable, pathogen-free protoplast-based assay to identify a set of RXLR effectors from Phytophthora infestans (PiRXLRs), the causal agent of potato and tomato light blight that manipulate early stages of flg22-triggered signaling. Of thirty-three tested PiRXLR effector candidates, eight, called Suppressor of early Flg22-induced Immune response (SFI), significantly suppressed flg22-dependent activation of a reporter gene under control of a typical MAMP-inducible promoter (pFRK1-Luc) in tomato protoplasts. We extended our analysis to Arabidopsis thaliana, a non-host plant species of P. infestans. From the aforementioned eight SFI effectors, three appeared to share similar functions in both Arabidopsis and tomato by suppressing transcriptional activation of flg22-induced marker genes downstream of post-translational MAP kinase activation. A further three effectors interfere with MAMP signaling at, or upstream of, the MAP kinase cascade in tomato, but not in Arabidopsis. Transient expression of the SFI effectors in Nicotiana benthamiana enhances susceptibility to P. infestans and, for the most potent effector, SFI1, nuclear localization is required for both suppression of MAMP signaling and virulence function. The present study provides a framework to decipher the molecular

  1. Phytophthora infestans RXLR Effector AVR1 Interacts with Exocyst Component Sec5 to Manipulate Plant Immunity1[OPEN

    PubMed Central

    Du, Yu; Mpina, Mohamed H.; Birch, Paul R.J.; Bouwmeester, Klaas; Govers, Francine

    2015-01-01

    Phytophthora infestans secretes numerous RXLR effectors that modulate host defense and thereby pave the way for successful invasion. Here, we show that the RXLR effector AVR1 is a virulence factor that promotes colonization and suppresses callose deposition, a hallmark of basal defense. To identify host targets of AVR1, we performed yeast two-hybrid screens and selected Sec5 as a candidate. Sec5 is a subunit of the exocyst, a protein complex that is involved in vesicle trafficking. AVR1-like (A-L), a close homolog of AVR1, also acts as a virulence factor, but unlike AVR1, A-L does not suppress CRINKLER2 (CRN2)-induced cell death or interact with Sec5. Compared with AVR1, A-L is shorter and lacks the carboxyl-terminal tail, the T-region that is crucial for CRN2-induced cell death suppression and Sec5 interaction. In planta analyses revealed that AVR1 and Sec5 are in close proximity, and coimmunoprecipitation confirmed the interaction. Sec5 is required for secretion of the pathogenesis-related protein PR-1 and callose deposition and also plays a role in CRN2-induced cell death. Our findings show that P. infestans manipulates an exocyst subunit and thereby potentially disturbs vesicle trafficking, a cellular process that is important for basal defense. This is a novel strategy that oomycete pathogens exploit to modulate host defense. PMID:26336092

  2. Mapping of avirulence genes in Phytophthora infestans with amplified fragment length polymorphism markers selected by bulked segregant analysis.

    PubMed Central

    van der Lee, T; Robold, A; Testa, A; van 't Klooster, J W; Govers, F

    2001-01-01

    In this study we investigated the genetic control of avirulence in the diploid oomycete pathogen Phytophthora infestans, the causal agent of late blight on potato. The dominant avirulence (Avr) genes matched six race-specific resistance genes introgressed in potato from a wild Solanum species. AFLP markers linked to Avr genes were selected by bulked segregant analysis and used to construct two high-density linkage maps, one containing Avr4 (located on linkage group A2-a) and the other containing a cluster of three tightly linked genes, Avr3, Avr10, and Avr11 (located on linkage group VIII). Bulked segregant analysis also resulted in a marker linked to Avr1 and this allowed positioning of Avr1 on linkage group IV. No bulked segregant analysis was performed for Avr2, but linkage to a set of random markers placed Avr2 on linkage group VI. Of the six Avr genes, five were located on the most distal part of the linkage group, possibly close to the telomere. The high-density mapping was initiated to facilitate future positional cloning of P. infestans Avr genes. PMID:11238385

  3. The Transcriptome of Compatible and Incompatible Interactions of Potato (Solanum tuberosum) with Phytophthora infestans Revealed by DeepSAGE Analysis

    PubMed Central

    Gyetvai, Gabor; Sønderkær, Mads; Göbel, Ulrike; Basekow, Rico; Ballvora, Agim; Imhoff, Maren; Kersten, Birgit; Nielsen, Kåre-Lehman; Gebhardt, Christiane

    2012-01-01

    Late blight, caused by the oomycete Phytophthora infestans, is the most important disease of potato (Solanum tuberosum). Understanding the molecular basis of resistance and susceptibility to late blight is therefore highly relevant for developing resistant cultivars, either by marker-assissted selection or by transgenic approaches. Specific P. infestans races having the Avr1 effector gene trigger a hypersensitive resistance response in potato plants carrying the R1 resistance gene (incompatible interaction) and cause disease in plants lacking R1 (compatible interaction). The transcriptomes of the compatible and incompatible interaction were captured by DeepSAGE analysis of 44 biological samples comprising five genotypes, differing only by the presence or absence of the R1 transgene, three infection time points and three biological replicates. 30.859 unique 21 base pair sequence tags were obtained, one third of which did not match any known potato transcript sequence. Two third of the tags were expressed at low frequency (<10 tag counts/million). 20.470 unitags matched to approximately twelve thousand potato transcribed genes. Tag frequencies were compared between compatible and incompatible interactions over the infection time course and between compatible and incompatible genotypes. Transcriptional changes were more numerous in compatible than in incompatible interactions. In contrast to incompatible interactions, transcriptional changes in the compatible interaction were observed predominantly for multigene families encoding defense response genes and genes functional in photosynthesis and CO2 fixation. Numerous transcriptional differences were also observed between near isogenic genotypes prior to infection with P. infestans. Our DeepSAGE transcriptome analysis uncovered novel candidate genes for plant host pathogen interactions, examples of which are discussed with respect to possible function. PMID:22328937

  4. A Recent Expansion of the RXLR Effector Gene Avrblb2 Is Maintained in Global Populations of Phytophthora infestans Indicating Different Contributions to Virulence.

    PubMed

    Oliva, Ricardo F; Cano, Liliana M; Raffaele, Sylvain; Win, Joe; Bozkurt, Tolga O; Belhaj, Khaoula; Oh, Sang-Keun; Thines, Marco; Kamoun, Sophien

    2015-08-01

    The introgression of disease resistance (R) genes encoding immunoreceptors with broad-spectrum recognition into cultivated potato appears to be the most promising approach to achieve sustainable management of late blight caused by the oomycete pathogen Phytophthora infestans. Rpi-blb2 from Solanum bulbocastanum shows great potential for use in agriculture based on preliminary potato disease trials. Rpi-blb2 confers immunity by recognizing the P. infestans avirulence effector protein AVRblb2 after it is translocated inside the plant cell. This effector belongs to the RXLR class of effectors and is under strong positive selection. Structure-function analyses revealed a key polymorphic amino acid (position 69) in AVRblb2 effector that is critical for activation of Rpi-blb2. In this study, we reconstructed the evolutionary history of the Avrblb2 gene family and further characterized its genetic structure in worldwide populations. Our data indicate that Avrblb2 evolved as a single-copy gene in a putative ancestral species of P. infestans and has recently expanded in the Phytophthora spp. that infect solanaceous hosts. As a consequence, at least four variants of AVRblb2 arose in P. infestans. One of these variants, with a Phe residue at position 69, evades recognition by the cognate resistance gene. Surprisingly, all Avrblb2 variants are maintained in pathogen populations. This suggests a potential benefit for the pathogen in preserving duplicated versions of AVRblb2, possibly because the variants may have different contributions to pathogen fitness in a diversified solanaceous host environment.

  5. First Report of Haplotype I-b of Phytophthora infestans in central Mexico

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Central Mexico is considered a center of genetic diversity for P. infestans based on a range of genotypic and phenotypic characteristics. Surprisingly, while mtDNA haplotypes I-a, II-a and II-b have been reported from central Mexico, haplotype I-b has not been found in central Mexico. Therefore a mo...

  6. The Full-Size ABCG Transporters Nb-ABCG1 and Nb-ABCG2 Function in Pre- and Postinvasion Defense against Phytophthora infestans in Nicotiana benthamiana.

    PubMed

    Shibata, Yusuke; Ojika, Makoto; Sugiyama, Akifumi; Yazaki, Kazufumi; Jones, David A; Kawakita, Kazuhito; Takemoto, Daigo

    2016-05-01

    The sesquiterpenoid capsidiol is the major phytoalexin produced by Nicotiana and Capsicum species. Capsidiol is produced in plant tissues attacked by pathogens and plays a major role in postinvasion defense by inhibiting pathogen growth. Using virus-induced gene silencing-based screening, we identified two Nicotiana benthamiana (wild tobacco) genes encoding functionally redundant full-size ABCG (PDR-type) transporters, Nb-ABCG1/PDR1 and Nb-ABCG2/PDR2, which are essential for resistance to the potato late blight pathogen Phytophthora infestans Silencing of Nb-ABCG1/2 compromised secretion of capsidiol, revealing Nb-ABCG1/2 as probable exporters of capsidiol. Accumulation of plasma membrane-localized Nb-ABCG1 and Nb-ABCG2 was observed at the site of pathogen penetration. Silencing of EAS (encoding 5-epi-aristolochene synthase), a gene for capsidiol biosynthesis, reduced resistance to P. infestans, but penetration by P. infestans was not affected. By contrast, Nb-ABCG1/2-silenced plants showed reduced penetration defense, indicating that Nb-ABCG1/2 are involved in preinvasion defense against P. infestans Plastidic GGPPS1 (geranylgeranyl diphosphate synthase) was also found to be required for preinvasion defense, thereby suggesting that plastid-produced diterpene(s) are the antimicrobial compounds active in preinvasion defense. These findings suggest that N. benthamiana ABCG1/2 are involved in the export of both antimicrobial diterpene(s) for preinvasion defense and capsidiol for postinvasion defense against P. infestans.

  7. Local adaptation to temperature in populations and clonal lineages of the Irish potato famine pathogen Phytophthora infestans.

    PubMed

    Mariette, Nicolas; Androdias, Annabelle; Mabon, Romain; Corbière, Roselyne; Marquer, Bruno; Montarry, Josselin; Andrivon, Didier

    2016-09-01

    Environmental factors such as temperature strongly impact microbial communities. In the current context of global warming, it is therefore crucial to understand the effects of these factors on human, animal, or plant pathogens. Here, we used a common-garden experiment to analyze the thermal responses of three life-history traits (latent period, lesion growth, spore number) in isolates of the potato late blight pathogen Phytophthora infestans from different climatic zones. We also used a fitness index (FI) aggregating these traits into a single parameter. The experiments revealed patterns of local adaptation to temperature for several traits and for the FI, both between populations and within clonal lineages. Local adaptation to temperature could result from selection for increased survival between epidemics, when isolates are exposed to more extreme climatic conditions than during epidemics. We also showed different thermal responses among two clonal lineages sympatric in western Europe, with lower performances of lineage 13_A2 compared to 6_A1, especially at low temperatures. These data therefore stress the importance of thermal adaptation in a widespread, invasive pathogen, where adaptation is usually considered almost exclusively with respect to host plants. This must now be taken into account to explain, and possibly predict, the global distribution of specific lineages and their epidemic potential. PMID:27648246

  8. Changes in the initial phase of lipid peroxidation induced by elicitor from Phytophthora infestans in Solanum species.

    PubMed

    Polkowska-Kowalczyk, Lidia; Montillet, Jean-Luc; Agnel, Jean-Pierre; Triantaphylidès, Christian; Wielgat, Bernard; Maciejewska, Urszula

    2008-12-01

    The initial phase of the lipid peroxidation process in leaves of Solanum nigrum var. gigantea, Solanum tuberosum cv Bzura and clone H-8105, which represent non-host resistance, field resistance and susceptibility, respectively, against Phytophthora infestans, was investigated. Based on quantitative and qualitative high-performance liquid chromatography (HPLC) analyses of free and esterified fatty acid hydroperoxides (FAHs), we characterized the lipid peroxidation process induced by the pathogen-derived elicitor, culture filtrate (CF), in leaves of the studied genotypes. In all plants, FAHs generated due to 13-lipoxygenase (LOX) action dominated over those from the non-enzymatic pathway. The FAHs derived from 9-LOX activity were found only in CF-treated leaves of the non-host resistant S. nigrum. However, experiments in vitro and in planta with exogenous linoleic acid (LA) as a substrate for LOX revealed high constitutive activity of 9-LOX in all genotypes, which increased in response to CF treatment. The time course changes in polyunsaturated fatty acid (PUFA) pools in the total lipid fractions as well as the degree of their oxidation suggested that CF-induced PUFA peroxidation was enhanced mostly in S. nigrum, less so in Bzura and least in the susceptible clone H-8105. The obtained results are discussed in light of the overall biochemical cell status of plants in the studied interactions.

  9. Four potato (Solanum tuberosum) ABCG transporters and their expression in response to abiotic factors and Phytophthora infestans infection.

    PubMed

    Ruocco, Michelina; Ambrosino, Patrizia; Lanzuise, Stefania; Woo, Sheridan Lois; Lorito, Matteo; Scala, Felice

    2011-12-15

    Pleiotropic drug resistant (PDR/ABCG) genes are involved in plant response to biotic and abiotic stresses. In this work, we cloned, from Solanum tuberosum, four PDR/ABCG transporter genes named StPDR1, StPDR2, StPDR3 and StPDR4, which were differentially expressed in plant tissues and cell cultures. A number of different chemically unrelated compounds were found to regulate the transcript levels of the four genes in cultured cells. In particular, StPDR2 was highly up-regulated in the presence of Botrytis cinerea cell walls, NaCl, 2,4-dichlorophenol, sclareol and α-solanin and biological compounds. The expression of the genes was also investigated by real time RT-PCR during infection by Phytophthora infestans. StPDR1 and StPDR2 were up-regulated about 13- and 37-fold at 48 h post-infection (hpi), StPDR3 was expressed (4-5-fold) at 24 and 48 hpi and then rapidly decreased, while StPDR4 RNA accumulation was stimulated (about 4-fold) at 12 and 24 hpi, decreased at 48 hpi and increased again at 96 hpi. We discuss the role of StPDR1-4 genes in response to pathogens and abiotic stresses.

  10. Quantitative label-free phosphoproteomics of six different life stages of the late blight pathogen Phytophthora infestans reveals abundant phosphorylation of members of the CRN effector family.

    PubMed

    Resjö, Svante; Ali, Ashfaq; Meijer, Harold J G; Seidl, Michael F; Snel, Berend; Sandin, Marianne; Levander, Fredrik; Govers, Francine; Andreasson, Erik

    2014-04-01

    The oomycete Phytophthora infestans is the causal agent of late blight in potato and tomato. Since the underlying processes that govern pathogenicity and development in P. infestans are largely unknown, we have performed a large-scale phosphoproteomics study of six different P. infestans life stages. We have obtained quantitative data for 2922 phosphopeptides and compared their abundance. Life-stage-specific phosphopeptides include ATP-binding cassette transporters and a kinase that only occurs in appressoria. In an extended data set, we identified 2179 phosphorylation sites and deduced 22 phosphomotifs. Several of the phosphomotifs matched consensus sequences of kinases that occur in P. infestans but not Arabidopsis. In addition, we detected tyrosine phosphopeptides that are potential targets of kinases resembling mammalian tyrosine kinases. Among the phosphorylated proteins are members of the RXLR and Crinkler effector families. The latter are phosphorylated in several life stages and at multiple positions, in sites that are conserved between different members of the Crinkler family. This indicates that proteins in the Crinkler family have functions beyond their putative role as (necrosis-inducing) effectors. This phosphoproteomics data will be instrumental for studies on oomycetes and host-oomycete interactions. The data sets have been deposited to ProteomeXchange (identifier PXD000433).

  11. U-box E3 ubiquitin ligase PUB17 acts in the nucleus to promote specific immune pathways triggered by Phytophthora infestans.

    PubMed

    He, Qin; McLellan, Hazel; Boevink, Petra C; Sadanandom, Ari; Xie, Conghua; Birch, Paul R J; Tian, Zhendong

    2015-06-01

    Ubiquitination regulates many processes in plants, including immunity. The E3 ubiquitin ligase PUB17 is a positive regulator of programmed cell death (PCD) triggered by resistance proteins CF4/9 in tomato. Its role in immunity to the potato late blight pathogen, Phytophthora infestans, was investigated here. Silencing StPUB17 in potato by RNAi and NbPUB17 in Nicotiana benthamiana by virus-induced gene silencing (VIGS) each enhanced P. infestans leaf colonization. PAMP-triggered immunity (PTI) transcriptional responses activated by flg22, and CF4/Avr4-mediated PCD were attenuated by silencing PUB17. However, silencing PUB17 did not compromise PCD triggered by P. infestans PAMP INF1, or co-expression of R3a/AVR3a, demonstrating that not all PTI- and PCD-associated responses require PUB17. PUB17 localizes to the plant nucleus and especially in the nucleolus. Transient over-expression of a dominant-negative StPUB17(V314I,V316I) mutant, which retained nucleolar localization, suppressed CF4-mediated cell death and enhanced P. infestans colonization. Exclusion of the StPUB17(V314I,V316I) mutant from the nucleus abolished its dominant-negative activity, demonstrating that StPUB17 functions in the nucleus. PUB17 is a positive regulator of immunity to late blight that acts in the nucleus to promote specific PTI and PCD pathways. PMID:25873665

  12. Alteration of pathogenicity-linked life-history traits by resistance of its host Solanum tuberosum impacts sexual reproduction of the plant pathogenic oomycete Phytophthora infestans.

    PubMed

    Clément, J A J; Magalon, H; Pellé, R; Marquer, B; Andrivon, D

    2010-12-01

    Although sexual reproduction implies a cost, it represents an evolutionary advantage for the adaptation and survival of facultative sexual pathogens. Understanding the maintenance of sex in pathogens requires to analyse how host resistance will impact their sexual reproduction through the alteration of their life-history traits. We explored this experimentally using potato (Solanum tuberosum) and one of its pathogens, the heterothallic oomycete Phytophthora infestans. Sexual reproduction was highest on hosts favouring asexual multiplication of the pathogen, suggesting similar nutritional requirements for both sexual and asexual sporulation. Sexual reproduction was also highest on hosts decreasing the latent period, probably because of a trade-off between growth and reproduction. Distinguishing host effects on each pathogenic trait remains however uneasy, as most life-history traits linked to pathogenicity were not independent of each other. We argue that sexual reproduction of P. infestans is an adaptation to survive when the host is susceptible and rapidly destroyed.

  13. Differences in intensity and specificity of hypersensitive response induction in Nicotiana spp. by INF1, INF2A, and INF2B of Phytophthora infestans.

    PubMed

    Huitema, Edgar; Vleeshouwers, Vivianne G A A; Cakir, Cahit; Kamoun, Sophien; Govers, Francine

    2005-03-01

    Elicitins form a family of structurally related proteins that induce the hypersensitive response (HR) in plants, particularly Nicotiana spp. The elicitin family is composed of several classes. Most species of the plant-pathogenic oomycete genus Phytophthora produce the well-characterized 10-kDa canonical elicitins (class I), such as INF1 of the potato and tomato pathogen Phytophthora infestans. Two genes, inf2A and inf2B, encoding a distinct class (class III) of elicitin-like proteins, also occur in P. infestans. Unlike secreted class I elicitins, class III elicitins are thought to be cell-surface-anchored polypeptides. Molecular characterization of the inf2 genes indicated that they are widespread in Phytophthora spp. and occur as a small gene family. In addition, Southern blot and Northern blot hybridizations using gene-specific probes showed that inf2A and inf2B genes and transcripts can be detected in 17 different P. infestans isolates. Functional secreted expression in plant cells of the elicitin domain of the infl and inf2 genes was conducted using a binary Potato virus X (PVX) vector (agroinfection) and Agrobacterium tumefaciens transient transformation assays (agroinfiltration), and resulted in HR-like necrotic symptoms and induction of defense response genes in tobacco. However, comparative analyses of elicitor activity of INF1, INF2A, and INF2B revealed significant differences in intensity, specificity, and consistency of HR induction. Whereas INF1 induced the HR in Nicotiana benthamiana, INF2A induced weak symptoms and INF2B induced no symptoms on this plant. Nonetheless, similar to INF1, HR induction by INF2A in N. benthamiana required the ubiquitin ligase-associated protein SGT1. Overall, these results suggest that variation in the resistance of Nicotiana spp. to P. infestans is shadowed by variation in the response to INF elicitins. The ability of tobacco, but not N. benthamiana, to respond to INF2B could explain differences in resistance to P

  14. cDNA cloning of sesquiterpene cyclase and squalene synthase, and expression of the genes in potato tuber infected with Phytophthora infestans.

    PubMed

    Yoshioka, H; Yamada, N; Doke, N

    1999-09-01

    Sesquiterpene cyclase and squalene synthase are key branch point enzymes in isoprenoid pathway for the synthesis of sesquiterpenoid phytoalexins and sterols/steroid glycoalkaloids, respectively. cDNA clones encoding these enzymes were isolated from potato. A phylogenetic tree showed that the sesquiterpene cyclase is vetispiradiene synthase. Infection of Phytophthora infestans with potato tubers caused transient increases in the transcript level of vetispiradiene synthase in a compatible and an incompatible interactions. On the other hand, wound-induced expression of the squalene synthase was suppressed in favor of the expression of vetispiradiene synthase regardless of inoculated races. PMID:10588069

  15. Polymorphisms in Phytophthora infestans: Four Mitochondrial Haplotypes Are Detected after PCR Amplification of DNA from Pure Cultures or from Host Lesions

    PubMed Central

    Griffith, Gareth W.; Shaw, David S.

    1998-01-01

    Four pairs of primers were designed for PCR amplification of known polymorphic regions of the mitochondrial genome of Phytophthora infestans. Digestion of the amplified products with restriction enzymes allows identification of previously identified haplotypes. Product P2 cut with MspI uniquely identifies haplotypes Ib and IIa, while types Ia and IIb are differentiated by digestion of product P4 with EcoRI. Digestion of products P1 and P3 gave results similar to that with digestion of P4, but amplification of these products was less robust. Thus, all four common haplotypes are identified by amplifying and digesting products P2 and P4. Identification of haplotypes was also possible from DNA extracted directly from small, late-blight lesions on both tomato and potato leaves, making isolation of the fungus unnecessary. A rapid and efficient method of monitoring changes in the pathogen population is facilitated. These PCR primers were also useful for differentiating other Phytophthora species. PMID:9758833

  16. Potato NPH3/RPT2-Like Protein StNRL1, Targeted by a Phytophthora infestans RXLR Effector, Is a Susceptibility Factor1[OPEN

    PubMed Central

    Yang, Lina; Naqvi, Shaista; Boevink, Petra C.; Armstrong, Miles; Giuliani, Licida M.; Zhan, Jiasui; Birch, Paul R.J.

    2016-01-01

    Plant pathogens deliver effectors to manipulate host processes. We know little about how fungal and oomycete effectors target host proteins to promote susceptibility, yet such knowledge is vital to understand crop disease. We show that either transient expression in Nicotiana benthamiana, or stable transgenic expression in potato (Solanum tuberosum), of the Phytophthora infestans RXLR effector Pi02860 enhances leaf colonization by the pathogen. Expression of Pi02860 also attenuates cell death triggered by the P. infestans microbe-associated molecular pattern INF1, indicating that the effector suppresses pattern-triggered immunity. However, the effector does not attenuate cell death triggered by Cf4/Avr4 coexpression, showing that it does not suppress all cell death activated by cell surface receptors. Pi02860 interacts in yeast two-hybrid assays with potato NPH3/RPT2-LIKE1 (NRL1), a predicted CULLIN3-associated ubiquitin E3 ligase. Interaction of Pi02860 in planta was confirmed by coimmunoprecipitation and bimolecular fluorescence complementation assays. Virus-induced gene silencing of NRL1 in N. benthamiana resulted in reduced P. infestans colonization and accelerated INF1-mediated cell death, indicating that this host protein acts as a negative regulator of immunity. Moreover, whereas NRL1 virus-induced gene silencing had no effect on the ability of the P. infestans effector Avr3a to suppress INF1-mediated cell death, such suppression by Pi02860 was significantly attenuated, indicating that this activity of Pi02860 is mediated by NRL1. Transient overexpression of NRL1 resulted in the suppression of INF1-mediated cell death and enhanced P. infestans leaf colonization, demonstrating that NRL1 acts as a susceptibility factor to promote late blight disease. PMID:26966171

  17. Potato NPH3/RPT2-Like Protein StNRL1, Targeted by a Phytophthora infestans RXLR Effector, Is a Susceptibility Factor.

    PubMed

    Yang, Lina; McLellan, Hazel; Naqvi, Shaista; He, Qin; Boevink, Petra C; Armstrong, Miles; Giuliani, Licida M; Zhang, Wei; Tian, Zhendong; Zhan, Jiasui; Gilroy, Eleanor M; Birch, Paul R J

    2016-05-01

    Plant pathogens deliver effectors to manipulate host processes. We know little about how fungal and oomycete effectors target host proteins to promote susceptibility, yet such knowledge is vital to understand crop disease. We show that either transient expression in Nicotiana benthamiana, or stable transgenic expression in potato (Solanum tuberosum), of the Phytophthora infestans RXLR effector Pi02860 enhances leaf colonization by the pathogen. Expression of Pi02860 also attenuates cell death triggered by the P. infestans microbe-associated molecular pattern INF1, indicating that the effector suppresses pattern-triggered immunity. However, the effector does not attenuate cell death triggered by Cf4/Avr4 coexpression, showing that it does not suppress all cell death activated by cell surface receptors. Pi02860 interacts in yeast two-hybrid assays with potato NPH3/RPT2-LIKE1 (NRL1), a predicted CULLIN3-associated ubiquitin E3 ligase. Interaction of Pi02860 in planta was confirmed by coimmunoprecipitation and bimolecular fluorescence complementation assays. Virus-induced gene silencing of NRL1 in N. benthamiana resulted in reduced P. infestans colonization and accelerated INF1-mediated cell death, indicating that this host protein acts as a negative regulator of immunity. Moreover, whereas NRL1 virus-induced gene silencing had no effect on the ability of the P. infestans effector Avr3a to suppress INF1-mediated cell death, such suppression by Pi02860 was significantly attenuated, indicating that this activity of Pi02860 is mediated by NRL1. Transient overexpression of NRL1 resulted in the suppression of INF1-mediated cell death and enhanced P. infestans leaf colonization, demonstrating that NRL1 acts as a susceptibility factor to promote late blight disease. PMID:26966171

  18. Potato NPH3/RPT2-Like Protein StNRL1, Targeted by a Phytophthora infestans RXLR Effector, Is a Susceptibility Factor.

    PubMed

    Yang, Lina; McLellan, Hazel; Naqvi, Shaista; He, Qin; Boevink, Petra C; Armstrong, Miles; Giuliani, Licida M; Zhang, Wei; Tian, Zhendong; Zhan, Jiasui; Gilroy, Eleanor M; Birch, Paul R J

    2016-05-01

    Plant pathogens deliver effectors to manipulate host processes. We know little about how fungal and oomycete effectors target host proteins to promote susceptibility, yet such knowledge is vital to understand crop disease. We show that either transient expression in Nicotiana benthamiana, or stable transgenic expression in potato (Solanum tuberosum), of the Phytophthora infestans RXLR effector Pi02860 enhances leaf colonization by the pathogen. Expression of Pi02860 also attenuates cell death triggered by the P. infestans microbe-associated molecular pattern INF1, indicating that the effector suppresses pattern-triggered immunity. However, the effector does not attenuate cell death triggered by Cf4/Avr4 coexpression, showing that it does not suppress all cell death activated by cell surface receptors. Pi02860 interacts in yeast two-hybrid assays with potato NPH3/RPT2-LIKE1 (NRL1), a predicted CULLIN3-associated ubiquitin E3 ligase. Interaction of Pi02860 in planta was confirmed by coimmunoprecipitation and bimolecular fluorescence complementation assays. Virus-induced gene silencing of NRL1 in N. benthamiana resulted in reduced P. infestans colonization and accelerated INF1-mediated cell death, indicating that this host protein acts as a negative regulator of immunity. Moreover, whereas NRL1 virus-induced gene silencing had no effect on the ability of the P. infestans effector Avr3a to suppress INF1-mediated cell death, such suppression by Pi02860 was significantly attenuated, indicating that this activity of Pi02860 is mediated by NRL1. Transient overexpression of NRL1 resulted in the suppression of INF1-mediated cell death and enhanced P. infestans leaf colonization, demonstrating that NRL1 acts as a susceptibility factor to promote late blight disease.

  19. Polyethylene mulch modifies greenhouse microclimate and reduces infection of phytophthora infestans in tomato and Pseudoperonospora cubensis in cucumber.

    PubMed

    Shtienberg, D; Elad, Y; Bornstein, M; Ziv, G; Grava, A; Cohen, S

    2010-01-01

    The individual and joint effects of covering the soil with polyethylene mulch before planting and fungicides commonly used by organic growers on tomato late blight (caused by Phytophthora infestans) were studied in three experiments conducted from 2002 to 2005. Application of fungicides resulted in inconsistent and insufficient late blight suppression (control efficacy +/- standard error of 34.5 +/- 14.3%) but the polyethylene mulch resulted in consistent, effective, and highly significant suppression (control efficacy of 83.6 +/- 5.5%) of the disease. The combined effect of the two measures was additive. In a second set of three experiments carried out between 2004 and 2006, it was found that the type of polyethylene mulch used (bicolor aluminized, clear, or black) did not affect the efficacy of late blight suppression (control efficacy of 60.1 to 95.8%) and the differences in the effects among the different polyethylene mulches used were insignificant. Next, the ability of the mulch to suppress cucumber downy mildew (caused by Pseudoperonospora cubensis) was studied in four experiments carried out between 2006 and 2008. The mulch effectively suppressed cucumber downy mildew but the effect was less substantial (control efficacy of 34.9 +/- 4.8%) than that achieved for tomato late blight. The disease-suppressing effect of mulch appeared to come from a reduction in leaf wetness duration, because mulching led to reductions in both the frequency of nights when dew formed and the number of dew hours per night when it formed. Mulching also reduced relative humidity in the canopy, which may have reduced sporulation.

  20. Genomic analyses of dominant U.S. clonal lineages of Phytophthora infestans reveals a shared common ancestry for clonal lineages US11 and US18 and a lack of recently shared ancestry among all other U.S. lineages

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The populations of the potato and tomato late blight pathogen, Phytophthora infestans, in the US are well known for emerging repeatedly as novel clonal lineages. These successions of dominant clones have historically been named US1-US24, in order of appearance, since their first characterization usi...

  1. Revealing the importance of meristems and roots for the development of hypersensitive responses and full foliar resistance to Phytophthora infestans in the resistant potato cultivar Sarpo Mira.

    PubMed

    Orłowska, Elzbieta; Basile, Alessio; Kandzia, Izabela; Llorente, Briardo; Kirk, Hanne Grethe; Cvitanich, Cristina

    2012-08-01

    The defence responses of potato against Phytophthora infestans were studied using the highly resistant Sarpo Mira cultivar. The effects of plant integrity, meristems, and roots on the hypersensitive response (HR), plant resistance, and the regulation of PR genes were analysed. Sarpo Mira shoots and roots grafted with the susceptible Bintje cultivar as well as non-grafted different parts of Sarpo Mira plants were inoculated with P. infestans. The progress of the infection and the number of HR lesions were monitored, and the regulation of PR genes was compared in detached and attached leaves. Additionally, the antimicrobial activity of plant extracts was assessed. The presented data show that roots are needed to achieve full pathogen resistance, that the removal of meristems in detached leaves inhibits the formation of HR lesions, that PR genes are differentially regulated in detached leaves compared with leaves of whole plants, and that antimicrobial compounds accumulate in leaves and roots of Sarpo Mira plants challenged with P. infestans. While meristems are necessary for the formation of HR lesions, the roots of Sarpo Mira plants participate in the production of defence-associated compounds that increase systemic resistance. Based on the literature and on the presented results, a model is proposed for mechanisms involved in Sarpo Mira resistance that may apply to other resistant potato cultivars.

  2. Nucleoporin 75 is involved in the ethylene-mediated production of phytoalexin for the resistance of Nicotiana benthamiana to Phytophthora infestans.

    PubMed

    Ohtsu, Mina; Shibata, Yusuke; Ojika, Makoto; Tamura, Kentaro; Hara-Nishimura, Ikuko; Mori, Hitoshi; Kawakita, Kazuhito; Takemoto, Daigo

    2014-12-01

    Mature Nicotiana benthamiana shows stable resistance to the oomycete pathogen Phytophthora infestans. Induction of phytoalexin (capsidiol) production is essential for the resistance, which is upregulated via a mitogen-activated protein kinase (MAPK) cascade (NbMEK2-WIPK/SIPK) followed by ethylene signaling. In this study, NbNup75 (encodes a nuclear pore protein Nucleoporin75) was identified as an essential gene for resistance of N. benthamiana to P. infestans. In NbNup75-silenced plants, initial events of elicitor-induced responses such as phosphorylation of MAPK and expression of defense-related genes were not affected, whereas induction of later defense responses such as capsidiol production and cell death induction was suppressed or delayed. Ethylene production induced by either INF1 or NbMEK2 was reduced in NbNup75-silenced plants, whereas the expression of NbEAS (a gene for capsidiol biosynthesis) induced by ethylene was not affected, indicating that Nup75 is required for the induction of ethylene production but not for ethylene signaling. Given that nuclear accumulation of polyA RNA was increased in NbNup75-silenced plants, efficient export of mRNA from nuclei via nuclear pores would be important for the timely upregulation of defense responses. Collectively, Nup75 is involved in the induction of a later stage of defense responses, including the ethylene-mediated production of phytoalexin for the resistance of N. benthamiana to P. infestans.

  3. Loop-mediated isothermal amplification for detection of the tomato and potato late blight pathogen, Phytophthora infestans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aims: To design and validate a colorimetric loop-mediated isothermal amplification assay for rapid detection of P. infestans DNA. Methods and Results: Two sets of LAMP primers were designed and evaluated for their sensitivity and specificity for P. infestans. ITSII primers targeted a portion of the ...

  4. [Regulation of peroxidase activity under the influence of signaling molecules and Bacillus subtilis 26D in potato plants infected with Phytophthora infestans].

    PubMed

    Maksimov, I V; Abizgil'dina, R R; Sorokan', A V; Burkhanova, G F

    2014-01-01

    The influence of sequential exposure of 5 x 10(-5) M salicylic acid (SA) or 1 x 10(-7) M jasmonic acid (JA) and endophytic bacterium Bacillus subtilis strain 26D on peroxidase activity, transcription of the M21334 isoperoxidase gene from potato (Solarium tuberosum L.), and the formation of resistance to the infective agent of potato blight Phytophthora infestans (Mont.) de Bary was studied. It was found that individual application of JA or Bacillus subtilis 26D and sequential application of SA and B. subtilis 26D were the most effective in protecting plants against pathogens, while sequential application of JA and B. subtilis 26D drastically suppressed plant resistance. The results suggest the need for strict compliance with regulations when using SA and JA, as well as biological products based on living bacteria as modern plant protection products with immunomodulatory properties that trigger specific signaling pathways, which often interfere with each other. PMID:25272739

  5. And the nasty ones lose in the end: foliar pathogenicity trades off with asexual transmission in the Irish famine pathogen Phytophthora infestans.

    PubMed

    Pasco, Claudine; Montarry, Josselin; Marquer, Bruno; Andrivon, Didier

    2016-01-01

    A trade-off between pathogenicity and transmission is often postulated to explain the persistence of pathogens over time. If demonstrated, it would help to predict the evolution of pathogenicity across cropping seasons, and to develop sustainable control strategies from this prediction. Unfortunately, experimental demonstration of such trade-offs in agricultural plant pathogens remains elusive. We measured asexual transmission of Phytophthora infestans isolates differing in pathogenicity in two sets of artificial infection experiments under controlled, semi-outdoor conditions. Higher foliar pathogenicity decreased mean daughter tuber weight, increased infection severity in daughter tubers, and increased stem mortality before emergence. The most pathogenic isolates thus suffer a double penalty for asexual transmission: a lower survival probability within small and severely infected tubers; and a lower infection probability of neighbouring healthy plants due to fewer infected stems produced by surviving tubers. Moderate tuber resistance favoured transmission of the least pathogenic isolates, while high levels of resistance almost abolished transmission of all isolates. These data demonstrate a trade-off between foliar pathogenicity and asexual transmission over seasons in P. infestans, which should stabilise pathogenicity over time in the potato late blight pathosystem and possibly favour clone replacement by less pathogenic lineages after demographic bottlenecks.

  6. Tomato I2 Immune Receptor Can Be Engineered to Confer Partial Resistance to the Oomycete Phytophthora infestans in Addition to the Fungus Fusarium oxysporum.

    PubMed

    Giannakopoulou, Artemis; Steele, John F C; Segretin, Maria Eugenia; Bozkurt, Tolga O; Zhou, Ji; Robatzek, Silke; Banfield, Mark J; Pais, Marina; Kamoun, Sophien

    2015-12-01

    Plants and animals rely on immune receptors, known as nucleotide-binding domain and leucine-rich repeat (NLR)-containing proteins, to defend against invading pathogens and activate immune responses. How NLR receptors respond to pathogens is inadequately understood. We previously reported single-residue mutations that expand the response of the potato immune receptor R3a to AVR3a(EM), a stealthy effector from the late blight oomycete pathogen Phytophthora infestans. I2, another NLR that mediates resistance to the will-causing fungus Fusarium oxysporum f. sp. lycopersici, is the tomato ortholog of R3a. We transferred previously identified R3a mutations to I2 to assess the degree to which the resulting I2 mutants have an altered response. We discovered that wild-type I2 protein responds weakly to AVR3a. One mutant in the N-terminal coiled-coil domain, I2(I141N), appeared sensitized and displayed markedly increased response to AVR3a. Remarkably, I2(I141N) conferred partial resistance to P. infestans. Further, I2(I141N) has an expanded response spectrum to F. oxysporum f. sp. lycopersici effectors compared with the wild-type I2 protein. Our results suggest that synthetic immune receptors can be engineered to confer resistance to phylogenetically divergent pathogens and indicate that knowledge gathered for one NLR could be exploited to improve NLR from other plant species.

  7. Increased frequency of self-fertile isolates in Phytophthora infestans may attribute to their higher fitness relative to the A1 isolates

    PubMed Central

    Zhu, Wen; Shen, Lin-Lin; Fang, Zhi-Guo; Yang, Li-Na; Zhang, Jia-Feng; Sun, Dan-Li; Zhan, Jiasui

    2016-01-01

    Knowledge of population dynamics of mating types is important for better understanding pathogen’s evolutionary potential and sustainable management of natural and chemical resources such as host resistances and fungicides. In this study, 2250 Phytophthora infestans isolates sampled from 61 fields across China were assayed for spatiotemporal dynamics of mating type frequency. Self-fertile isolates dominated in ~50% of populations and all but one cropping region with an average frequency of 0.64 while no A2 isolates were detected. Analyses of 140 genotypes consisting of 82 self-fertile and 58 A1 isolates indicated that on average self-fertile isolates grew faster, demonstrated higher aggressiveness and were more tolerant to fungicides than A1 isolates; Furthermore, pattern of association between virulence complexity (defined as the number of differential cultivars on which an isolate can induce disease) and frequency was different in the two mating types. In A1 isolates, virulence complexity was negatively correlated (r = −0.515, p = 0.043) with frequency but this correlation was positive (r = 0.532, p = 0.037) in self-fertile isolates. Our results indicate a quick increase of self-fertile isolates possibly attributable to their higher fitness relative to A1 mating type counterpart in the field populations of P. infestans in China. PMID:27384813

  8. Identification of three elicitins and a galactan-based complex polysaccharide from a concentrated culture filtrate of Phytophthora infestans efficient against Pectobacterium atrosepticum.

    PubMed

    Saubeau, Guillaume; Gaillard, Fanny; Legentil, Laurent; Nugier-Chauvin, Caroline; Ferrières, Vincent; Andrivon, Didier; Val, Florence

    2014-01-01

    The induction of plant immunity by Pathogen Associated Molecular Patterns (PAMPs) constitutes a powerful strategy for crop protection. PAMPs indeed induce general defense responses in plants and thus increase plant resistance to pathogens. Phytophthora infestans culture filtrates (CCFs) are known to induce defense responses and decrease the severity of soft rot due to Pectobacterium atrosepticum in potato tubers. The aim of this study was to identify and characterize the active compounds from P. infestans filtrate. The filtrate was fractionated by gel filtration, and the protection effects against P. atrosepticum and the ability to induce PAL activity were tested for each fraction. The fraction active in protection (F1) also induced PAL activity, as did the whole filtrate. Three elicitins (INF1, INF4 and INF5) were identified in F1b, subfraction of F1, by MALDI-TOF-MS and MS/MS analyses. However, deproteinized F1b still showed biological activity against the bacterium, revealing the presence of an additional active compound. GC-MS analyses of the deproteinized fraction highlighted the presence of a galactan-based complex polysaccharide. These experiments demonstrate that the biological activity of the CCF against P. atrosepticum results from a combined action of three elicitins and a complex polysaccharide, probably through the activation of general defense responses. PMID:25264828

  9. Phytophthora infestans RXLR-WY Effector AVR3a Associates with Dynamin-Related Protein 2 Required for Endocytosis of the Plant Pattern Recognition Receptor FLS2.

    PubMed

    Chaparro-Garcia, Angela; Schwizer, Simon; Sklenar, Jan; Yoshida, Kentaro; Petre, Benjamin; Bos, Jorunn I B; Schornack, Sebastian; Jones, Alexandra M E; Bozkurt, Tolga O; Kamoun, Sophien

    2015-01-01

    Pathogens utilize effectors to suppress basal plant defense known as PTI (Pathogen-associated molecular pattern-triggered immunity). However, our knowledge of PTI suppression by filamentous plant pathogens, i.e. fungi and oomycetes, remains fragmentary. Previous work revealed that the co-receptor BAK1/SERK3 contributes to basal immunity against the potato pathogen Phytophthora infestans. Moreover BAK1/SERK3 is required for the cell death induced by P. infestans elicitin INF1, a protein with characteristics of PAMPs. The P. infestans host-translocated RXLR-WY effector AVR3a is known to supress INF1-mediated cell death by binding the plant E3 ligase CMPG1. In contrast, AVR3aKI-Y147del, a deletion mutant of the C-terminal tyrosine of AVR3a, fails to bind CMPG1 and does not suppress INF1-mediated cell death. Here, we studied the extent to which AVR3a and its variants perturb additional BAK1/SERK3-dependent PTI responses in N. benthamiana using the elicitor/receptor pair flg22/FLS2 as a model. We found that all tested variants of AVR3a suppress defense responses triggered by flg22 and reduce internalization of activated FLS2. Moreover, we discovered that AVR3a associates with the Dynamin-Related Protein 2 (DRP2), a plant GTPase implicated in receptor-mediated endocytosis. Interestingly, silencing of DRP2 impaired ligand-induced FLS2 internalization but did not affect internalization of the growth receptor BRI1. Our results suggest that AVR3a associates with a key cellular trafficking and membrane-remodeling complex involved in immune receptor-mediated endocytosis. We conclude that AVR3a is a multifunctional effector that can suppress BAK1/SERK3-mediated immunity through at least two different pathways. PMID:26348328

  10. Utilizing “Omic” Technologies to Identify and Prioritize Novel Sources of Resistance to the Oomycete Pathogen Phytophthora infestans in Potato Germplasm Collections

    PubMed Central

    Van Weymers, Pauline S. M.; Baker, Katie; Chen, Xinwei; Harrower, Brian; Cooke, David E. L.; Gilroy, Eleanor M.; Birch, Paul R. J.; Thilliez, Gaëtan J. A.; Lees, Alison K.; Lynott, James S.; Armstrong, Miles R.; McKenzie, Gaynor; Bryan, Glenn J.; Hein, Ingo

    2016-01-01

    The greatest threat to potato production world-wide is late blight, caused by the oomycete pathogen Phytophthora infestans. A screen of 126 wild diploid Solanum accessions from the Commonwealth Potato Collection (CPC) with P. infestans isolates belonging to the genotype 13-A2 identified resistances in the species S. bulbocastanum, S. capsicibaccatum, S. microdontum, S. mochiquense, S. okadae, S. pinnatisectum, S. polyadenium, S. tarijense, and S. verrucosum. Effector-omics, allele mining, and diagnostic RenSeq (dRenSeq) were utilized to investigate the nature of resistances in S. okadae accessions. dRenSeq in resistant S. okadae accessions 7129, 7625, 3762, and a bulk of 20 resistant progeny confirmed the presence of full-length Rpi-vnt1.1 under stringent mapping conditions and corroborated allele mining results in the accessions 7129 and 7625 as well as Avr-vnt1 recognition in transient expression assays. In contrast, susceptible S. okadae accession 3761 and a bulk of 20 susceptible progeny lacked sequence homology in the 5′ end compared to the functional Rpi-vnt1.1 gene. Further evaluation of S. okadae accessions with P. infestans isolates that have a broad spectrum of virulence demonstrated that, although S. okadae accessions 7129, 7625, and 7629 contain functional Rpi-vnt1.1, they also carry a novel resistance gene. We provide evidence that existing germplasm collections are important sources of novel resistances and that “omic” technologies such as dRenSeq-based genomics and effector-omics are efficacious tools to rapidly explore the diversity within these collections. PMID:27303410

  11. Phytophthora infestans RXLR-WY Effector AVR3a Associates with Dynamin-Related Protein 2 Required for Endocytosis of the Plant Pattern Recognition Receptor FLS2

    PubMed Central

    Chaparro-Garcia, Angela; Schwizer, Simon; Sklenar, Jan; Yoshida, Kentaro; Petre, Benjamin; Bos, Jorunn I. B.; Schornack, Sebastian; Jones, Alexandra M. E.; Bozkurt, Tolga O.; Kamoun, Sophien

    2015-01-01

    Pathogens utilize effectors to suppress basal plant defense known as PTI (Pathogen-associated molecular pattern-triggered immunity). However, our knowledge of PTI suppression by filamentous plant pathogens, i.e. fungi and oomycetes, remains fragmentary. Previous work revealed that the co-receptor BAK1/SERK3 contributes to basal immunity against the potato pathogen Phytophthora infestans. Moreover BAK1/SERK3 is required for the cell death induced by P. infestans elicitin INF1, a protein with characteristics of PAMPs. The P. infestans host-translocated RXLR-WY effector AVR3a is known to supress INF1-mediated cell death by binding the plant E3 ligase CMPG1. In contrast, AVR3aKI-Y147del, a deletion mutant of the C-terminal tyrosine of AVR3a, fails to bind CMPG1 and does not suppress INF1-mediated cell death. Here, we studied the extent to which AVR3a and its variants perturb additional BAK1/SERK3-dependent PTI responses in N. benthamiana using the elicitor/receptor pair flg22/FLS2 as a model. We found that all tested variants of AVR3a suppress defense responses triggered by flg22 and reduce internalization of activated FLS2. Moreover, we discovered that AVR3a associates with the Dynamin-Related Protein 2 (DRP2), a plant GTPase implicated in receptor-mediated endocytosis. Interestingly, silencing of DRP2 impaired ligand-induced FLS2 internalization but did not affect internalization of the growth receptor BRI1. Our results suggest that AVR3a associates with a key cellular trafficking and membrane-remodeling complex involved in immune receptor-mediated endocytosis. We conclude that AVR3a is a multifunctional effector that can suppress BAK1/SERK3-mediated immunity through at least two different pathways. PMID:26348328

  12. Utilizing "Omic" Technologies to Identify and Prioritize Novel Sources of Resistance to the Oomycete Pathogen Phytophthora infestans in Potato Germplasm Collections.

    PubMed

    Van Weymers, Pauline S M; Baker, Katie; Chen, Xinwei; Harrower, Brian; Cooke, David E L; Gilroy, Eleanor M; Birch, Paul R J; Thilliez, Gaëtan J A; Lees, Alison K; Lynott, James S; Armstrong, Miles R; McKenzie, Gaynor; Bryan, Glenn J; Hein, Ingo

    2016-01-01

    The greatest threat to potato production world-wide is late blight, caused by the oomycete pathogen Phytophthora infestans. A screen of 126 wild diploid Solanum accessions from the Commonwealth Potato Collection (CPC) with P. infestans isolates belonging to the genotype 13-A2 identified resistances in the species S. bulbocastanum, S. capsicibaccatum, S. microdontum, S. mochiquense, S. okadae, S. pinnatisectum, S. polyadenium, S. tarijense, and S. verrucosum. Effector-omics, allele mining, and diagnostic RenSeq (dRenSeq) were utilized to investigate the nature of resistances in S. okadae accessions. dRenSeq in resistant S. okadae accessions 7129, 7625, 3762, and a bulk of 20 resistant progeny confirmed the presence of full-length Rpi-vnt1.1 under stringent mapping conditions and corroborated allele mining results in the accessions 7129 and 7625 as well as Avr-vnt1 recognition in transient expression assays. In contrast, susceptible S. okadae accession 3761 and a bulk of 20 susceptible progeny lacked sequence homology in the 5' end compared to the functional Rpi-vnt1.1 gene. Further evaluation of S. okadae accessions with P. infestans isolates that have a broad spectrum of virulence demonstrated that, although S. okadae accessions 7129, 7625, and 7629 contain functional Rpi-vnt1.1, they also carry a novel resistance gene. We provide evidence that existing germplasm collections are important sources of novel resistances and that "omic" technologies such as dRenSeq-based genomics and effector-omics are efficacious tools to rapidly explore the diversity within these collections. PMID:27303410

  13. Targeted and Untargeted Approaches Unravel Novel Candidate Genes and Diagnostic SNPs for Quantitative Resistance of the Potato (Solanum tuberosum L.) to Phytophthora infestans Causing the Late Blight Disease.

    PubMed

    Mosquera, Teresa; Alvarez, Maria Fernanda; Jiménez-Gómez, José M; Muktar, Meki Shehabu; Paulo, Maria João; Steinemann, Sebastian; Li, Jinquan; Draffehn, Astrid; Hofmann, Andrea; Lübeck, Jens; Strahwald, Josef; Tacke, Eckhard; Hofferbert, Hans-Reinhardt; Walkemeier, Birgit; Gebhardt, Christiane

    2016-01-01

    The oomycete Phytophthora infestans causes late blight of potato, which can completely destroy the crop. Therefore, for the past 160 years, late blight has been the most important potato disease worldwide. The identification of cultivars with high and durable field resistance to P. infestans is an objective of most potato breeding programs. This type of resistance is polygenic and therefore quantitative. Its evaluation requires multi-year and location trials. Furthermore, quantitative resistance to late blight correlates with late plant maturity, a negative agricultural trait. Knowledge of the molecular genetic basis of quantitative resistance to late blight not compromised by late maturity is very limited. It is however essential for developing diagnostic DNA markers that facilitate the efficient combination of superior resistance alleles in improved cultivars. We used association genetics in a population of 184 tetraploid potato cultivars in order to identify single nucleotide polymorphisms (SNPs) that are associated with maturity corrected resistance (MCR) to late blight. The population was genotyped for almost 9000 SNPs from three different sources. The first source was candidate genes specifically selected for their function in the jasmonate pathway. The second source was novel candidate genes selected based on comparative transcript profiling (RNA-Seq) of groups of genotypes with contrasting levels of quantitative resistance to P. infestans. The third source was the first generation 8.3k SolCAP SNP genotyping array available in potato for genome wide association studies (GWAS). Twenty seven SNPs from all three sources showed robust association with MCR. Some of those were located in genes that are strong candidates for directly controlling quantitative resistance, based on functional annotation. Most important were: a lipoxygenase (jasmonate pathway), a 3-hydroxy-3-methylglutaryl coenzyme A reductase (mevalonate pathway), a P450 protein (terpene biosynthesis

  14. Targeted and Untargeted Approaches Unravel Novel Candidate Genes and Diagnostic SNPs for Quantitative Resistance of the Potato (Solanum tuberosum L.) to Phytophthora infestans Causing the Late Blight Disease

    PubMed Central

    Jiménez-Gómez, José M.; Muktar, Meki Shehabu; Paulo, Maria João; Steinemann, Sebastian; Li, Jinquan; Draffehn, Astrid; Hofmann, Andrea; Lübeck, Jens; Strahwald, Josef; Tacke, Eckhard; Hofferbert, Hans-Reinhardt; Walkemeier, Birgit; Gebhardt, Christiane

    2016-01-01

    The oomycete Phytophthora infestans causes late blight of potato, which can completely destroy the crop. Therefore, for the past 160 years, late blight has been the most important potato disease worldwide. The identification of cultivars with high and durable field resistance to P. infestans is an objective of most potato breeding programs. This type of resistance is polygenic and therefore quantitative. Its evaluation requires multi-year and location trials. Furthermore, quantitative resistance to late blight correlates with late plant maturity, a negative agricultural trait. Knowledge of the molecular genetic basis of quantitative resistance to late blight not compromised by late maturity is very limited. It is however essential for developing diagnostic DNA markers that facilitate the efficient combination of superior resistance alleles in improved cultivars. We used association genetics in a population of 184 tetraploid potato cultivars in order to identify single nucleotide polymorphisms (SNPs) that are associated with maturity corrected resistance (MCR) to late blight. The population was genotyped for almost 9000 SNPs from three different sources. The first source was candidate genes specifically selected for their function in the jasmonate pathway. The second source was novel candidate genes selected based on comparative transcript profiling (RNA-Seq) of groups of genotypes with contrasting levels of quantitative resistance to P. infestans. The third source was the first generation 8.3k SolCAP SNP genotyping array available in potato for genome wide association studies (GWAS). Twenty seven SNPs from all three sources showed robust association with MCR. Some of those were located in genes that are strong candidates for directly controlling quantitative resistance, based on functional annotation. Most important were: a lipoxygenase (jasmonate pathway), a 3-hydroxy-3-methylglutaryl coenzyme A reductase (mevalonate pathway), a P450 protein (terpene biosynthesis

  15. Targeted and Untargeted Approaches Unravel Novel Candidate Genes and Diagnostic SNPs for Quantitative Resistance of the Potato (Solanum tuberosum L.) to Phytophthora infestans Causing the Late Blight Disease.

    PubMed

    Mosquera, Teresa; Alvarez, Maria Fernanda; Jiménez-Gómez, José M; Muktar, Meki Shehabu; Paulo, Maria João; Steinemann, Sebastian; Li, Jinquan; Draffehn, Astrid; Hofmann, Andrea; Lübeck, Jens; Strahwald, Josef; Tacke, Eckhard; Hofferbert, Hans-Reinhardt; Walkemeier, Birgit; Gebhardt, Christiane

    2016-01-01

    The oomycete Phytophthora infestans causes late blight of potato, which can completely destroy the crop. Therefore, for the past 160 years, late blight has been the most important potato disease worldwide. The identification of cultivars with high and durable field resistance to P. infestans is an objective of most potato breeding programs. This type of resistance is polygenic and therefore quantitative. Its evaluation requires multi-year and location trials. Furthermore, quantitative resistance to late blight correlates with late plant maturity, a negative agricultural trait. Knowledge of the molecular genetic basis of quantitative resistance to late blight not compromised by late maturity is very limited. It is however essential for developing diagnostic DNA markers that facilitate the efficient combination of superior resistance alleles in improved cultivars. We used association genetics in a population of 184 tetraploid potato cultivars in order to identify single nucleotide polymorphisms (SNPs) that are associated with maturity corrected resistance (MCR) to late blight. The population was genotyped for almost 9000 SNPs from three different sources. The first source was candidate genes specifically selected for their function in the jasmonate pathway. The second source was novel candidate genes selected based on comparative transcript profiling (RNA-Seq) of groups of genotypes with contrasting levels of quantitative resistance to P. infestans. The third source was the first generation 8.3k SolCAP SNP genotyping array available in potato for genome wide association studies (GWAS). Twenty seven SNPs from all three sources showed robust association with MCR. Some of those were located in genes that are strong candidates for directly controlling quantitative resistance, based on functional annotation. Most important were: a lipoxygenase (jasmonate pathway), a 3-hydroxy-3-methylglutaryl coenzyme A reductase (mevalonate pathway), a P450 protein (terpene biosynthesis

  16. Population dynamics of Phytophthora infestans in the Netherlands reveals expansion and spread of dominant clonal lineages and virulence in sexual offspring.

    PubMed

    Li, Y; van der Lee, T A J; Evenhuis, A; van den Bosch, G B M; van Bekkum, P J; Förch, M G; van Gent-Pelzer, M P E; van Raaij, H M G; Jacobsen, E; Huang, S W; Govers, F; Vleeshouwers, V G A A; Kessel, G J T

    2012-12-01

    For a comprehensive survey of the structure and dynamics of the Dutch Phytophthora infestans population, 652 P. infestans isolates were collected from commercial potato fields in the Netherlands during the 10-year period 2000-2009. Genotyping was performed using 12 highly informative microsatellite markers and mitochondrial haplotypes. In addition, for each isolate, the mating type was determined. STRUCTURE analysis grouped the 322 identified genotypes in three clusters. Cluster 1 consists of a single clonal lineage NL-001, known as "Blue_13"; all isolates in this cluster have the A2 mating type and the Ia mitochondrial haplotype. Clusters 2 and 3 display a more elaborate substructure containing many unique genotypes. In Cluster 3, several distinct clonal lineages were also identified. This survey witnesses that the Dutch population underwent dramatic changes in the 10 years under study. The most notable change was the emergence and spread of A2 mating type strain NL-001 (or "Blue_13"). The results emphasize the importance of the sexual cycle in generating genetic diversity and the importance of the asexual cycle as the propagation and dispersal mechanism for successful genotypes. Isolates were also screened for absence of the Avrblb1/ipiO class I gene, which is indicative for virulence on Rpi-blb1. This is also the first report of Rpi-blb1 breakers in the Netherlands. Superimposing the virulence screening on the SSR genetic backbone indicates that lack the Avrblb1/ipiO class I gene only occurred in sexual progeny. So far, the asexual spread of the virulent isolates identified has been limited.

  17. Selection and validation of potato candidate genes for maturity corrected resistance to Phytophthora infestans based on differential expression combined with SNP association and linkage mapping

    PubMed Central

    Muktar, Meki S.; Lübeck, Jens; Strahwald, Josef; Gebhardt, Christiane

    2015-01-01

    Late blight of potato (Solanum tuberosum L.) caused by the oomycete Phytophthora infestans (Mont.) de Bary, is one of the most important bottlenecks of potato production worldwide. Cultivars with high levels of durable, race unspecific, quantitative resistance are part of a solution to this problem. However, breeding for quantitative resistance is hampered by the correlation between resistance and late plant maturity, which is an undesirable agricultural attribute. The objectives of our research are (i) the identification of genes that condition quantitative resistance to P. infestans not compromised by late plant maturity and (ii) the discovery of diagnostic single nucleotide polymorphism (SNP) markers to be used as molecular tools to increase efficiency and precision of resistance breeding. Twenty two novel candidate genes were selected based on comparative transcript profiling by SuperSAGE (serial analysis of gene expression) in groups of plants with contrasting levels of maturity corrected resistance (MCR). Reproducibility of differential expression was tested by quantitative real time PCR and allele specific pyrosequencing in four new sets of genotype pools with contrasting late blight resistance levels, at three infection time points and in three independent infection experiments. Reproducibility of expression patterns ranged from 28 to 97%. Association mapping in a panel of 184 tetraploid cultivars identified SNPs in five candidate genes that were associated with MCR. These SNPs can be used in marker-assisted resistance breeding. Linkage mapping in two half-sib families (n = 111) identified SNPs in three candidate genes that were linked with MCR. The differentially expressed genes that showed association and/or linkage with MCR putatively function in phytosterol synthesis, fatty acid synthesis, asparagine synthesis, chlorophyll synthesis, cell wall modification, and in the response to pathogen elicitors. PMID:26442110

  18. Population Dynamics of Phytophthora infestans in the Netherlands Reveals Expansion and Spread of Dominant Clonal Lineages and Virulence in Sexual Offspring

    PubMed Central

    Li, Y.; van der Lee, T. A. J.; Evenhuis, A.; van den Bosch, G. B. M.; van Bekkum, P. J.; Förch, M. G.; van Gent-Pelzer, M. P. E; van Raaij, H. M. G.; Jacobsen, E.; Huang, S. W.; Govers, F.; Vleeshouwers, V. G. A. A.; Kessel, G. J. T.

    2012-01-01

    For a comprehensive survey of the structure and dynamics of the Dutch Phytophthora infestans population, 652 P. infestans isolates were collected from commercial potato fields in the Netherlands during the 10-year period 2000–2009. Genotyping was performed using 12 highly informative microsatellite markers and mitochondrial haplotypes. In addition, for each isolate, the mating type was determined. STRUCTURE analysis grouped the 322 identified genotypes in three clusters. Cluster 1 consists of a single clonal lineage NL-001, known as “Blue_13”; all isolates in this cluster have the A2 mating type and the Ia mitochondrial haplotype. Clusters 2 and 3 display a more elaborate substructure containing many unique genotypes. In Cluster 3, several distinct clonal lineages were also identified. This survey witnesses that the Dutch population underwent dramatic changes in the 10 years under study. The most notable change was the emergence and spread of A2 mating type strain NL-001 (or “Blue_13”). The results emphasize the importance of the sexual cycle in generating genetic diversity and the importance of the asexual cycle as the propagation and dispersal mechanism for successful genotypes. Isolates were also screened for absence of the Avrblb1/ipiO class I gene, which is indicative for virulence on Rpi-blb1. This is also the first report of Rpi-blb1 breakers in the Netherlands. Superimposing the virulence screening on the SSR genetic backbone indicates that lack the Avrblb1/ipiO class I gene only occurred in sexual progeny. So far, the asexual spread of the virulent isolates identified has been limited. PMID:23275876

  19. Knowing your Phytophthora

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora pathogens are known as some of the most important plant killers known to man. Two particularly notorious killers include the Irish potato famine pathogen P. infestans affecting potato and tomato and the sudden oak death P. ramorum affecting woody ornamentals and trees. Phytophthora path...

  20. Novel core promoter elements in the oomycete pathogen Phytophthora infestans and their influence on expression detected by genome-wide analysis

    PubMed Central

    2013-01-01

    Background The core promoter is the region flanking the transcription start site (TSS) that directs formation of the pre-initiation complex. Core promoters have been studied intensively in mammals and yeast, but not in more diverse eukaryotes. Here we investigate core promoters in oomycetes, a group within the Stramenopile kingdom that includes important plant and animal pathogens. Prior studies of a small collection of genes proposed that oomycete core promoters contain a 16 to 19 nt motif bearing an Initiator-like sequence (INR) flanked by a novel sequence named FPR, but this has not been extended to whole-genome analysis. Results We used expectation maximization to find over-represented motifs near TSSs of Phytophthora infestans, the potato blight pathogen. The motifs corresponded to INR, FPR, and a new element found about 25 nt downstream of the TSS called DPEP. TATA boxes were not detected. Assays of DPEP function by mutagenesis were consistent with its role as a core motif. Genome-wide searches found a well-conserved combined INR+FPR in only about 13% of genes after correcting for false discovery, which contradicted prior reports that INR and FPR are found together in most genes. INR or FPR were found alone near TSSs in 18% and 7% of genes, respectively. Promoters lacking the motifs had pyrimidine-rich regions near the TSS. The combined INR+FPR motif was linked to higher than average mRNA levels, developmentally-regulated transcription, and functions related to plant infection, while DPEP and FPR were over-represented in constitutively-expressed genes. The INR, FPR, and combined INR+FPR motifs were detected in other oomycetes including Hyaloperonospora arabidopsidis, Phytophthora sojae, Pythium ultimum, and Saprolegnia parasitica, while DPEP was found in all but S. parasitica. Only INR seemed present in a non-oomycete stramenopile. Conclusions The absence of a TATA box and presence of novel motifs show that the oomycete core promoter is diverged from that of

  1. Curdlan β-1,3-glucooligosaccharides induce the defense responses against Phytophthora infestans infection of potato (Solanum tuberosum L. cv. McCain G1) leaf cells.

    PubMed

    Li, Jing; Zhu, Li; Lu, Guangxing; Zhan, Xiao-Bei; Lin, Chi-Chung; Zheng, Zhi-Yong

    2014-01-01

    Activation of the innate immune system before the invasion of pathogens is a promising way to improve the resistance of plant against infection while reducing the use of agricultural chemicals. Although several elicitors were used to induce the resistance of potato plant to microbial pathogen infection, the role of curdlan oligosaccharide (CurdO) has not been established. In the current study, the defense responses were investigated at biochemical and proteomic levels to elucidate the elicitation effect of CurdOs in foliar tissues of potato (Solanum tuberosum L. cv. McCain G1). The results indicate that the CurdOs exhibit activation effect on the early- and late-defense responses in potato leaves. In addition, glucopentaose was proved to be the shortest active curdlan molecule based on the accumulation of H₂O₂ and salicylic acid and the activities of phenylalanine amino-lyase, β-1,3-glucanase and chitinase. The 2D-PAGE analysis reveals that CurdOs activate the integrated response reactions in potato cells, as a number of proteins with various functions are up-regulated including disease/defense, metabolism, transcription, and cell structure. The pathogenesis assay shows that the ratio of lesion area of potato leaf decreased from 15.82%±5.44% to 7.79%±3.03% when the plants were treated with CurdOs 1 day before the infection of Phytophthora infestans. Furthermore, the results on potato yield and induction reactions indicate that the defense responses induced by CurdOs lasted for short period of time but disappeared gradually.

  2. Interaction between the moss Physcomitrella patens and Phytophthora: a novel pathosystem for live-cell imaging of subcellular defence.

    PubMed

    Overdijk, Elysa J R; DE Keijzer, Jeroen; DE Groot, Deborah; Schoina, Charikleia; Bouwmeester, Klaas; Ketelaar, Tijs; Govers, Francine

    2016-08-01

    Live-cell imaging of plant-pathogen interactions is often hampered by the tissue complexity and multicell layered nature of the host. Here, we established a novel pathosystem with the moss Physcomitrella patens as host for Phytophthora. The tip-growing protonema cells of this moss are ideal for visualizing interactions with the pathogen over time using high-resolution microscopy. We tested four Phytophthora species for their ability to infect P. patens and showed that P. sojae and P. palmivora were only rarely capable to infect P. patens. In contrast, P. infestans and P. capsici frequently and successfully penetrated moss protonemal cells, showed intracellular hyphal growth and formed sporangia. Next to these successful invasions, many penetration attempts failed. Here the pathogen was blocked by a barrier of cell wall material deposited in papilla-like structures, a defence response that is common in higher plants. Another common response is the upregulation of defence-related genes upon infection and also in moss we observed this upregulation in tissues infected with Phytophthora. For more advanced analyses of the novel pathosystem we developed a special set-up that allowed live-cell imaging of subcellular defence processes by high-resolution microscopy. With this set-up, we revealed that Phytophthora infection of moss induces repositioning of the nucleus, accumulation of cytoplasm and rearrangement of the actin cytoskeleton, but not of microtubules.

  3. Phytophthora infestans in the USA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This book chapter is specific to late blight in the United States and will include a review and discussion of the history of late blight on potato and tomato crops, changes in grower attitudes towards late blight, present status of the disease in the US, methods for identification, management, and c...

  4. Rpi-blb2-Mediated Hypersensitive Cell Death Caused by Phytophthora infestans AVRblb2 Requires SGT1, but not EDS1, NDR1, Salicylic Acid-, Jasmonic Acid-, or Ethylene-Mediated Signaling.

    PubMed

    Oh, Sang-Keun; Kwon, Suk-Yoon; Choi, Doil

    2014-09-01

    Potato Rpi-blb2 encodes a protein with a coiled-coil-nucleotide binding site and leucine-rich repeat (CC-NBS-LRR) motif that recognizes the Phytophthora infestans AVRblb2 effector and triggers hypersensitive cell death (HCD). To better understand the components required for Rpi-blb2-mediated HCD in plants, we used virus-induced gene silencing to repress candidate genes in Rpi-blb2-transgenic Nicotiana benthamiana plants and assayed the plants for AVRblb2 effector. Rpi-blb2 triggers HCD through NbSGT1-mediated pathways, but not NbEDS1- or NbNDR1-mediated pathways. In addition, the role of salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) in Rpi-blb2-mediated HCD were analyzed by monitoring of the responses of NbICS1-, NbCOI1-, or NbEIN2-silenced or Rpi-blb2::NahG-transgenic plants. Rpi-blb2-mediated HCD in response to AVRblb2 was not associated with SA accumulation. Thus, SA affects Rpi-blb2-mediated resistance against P. infestans, but not Rpi-blb2-mediated HCD in response to AVRblb2. Additionally, JA and ET signaling were not required for Rpi-blb2-mediated HCD in N. benthamiana. Taken together, these findings suggest that NbSGT1 is a unique positive regulator of Rpi-blb2-mediated HCD in response to AVRblb2, but EDS1, NDR1, SA, JA, and ET are not required.

  5. Sporulation capacity of Phytophthora ramorum on northern red oak and chestnut oak

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Branches from six 2 to 3-year old northern red and chestnut oak seedlings were dip-inoculated with ca. 5,000 sporangia per milliliter of Phytophthora ramorum isolate Pr-6 and incubated at 100 percent relative humidity in dew chambers for 6 days. Three plants were then used to assess sporangia produ...

  6. Infectivity and sporulation of Phytophthora ramorum on northern red oak and chestnut oak

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Branches from northern red and chestnut oak seedlings were dip-inoculated with 5,000 sporangia per milliliter of Phytophthora ramorum and incubated at 100 percent relative humidity in dew chambers for 6 days. Three plants were then used to assess sporangia production, while the other three plants w...

  7. Transient Induction of Phenylalanine Ammonia-Lyase and 4-Coumarate: CoA Ligase mRNAs in Potato Leaves Infected with Virulent or Avirulent Races of Phytophthora infestans1

    PubMed Central

    Fritzemeier, Karl-Heinrich; Cretin, Claude; Kombrink, Erich; Rohwer, Frauke; Taylor, Janet; Scheel, Dierk; Hahlbrock, Klaus

    1987-01-01

    Infection of potato leaves with the fungal pathogen Phytophthora infestans (Pi) resulted in the rapid stimulation of phenylpropanoid metabolism. Increases in the activities of several mRNAs, including those encoding phenylalanine ammonia-lyase (PAL) and 4-coumarate:CoA ligase (4CL), were detectable within a few hours postinoculation, as demonstrated by two-dimensional gel electrophoresis of proteins synthesized in vitro. This effect was closely mimicked by application of Pi culture filtrate through cut leaf stems. PAL and 4CL mRNA activities were also rapidly and transiently induced in potato cell suspension cultures by treatments with Pi culture filtrate or arachidonic acid. This induction was exploited to generate cDNA probes complementary to PAL and 4CL mRNAs. Blot hybridizations using these probes revealed almost immediate, transient and coordinate increases in the transcription rates and subsequent changes in the amounts of PAL and 4CL mRNAs in leaves treated with Pi culture filtrate. Similar changes in the mRNA amounts were found in infected leaves of potato cultivars carrying resistance genes R1 (cv Datura) or R4 (cv Isola), independent of whether a virulent or an avirulent Pi pathotype was used for inoculation. These results are discussed in relation to recent cytological observations with the same potato cultivars and Pi pathotypes. Images Fig. 1 Fig. 4 Fig. 5 Fig. 6 PMID:16665678

  8. Two novel species representing a new clade and cluster of Phytophthora.

    PubMed

    Yang, Xiao; Copes, Warren E; Hong, Chuanxue

    2014-01-01

    Phytophthora stricta sp. nov. and Phytophthora macilentosa sp. nov. are described based on morphological, physiological and molecular characters in this study. Phytophthora stricta represents a previously unknown clade in the rRNA internal transcribed spacer (ITS)-based phylogeny. Phytophthora macilentosa, along with nine other species, consistently forms a high temperature-tolerant cluster within ITS clade 9. These observations are supported by the sequence analysis of the mitochondrial cytochrome c oxidase 1 gene. Both species are heterothallic and all examined isolates are A1 mating type. Phytophthora stricta produces nonpapillate and slightly caducous sporangia. This species is named after its characteristic constrictions on sporangiophores. Phytophthora macilentosa produces nonpapillate and noncaducous sporangia, which are mostly elongated obpyriform with a high length to breadth ratio. Both species were recovered from irrigation water of an ornamental plant nursery in Mississippi, USA and P. stricta was also recovered from stream water in Virginia, USA.

  9. Phytophthora irrigata, a new species isolated from irrigation reservoirs and rivers in Eastern United States of America.

    PubMed

    Hong, Chuanxue; Gallegly, Mannon E; Richardson, Patricia A; Kong, Ping; Moorman, Gary W

    2008-08-01

    A new species of Phytophthora, previously referred to as the 'Dre I' taxon, is named Phytophthora irrigata. Isolates of P. irrigata morphologically and physiologically resemble Phytophthora drechsleri. They are heterothallic, produce nonpapillate sporangia, and grow well at 35 degrees C. The above two species differ significantly in uniformity of mycelium, presence of chlamydospores, DNA fingerprint, and sequences of multiple nuclear and mtDNA regions. Phytophthora irrigata produces smaller sporangia and a distinct DNA fingerprint. Sequence alignments in the ribosomal DNA internal transcribed spacer regions place Phytophthora fallax and Phytophthora captisoa as its closest relatives. The optimal temperature for culture growth is above 30 degrees C and the maximum temperature is 40 degrees C. This new species is abundant in irrigation reservoirs and natural waterways in Virginia and was also isolated in Pennsylvania.

  10. Examination of some morphologically unusual cultures of Phytophthora species using a mitochondrial DNA miniprep technique and a standardised sporangium caducity assessment.

    PubMed

    Hall, G S

    Using the mitochondrial DNA miniprep technique, the identity of sixteen morphologically unusual cultures allocated to Phytophthora nicotianae, Phytophthora mexicana or Phytophthora porri was determined by comparison with a library of mtDNA band patterns obtained from reference cultures. Seven cultures were identified as Phytophthora nicotianae (including those assigned to Phytophthora mexicana and Phytophthora porri), six as strains of Phytophthora palmivora with small, ovoid, weakly caducous sporangia, and one as Phytophthora citrophthora. Some cultures of P. nicotianae had a low percentage of caducous sporangia. Percentage sporangium caducity, but not sporangium L:B ratio, is considered a useful taxonomic criterion for separating species morphologically similar to Phytophthora nicotianae. One culture from tobacco in New Zealand had a highly unusual morphology and a unique DNA band pattern, but was not identifiable. One culture from Acacia mearnsii in South Africa had a unique DNA band pattern which was identical to that of an isolate from Annona squamosa from Australia previously identified as Phytophthora palmivora, the precise identity of which is still unclear. The identity of most isolates from diseased durian was found to be Phytophthora palmivora, confirming its role as the main pathogen, but P. nicotianae was also identified from this host.

  11. New role for Cdc14 phosphatase: localization to basal bodies in the oomycete phytophthora and its evolutionary coinheritance with eukaryotic flagella.

    PubMed

    Ah-Fong, Audrey M V; Judelson, Howard S

    2011-01-01

    Cdc14 protein phosphatases are well known for regulating the eukaryotic cell cycle, particularly during mitosis. Here we reveal a distinctly new role for Cdc14 based on studies of the microbial eukaryote Phytophthora infestans, the Irish potato famine agent. While Cdc14 is transcribed constitutively in yeast and animal cells, the P. infestans ortholog is expressed exclusively in spore stages of the life cycle and not in vegetative hyphae where the bulk of mitosis takes place. PiCdc14 expression is first detected in nuclei at sporulation, and during zoospore formation the protein accumulates at the basal body, which is the site from which flagella develop. The association of PiCdc14 with basal bodies was supported by co-localization studies with the DIP13 basal body protein and flagellar β-tubulin, and by demonstrating the enrichment of PiCdc14 in purified flagella-basal body complexes. Overexpressing PiCdc14 did not cause defects in growth or mitosis in hyphae, but interfered with cytoplasmic partitioning during zoosporogenesis. This cytokinetic defect might relate to its ability to bind microtubules, which was shown using an in vitro cosedimentation assay. The use of gene silencing to reveal the precise function of PiCdc14 in flagella is not possible since we showed previously that silencing prevents the formation of the precursor stage, sporangia. Nevertheless, the association of Cdc14 with flagella and basal bodies is consistent with their phylogenetic distribution in eukaryotes, as species that lack the ability to produce flagella generally also lack Cdc14. An ancestral role of Cdc14 in the flagellar stage of eukaryotes is thereby proposed.

  12. Production and release of asexual sporangia in Plasmopara viticola.

    PubMed

    Caffi, Tito; Gilardi, Giovanna; Monchiero, Matteo; Rossi, Vittorio

    2013-01-01

    To study the influence of environmental conditions on sporulation of Plasmopara viticola lesions under vineyard's conditions, unsprayed vines were inspected every second or third day and the numbers of sporulating and nonsporulating lesions were counted in two North Italy vineyards in 2008 to 2010. Infected leaves were removed so that only fresh lesions were assessed at each field assessment. Sporulation was studied at two scales, across field assessments and across the seasonal population of lesions. Frequencies of sporulating lesions were positively correlated with the numbers of moist hours in the preceding dark period (i.e., the number of hours between 8:00 p.m. and 7:00 a.m. with relative humidity ≥80%, rainfall >0 mm, or wetness duration >30 min). In a receiver operating characteristic analysis, predicted sporulation based on the occurrence of ≥3 moist hours at night provided overall accuracy of 0.85. To study the time course of sporulation on lesions which were not washed by rainfall, numbers of sporangia produced per square millimeter of lesion were estimated on individual cohorts of lesions over the whole infectious period. The numbers of sporangia per square millimeter of lesion increased rapidly during the first 4 days after the beginning of sporulation and then tapered off prior to a halt; the time course of cumulative sporangia production by a lesion followed a monomolecular growth model (R(2) = 0.97). The total number of sporangia produced by a square millimeter of lesion increased as the maximum temperature decreased and moist hours in the dark increased. To study the release pattern of the sporangia, spore samplers were placed near grapevines with sporulating lesions. Airborne sporangia were caught in 91.2% of the days over a wide range of weather conditions, including rainless periods. The results of this study provide quantitative information on production of P. viticola sporangia that may help refine epidemiological models used as decision

  13. Effects of inoculum density and wounding on stem infection of three Eastern U.S. forest species by Phytophthora ramorum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Seedlings of three Eastern US forest species (red maple, northern red oak, and chestnut oak) were inoculated by applying Phytophthora ramorum sporangia to stems at different inoculum densities with and without wounding. Disease occurred in all treatments involving wounds, and no disease was observe...

  14. Defining species boundaries in the genus Phytophthora: the case of Phytophthora andina. A response to “Phytophthora andina sp. nov., a newly identified heterothallic pathogen of solanaceous hosts in the Andean highlands

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The newly described species Phytophthora andina is a relative of the potato late blight pathogen P. infestans. The formal P. andina species description is based on three types of evidence. First, the fact that these Ecuadorian isolates were found causing disease on different wild Solanum spp. that a...

  15. Development of PCR primers from internal transcribed spacer region 2 for detection of Phytophthora species infecting potatoes.

    PubMed Central

    Tooley, P W; Bunyard, B A; Carras, M M; Hatziloukas, E

    1997-01-01

    We developed PCR primers and assay methods to detect and differentiate three Phytophthora species which infect potatoes and cause late blight (Phytophthora infestans) and pink rot (P. erythroseptica and P. nicotianae) diseases. Primers based on sequence analysis of internal transcribed spacer region 2 of ribosomal DNA produced PCR products of 456 bp (P. infestans), 136 bp (P. erythroseptica), and 455 bp (P. nicotianae) and were used to detect the pathogens in potato leaf (P. infestans) and tuber (P. infestans, P. erythroseptica, and P. nicotianae) tissue with a sensitivity of 1 to 10 pg of DNA. Leaf and tuber tissue were processed for PCR by a rapid NaOH method as well as a method based on the use of commercially available ion-exchange columns of P. infestans primers and the rapid NaOH extraction method were used to detect late blight in artificially and naturally infected tubers of potato cultivar Red LaSoda. In sampling studies, P. infestans was detected by PCR from artificially infected tubers at 4 days postinoculation, before any visible symptoms were present. The PCR assay and direct tissue extraction methods provide tools which may be used to detect Phytophthora pathogens in potato seedlots and storages and thus limit the transmission and spread of new, aggressive strains of P. infestans in U.S. potato-growing regions. PMID:9097445

  16. Phytophthora ramorum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora ramorum is a recently emerged plant pathogen and causal agent of one of the most destructive and devastating diseases currently affecting US horticulture and forests. Formally described in 2001, P. ramorum is a filamentous, diploid protozoan that is one of 117 currently recognized Phyto...

  17. Detection of a quantitative trait locus for both foliage and tuber resistance to late blight [Phytophthora infestans (Mont.) de Bary] on chromosome 4 of a dihaploid potato clone (Solanum tuberosum subsp. tuberosum).

    PubMed

    Bradshaw, John E; Hackett, Christine A; Lowe, Robert; McLean, Karen; Stewart, Helen E; Tierney, Irene; Vilaro, Marco D R; Bryan, Glenn J

    2006-09-01

    Linkage analysis, Kruskal-Wallis analysis, interval mapping and graphical genotyping were performed on a potato diploid backcross family comprising 120 clones segregating for resistance to late blight. A hybrid between the Solanum tuberosum dihaploid clone PDH247 and the long-day-adapted S. phureja clone DB226(70) had been crossed to DB226(70) to produce the backcross family. Eighteen AFLP primer combinations provided 186 and 123 informative maternal and paternal markers respectively, with 63 markers in common to both parents. Eleven microsatellite (SSR) markers proved useful for identifying chromosomes. Linkage maps of both backcross parents were constructed. The results of a Kruskal-Wallis analysis, interval mapping and graphical genotyping were all consistent with a QTL or QTLs for blight resistance between two AFLP markers 30 cM apart on chromosome 4, which was identified by a microsatellite marker. The simplest explanation of the results is a single QTL with an allele from the dihaploid parent conferring resistance to race 1, 4 of P. infestans in the foliage in the glasshouse and to race 1, 2, 3, 4, 6, 7 in the foliage in the field and in tubers from glasshouse raised plants. The QTL was of large effect, and explained 78 and 51% of the variation in phenotypic scores for foliage blight in the glasshouse and field respectively, as well as 27% of the variation in tuber blight. Graphical genotyping and the differences in blight scores between the parental clones showed that all of the foliage blight resistance is accounted for by chromosome 4, whereas undetected QTLs for tuber resistance probably exist on other chromosomes. Graphical genotyping also explained the lack of precision in mapping the QTL(s) in terms of lack of appropriate recombinant chromosomes.

  18. Mitochondrial genome sequences reveal evolutionary relationships of the Phytophthora 1c clade species.

    PubMed

    Lassiter, Erica S; Russ, Carsten; Nusbaum, Chad; Zeng, Qiandong; Saville, Amanda C; Olarte, Rodrigo A; Carbone, Ignazio; Hu, Chia-Hui; Seguin-Orlando, Andaine; Samaniego, Jose A; Thorne, Jeffrey L; Ristaino, Jean B

    2015-11-01

    Phytophthora infestans is one of the most destructive plant pathogens of potato and tomato globally. The pathogen is closely related to four other Phytophthora species in the 1c clade including P. phaseoli, P. ipomoeae, P. mirabilis and P. andina that are important pathogens of other wild and domesticated hosts. P. andina is an interspecific hybrid between P. infestans and an unknown Phytophthora species. We have sequenced mitochondrial genomes of the sister species of P. infestans and examined the evolutionary relationships within the clade. Phylogenetic analysis indicates that the P. phaseoli mitochondrial lineage is basal within the clade. P. mirabilis and P. ipomoeae are sister lineages and share a common ancestor with the Ic mitochondrial lineage of P. andina. These lineages in turn are sister to the P. infestans and P. andina Ia mitochondrial lineages. The P. andina Ic lineage diverged much earlier than the P. andina Ia mitochondrial lineage and P. infestans. The presence of two mitochondrial lineages in P. andina supports the hybrid nature of this species. The ancestral state of the P. andina Ic lineage in the tree and its occurrence only in the Andean regions of Ecuador, Colombia and Peru suggests that the origin of this species hybrid in nature may occur there.

  19. Phytophthora species, new threats to the plant health in Korea.

    PubMed

    Hyun, Ik-Hwa; Choi, Woobong

    2014-12-01

    Given the lack of a resistant genetic pool in host plants, the introduction of exotic invasive pathogens can result in epidemics that affect a specific ecosystem and economy. Plant quarantine, which is designed to protect endemic plant resources, is a highly invaluable safeguard that should keep biosecurity with increasing international trade and global transportation. A total of 34 species of plant pathogens including Phytophthora infestans were documented as introduced from other countries into Korea from 1900 to 2010. The genus Phytophthora, classified in oomycetes, includes more than 120 species that are mostly recognized worldwide as highly invasive plant pathogens. After 2000, over 50 new species of Phytophthora were identified internationally as plant pathogens occurring in crops and forest trees. In Korea, Phytophthora is also one of the most serious plant pathogens. To date, 22 species (about one-fifth of known species) of the genus have been identified and reported as plant pathogens in the country. The likelihood of new exotic Phytophthora species being introduced into Korea continues to increase, thus necessitating intensive plant quarantine inspections. As new potential threats to plant health in Korea, six Phytophthora species, namely, P. alni, P. inundata, P. kernoviae, P. pinifolia, P. quercina, and P. ramorum, are discussed in this review with focus on history, disease, biology, management, and plant quarantine issues.

  20. Phytophthora Species, New Threats to the Plant Health in Korea

    PubMed Central

    Hyun, Ik-Hwa; Choi, Woobong

    2014-01-01

    Given the lack of a resistant genetic pool in host plants, the introduction of exotic invasive pathogens can result in epidemics that affect a specific ecosystem and economy. Plant quarantine, which is designed to protect endemic plant resources, is a highly invaluable safeguard that should keep biosecurity with increasing international trade and global transportation. A total of 34 species of plant pathogens including Phytophthora infestans were documented as introduced from other countries into Korea from 1900 to 2010. The genus Phytophthora, classified in oomycetes, includes more than 120 species that are mostly recognized worldwide as highly invasive plant pathogens. After 2000, over 50 new species of Phytophthora were identified internationally as plant pathogens occurring in crops and forest trees. In Korea, Phytophthora is also one of the most serious plant pathogens. To date, 22 species (about one-fifth of known species) of the genus have been identified and reported as plant pathogens in the country. The likelihood of new exotic Phytophthora species being introduced into Korea continues to increase, thus necessitating intensive plant quarantine inspections. As new potential threats to plant health in Korea, six Phytophthora species, namely, P. alni, P. inundata, P. kernoviae, P. pinifolia, P. quercina, and P. ramorum, are discussed in this review with focus on history, disease, biology, management, and plant quarantine issues. PMID:25506298

  1. Evaluation of Watermelon Germplasm for Resistance to Phytophthora Blight Caused by Phytophthora capsici

    PubMed Central

    Kim, Min-Jeong; Shim, Chang-Ki; Kim, Yong-Ki; Jee, Hyeong-Jin; Hong, Sung-Jun; Park, Jong-Ho; Han, Eun-Jung

    2013-01-01

    This study was conducted to determine the Phytophthora rot resistance of 514 accessions of watermelon germplasm, Citrullus lanatus var lanatus. About 46% of the 514 accessions tested were collections from Uzbekistan, Turkey, China, U.S.A., and Ukraine. Phytophthora capsici was inoculated to 45-day-old watermelon seedlings by drenching with 5 ml of sporangial suspension (106 sporangia/ml). At 7 days after inoculation, 21 accessions showed no disease symptoms while 291 accessions of susceptible watermelon germplasm showed more than 60.1% disease severity. A total of 510 accessions of watermelon germplasm showed significant disease symptoms and were rated as susceptible to highly susceptible 35 days after inoculation. The highly susceptible watermelon germplasm exhibited white fungal hyphae on the lesion or damping off with water-soaked and browning symptoms. One accession (IT032840) showed moderate resistance and two accessions (IT185446 and IT187904) were resistant to P. capsici. Results suggest that these two resistant germplasm can be used as a rootstock and as a source of resistance in breeding resistant watermelon varieties against Phytophthora. PMID:25288932

  2. Phytophthora terminalis sp. nov. and Phytophthora occultans sp. nov., two invasive pathogens of ornamental plants in Europe.

    PubMed

    Man In 't Veld, Willem A; Rosendahl, Karin C H M; van Rijswick, Patricia C J; Meffert, Johan P; Westenberg, Marcel; van de Vossenberg, Bart T L H; Denton, Geoff; van Kuik, Fons A J

    2015-01-01

    In the past decade several Phytophthora strains were isolated from diseased Pachysandra terminalis plants suffering stem base and root rot, originating from the Netherlands and Belgium. All isolates were homothallic and had a felt-like colony pattern, produced semi-papillate sporangia, globose oogonia and had a maximum growth at ~ 27 C. Several additional Phytophthora strains were isolated from diseased Buxus sempervirens plants, originating from the Netherlands and Belgium, which had sustained stem base and root rot; similar strains also were isolated from Acer palmatum, Choisya ternata and Taxus in the United Kingdom. All isolates were homothallic and had a stellate colony pattern, produced larger semi-papillate sporangia and smaller globose oogonia than the isolates from Pa. terminalis and had a maximum growth temperature of ~ 30 C. Phylogenetic analyses of both species using the internal transcribed spacer region of the nuc rDNA (ITS), mt cytochrome oxidases subunit I gene (CoxI) and nuc translation elongation factor 1-α gene (TEF1α) revealed that all sequences of each species were identical at each locus and unique to that species, forming two distinct clusters in subclade 2a. Sequence analysis of partial β-tubulin genes showed that both taxa share an identical sequence that is identical to that of Ph. himalsilva, a species originating from Asia, suggesting a common Asian origin. Pathogenicity trials demonstrated disease symptoms on their respective hosts, and re-isolation and re-identification of the inoculated pathogens confirmed Koch's postulates.

  3. Five reasons to consider Phytophthora infestans a reemerging pathogen

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Late blight disease of potato and tomato (and several other Solanaceous plants) has emerged and re-emerged so many times, that it might be logical to conclude that nothing new could be said about this disease. And yet, much continues to be said (and written). In the year 2007, a search for Phyto...

  4. Phytophthora suppressor of RNA silencing 2 is a conserved RxLR effector that promotes infection in soybean and Arabidopsis thaliana.

    PubMed

    Xiong, Qin; Ye, Wenwu; Choi, Duseok; Wong, James; Qiao, Yongli; Tao, Kai; Wang, Yuanchao; Ma, Wenbo

    2014-12-01

    The genus Phytophthora consists of notorious and emerging pathogens of economically important crops. Each Phytophthora genome encodes several hundreds of cytoplasmic effectors, which are believed to manipulate plant immune response inside the host cells. However, the majority of Phytophthora effectors remain functionally uncharacterized. We recently discovered two effectors from the soybean stem and root rot pathogen Phytophthora sojae with the activity to suppress RNA silencing in plants. These effectors are designated Phytophthora suppressor of RNA silencing (PSRs). Here, we report that the P. sojae PSR2 (PsPSR2) belongs to a conserved and widespread effector family in Phytophthora. A PsPSR2-like effector produced by P. infestans (PiPSR2) can also suppress RNA silencing in plants and promote Phytophthora infection, suggesting that the PSR2 family effectors have conserved functions in plant hosts. Using Agrobacterium rhizogenes-mediated hairy roots induction, we demonstrated that the expression of PsPSR2 rendered hypersusceptibility of soybean to P. sojae. Enhanced susceptibility was also observed in PsPSR2-expressing Arabidopsis thaliana plants during Phytophthora but not bacterial infection. These experiments provide strong evidence that PSR2 is a conserved Phytophthora effector family that performs important virulence functions specifically during Phytophthora infection of various plant hosts.

  5. Phytophthora suppressor of RNA silencing 2 is a conserved RxLR effector that promotes infection in soybean and Arabidopsis thaliana.

    PubMed

    Xiong, Qin; Ye, Wenwu; Choi, Duseok; Wong, James; Qiao, Yongli; Tao, Kai; Wang, Yuanchao; Ma, Wenbo

    2014-12-01

    The genus Phytophthora consists of notorious and emerging pathogens of economically important crops. Each Phytophthora genome encodes several hundreds of cytoplasmic effectors, which are believed to manipulate plant immune response inside the host cells. However, the majority of Phytophthora effectors remain functionally uncharacterized. We recently discovered two effectors from the soybean stem and root rot pathogen Phytophthora sojae with the activity to suppress RNA silencing in plants. These effectors are designated Phytophthora suppressor of RNA silencing (PSRs). Here, we report that the P. sojae PSR2 (PsPSR2) belongs to a conserved and widespread effector family in Phytophthora. A PsPSR2-like effector produced by P. infestans (PiPSR2) can also suppress RNA silencing in plants and promote Phytophthora infection, suggesting that the PSR2 family effectors have conserved functions in plant hosts. Using Agrobacterium rhizogenes-mediated hairy roots induction, we demonstrated that the expression of PsPSR2 rendered hypersusceptibility of soybean to P. sojae. Enhanced susceptibility was also observed in PsPSR2-expressing Arabidopsis thaliana plants during Phytophthora but not bacterial infection. These experiments provide strong evidence that PSR2 is a conserved Phytophthora effector family that performs important virulence functions specifically during Phytophthora infection of various plant hosts. PMID:25387135

  6. Comparative structural and functional analysis of genes encoding pectin methylesterases in Phytophthora spp.

    PubMed

    Mingora, Christina; Ewer, Jason; Ospina-Giraldo, Manuel

    2014-03-15

    We have scanned the Phytophthora infestans, P. ramorum, and P. sojae genomes for the presence of putative pectin methylesterase genes and conducted a sequence analysis of all gene models found. We also searched for potential regulatory motifs in the promoter region of the proposed P. infestans models, and investigated the gene expression levels throughout the course of P. infestans infection on potato plants, using in planta and detached leaf assays. We found that genes located on contiguous chromosomal regions contain similar motifs in the promoter region, indicating the possibility of a shared regulatory mechanism. Results of our investigations also suggest that, during the pathogenicity process, the expression levels of some of the analyzed genes vary considerably when compared to basal expression observed in in vitro cultures of non-sporulating mycelium. These results were observed both in planta and in detached leaf assays.

  7. Carbohydrate-related enzymes of important Phytophthora plant pathogens.

    PubMed

    Brouwer, Henk; Coutinho, Pedro M; Henrissat, Bernard; de Vries, Ronald P

    2014-11-01

    Carbohydrate-Active enZymes (CAZymes) form particularly interesting targets to study in plant pathogens. Despite the fact that many CAZymes are pathogenicity factors, oomycete CAZymes have received significantly less attention than effectors in the literature. Here we present an analysis of the CAZymes present in the Phytophthora infestans, Ph. ramorum, Ph. sojae and Pythium ultimum genomes compared to growth of these species on a range of different carbon sources. Growth on these carbon sources indicates that the size of enzyme families involved in degradation of cell-wall related substrates like cellulose, xylan and pectin is not always a good predictor of growth on these substrates. While a capacity to degrade xylan and cellulose exists the products are not fully saccharified and used as a carbon source. The Phytophthora genomes encode larger CAZyme sets when compared to Py. ultimum, and encode putative cutinases, GH12 xyloglucanases and GH10 xylanases that are missing in the Py. ultimum genome. Phytophthora spp. also encode a larger number of enzyme families and genes involved in pectin degradation. No loss or gain of complete enzyme families was found between the Phytophthora genomes, but there are some marked differences in the size of some enzyme families.

  8. Occurrence and characterization of a Phytophthora sp. pathogenic to asparagus (Asparagus officinalis) in Michigan.

    PubMed

    Saude, C; Hurtado-Gonzales, O P; Lamour, K H; Hausbeck, M K

    2008-10-01

    A homothallic Phytophthora sp. was recovered from asparagus (Asparagus officinalis) spears, storage roots, crowns, and stems in northwest and central Michigan in 2004 and 2005. Isolates (n = 131) produced ovoid, nonpapillate, noncaducous sporangia 45 microm long x 26 microm wide and amphigynous oospores of 25 to 30 microm diameter. Mycelial growth was optimum at 25 degrees C with no growth at 5 and 30 degrees C. All isolates were sensitive to 100 ppm mefenoxam. Pathogenicity studies confirmed the ability of the isolates to infect asparagus as well as cucurbits. Amplified fragment length polymorphism analysis of 99 isolates revealed identical fingerprints, with 12 clearly resolved fragments present and no clearly resolved polymorphic fragments, suggesting a single clonal lineage. The internal transcribed spacer regions of representative isolates were homologous with a Phytophthora sp. isolated from diseased asparagus in France and a Phytophthora sp. from agave in Australia. Phylogenetic analysis supports the conclusion that the Phytophthora sp. isolated from asparagus in Michigan is a distinct species, and has been named Phytophthora asparagi.

  9. Occurrence and characterization of a Phytophthora sp. pathogenic to asparagus (Asparagus officinalis) in Michigan.

    PubMed

    Saude, C; Hurtado-Gonzales, O P; Lamour, K H; Hausbeck, M K

    2008-10-01

    A homothallic Phytophthora sp. was recovered from asparagus (Asparagus officinalis) spears, storage roots, crowns, and stems in northwest and central Michigan in 2004 and 2005. Isolates (n = 131) produced ovoid, nonpapillate, noncaducous sporangia 45 microm long x 26 microm wide and amphigynous oospores of 25 to 30 microm diameter. Mycelial growth was optimum at 25 degrees C with no growth at 5 and 30 degrees C. All isolates were sensitive to 100 ppm mefenoxam. Pathogenicity studies confirmed the ability of the isolates to infect asparagus as well as cucurbits. Amplified fragment length polymorphism analysis of 99 isolates revealed identical fingerprints, with 12 clearly resolved fragments present and no clearly resolved polymorphic fragments, suggesting a single clonal lineage. The internal transcribed spacer regions of representative isolates were homologous with a Phytophthora sp. isolated from diseased asparagus in France and a Phytophthora sp. from agave in Australia. Phylogenetic analysis supports the conclusion that the Phytophthora sp. isolated from asparagus in Michigan is a distinct species, and has been named Phytophthora asparagi. PMID:18943453

  10. Use of genome sequence data in the design and testing of SSR markers for Phytophthora species

    PubMed Central

    Schena, Leonardo; Cardle, Linda; Cooke, David EL

    2008-01-01

    Background Microsatellites or single sequence repeats (SSRs) are a powerful choice of marker in the study of Phytophthora population biology, epidemiology, ecology, genetics and evolution. A strategy was tested in which the publicly available unigene datasets extracted from genome sequences of P. infestans, P. sojae and P. ramorum were mined for candidate SSR markers that could be applied to a wide range of Phytophthora species. Results A first approach, aimed at the identification of polymorphic SSR loci common to many Phytophthora species, yielded 171 reliable sequences containing 211 SSRs. Microsatellites were identified from 16 target species representing the breadth of diversity across the genus. Repeat number ranged from 3 to 16 with most having seven repeats or less and four being the most commonly found. Trinucleotide repeats such as (AAG)n, (AGG)n and (AGC)n were the most common followed by pentanucleotide, tetranucleotide and dinucleotide repeats. A second approach was aimed at the identification of useful loci common to a restricted number of species more closely related to P. sojae (P. alni, P. cambivora, P. europaea and P. fragariae). This analysis yielded 10 trinucleotide and 2 tetranucleotide SSRs which were repeated 4, 5 or 6 times. Conclusion Key studies on inter- and intra-specific variation of selected microsatellites remain. Despite the screening of conserved gene coding regions, the sequence diversity between species was high and the identification of useful SSR loci applicable to anything other than the most closely related pairs of Phytophthora species was challenging. That said, many novel SSR loci for species other than the three 'source species' (P. infestans, P. sojae and P. ramorum) are reported, offering great potential for the investigation of Phytophthora populations. In addition to the presence of microsatellites, many of the amplified regions may represent useful molecular marker regions for other studies as they are highly variable

  11. Phenotypic diversification by gene silencing in Phytophthora plant pathogens

    PubMed Central

    Vetukuri, Ramesh R; Åsman, Anna KM; Jahan, Sultana N; Avrova, Anna O; Whisson, Stephen C; Dixelius, Christina

    2013-01-01

    Advances in genome sequencing technologies have enabled generation of unprecedented information on genome content and organization. Eukaryote genomes in particular may contain large populations of transposable elements (TEs) and other repeated sequences. Active TEs can result in insertional mutations, altered transcription levels and ectopic recombination of DNA. The genome of the oomycete plant pathogen, Phytophthora infestans, contains vast numbers of TE sequences. There are also hundreds of predicted disease-promoting effector proteins, predominantly located in TE-rich genomic regions. Expansion of effector gene families is also a genomic signature of related oomycetes such as P. sojae. Deep sequencing of small RNAs (sRNAs) from P. infestans has identified sRNAs derived from all families of transposons, highlighting the importance of RNA silencing for maintaining these genomic invaders in an inactive form. Small RNAs were also identified from specific effector encoding genes, possibly leading to RNA silencing of these genes and variation in pathogenicity and virulence toward plant resistance genes. Similar findings have also recently been made for the distantly related species, P. sojae. Small RNA “hotspots” originating from arrays of amplified gene sequences, or from genes displaying overlapping antisense transcription, were also identified in P. infestans. These findings suggest a major role for RNA silencing processes in the adaptability and diversification of these economically important plant pathogens. Here we review the latest progress and understanding of gene silencing in oomycetes with emphasis on transposable elements and sRNA-associated events. PMID:24563702

  12. Species from within the Phytophthora cryptogea complex and related species, P. erythroseptica and P. sansomeana, readily hybridize.

    PubMed

    Safaiefarahani, Banafsheh; Mostowfizadeh-Ghalamfarsa, Reza; Hardy, Giles E St J; Burgess, Treena I

    2016-08-01

    During a study on the phylogenetic relationships between species in the Phytophthora cryptogea complex and related species, Phytophthora erythroseptica and Phytophthora sansomeana, 19 hybrid isolates with multiple polymorphisms in the nuclear sequences were observed. Molecular characterization of hybrids was achieved by sequencing three nuclear (internal transcribed spacers, β-tubulin (TUB), heat shock protein 90) and two mitochondrial (cytochrome c oxidase subunit I (coxI), NADH dehydrogenase subunit I (NADH)) gene regions and cloning of the single-copy nuclear gene, TUB. Based on the molecular studies the hybrid isolates belonged to six distinct groups between P. cryptogea, P. erythroseptica, Phytophthora pseudocryptogea, P. sansomeana, and Phytophthora sp. kelmania. In all cases, only a single coxI and NADH allele was detected and nuclear genes were biparentally inherited, suggesting that the hybrids arose from sexual recombination events. Colony morphology, growth rate, cardinal temperatures, breeding system, and morphology of sporangia, oogonia, oospores, and antheridia were also determined. Some morphological differences between the hybrids and the parental species were noted; however, they were not sufficient to reliably distinguish the taxa and DNA markers from nuclear and mitochondrial genes will to be necessary for their identification. The parental species are all important pathogens of agricultural fields that have been transported globally. With the apparent ease of hybridization within this group there is ample opportunity for virulent hybrids to form, perhaps with extended host ranges. PMID:27521629

  13. Phytophthora gallica sp. nov., a new species from rhizosphere soil of declining oak and reed stands in France and Germany.

    PubMed

    Jung, Thomas; Nechwatal, Jan

    2008-10-01

    A non-papillate, slow-growing Phytophthora species, which could not be assigned to any existing taxon, was isolated from rhizosphere soil of a declining oak in Northeast France, and from the rhizosphere of Phragmites australis at Lake Constance in south-west Germany in 1998 and 2004, respectively. We describe this species, previously informally designated Phytophthora taxon 'G', as Phytophthora gallica sp. nov. Morphology, growth rates, and pathogenicity against cuttings of riparian tree species and leaves of reed are described and compared with those of morphologically and phylogenetically similar Phytophthora species. P. gallica produces colonies with limited aerial mycelium and variable growth patterns. Gametangia are not formed in single or mixed cultures with tester strains of known mating types. P. gallica produces globose and elongated irregular chlamydospores, of which a high proportion is abortive. In water culture irregular hyphal swellings and non-papillate persistent sporangia are formed abundantly. P. gallica is moderately aggressive to Alnus glutinosa and Fagus sylvatica, weakly aggressive to Quercus robur and Salix alba and non-pathogenic to Fraxinus excelsior and Phragmites australis. According to ITS and mtDNA sequence data P. gallica belongs to a distinct Phytophthora clade, with P. boehmeriae and P. kernoviae being the closest relatives. The origin of P. gallica and its ecological role in wet ecosystems remain unclear.

  14. Fighting phytophthora in blueberries

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora cinnamomi Rands is a ubiquitous soilborne pathogen associated with root rot in many woody perennial plant species, including highbush blueberry (Vaccinium sp.). To identify genotypes with resistance to the pathogen, cultivars and advanced selections of highbush blueberry were grown in a...

  15. Direct measurement of dispersal of Dictyocaulus viviparus in sporangia of Pilobolus species.

    PubMed

    Eysker, M

    1991-01-01

    Dispersal of Dictyocaulus viviparus larvae by Pilobolus sporangia was studied on 29 faecal pats deposited between the end of June and late October 1988. Faecal pats were covered daily from day 3 to 4 after deposition with a large petri dish to measure the numbers of sporangia released and the numbers of larvae carried. The yield of both was variable. Dispersal of lungworm larvae was lowest on over-grazed pasture or when Pilobolus growth was very poor. When faecal pats were sheltered by a long sward, 17 per cent or more of larvae present at deposition were transported in this manner. In July and August, peak dispersal of lungworm larvae was on day 5, in September on day 6 and in October on day 7, the increasing time intervals being probably associated with decreasing temperature. PMID:2047589

  16. A novel workflow correlating RNA-seq data to Phythophthora infestans resistance levels in wild Solanum species and potato clones

    PubMed Central

    Frades, Itziar; Abreha, Kibrom B.; Proux-Wéra, Estelle; Lankinen, Åsa; Andreasson, Erik; Alexandersson, Erik

    2015-01-01

    Comparative transcriptomics between species can provide valuable understanding of plant-pathogen interactions. Here, we focus on wild Solanum species and potato clones with varying degree of resistance against Phytophthora infestans, which causes the devastating late blight disease in potato. The transcriptomes of three wild Solanum species native to Southern Sweden, Solanum dulcamara, Solanum nigrum, and Solanum physalifolium were compared to three potato clones, Desiree (cv.), SW93-1015 and Sarpo Mira. Desiree and S. physalifolium are susceptible to P. infestans whereas the other four have different degrees of resistance. By building transcript families based on de novo assembled RNA-seq across species and clones and correlating these to resistance phenotypes, we created a novel workflow to identify families with expanded or depleted number of transcripts in relation to the P. infestans resistance level. Analysis was facilitated by inferring functional annotations based on the family structure and semantic clustering. More transcript families were expanded in the resistant clones and species and the enriched functions of these were associated to expected gene ontology (GO) terms for resistance mechanisms such as hypersensitive response, host programmed cell death and endopeptidase activity. However, a number of unexpected functions and transcripts were also identified, for example transmembrane transport and protein acylation expanded in the susceptible group and a cluster of Zinc knuckle family proteins expanded in the resistant group. Over 400 expressed putative resistance (R-)genes were identified and resistant clones Sarpo Mira and SW93-1015 had ca 25% more expressed putative R-genes than susceptible cultivar Desiree. However, no differences in numbers of susceptibility (S-)gene homologs were seen between species and clones. In addition, we identified P. infestans transcripts including effectors in the early stages of P. infestans-Solanum interactions. PMID

  17. The phytophthora genome initiative database: informatics and analysis for distributed pathogenomic research.

    PubMed

    Waugh, M; Hraber, P; Weller, J; Wu, Y; Chen, G; Inman, J; Kiphart, D; Sobral, B

    2000-01-01

    The Phytophthora Genome Initiative (PGI) is a distributed collaboration to study the genome and evolution of a particularly destructive group of plant pathogenic oomycete, with the goal of understanding the mechanisms of infection and resistance. NCGR provides informatics support for the collaboration as well as a centralized data repository. In the pilot phase of the project, several investigators prepared Phytophthora infestans and Phytophthora sojae EST and Phytophthora sojae BAC libraries and sent them to another laboratory for sequencing. Data from sequencing reactions were transferred to NCGR for analysis and curation. An analysis pipeline transforms raw data by performing simple analyses (i.e., vector removal and similarity searching) that are stored and can be retrieved by investigators using a web browser. Here we describe the database and access tools, provide an overview of the data therein and outline future plans. This resource has provided a unique opportunity for the distributed, collaborative study of a genus from which relatively little sequence data are available. Results may lead to insight into how better to control these pathogens. The homepage of PGI can be accessed at http:www.ncgr.org/pgi, with database access through the database access hyperlink.

  18. Phytophthora pseudosyringae sp. nov., a new species causing root and collar rot of deciduous tree species in Europe.

    PubMed

    Jung, Thomas; Nechwatal, Jan; Cooke, David E L; Hartmann, Günther; Blaschke, Markus; Osswald, Wolfgang F; Duncan, James M; Delatour, Claude

    2003-07-01

    In several studies of oak decline in Europe, a semi-papillate homothallic Phytophthora taxon was consistently isolated, together with other Phytophthora species, from rhizosphere soil samples. It was also found associated with necrotic fine roots and stem necroses of Fagus sylvatica and Alnus glutinosa. Due to morphological and physiological similarities, the semi-papillate isolates were previously identified as P. syringae by various authors. The morphology, physiology and pathogenicity against fine roots of Quercus robur, Q. petraea and F. sylvatica, bark of A. glutinosa, leaves of Ilex aquifolium and apple fruits of this Phytophthora species are described and compared with those of related and similar Phytophthora species, namely P. ilicis, P. psychrophila, P. quercina, P. citricola and P. syringae. The phylogenetic placement on the basis of ITS and mtDNA sequence data was also examined. Isolates of this taxon produce colonies with stellate to rosaceous growth patterns and limited aerial mycelium on various agar media. Antheridia are predominantly paragynous. In water culture catenulate hyphal swellings and semi-papillate caducous sporangia, that are usually limoniform, ellipsoid or ovoid, are formed abundandly, mostly in lax or dense sympodia. This taxon is a moderately slow growing, low temperature species with optimum and maximum temperatures around 20 and 25 degrees C, respectively. Tested isolates are moderately aggressive to fine roots of oaks and beech, highly aggressive to holly leaves and apple fruits, and slightly pathogenic to alder bark. Thirteen tested isolates had an identical and distinct ITS sequence which was more similar to that of P. ilicis and P. psychrophila than any other known taxa. On the basis of their unique combination of morphological characters, colony growth patterns, cardinal temperatures for growth, growth rates, pathogenicity to oaks, beech, alder, apple and holly, their host range, and ITS and mtDNA sequences the semi

  19. Effects of microgravity on the susceptibility of soybean to Phytophthora sojae.

    PubMed

    Nedukha, O M; Leach, J E; Ryba-White, M; Hilaire, E; Guikema, J; Kordyum, E L

    1998-07-01

    The study of pathogenicity of higher plants under conditions of microgravity is of great importance for the future production of food in space. Previous work suggests that microgravity affects both microbes and plants. Bacterial numbers increased after 17 days in an algae-bacterium association on the biosatellite "Kosmos-1887". This was speculated to result from an increase in the multiplication rate of the bacteria. Sporangia of both Actinomices brevis, in the shuttles "Soyuz-19" and "Appolon", and Phycomyces blakes, in biosatellite "Kosmos-936", formed after 10 days in microgravity. Sporangia did not form in the ground controls in the same time suggesting that the rate of fungal development is enhanced in microgravity. Plant responses to pathogens in microgravity have not been studied, however, microgravity profoundly impacts plant cell development, cytology, and physiology. In microgravity, developing cell walls are thinner and contain less lignin than ground-grown plants. The demonstrated effects of microgravity on both plants and microbes lead us to hypothesize that plants may be more susceptible to pathogens under conditions of microgravity. The aim of this study was to determine the influence of microgravity on the susceptibility of soybean to the fungal root rot pathogen, Phytophthora sojae.

  20. L-type lectin receptor kinases in Nicotiana benthamiana and tomato and their role in Phytophthora resistance

    PubMed Central

    Wang, Yan; Weide, Rob; Govers, Francine; Bouwmeester, Klaas

    2015-01-01

    Membrane-bound receptors play crucial roles as sentinels of plant immunity against a large variety of invading microbes. One class of receptors known to be involved in self/non-self-surveillance and plant resistance comprises the L-type lectin receptor kinases (LecRKs). Previously, we reported that several Arabidopsis LecRKs play a role in resistance to Phytophthora pathogens. In this study, we determined whether homologues of these LecRKs from the Solanaceous plants Nicotiana benthamiana and tomato (Solanum lycopersicum) play similar roles in defence against Phytophthora. In genome-wide screenings, a total of 38 (Nb)LecRKs were identified in N. benthamiana and 22 (Sl)LecRKs in tomato, each consisting of both a lectin and a kinase domain. Phylogenetic analysis revealed that, in contrast to Arabidopsis, which has a LecRK family comprising nine clades, Solanaceous species have just five of these nine clades (i.e. IV, VI, VII, VIII, and IX), plus four additional clades that lack Arabidopsis homologues. Several of the Solanaceous LecRKs were selected for functional analysis using virus-induced gene silencing. Infection assays with Phytophthora capsici and Phytophthora infestans on LecRK-silenced plants revealed that N. benthamiana and tomato homologues in clade IX play a role in Phytophthora resistance similar to the two Arabidopsis LecRKs in this clade, suggesting conserved functions of clade IX LecRKs across different plant families. This study provides a first insight into the diversity of Solanaceous LecRKs and their role in plant immunity, and shows the potential of LecRKs for Phytophthora resistance breeding. PMID:26248665

  1. L-type lectin receptor kinases in Nicotiana benthamiana and tomato and their role in Phytophthora resistance.

    PubMed

    Wang, Yan; Weide, Rob; Govers, Francine; Bouwmeester, Klaas

    2015-11-01

    Membrane-bound receptors play crucial roles as sentinels of plant immunity against a large variety of invading microbes. One class of receptors known to be involved in self/non-self-surveillance and plant resistance comprises the L-type lectin receptor kinases (LecRKs). Previously, we reported that several Arabidopsis LecRKs play a role in resistance to Phytophthora pathogens. In this study, we determined whether homologues of these LecRKs from the Solanaceous plants Nicotiana benthamiana and tomato (Solanum lycopersicum) play similar roles in defence against Phytophthora. In genome-wide screenings, a total of 38 (Nb)LecRKs were identified in N. benthamiana and 22 (Sl)LecRKs in tomato, each consisting of both a lectin and a kinase domain. Phylogenetic analysis revealed that, in contrast to Arabidopsis, which has a LecRK family comprising nine clades, Solanaceous species have just five of these nine clades (i.e. IV, VI, VII, VIII, and IX), plus four additional clades that lack Arabidopsis homologues. Several of the Solanaceous LecRKs were selected for functional analysis using virus-induced gene silencing. Infection assays with Phytophthora capsici and Phytophthora infestans on LecRK-silenced plants revealed that N. benthamiana and tomato homologues in clade IX play a role in Phytophthora resistance similar to the two Arabidopsis LecRKs in this clade, suggesting conserved functions of clade IX LecRKs across different plant families. This study provides a first insight into the diversity of Solanaceous LecRKs and their role in plant immunity, and shows the potential of LecRKs for Phytophthora resistance breeding.

  2. L-type lectin receptor kinases in Nicotiana benthamiana and tomato and their role in Phytophthora resistance.

    PubMed

    Wang, Yan; Weide, Rob; Govers, Francine; Bouwmeester, Klaas

    2015-11-01

    Membrane-bound receptors play crucial roles as sentinels of plant immunity against a large variety of invading microbes. One class of receptors known to be involved in self/non-self-surveillance and plant resistance comprises the L-type lectin receptor kinases (LecRKs). Previously, we reported that several Arabidopsis LecRKs play a role in resistance to Phytophthora pathogens. In this study, we determined whether homologues of these LecRKs from the Solanaceous plants Nicotiana benthamiana and tomato (Solanum lycopersicum) play similar roles in defence against Phytophthora. In genome-wide screenings, a total of 38 (Nb)LecRKs were identified in N. benthamiana and 22 (Sl)LecRKs in tomato, each consisting of both a lectin and a kinase domain. Phylogenetic analysis revealed that, in contrast to Arabidopsis, which has a LecRK family comprising nine clades, Solanaceous species have just five of these nine clades (i.e. IV, VI, VII, VIII, and IX), plus four additional clades that lack Arabidopsis homologues. Several of the Solanaceous LecRKs were selected for functional analysis using virus-induced gene silencing. Infection assays with Phytophthora capsici and Phytophthora infestans on LecRK-silenced plants revealed that N. benthamiana and tomato homologues in clade IX play a role in Phytophthora resistance similar to the two Arabidopsis LecRKs in this clade, suggesting conserved functions of clade IX LecRKs across different plant families. This study provides a first insight into the diversity of Solanaceous LecRKs and their role in plant immunity, and shows the potential of LecRKs for Phytophthora resistance breeding. PMID:26248665

  3. The plant pathogen Phytophthora andina emerged via hybridization of an unknown Phytophthora species and the Irish famine pathogen, P. infestans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The global movement of plant pathogens threatens natural ecosystems, food security, and commercial interests. Introduction of a plant pathogen to new geographic regions has been the primary mechanism by which new pathogens have emerged. Another documented mechanism for the emergence of plant pathoge...

  4. Molecular Characterization of Natural Hybrids Formed between Five Related Indigenous Clade 6 Phytophthora Species

    PubMed Central

    Burgess, Treena I.

    2015-01-01

    Most Phytophthora hybrids characterized to date have emerged from nurseries and managed landscapes, most likely generated as a consequence of biological invasions associated with the movement of living plants and germplasm for ornamental, horticultural and agricultural purposes. Presented here is evidence for natural hybridization among a group of five closely related indigenous clade 6 Phytophthora species isolated from waterways and riparian ecosystems in Western Australia. Molecular characterization of hybrids consisted of cloning and sequencing two nuclear genes (ITS and ASF), sequencing of two further nuclear loci (BT and HSP) and of two mitochondrial loci (COI and NADH). Additionally, phenotypic traits including morphology of sporangia and optima and maxima temperatures for growth were also determined. In most cases the nuclear genes were biparentally and in all cases the mtDNA were uniparentally inherited, indicating hybrid formation through sexual crosses. Some isolates bear the molecular signature of three parents suggesting additional hybrid events, although it cannot be determined from the data if these were sequential or simultaneous. These species and their hybrids co-exist in riparian ecosystems and waterways where their ability for rapid asexual proliferation would enable them to rapidly colonize green plant litter. The apparent ease of hybridization could eventually lead to the merging of species through introgression. However, at this point in time, species integrity has been maintained and a more likely scenario is that the hybrids are not stable evolutionary lineages, but rather transient hybrid clones. PMID:26248187

  5. The Anti-Phytophthora Effect of Selected Potato-Associated Pseudomonas Strains: From the Laboratory to the Field

    PubMed Central

    Guyer, Anouk; De Vrieze, Mout; Bönisch, Denise; Gloor, Ramona; Musa, Tomke; Bodenhausen, Natacha; Bailly, Aurélien; Weisskopf, Laure

    2015-01-01

    Late blight, caused by the oomycete Phytophthora infestans, is the most devastating disease of potato. In organic farming, late blight is controlled by repeated applications of copper-based products, which negatively impact the environment. To find alternative solutions for late blight management, we have previously isolated a large collection of bacteria from the phyllosphere and the rhizosphere of potatoes. Here we report the antagonistic potential of these strains when co-cultivated with P. infestans as well as with other potato pathogens. We then focused on three Pseudomonas strains and compared their protective impact against late blight to that of well-known biocontrol strains in planta using a high-throughput leaf disk assay with automated picture analysis. When sprayed on the leaves of potatoes in the greenhouse, the strains were able to survive for at least 15 days. Under field conditions, populations decreased faster but all tested strains could still be retrieved after 8 days. The most active strain in vitro, P. chlororaphis R47, was also the best protectant on leaf disks from plants grown in the greenhouse experiment, but its protection potential could not be verified in the field due to unfavorable infection conditions. However, its protective effect against P. infestans in planta, its survival in the phyllosphere as well as its ability to colonize the potato rhizosphere in very high population densities, suggest a potential for field application, e.g., in the form of tuber treatment or leaf spray. PMID:26640460

  6. A Strategy for the Proliferation of Ulva prolifera, Main Causative Species of Green Tides, with Formation of Sporangia by Fragmentation

    PubMed Central

    Gao, Shan; Chen, Xiaoyuan; Yi, Qianqian; Wang, Guangce; Pan, Guanghua; Lin, Apeng; Peng, Guang

    2010-01-01

    Ulva prolifera, a common green seaweed, is one of the causative species of green tides that occurred frequently along the shores of Qingdao in 2008 and had detrimental effects on the preparations for the 2008 Beijing Olympic Games sailing competition, since more than 30 percent of the area of the games was invaded. In view of the rapid accumulation of the vast biomass of floating U. prolifera in green tides, we investigated the formation of sporangia in disks of different diameters excised from U. prolifera, changes of the photosynthetic properties of cells during sporangia formation, and development of spores. The results suggested that disks less than 1.00 mm in diameter were optimal for the formation of sporangia, but there was a small amount of spore release in these. The highest percentage of area of spore release occurred in disks that were 2.50 mm in diameter. In contrast, sporangia were formed only at the cut edges of larger disks (3.00 mm, 3.50 mm, and 4.00 mm in diameter). Additionally, the majority of spores liberated from the disks appeared vigorous and developed successfully into new individuals. These results implied that fragments of the appropriate size from the U. prolifera thalli broken by a variety of factors via producing spores gave rise to the rapid proliferation of the seaweed under field conditions, which may be one of the most important factors to the rapid accumulation of the vast biomass of U. prolifera in the green tide that occurred in Qingdao, 2008. PMID:20052408

  7. Simultaneous quantification of sporangia and zoospores in a biotrophic oomycete with an automatic particle analyzer: disentangling dispersal and infection potentials.

    PubMed

    Delmas, Chloé E L; Mazet, Isabelle D; Jolivet, Jérôme; Delière, Laurent; Delmotte, François

    2014-12-01

    Quantitative pathogenicity traits drive the fitness and dynamics of pathogens in agricultural ecosystems and are key determinants of the correct management of crop production over time. However, traits relating to infection potential (i.e. zoospore production) have been less thoroughly investigated in oomycetes than traits relating to dispersal (i.e. sporangium production). We simultaneously quantified sporangium and zoospore production in a biotrophic oomycete, for the joint assessment of life-cycle traits relating to dispersal and infection potentials. We used an automatic particle analyzer to count and size the sporangia and/or zoospores produced at t = 0 min (no zoospore release) and t = 100 min (zoospore release) in 43 Plasmopara viticola isolates growing on the susceptible Vitis vinifera cv. Cabernet Sauvignon. We were able to differentiate and quantify three types of propagules from different stages of the pathogen life cycle: full sporangia, empty sporangia and zoospores. The method was validated by comparing the sporangium and zoospore counts obtained with an automatic particle analyzer and under a stereomicroscope (manual counting). Each isolate produced a mean of 5.8 ± 1.9 (SD) zoospores per sporangium. Significant relationships were found between sporangium production and sporangium size (negative) and between sporangium size and the number of zoospores produced per sporangium (positive). However, there was a significant positive correlation between total sporangium production and total zoospore production. This procedure can provide a valid quantification of the production of both sporangia and zoospores by oomycetes in large numbers of samples, facilitating joint estimation of the dispersal and infection potentials of plant pathogens in various agro-ecological contexts.

  8. Using effectors of Phytophthora infestans to teach pathogenesis: Our attempt to provide a more comprehensive education

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The topic of pathogenesis mechanisms (R/avirulence genes, effectors, and hypersensitive response) has proved challenging for students in our introductory plant pathology course. An apparent gap exists in the curriculum between this introductory course and higher level plant-microbe interaction cours...

  9. Inhibition of Phytophthora parasitica and P. capsici by Silver Nanoparticles Synthesized Using Aqueous Extract of Artemisia absinthium.

    PubMed

    Ali, Mohammad; Kim, Bosung; Belfield, Kevin D; Norman, David; Brennan, Mary; Ali, Gul Shad

    2015-09-01

    Application of nanoparticles for controlling plant pathogens is a rapidly emerging area in plant disease management, and nanoparticles synthesis methods that are economical and ecofriendly are extensively investigated. In this project, we investigated the potential of silver nanoparticles (AgNPs) synthesized with aqueous extract of Artemisia absinthium against several Phytophthora spp., which cause many economically important crop diseases. In in vitro dose-response tests conducted in microtiter plates, 10 µg ml⁻¹ of AgNPs inhibited mycelial growth of P. parasitica, P. infestans, P. palmivora, P. cinnamomi, P. tropicalis, P. capsici, and P. katsurae. Detailed in vitro dose-response analyses conducted with P. parasitica and P. capsici revealed that AgNPs synthesized with A. absinthium extract were highly potent (IC50: 2.1 to 8.3 µg ml⁻¹) and efficacious (100%) in inhibiting mycelial growth, zoospore germination, germ tube elongation, and zoospore production. Interestingly, AgNP treatment accelerated encystment of zoospores. Consistent with in vitro results, in planta experiments conducted in a greenhouse revealed that AgNP treatments prevented Phytophthora infection and improved plant survival. Moreover, AgNP in in planta experiments did not produce any adverse effects on plant growth. These investigations provide a simple and economical method for controlling Phytophthora with AgNP without affecting normal plant physiology.

  10. Leaf surface chemicals fromNicotiana affecting germination ofPeronospora tabacina (adam) sporangia.

    PubMed

    Kennedy, B S; Nielsen, M T; Severson, R F; Sisson, V A; Stephenson, M K; Jackson, D M

    1992-09-01

    A bioassay was used to evaluate the effects of cuticular leaf components, isolated fromN. tabacum, N. glutinosa (accessions 24 and 24a), and 23other Nicotiana species, on germinationof P. tabacina (blue mold). The leaf surface compounds includedα- andβ-4,8,13,-duvatriene-l,3-diols (DVT-diols), (13-E)-labda-13-ene-8α-,15-diol (labdenediol), (12-Z)-labda-12,14-diene-8α-ol (cis-abienol), (13-R)-labda-8,14-diene-13-ol (manool), 2-hydroxymanool, a mixture of (13-R)-labda-14-ene-8α,13-diol (sclareol), and (13-S)-labda-14-ene-8α,13-diol (episclareol), and various glucose and/or sucrose ester isolates. The above in acetone were applied onto leaf disks of the blue moldsusceptibleN. tabacum cv. TI 1406, which was then inoculated with blue mold sporangia. Estimated IC50 values (inhibitory concentration) were 3.0μg/cm(2) forα-DVT-diol, 2.9μ/cm(2) forβ-DVT-diol, 0.4μg/cm(2) for labdenediol and 4.7μg/cm(2) for the sclareol mixture. Manool, 2-hydroxymanool, andcis-abienol at application rates up to 30μg/cm(2) had little or no effect on sporangium germination. Glucose and/or sucrose ester isolates from the cuticular leaf extracts of 23Nicotiana species and three different fractions fromN. bigelovii were also evaluated for antimicrobial activity at a concentration of 30μg/cm(2). Germination was inhibited by >20% when exposed to sugar esters isolated fromN. acuminata, N. benthamiana, N. attenuata, N. clevelandii, andN. miersii, and accessions 10 and 12 ofN. bigelovii. These results imply that a number of compounds may influence resistance to blue mold in tobacco. PMID:24254279

  11. A high-temperature tolerant species in clade 9 of the genus Phytophthora: P. hydrogena sp. nov.

    PubMed

    Yang, Xiao; Gallegly, Mannon E; Hong, Chuanxue

    2014-01-01

    A previously unknown Phytophthora species was isolated from irrigation water in Virginia, USA. This novel species produces abundant noncaducous and nonpapillate sporangia in soil water extract solution. It sometimes produces chlamydospores and hyphal swellings in aged cultures and in Petri's solution. This species has optimum vegetative growth at 30 C and grows well at 35 C. The lowest and highest temperatures for growth are 5 and 40 C. All isolates examined in this study are compatibility type A1 and produce mostly plerotic oospores when paired with an A2 mating-type tester of P. cinnamomi. Sequence analyses of the rDNA internal transcribed spacer (ITS) regions and the mitochondrially encoded cytochrome c oxidase 1 (cox 1) gene placed this species in clade 9 of the genus Phytophthora. These characteristics support the description of this taxon as a new species for which we propose the name P. hydrogena sp. nov. Further phylogenetic and physiological investigations of clade 9 species revealed a high-temperature tolerant cluster including P. hydrogena, P. aquimorbida, P. hydropathica, P. irrigata, P. chrysanthemi, P. insolita, P. polonica and P. parsiana. These species all grow well at 35 C. The monophyly of the species in this heat-tolerant cluster except P. insolita and P. polonica is highly supported by the maximum-likelihood analyses of the ITS and cox 1 sequences.

  12. Phytophthora Have Distinct Endogenous Small RNA Populations That Include Short Interfering and microRNAs

    PubMed Central

    Fahlgren, Noah; Bollmann, Stephanie R.; Kasschau, Kristin D.; Cuperus, Josh T.; Press, Caroline M.; Sullivan, Christopher M.; Chapman, Elisabeth J.; Hoyer, J. Steen; Gilbert, Kerrigan B.; Grünwald, Niklaus J.; Carrington, James C.

    2013-01-01

    In eukaryotes, RNA silencing pathways utilize 20-30-nucleotide small RNAs to regulate gene expression, specify and maintain chromatin structure, and repress viruses and mobile genetic elements. RNA silencing was likely present in the common ancestor of modern eukaryotes, but most research has focused on plant and animal RNA silencing systems. Phytophthora species belong to a phylogenetically distinct group of economically important plant pathogens that cause billions of dollars in yield losses annually as well as ecologically devastating outbreaks. We analyzed the small RNA-generating components of the genomes of P. infestans, P. sojae and P. ramorum using bioinformatics, genetic, phylogenetic and high-throughput sequencing-based methods. Each species produces two distinct populations of small RNAs that are predominantly 21- or 25-nucleotides long. The 25-nucleotide small RNAs were primarily derived from loci encoding transposable elements and we propose that these small RNAs define a pathway of short-interfering RNAs that silence repetitive genetic elements. The 21-nucleotide small RNAs were primarily derived from inverted repeats, including a novel microRNA family that is conserved among the three species, and several gene families, including Crinkler effectors and type III fibronectins. The Phytophthora microRNA is predicted to target a family of amino acid/auxin permeases, and we propose that 21-nucleotide small RNAs function at the post-transcriptional level. The functional significance of microRNA-guided regulation of amino acid/auxin permeases and the association of 21-nucleotide small RNAs with Crinkler effectors remains unclear, but this work provides a framework for testing the role of small RNAs in Phytophthora biology and pathogenesis in future work. PMID:24204767

  13. Impregnated Netting Slows Infestation by Triatoma infestans

    PubMed Central

    Levy, Michael Z.; Quíspe-Machaca, Victor R.; Ylla-Velasquez, Jose L.; Waller, Lance A.; Richards, Jean M.; Rath, Bruno; Borrini-Mayori, Katty; del Carpio, Juan G. Cornejo; Cordova-Benzaquen, Eleazar; McKenzie, F. Ellis; Wirtz, Robert A.; Maguire, James H.; Gilman, Robert H.; Bern, Caryn

    2008-01-01

    We used sentinel animal enclosures to measure the rate of infestation by the Chagas disease vector, Triatoma infestans, in an urban community of Arequipa, Peru, and to evaluate the effect of deltamethrin-impregnated netting on that rate. Impregnated netting decreased the rate of infestation of sentinel enclosures (rate ratio, 0.23; 95% confidence interval, 0.13–0.38; P < 0.001), controlling for the density of surrounding vector populations and the distance of these to the sentinel enclosures. Most migrant insects were early-stage nymphs, which are less likely to carry the parasitic agent of Chagas disease, Trypanosoma cruzi. Spread of the vector in the city therefore likely precedes spread of the parasite. Netting was particularly effective against adult insects and late-stage nymphs; taking into account population structure, netting decreased the reproductive value of migrant populations from 443.6 to 40.5. Impregnated netting can slow the spread of T. infestans and is a potentially valuable tool in the control of Chagas disease. PMID:18840739

  14. Impregnated netting slows infestation by Triatoma infestans.

    PubMed

    Levy, Michael Z; Quíspe-Machaca, Victor R; Ylla-Velasquez, Jose L; Waller, Lance A; Richards, Jean M; Rath, Bruno; Borrini-Mayori, Katty; del Carpio, Juan G Cornejo; Cordova-Benzaquen, Eleazar; McKenzie, F Ellis; Wirtz, Robert A; Maguire, James H; Gilman, Robert H; Bern, Caryn

    2008-10-01

    We used sentinel animal enclosures to measure the rate of infestation by the Chagas disease vector, Triatoma infestans, in an urban community of Arequipa, Peru, and to evaluate the effect of deltamethrin-impregnated netting on that rate. Impregnated netting decreased the rate of infestation of sentinel enclosures (rate ratio, 0.23; 95% confidence interval, 0.13-0.38; P < 0.001), controlling for the density of surrounding vector populations and the distance of these to the sentinel enclosures. Most migrant insects were early-stage nymphs, which are less likely to carry the parasitic agent of Chagas disease, Trypanosoma cruzi. Spread of the vector in the city therefore likely precedes spread of the parasite. Netting was particularly effective against adult insects and late-stage nymphs; taking into account population structure, netting decreased the reproductive value of migrant populations from 443.6 to 40.5. Impregnated netting can slow the spread of T. infestans and is a potentially valuable tool in the control of Chagas disease.

  15. [Ontogeny of strobili, sporangia development and sporogenesis in Equisetum giganteum (Equisetaceae) from the Colombian Andes].

    PubMed

    Rincón Barón, Edgar Javier; Forero Ballesteros, Helkin Giovani; Gélvez Landazábal, Leidy Viviana; Andrés Torres, Gerardo; Hilda Rolleri, Cristina

    2011-12-01

    Studies on the ontogeny of the strobilus, sporangium and reproductive biology of this group of ferns are scarce. Here we describe the ontogeny of the strobilus and sporangia, and the process of sporogenesis using specimens of E. giganteum from Colombia collected along the Rio Frio, Distrito de Sevilla, Piedecuesta, Santander, at 2200m altitude. The strobili in different stages of development were fixed, dehydrated, embedded in paraffin, sectioned using a rotatory microtome and stained with the safranin O and fast green technique. Observations were made using differential interference contrast microscopy (DIC) or Nomarski microscopy, an optical microscopy illumination technique that enhances the contrast in unstained, transparent. Strobili arise and begin to develop in the apical meristems of the main axis and lateral branches, with no significant differences in the ontogeny of strobili of one or other axis. Successive processes of cell division and differentiation lead to the growth of the strobilus and the formation of sporangiophores. These are formed by the scutellum, the manubrium or pedicel-like, basal part of the sporangiophore, and initial cells of sporangium, which differentiate to form the sporangium wall, the sporocytes and the tapetum. There is not formation of a characteristic arquesporium, as sporocytes quickly undergo meiosis originating tetrads of spores. The tapetum retains its histological integrity, but subsequently the cell walls break down and form a plasmodium that invades the sporangial cavity, partially surrounding the tetrads, and then the spores. Towards the end of the sporogenesis the tapetum disintegrates leaving spores with elaters free within the sporangial cavity. Two layers finally form the sporangium wall: the sporangium wall itself, with thickened, lignified cell walls and an underlying pyknotic layer. The mature spores are chlorofilous, morphologically similar and have exospore, a thin perispore and two elaters. This study of the

  16. Phytophthora niederhauserii sp. nov., a polyphagous species associated with ornamentals, fruit trees and native plants in 13 countries.

    PubMed

    Abad, Z Gloria; Abad, Jorge A; Cacciola, Santa Olga; Pane, Antonella; Faedda, Roberto; Moralejo, Eduardo; Pérez-Sierra, Ana; Abad-Campos, Paloma; Alvarez-Bernaola, Luis A; Bakonyi, József; Józsa, András; Herrero, Maria Luz; Burgess, Treena I; Cunnington, James H; Smith, Ian W; Balci, Yilmaz; Blomquist, Cheryl; Henricot, Béatrice; Denton, Geoffrey; Spies, Chris; Mcleod, Adele; Belbahri, Lassaad; Cooke, David; Kageyama, Koji; Uematsu, Seiji; Kurbetli, Ilker; Değirmenci, Kemal

    2014-01-01

    A non-papillate, heterothallic Phytophthora species first isolated in 2001 and subsequently from symptomatic roots, crowns and stems of 33 plant species in 25 unrelated botanical families from 13 countries is formally described here as a new species. Symptoms on various hosts included crown and stem rot, chlorosis, wilting, leaf blight, cankers and gumming. This species was isolated from Australia, Hungary, Israel, Italy, Japan, the Netherlands, Norway, South Africa, Spain, Taiwan, Turkey, the United Kingdom and United States in association with shrubs and herbaceous ornamentals grown mainly in greenhouses. The most prevalent hosts are English ivy (Hedera helix) and Cistus (Cistus salvifolius). The association of the species with acorn banksia (Banksia prionotes) plants in natural ecosystems in Australia, in affected vineyards (Vitis vinifera) in South Africa and almond (Prunus dulcis) trees in Spain and Turkey in addition to infection of shrubs and herbaceous ornamentals in a broad range of unrelated families are a sign of a wide ecological adaptation of the species and its potential threat to agricultural and natural ecosystems. The morphology of the persistent non-papillate ellipsoid sporangia, unique toruloid lobate hyphal swellings and amphigynous antheridia does not match any of the described species. Phylogenetic analysis based on sequences of the ITS rDNA, EF-1α, and β-tub supported that this organism is a hitherto unknown species. It is closely related to species in ITS clade 7b with the most closely related species being P. sojae. The name Phytophthora niederhauserii has been used in previous studies without the formal description of the holotype. This name is validated in this manuscript with the formal description of Phytophthora niederhauserii Z.G. Abad et J.A. Abad, sp. nov. The name is coined to honor Dr John S. Niederhauser, a notable plant pathologist and the 1990 World Food Prize laureate.

  17. Phytophthora niederhauserii sp. nov., a polyphagous species associated with ornamentals, fruit trees and native plants in 13 countries.

    PubMed

    Abad, Z Gloria; Abad, Jorge A; Cacciola, Santa Olga; Pane, Antonella; Faedda, Roberto; Moralejo, Eduardo; Pérez-Sierra, Ana; Abad-Campos, Paloma; Alvarez-Bernaola, Luis A; Bakonyi, József; Józsa, András; Herrero, Maria Luz; Burgess, Treena I; Cunnington, James H; Smith, Ian W; Balci, Yilmaz; Blomquist, Cheryl; Henricot, Béatrice; Denton, Geoffrey; Spies, Chris; Mcleod, Adele; Belbahri, Lassaad; Cooke, David; Kageyama, Koji; Uematsu, Seiji; Kurbetli, Ilker; Değirmenci, Kemal

    2014-01-01

    A non-papillate, heterothallic Phytophthora species first isolated in 2001 and subsequently from symptomatic roots, crowns and stems of 33 plant species in 25 unrelated botanical families from 13 countries is formally described here as a new species. Symptoms on various hosts included crown and stem rot, chlorosis, wilting, leaf blight, cankers and gumming. This species was isolated from Australia, Hungary, Israel, Italy, Japan, the Netherlands, Norway, South Africa, Spain, Taiwan, Turkey, the United Kingdom and United States in association with shrubs and herbaceous ornamentals grown mainly in greenhouses. The most prevalent hosts are English ivy (Hedera helix) and Cistus (Cistus salvifolius). The association of the species with acorn banksia (Banksia prionotes) plants in natural ecosystems in Australia, in affected vineyards (Vitis vinifera) in South Africa and almond (Prunus dulcis) trees in Spain and Turkey in addition to infection of shrubs and herbaceous ornamentals in a broad range of unrelated families are a sign of a wide ecological adaptation of the species and its potential threat to agricultural and natural ecosystems. The morphology of the persistent non-papillate ellipsoid sporangia, unique toruloid lobate hyphal swellings and amphigynous antheridia does not match any of the described species. Phylogenetic analysis based on sequences of the ITS rDNA, EF-1α, and β-tub supported that this organism is a hitherto unknown species. It is closely related to species in ITS clade 7b with the most closely related species being P. sojae. The name Phytophthora niederhauserii has been used in previous studies without the formal description of the holotype. This name is validated in this manuscript with the formal description of Phytophthora niederhauserii Z.G. Abad et J.A. Abad, sp. nov. The name is coined to honor Dr John S. Niederhauser, a notable plant pathologist and the 1990 World Food Prize laureate. PMID:24871599

  18. A novel method for efficient and abundant production of Phytophthora brassicae zoospores on Brussels sprout leaf discs

    PubMed Central

    Bouwmeester, Klaas; Govers, Francine

    2009-01-01

    Background Phytophthora species are notorious oomycete pathogens that cause diseases on a wide range of plants. Our understanding how these pathogens are able to infect their host plants will benefit greatly from information obtained from model systems representative for plant-Phytophthora interactions. One attractive model system is the interaction between Arabidopsis and Phytophthora brassicae. Under laboratory conditions, Arabidopsis can be easily infected with mycelial plugs as inoculum. In the disease cycle, however, sporangia or zoospores are the infectious propagules. Since the current P. brassicae zoospore isolation methods are generally regarded as inefficient, we aimed at developing an alternative method for obtaining high concentrations of P. brassicae zoospores. Results P. brassicae isolates were tested for pathogenicity on Brussels sprout plants (Brassica oleracea var. gemmifera). Microscopic examination of leaves, stems and roots infected with a GFP-tagged transformant of P. brassicae clearly demonstrated the susceptibility of the various tissues. Leaf discs were cut from infected Brussels sprout leaves, transferred to microwell plates and submerged in small amounts of water. In the leaf discs the hyphae proliferated and abundant formation of zoosporangia was observed. Upon maturation the zoosporangia released zoospores in high amounts and zoospore production continued during a period of at least four weeks. The zoospores were shown to be infectious on Brussels sprouts and Arabidopsis. Conclusion The in vitro leaf disc method established from P. brassicae infected Brussels sprout leaves facilitates convenient and high-throughput production of infectious zoospores and is thus suitable to drive small and large scale inoculation experiments. The system has the advantage that zoospores are produced continuously over a period of at least one month. PMID:19698127

  19. Dual RNA-Sequencing of Eucalyptus nitens during Phytophthora cinnamomi Challenge Reveals Pathogen and Host Factors Influencing Compatibility

    PubMed Central

    Meyer, Febé E.; Shuey, Louise S.; Naidoo, Sitha; Mamni, Thandekile; Berger, Dave K.; Myburg, Alexander A.; van den Berg, Noëlani; Naidoo, Sanushka

    2016-01-01

    Damage caused by Phytophthora cinnamomi Rands remains an important concern on forest tree species. The pathogen causes root and collar rot, stem cankers, and dieback of various economically important Eucalyptus spp. In South Africa, susceptible cold tolerant Eucalyptus plantations have been affected by various Phytophthora spp. with P. cinnamomi considered one of the most virulent. The molecular basis of this compatible interaction is poorly understood. In this study, susceptible Eucalyptus nitens plants were stem inoculated with P. cinnamomi and tissue was harvested five days post inoculation. Dual RNA-sequencing, a technique which allows the concurrent detection of both pathogen and host transcripts during infection, was performed. Approximately 1% of the reads mapped to the draft genome of P. cinnamomi while 78% of the reads mapped to the Eucalyptus grandis genome. The highest expressed P. cinnamomi gene in planta was a putative crinkler effector (CRN1). Phylogenetic analysis indicated the high similarity of this P. cinnamomi CRN1 to that of Phytophthora infestans. Some CRN effectors are known to target host nuclei to suppress defense. In the host, over 1400 genes were significantly differentially expressed in comparison to mock inoculated trees, including suites of pathogenesis related (PR) genes. In particular, a PR-9 peroxidase gene with a high similarity to a Carica papaya PR-9 ortholog previously shown to be suppressed upon infection by Phytophthora palmivora was down-regulated two-fold. This PR-9 gene may represent a cross-species effector target during P. cinnamomi infection. This study identified pathogenicity factors, potential manipulation targets, and attempted host defense mechanisms activated by E. nitens that contributed to the susceptible outcome of the interaction. PMID:26973660

  20. Dual RNA-Sequencing of Eucalyptus nitens during Phytophthora cinnamomi Challenge Reveals Pathogen and Host Factors Influencing Compatibility.

    PubMed

    Meyer, Febé E; Shuey, Louise S; Naidoo, Sitha; Mamni, Thandekile; Berger, Dave K; Myburg, Alexander A; van den Berg, Noëlani; Naidoo, Sanushka

    2016-01-01

    Damage caused by Phytophthora cinnamomi Rands remains an important concern on forest tree species. The pathogen causes root and collar rot, stem cankers, and dieback of various economically important Eucalyptus spp. In South Africa, susceptible cold tolerant Eucalyptus plantations have been affected by various Phytophthora spp. with P. cinnamomi considered one of the most virulent. The molecular basis of this compatible interaction is poorly understood. In this study, susceptible Eucalyptus nitens plants were stem inoculated with P. cinnamomi and tissue was harvested five days post inoculation. Dual RNA-sequencing, a technique which allows the concurrent detection of both pathogen and host transcripts during infection, was performed. Approximately 1% of the reads mapped to the draft genome of P. cinnamomi while 78% of the reads mapped to the Eucalyptus grandis genome. The highest expressed P. cinnamomi gene in planta was a putative crinkler effector (CRN1). Phylogenetic analysis indicated the high similarity of this P. cinnamomi CRN1 to that of Phytophthora infestans. Some CRN effectors are known to target host nuclei to suppress defense. In the host, over 1400 genes were significantly differentially expressed in comparison to mock inoculated trees, including suites of pathogenesis related (PR) genes. In particular, a PR-9 peroxidase gene with a high similarity to a Carica papaya PR-9 ortholog previously shown to be suppressed upon infection by Phytophthora palmivora was down-regulated two-fold. This PR-9 gene may represent a cross-species effector target during P. cinnamomi infection. This study identified pathogenicity factors, potential manipulation targets, and attempted host defense mechanisms activated by E. nitens that contributed to the susceptible outcome of the interaction. PMID:26973660

  1. Phytophthora sojae TatD nuclease positively regulates sporulation and negatively regulates pathogenesis.

    PubMed

    Chen, Linlin; Shen, Danyu; Sun, Nannan; Xu, Jing; Wang, Wen; Dou, Daolong

    2014-10-01

    During pathogenic interactions, both the host and pathogen are exposed to conditions that induce programmed cell death (PCD). Certain aspects of PCD have been recently examined in eukaryotic microbes but not in oomycetes. Here, we identified conserved TatD proteins in Phytophthora sojae; the proteins are key components of DNA degradation in apoptosis. We selected PsTatD4 for further investigation because the enzyme is unique to the oomycete branch of the phylogenetic tree. The purified protein exhibited DNase activity in vitro. Its expression was upregulated in sporangia and later infective stages but downregulated in cysts and during early infection. Functional analysis revealed that the gene was required for sporulation and zoospore production, and the expression levels were associated with the numbers of hydrogen-peroxide-induced terminal dUTP nick end-labeling-positive cells. Furthermore, overexpression of PsTatD4 gene reduced the virulence in a susceptible soybean cultivar. Together, these data suggest that apoptosis may play different roles in the early and late infective stages of P. sojae, and that PsTatD4 is a key regulator of infection. The association of PsTatD4 and apoptosis will lay a foundation to understanding the basic biology of apoptosis and its roles in P. sojae disease cycle.

  2. Quantification of induced resistance against Phytophthora species expressing GFP as a vital marker: beta-aminobutyric acid but not BTH protects potato and Arabidopsis from infection.

    PubMed

    Si-Ammour, Azeddine; Mauch-Mani, Brigitte; Mauch, Felix

    2003-07-01

    SUMMARY Induced resistance was studied in the model pathosystem Arabidopsis-Phytophthora brassicae (formerly P. porri) in comparison with the agronomically important late blight disease of potato caused by Phytophthora infestans. For the quantification of disease progress, both Phytophthora species were transformed with the vector p34GFN carrying the selectable marker gene neomycine phosphotransferase (nptII) and the reporter gene green fluorescent protein (gfp). Eighty five per cent of the transformants of P. brassicae and P. infestans constitutively expressed GFP at high levels at all developmental stages both in vitro and in planta. Transformants with high GFP expression and normal in vitro growth and virulence were selected to quantify pathogen growth by measuring the in planta emitted GFP fluorescence. This non-destructive monitoring of the infection process was applied to analyse the efficacy of two chemical inducers of disease resistance, a functional SA-analogue, benzothiadiazole (BTH), and beta-aminobutyric acid (BABA) which is involved in priming mechanisms of unknown nature. BABA pre-treatment (300 microm) via soil drench applied 24 h before inoculation completely protected the susceptible Arabidopsis accession Landsberg erecta (Ler) from infection with P. brassicae. A similar treatment with BTH (330 microm) did not induce resistance. Spraying the susceptible potato cultivar Bintje with BABA (1 mm) 2 days before inoculation resulted in a phenocopy of the incompatible interaction shown by the resistant potato cultivar Matilda while BTH (1.5 mm) did not protect Bintje from severe infection. Thus, in both pathosystems, the mechanisms of induced resistance appeared to be similar, suggesting that the Arabidopsis-P. brassicae pathosystem is a promising model for the molecular analysis of induced resistance mechanisms of potato against the late blight disease.

  3. Phytophthora Database: A forensic database supporting the identification and monitoring of Phytophthora

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Due to their high virulence and ability to spread rapidly, Phytophthora species represent a serious threat to agricultural production and ecological systems. Many novel Phytophthora species have been reported in recent years, indicative of our limited understanding of the ecology and diversity of Ph...

  4. Genome sequences of two Phytophthora species responsible for Sudden Oak Death and Soybean Root Rot provide novel insights into their evolutionary origins and mechanisms of pathogenesis

    SciTech Connect

    Tyler, Brett M.; Tripathi, Sucheta; Aerts, Andrea; Bensasson, Douda; Dehal, Paramvir; Dubchak, Inna; Garbelotto, Matteo; Gijzen, Mark; Huang, Wayne; Ivors, Kelly; Jiang, Rays; Kamoun, Sophien; Krampis, Konstantinos; Lamour, Kurt; McDonald, Hayes; Medina, Monica; Morris, Paul; Putnam, Nik; Rash, Sam; Salamov, Asaf; Smith, Brian; Smith, Joe; Terry, Astrid; Torto, Trudy; Grigoriev, Igor; Rokhsar, Daniel; Boore, Jeffrey

    2005-12-01

    The approximately 60 species of Phytophthora are all destructive pathogens, causing rots of roots, stems, leaves and fruits of a wide range of agriculturally and ornamentally important plants (1). Some species, such as P. cinnamomi, P. parasitica and P. cactorum, each attack hundreds of different plant host species, whereas others are more restricted. Some of the crops where Phytophthora infections cause the greatest financial losses include potato, soybean, tomato, alfalfa, tobacco, peppers, cucurbits, pineapple, strawberry, raspberry and a wide range of perennial tree crops, especially citrus, avocado, almonds, walnuts, apples and cocoa, and they also heavily affect the ornamental, nursery and forestry industries. The economic damage overall to crops in the United States by Phytophthora species is estimated in the tens of billions of dollars, including the costs of control measures, and worldwide it is many times this amount (1). In the northern midwest of the U.S., P. sojae causes $200 million in annual losses to soybean alone, and worldwide causes around $1-2 billion in losses per year. P. infestans infections resulted in the Irish potato famine last century and continues to be a difficult and worsening problem for potato and tomato growers worldwide, with worldwide costs estimated at $5 billion per year.

  5. Diagnostics of Tree Diseases Caused by Phytophthora austrocedri Species.

    PubMed

    Mulholland, Vincent; Elliot, Matthew; Green, Sarah

    2015-01-01

    We present methods for the detection and quantification of four Phytophthora species which are pathogenic on trees; Phytophthora ramorum, Phytophthora kernoviae, Phytophthora lateralis, and Phytophthora austrocedri. Nucleic acid extraction methods are presented for phloem tissue from trees, soil, and pure cultures on agar plates. Real-time PCR methods are presented and include primer and probe sets for each species, general advice on real-time PCR setup and data analysis. A method for sequence-based identification, useful for pure cultures, is also included.

  6. Phytophthora parasitica: a model oomycete plant pathogen

    PubMed Central

    Meng, Yuling; Zhang, Qiang; Ding, Wei; Shan, Weixing

    2014-01-01

    Oomycetes are eukaryotic microorganisms morphologically similar to but phylogenetically distant from true fungi. Most species in the genus Phytophthora of oomycetes are devastating plant pathogens, causing damages to both agricultural production and natural ecosystems. Tremendous progress has been achieved in recent years in diversity, evolution and lifestyles of oomycete plant pathogens, as well as on the understanding of genetic and molecular basis of oomycete-plant interactions. Phytophthora parasitica is a soilborne pathogen with a wide range of host plants and represents most species in the genus Phytophthora. In this review, we present some recent progress of P. parasitica research by highlighting important features that make it emerge as a model species of oomycete pathogens. The emerged model pathogen will facilitate improved understanding of oomycete biology and pathology that are crucial to the development of novel disease-control strategies and improved disease-control measures. PMID:24999436

  7. Phytophthora parasitica: a model oomycete plant pathogen.

    PubMed

    Meng, Yuling; Zhang, Qiang; Ding, Wei; Shan, Weixing

    2014-06-01

    Oomycetes are eukaryotic microorganisms morphologically similar to but phylogenetically distant from true fungi. Most species in the genus Phytophthora of oomycetes are devastating plant pathogens, causing damages to both agricultural production and natural ecosystems. Tremendous progress has been achieved in recent years in diversity, evolution and lifestyles of oomycete plant pathogens, as well as on the understanding of genetic and molecular basis of oomycete-plant interactions. Phytophthora parasitica is a soilborne pathogen with a wide range of host plants and represents most species in the genus Phytophthora. In this review, we present some recent progress of P. parasitica research by highlighting important features that make it emerge as a model species of oomycete pathogens. The emerged model pathogen will facilitate improved understanding of oomycete biology and pathology that are crucial to the development of novel disease-control strategies and improved disease-control measures. PMID:24999436

  8. Bud Rot Caused by Phytophthora palmivora: A Destructive Emerging Disease of Oil Palm.

    PubMed

    Torres, G A; Sarria, G A; Martinez, G; Varon, F; Drenth, A; Guest, D I

    2016-04-01

    Oomycetes from the genus Phytophthora are among the most important plant pathogens in agriculture. Epidemics caused by P. infestans precipitated the great Irish famine and had a major impact on society and human history. In the tropics, P. palmivora is a pathogen of many plant species including cacao (Theobroma cacao), citrus (Citrus sp.), durian (Durio zibethines), jackfruit (Artrocarpus heterophyllus), rubber (Hevea brasiliensis), and several palm species including coconut (Cocos nucifera), and the African oil palm (Elaeis guineensis) as determined recently. The first localized epidemics of bud rot in oil palm in Colombia were reported in 1964. However, recent epidemics of bud rot have destroyed more than 70,000 ha of oil palm in the Western and Central oil palm growing regions of Colombia. The agricultural, social, and economic implications of these outbreaks have been significant in Colombia. Identification of the pathogen after 100 years of investigating the disease in the world enabled further understanding of infection, expression of a range of symptoms, and epidemiology of the disease. This review examines the identification of P. palmivora as the cause of bud rot in Colombia, its epidemiology, and discusses the importance of P. palmivora as a major threat to oil palm plantings globally.

  9. Mitochondrial genome sequences and comparative genomics ofPhytophthora ramorum and P. sojae

    SciTech Connect

    Martin, Frank N.; Douda, Bensasson; Tyler, Brett M.; Boore,Jeffrey L.

    2007-01-01

    The complete sequences of the mitochondrial genomes of theoomycetes of Phytophthora ramorum and P. sojae were determined during thecourse of their complete nuclear genome sequencing (Tyler, et al. 2006).Both are circular, with sizes of 39,314 bp for P. ramorum and 42,975 bpfor P. sojae. Each contains a total of 37 identifiable protein-encodinggenes, 25 or 26 tRNAs (P. sojae and P. ramorum, respectively)specifying19 amino acids, and a variable number of ORFs (7 for P. ramorum and 12for P. sojae) which are potentially additional functional genes.Non-coding regions comprise approximately 11.5 percent and 18.4 percentof the genomes of P. ramorum and P. sojae, respectively. Relative to P.sojae, there is an inverted repeat of 1,150 bp in P. ramorum thatincludes an unassigned unique ORF, a tRNA gene, and adjacent non-codingsequences, but otherwise the gene order in both species is identical.Comparisons of these genomes with published sequences of the P. infestansmitochondrial genome reveals a number of similarities, but the gene orderin P. infestans differs in two adjacent locations due to inversions.Sequence alignments of the three genomes indicated sequence conservationranging from 75 to 85 percent and that specific regions were morevariable than others.

  10. Bud Rot Caused by Phytophthora palmivora: A Destructive Emerging Disease of Oil Palm.

    PubMed

    Torres, G A; Sarria, G A; Martinez, G; Varon, F; Drenth, A; Guest, D I

    2016-04-01

    Oomycetes from the genus Phytophthora are among the most important plant pathogens in agriculture. Epidemics caused by P. infestans precipitated the great Irish famine and had a major impact on society and human history. In the tropics, P. palmivora is a pathogen of many plant species including cacao (Theobroma cacao), citrus (Citrus sp.), durian (Durio zibethines), jackfruit (Artrocarpus heterophyllus), rubber (Hevea brasiliensis), and several palm species including coconut (Cocos nucifera), and the African oil palm (Elaeis guineensis) as determined recently. The first localized epidemics of bud rot in oil palm in Colombia were reported in 1964. However, recent epidemics of bud rot have destroyed more than 70,000 ha of oil palm in the Western and Central oil palm growing regions of Colombia. The agricultural, social, and economic implications of these outbreaks have been significant in Colombia. Identification of the pathogen after 100 years of investigating the disease in the world enabled further understanding of infection, expression of a range of symptoms, and epidemiology of the disease. This review examines the identification of P. palmivora as the cause of bud rot in Colombia, its epidemiology, and discusses the importance of P. palmivora as a major threat to oil palm plantings globally. PMID:26714102

  11. Phytophthora Genome Sequences Uncover Evolutionary Origins and Mechanisms of Pathogenesis

    SciTech Connect

    Lamour, Kurt H; McDonald, W Hayes; Savidor, Alon

    2006-01-01

    Genome sequences of the soybean pathogen, Phytophthora sojae, and the sudden oak death pathogen, Phytophthora ramorum, suggest a photosynthetic past and reveal recent massive expansion and diversification of potential pathogenicity gene families. Abstract: Draft genome sequences of the soybean pathogen, Phytophthora sojae, and the sudden oak death pathogen, Phytophthora ramorum, have been determined. O mycetes such as these Phytophthora species share the kingdom Stramenopila with photosynthetic algae such as diatoms and the presence of many Phytophthora genes of probable phototroph origin support a photosynthetic ancestry for the stramenopiles. Comparison of the two species' genomes reveals a rapid expansion and diversification of many protein families associated with plant infection such as hydrolases, ABC transporters, protein toxins, proteinase inhibitors and, in particular, a superfamily of 700 proteins with similarity to known o mycete avirulence genes.

  12. Phytophthora Genome Sequences Uncover Evolutionary Origins and Mechanisms of Pathogenesis

    SciTech Connect

    Tyler, Brett M.; Tripathy, Sucheta; Zhang, Xuemin; Dehal, Paramvir; Jiang, Rays H. Y.; Aerts, Andrea; Arredondo, Felipe D.; Baxter, Laura; Bensasson, Douda; Beynon, JIm L.; Chapman, Jarrod; Damasceno, Cynthia M. B.; Dorrance, Anne E.; Dou, Daolong; Dickerman, Allan W.; Dubchak, Inna L.; Garbelotto, Matteo; Gijzen, Mark; Gordon, Stuart G.; Govers, Francine; Grunwald, NIklaus J.; Huang, Wayne; Ivors, Kelly L.; Jones, Richard W.; Kamoun, Sophien; Krampis, Konstantinos; Lamour, Kurt H.; Lee, Mi-Kyung; McDonald, W. Hayes; Medina, Monica; Meijer, Harold J. G.; Nordberg, Erik K.; Maclean, Donald J.; Ospina-Giraldo, Manuel D.; Morris, Paul F.; Phuntumart, Vipaporn; Putnam, Nicholas J.; Rash, Sam; Rose, Jocelyn K. C.; Sakihama, Yasuko; Salamov, Asaf A.; Savidor, Alon; Scheuring, Chantel F.; Smith, Brian M.; Sobral, Bruno W. S.; Terry, Astrid; Torto-Alalibo, Trudy A.; Win, Joe; Xu, Zhanyou; Zhang, Hongbin; Grigoriev, Igor V.; Rokhsar, Daniel S.; Boore, Jeffrey L.

    2006-04-17

    Draft genome sequences have been determined for the soybean pathogen Phytophthora sojae and the sudden oak death pathogen Phytophthora ramorum. Oömycetes such as these Phytophthora species share the kingdom Stramenopila with photosynthetic algae such as diatoms, and the presence of many Phytophthora genes of probable phototroph origin supports a photosynthetic ancestry for the stramenopiles. Comparison of the two species' genomes reveals a rapid expansion and diversification of many protein families associated with plant infection such as hydrolases, ABC transporters, protein toxins, proteinase inhibitors, and, in particular, a superfamily of 700 proteins with similarity to known oömycete avirulence genes.

  13. Secondary Kill Effect of Deltamethrin on Triatoma infestans

    PubMed Central

    MALONEY, KATHLEEN M.; ANCCA-JUAREZ, JENNY; SALAZAR, RENZO; BORRINI-MAYORI, KATTY; PAMO-TITO, DANITZA; KEATING, JOSEPH A.; LEVY, MICHAEL Z.

    2012-01-01

    Control of the Chagas disease vector, Triatoma infestans, relies on the application of pyrethroid insecticides, especially deltamethrin. We performed laboratory studies to determine whether a T. infestans nymph that comes into contact with a deltamethrin-treated surface horizontally transfers the insecticide to subsequent triatomines. We found that a triatomine that walks on a deltamethrin-treated surface for a short period of time has the ability to transport the insecticide in concentrations sufficient to kill other triatomines with which it comes into contact. The effect was limited to high-density environments, and mortality as a result of secondary exposure was greater among second-instar nymphs compared with fifth-instar nymphs. Our results suggest that deltamethrin could be killing triatomines through both direct and indirect contact, although it remains unclear whether the phenomenon occurs in natural conditions. PMID:21845956

  14. Urban ecology of Triatoma infestans in San Juan, Argentina.

    PubMed

    Vallvé, S L; Rojo, H; Wisnivesky-Colli, C

    1996-01-01

    This study was performed in an urban neighborhood of the capital city of the province of San Juan, Argentina. Erected as a housing complex, the place consists of 768 flats distributed in buildings of three and seven floors each. A survey was carried out in 33% of the dwellings, enquiring about the number of Triatoma infestans found indoors, stage of the bug development-nymph or adult- and how these insects had entered their homes. Adult T.infestans were found on all floors; 163 people (64%) had found them at least once, and 130 (51%) several times. Dispersal flight seems to have been the main mechanism of infestation by adult bugs in this area, and a total of 51% of the surveyed inhabitants reported that the insects had flown into their flats. PMID:9070399

  15. The effect of pyramiding Phytophthora infestans resistance genes RPi-mcd1 and RPi-ber in potato

    PubMed Central

    Tan, M. Y. Adillah; Hutten, Ronald C. B.; Visser, Richard G. F.

    2010-01-01

    Despite efforts to control late blight in potatoes by introducing Rpi-genes from wild species into cultivated potato, there are still concerns regarding the durability and level of resistance. Pyramiding Rpi-genes can be a solution to increase both durability and level of resistance. In this study, two resistance genes, RPi-mcd1 and RPi-ber, introgressed from the wild tuber-bearing potato species Solanum microdontum and S. berthaultii were combined in a diploid S. tuberosum population. Individual genotypes from this population were classified after four groups, carrying no Rpi-gene, with only RPi-mcd1, with only RPi-ber, and a group with the pyramided RPi-mcd1 and RPi-ber by means of tightly linked molecular markers. The levels of resistance between the groups were compared in a field experiment in 2007. The group with RPi-mcd1 showed a significant delay to reach 50% infection of the leaf area of 3 days. The group with RPi-ber showed a delay of 3 weeks. The resistance level in the pyramid group suggested an additive effect of RPi-mcd1 with RPi-ber. This suggests that potato breeding can benefit from combining individual Rpi-genes, irrespective of the weak effect of RPi-mcd1 or the strong effect of RPi-ber. PMID:20204320

  16. Wild Triatoma infestans, a potential threat that needs to be monitored.

    PubMed

    Noireau, François

    2009-07-01

    The current persistence of Triatoma infestans, and therefore of Chagas disease transmission, in the Andean valleys of Bolivia and the Gran Chaco (precisely where wild populations of the vector are widespread), indicates a possible relationship between these two occurrences. This paper provides an overview of the current knowledge regarding wild T. infestans in Bolivia. The different morphs of the wild vector, their known distributions and some traits of their biology and ecology are presented. Particularly interesting is the considerable behavioural and chromatic plasticity that is displayed by wild T. infestans. According to the biogeographic region, different morphs of the vector occur in rupicolous habitats (common form and Mataral morph in Andean wild T. infestans) or arboreal ones ('dark morph' populations from the Chaco). The high genetic variability found at the microgeographical scale in Andean wild T. infestans favours the hypothesis that the Andes were the centre of origin and dispersal of T. infestans throughout South America. The relevant question regarding the origin of domestic populations is also addressed. Finally, current considerations of the epidemiological significance of wild T. infestans are discussed in the context of recent discoveries. Even if several observations support the epidemiological risk represented by wild T. infestans, the climatic and environmental conditions of their distribution areas would not favour the continued flow of triatomines between sylvatic refuges and domestic environments.

  17. Phytophthora Database 2.0: update and future direction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The online community resource Phytophthora Database (PD) was developed to support accurate and rapid identification of Phytophthora and to help characterize and catalog the diversity and evolutionary relationships within the genus. Since its release in 2008, its sequence database has grown to cover ...

  18. Phytophthora root rot resistance in soybean E00003

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora root rot (PRR), caused by the oomycete Phytophthora sojae, is a devastating disease in soybean production. Using resistant cultivars has been suggested as the best solution for disease management. Michigan elite soybean E00003 is resistant to P. sojae and has been used as a PRR resist...

  19. Sylvatic Triatoma infestans (Reduviidae, Triatominae) in the Andean valleys of Bolivia.

    PubMed

    Cortez, Mirko Rojas; Emperaire, Laure; Piccinali, Romina V; Gürtler, Ricardo E; Torrico, Faustino; Jansen, Ana Maria; Noireau, François

    2007-04-01

    Triatoma infestans is the main vector of Chagas disease in the Southern Cone countries. Wild populations of T. infestans appear widespread throughout the Andean valleys of Bolivia. In Cotapachi (2750 m asl), all sorts of rocky outcrops, regardless of their size, provided good refuges for T. infestans. Of the 1120 ecotopes investigated, 330 (29.5%) contained triatomines and 92% of the collected insects were nymphal instars. In the cold season, triatomine densities were similar in small and large outcrops. During the hot season, bug densities were higher in the larger outcrops, particularly in those located in peridomestic sites. T. infestans populations apparently produced one generation per year. Over half the sampled bugs were positive for T. cruzi infection. At Mataral (1750 m asl), a site located in the inter-Andean Chaco, a new morph of T. infestans was detected in a sylvatic environment.

  20. Genome sequences of six Phytophthora species associated with forests in New Zealand.

    PubMed

    Studholme, D J; McDougal, R L; Sambles, C; Hansen, E; Hardy, G; Grant, M; Ganley, R J; Williams, N M

    2016-03-01

    In New Zealand there has been a long association of Phytophthora diseases in forests, nurseries, remnant plantings and horticultural crops. However, new Phytophthora diseases of trees have recently emerged. Genome sequencing has been performed for 12 Phytophthora isolates, from six species: Phytophthora pluvialis, Phytophthora kernoviae, Phytophthora cinnamomi, Phytophthora agathidicida, Phytophthora multivora and Phytophthora taxon Totara. These sequences will enable comparative analyses to identify potential virulence strategies and ultimately facilitate better control strategies. This Whole Genome Shotgun data have been deposited in DDBJ/ENA/GenBank under the accession numbers LGTT00000000, LGTU00000000, JPWV00000000, JPWU00000000, LGSK00000000, LGSJ00000000, LGTR00000000, LGTS00000000, LGSM00000000, LGSL00000000, LGSO00000000, and LGSN00000000.

  1. Genome sequences of six Phytophthora species associated with forests in New Zealand

    PubMed Central

    Studholme, D.J.; McDougal, R.L.; Sambles, C.; Hansen, E.; Hardy, G.; Grant, M.; Ganley, R.J.; Williams, N.M.

    2015-01-01

    In New Zealand there has been a long association of Phytophthora diseases in forests, nurseries, remnant plantings and horticultural crops. However, new Phytophthora diseases of trees have recently emerged. Genome sequencing has been performed for 12 Phytophthora isolates, from six species: Phytophthora pluvialis, Phytophthora kernoviae, Phytophthora cinnamomi, Phytophthora agathidicida, Phytophthora multivora and Phytophthora taxon Totara. These sequences will enable comparative analyses to identify potential virulence strategies and ultimately facilitate better control strategies. This Whole Genome Shotgun data have been deposited in DDBJ/ENA/GenBank under the accession numbers LGTT00000000, LGTU00000000, JPWV00000000, JPWU00000000, LGSK00000000, LGSJ00000000, LGTR00000000, LGTS00000000, LGSM00000000, LGSL00000000, LGSO00000000, and LGSN00000000. PMID:26981359

  2. The importin α subunit PsIMPA1 mediates the oxidative stress response and is required for the pathogenicity of Phytophthora sojae.

    PubMed

    Yang, Xinyu; Ding, Fa; Zhang, Lei; Sheng, Yuting; Zheng, Xiaobo; Wang, Yuanchao

    2015-09-01

    The sensing of extracellular signals and their transduction into an appropriate response are crucial for the survival and virulence of plant pathogens. Eukaryotic plant pathogens must overcome the obstacles posed by nuclear membranes to manipulate gene expression to adapt to the host challenge. A highly sophisticated mechanism is the use of importins to transport proteins into the nucleus. In this study, we identified a conserved importin α gene, PsIMPA1, in Phytophthora sojae that was differentially expressed during the life cycle of this soybean pathogen. PsIMPA1 expression was lowest in zoospores and cysts but relatively consistent during the other life cycle stages, except for a slight increase at 6h post infection. Silenced mutants Psimpa1 had a decreased growth rate, an aberrant mycelial morphology, and a severely impaired ability to form oospores and sporangia. In addition, the Psimpa1 mutants exhibited reduced pathogenicity compared to the wild type. 3,3-Diaminobenzidine (DAB) staining and in vitro hydrogen peroxide tolerance assays showed that the scavenging of reactive oxygen species by these mutants was significantly impaired. Taken together, these results indicate that PsIMPA1 regulates multiple processes during the life cycle of P. sojae.

  3. Genome sequencing and mapping reveal loss of heterozygosity as a mechanism for rapid adaptation in the vegetable pathogen Phytophthora capsici

    PubMed Central

    Lamour, Kurt H.; Mudge, Joann; Gobena, Daniel; Hurtado-Gonzales, Oscar P.; Schmutz, Jeremy; Kuo, Alan; Miller, Neil A.; Rice, Brandon J.; Raffaele, Sylvain; Cano, Liliana M.; Bharti, Arvind K.; Donahoo, Ryan S.; Finley, Sabra; Huitema, Edgar; Hulvey, Jon; Platt, Darren; Salamov, Asaf; Savidor, Alon; Sharma, Rahul; Stam, Remco; Storey, Dylan; Thines, Marco; Win, Joe; Haas, Brian J.; Dinwiddie, Darrell L.; Jenkins, Jerry; Knight, James R.; Affourtit, Jason P.; Han, Cliff S.; Chertkov, Olga; Lindquist, Erika A.; Detter, Chris; Grigoriev, Igor V.; Kamoun, Sophien; Kingsmore, Stephen F.

    2013-01-01

    The oomycete vegetable pathogen Phytophthora capsici has shown remarkable adaptation to fungicides and new hosts. Like other members of this destructive genus, P. capsici has an explosive epidemiology, rapidly producing massive numbers of asexual spores on infected hosts. In addition, P. capsici can remain dormant for years as sexually-recombined oospores, making it difficult to produce crops at infested sites, and allowing outcrossing populations to maintain significant genetic variation. Genome sequencing, development of a high-density genetic map, and integrative genomic/genetic characterization of P. capsici field isolates and intercross progeny revealed significant mitotic loss of heterozygosity (LOH) and higher levels of SNVs than those reported for humans, plants, and P. infestans. LOH was detected in clonally propagated field isolates and sexual progeny, cumulatively affecting >30% of the genome. LOH altered genotypes for more than 11,000 single nucleotide variant (SNV) sites and showed a strong association with changes in mating type and pathogenicity. Overall, it appears that LOH may provide a rapid mechanism for fixing alleles and may be an important component of adaptability for P. capsici. PMID:22712506

  4. BABA and Phytophthora nicotianae Induce Resistance to Phytophthora capsici in Chile Pepper (Capsicum annuum)

    PubMed Central

    Stamler, Rio A.; Holguin, Omar; Dungan, Barry; Schaub, Tanner; Sanogo, Soumaila; Goldberg, Natalie; Randall, Jennifer J.

    2015-01-01

    Induced resistance in plants is a systemic response to certain microorganisms or chemicals that enhances basal defense responses during subsequent plant infection by pathogens. Inoculation of chile pepper with zoospores of non-host Phytophthora nicotianae or the chemical elicitor beta-aminobutyric acid (BABA) significantly inhibited foliar blight caused by Phytophthora capsici. Tissue extract analyses by GC/MS identified conserved change in certain metabolite concentrations following P. nicotianae or BABA treatment. Induced chile pepper plants had reduced concentrations of sucrose and TCA cycle intermediates and increased concentrations of specific hexose-phosphates, hexose-disaccharides and amino acids. Galactose, which increased significantly in induced chile pepper plants, was shown to inhibit growth of P. capsici in a plate assay. PMID:26020237

  5. BABA and Phytophthora nicotianae Induce Resistance to Phytophthora capsici in Chile Pepper (Capsicum annuum).

    PubMed

    Stamler, Rio A; Holguin, Omar; Dungan, Barry; Schaub, Tanner; Sanogo, Soumaila; Goldberg, Natalie; Randall, Jennifer J

    2015-01-01

    Induced resistance in plants is a systemic response to certain microorganisms or chemicals that enhances basal defense responses during subsequent plant infection by pathogens. Inoculation of chile pepper with zoospores of non-host Phytophthora nicotianae or the chemical elicitor beta-aminobutyric acid (BABA) significantly inhibited foliar blight caused by Phytophthora capsici. Tissue extract analyses by GC/MS identified conserved change in certain metabolite concentrations following P. nicotianae or BABA treatment. Induced chile pepper plants had reduced concentrations of sucrose and TCA cycle intermediates and increased concentrations of specific hexose-phosphates, hexose-disaccharides and amino acids. Galactose, which increased significantly in induced chile pepper plants, was shown to inhibit growth of P. capsici in a plate assay.

  6. Ultrastructure of laevigate hilate spores in sporangia and spore masses from the Upper Silurian and Lower Devonian of the Welsh Borderland

    PubMed Central

    H.Wellman, C.

    1998-01-01

    Spore masses and isolated sporangia, containing laevigate hilate cryptospores attributable to the dispersed taxon Laevolancis divellomedia sensu lato, have been recovered on bulk maceration of Upper Silurian (Pridoli) and Lower Devonian (Lochkovian) deposits from the Welsh Borderland. Detailed morphological, anatomical and ultrastructural analysis, using light microscope, scanning electron microscope and transmission electron microscope techniques, reveals subtle differences between the specimens and they can be grouped into five distinct types. The different groups are distinguished principally by using sporangia-spore mass characteristics, presence or absence of extra-exosporal material and nature of spore-wall ultrastructure. Of the groups, one has a uniformly homogeneous exospore and the other four groups have a bilayered exospore. In the former the spores lack extra-exosporal material and occur in a discoidal sporangium. Of the bilayered groups, two have exospores of homogeneous composition but with the two layers differing in electron density. They occur in discoidal sporangia and spore masses and are distinguished on the presence or absence of extra-exosporal material and differences in the widths of the two layers. Finally, two bilayered groups possess a lamellate inner layer, but vary in presumed sporangial shape. Elongate sporangia have spores with concentric continuous lamellae, lacking further ultrastructure. In contrast, spores from a discoidal spore mass have white-line-centred, presumably tripartite, lamellae which are laterally discontinuous, overlapping and irregularly spaced. These findings, which suggest that morphologically similar spores were produced by a number of plant taxa, have important implications regarding the assessment of early land-plant diversity. The affinities of hilate cryptospore-producing plants are unknown and problematic, particularly as no extant non-angiosperm plants produce dyads, other than through meiotic irregularity

  7. First Finding of Melanic Sylvatic Triatoma infestans (Hemiptera: Reduviidae) Colonies in the Argentine Chaco

    PubMed Central

    CEBALLOS, L. A.; PICCINALI, R. V.; BERKUNSKY, I.; KITRON, U.; GÜRTLER, R. E.

    2009-01-01

    Triatoma infestans (Klug), the most important vector of Chagas disease in southern South America, is a highly domiciliated species with well-known sylvatic foci only in the Bolivian Andean valleys and in the Bolivian Chaco, where melanic insects designated as “dark morphs” were found. After the tentative identification of two melanic bugs collected from parrot nests in a forest reserve in the Argentine Chaco as T. infestans, we conducted an intensive search there using mouse-baited sticky traps in summer 2006 and 2007. Four live T. infestans bugs were collected in trees without parrot nests in 288 trap-nights, whereas no bug was collected from inside trees with active parrot nests in 51 trap-nights. To increase bug captures, hollow tree trunks that recently had had Amazona aestiva (Berlepsch) and Aratinga acuticaudata (Vieillot) parrot nests were treated with insecticide fumigant canisters exhibiting strong knockdown power. Four (22%) of 18 trees were positive for T. infestans with a dark phenotype. A fragment of the mitochondrial gene COI of 8 of the 14 triatomine bugs collected was successfully sequenced and confirmed as T. infestans. Most of the bugs were captured from Aspidosperma quebracho-blanco (Schlechter) hollow tree trunks harboring parrot nests. All of the T. infestans collected from the nearest house located at 10 km from the sylvatic foci displayed normal chromatic characters. The repeated finding of T. infestans in sylvatic habitats, albeit at very low density, shows that this species is capable of maintaining viable sylvatic foci in the absence of human hosts and immigration from domestic populations. These are the first confirmed findings of sylvatic T. infestans colonies in Argentina and of dark morphs in the Argentine Chaco. PMID:19769054

  8. Population structure of the Chagas disease vector, Triatoma infestans, at the urban-rural interface

    PubMed Central

    Foley, Erica A.; Khatchikian, Camilo E.; Hwang, Josephine; Ancca-Juárez, Jenny; Borrini-Mayori, Katty; Quıspe-Machaca, Victor R.; Levy, Michael Z.; Brisson, Dustin

    2013-01-01

    The increasing rate of biological invasions resulting from human transport or human-mediated changes to the environment have had devastating ecologic and public health consequences. The kissing bug, Triatoma infestans, has dispersed through the Peruvian city of Arequipa. The biological invasion of this insect has resulted in a public health crisis, putting thousands of residents of this city at risk of infection by Trypanosoma cruzi and subsequent development of Chagas disease. Here we show that populations of Tria. Infestans in geographically distinct districts within and around this urban center share a common recent evolutionary history although current gene flow is restricted even between proximal sites. The population structure among the Tria. Infestans in different districts is not correlated with the geographic distance between districts. These data suggest that migration among the districts is mediated by factors beyond the short-range migratory capabilities of Tria. Infestans and that human movement has played a significant role in the structuring of the Tria. Infestans population in the region. Rapid urbanization across southern South America will continue to create suitable environments for Tria. Infestans and knowledge of its urban dispersal patterns may play a fundamental role in mitigating human disease risk. PMID:24103030

  9. Population structure of the Chagas disease vector, Triatoma infestans, at the urban-rural interface.

    PubMed

    Foley, Erica A; Khatchikian, Camilo E; Hwang, Josephine; Ancca-Juárez, Jenny; Borrini-Mayori, Katty; Quıspe-Machaca, Victor R; Levy, Michael Z; Brisson, Dustin

    2013-10-01

    The increasing rate of biological invasions resulting from human transport or human-mediated changes to the environment has had devastating ecological and public health consequences. The kissing bug, Triatoma infestans, has dispersed through the Peruvian city of Arequipa. The biological invasion of this insect has resulted in a public health crisis, putting thousands of residents of this city at risk of infection by Trypanosoma cruzi and subsequent development of Chagas disease. Here, we show that populations of Tria. infestans in geographically distinct districts within and around this urban centre share a common recent evolutionary history although current gene flow is restricted even between proximal sites. The population structure among the Tria. infestans in different districts is not correlated with the geographical distance between districts. These data suggest that migration among the districts is mediated by factors beyond the short-range migratory capabilities of Tria. infestans and that human movement has played a significant role in the structuring of the Tria. infestans population in the region. Rapid urbanization across southern South America will continue to create suitable environments for Tria. infestans, and knowledge of its urban dispersal patterns may play a fundamental role in mitigating human disease risk. PMID:24103030

  10. Molecular phylogeography of the Chagas' disease vector Triatoma infestans in Argentina

    PubMed Central

    Pérez de Rosas, A R; Segura, E L; García, B A

    2011-01-01

    Triatoma infestans is the main vector of Chagas' disease in South America between latitudes 10°S and 46°S. A multilocus microsatellite data set of 836 individuals from 27 populations of T. infestans, from all its range of distribution in Argentina, was analyzed. Our results favor the hypothesis of two independent migration events of colonization in Argentina and secondary contacts. The majority of the populations of the western provinces of Catamarca, La Rioja, San Juan and the west of Cordoba province, had almost no shared ancestry with the rest of the populations analyzed. Probably those populations, belonging to localities close to the Andean region, could have been established by the dispersal line of T. infestans that would have arrived to Argentina through the Andes, whereas most of the rest of the populations analyzed may have derived from the dispersal line of T. infestans in non-Andean lowlands. Among them, those from the provinces of Formosa, Chaco, Santiago del Estero and Santa Fe shared different percentages of ancestry and presented lower degree of genetic differentiation. The migratory movement linked to regional economies and possibly associated with passive dispersal, would allow a higher genetic exchange among these populations of T. infestans. This study, using microsatellite markers, provides a new approach for evaluating the validity of the different hypotheses concerning the evolutionary history of this species. Two major lineages of T. infestans, an Andean and non-Andean, are suggested. PMID:21224874

  11. Eye pigments of the blood-sucking insect, Triatoma infestans Klug (Hemiptera, Reduviidae).

    PubMed

    Moraes, A S; Pimentel, E R; Rodrigues, V L C C; Mello, M L S

    2005-08-01

    The pigmentation of black (wild) and red (mutant) eyes of Triatoma infestans was studied spectrophotometrically and compared with red-eyed (wild) and white-eyed (mutant) forms of Drosophila melanogaster. The spectral absorption profiles of the black and red eye pigments of T. infestans were similar to each other and to that of the wild-type eyes of D. melanogaster. The similarity to the wild form of D. melanogaster indicated that both eye forms of T. infestans contained ommochromes of the xanthommatin type, a finding confirmed by ascending paper chromatography. Pteridines, melanins, and ommins were not detected as eye pigments in T. infestans. The eye color difference in T. infestans was assumed to be a function of the xanthommatin concentration, with a smaller content of ommochrome in red eyes, although this probably did not affect the insect's visual acuity. These data support other findings regarding the similarities between black- and red-eyed specimens of T. infestans for other characteristics. PMID:16341426

  12. Mapping the Progression of Phytophthora Ramorum

    NASA Astrophysics Data System (ADS)

    Banh, T.; Li, J.; El-Askary, H. M.

    2013-12-01

    There has been a plant pathogen, Phytophthora Ramorum that has been causing trouble for the plant species in the forests of California and Oregon. Phytophthora is essentially a water mold that infects oak species like California black oak, coast live oak as well as California bay laurel (Lamsal). What this project aims to accomplish is to observe any changes in NDVI values between the years of 2002 and 1994. What the project hopes to observe is a decline of NDVI values between the two years because the infection of Phytophtora Ramorum will cause stress to the plant or kill the plant, which will lower the values of NDVI. The project will utilizes satellite data to create NDVI images over the study area and two types of change detection methods to observe the differences between the NDVI values of the two years. Preliminary results for the project, data obtained from Landsat 7 ETM+ with a resolution of 240 meters, was not able to observe any significant changes. A finer resolution to differentiate the NDVI values would be needed. In addition the best way to keep the pathogen from getting out of control is with ground level management, or complete eradication of the pathogen. These eradication methods include burning the infected host plants and spreading herbicide (Alexander). With that in mind it would be ideal to have an early detection of the pathogen infestation. Therefore another goal of the project is to continue to research if remote sensing could play a role in an early detection method for the presence of Phytophtora Ramorum.

  13. 75 FR 44936 - Notice of Request for Extension of Approval of an Information Collection; Phytophthora Ramorum...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-30

    ... Collection; Phytophthora Ramorum; Quarantine and Regulations AGENCY: Animal and Plant Health Inspection... Phytophthora ramorum. DATES: We will consider all comments that we receive on or before September 28, 2010... prevent the spread of Phytophthora ramorum, contact Mr. Prakash Hebbar, Program Manager, Emergency...

  14. 40 CFR 180.1057 - Phytophthora palmivora; exemption from requirement of tolerance.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 24 2014-07-01 2014-07-01 false Phytophthora palmivora; exemption from... From Tolerances § 180.1057 Phytophthora palmivora; exemption from requirement of tolerance. Phytophthora palmivora is exempted from the requirement of a tolerance in or on the raw agricultural...

  15. 40 CFR 180.1057 - Phytophthora palmivora; exemption from requirement of tolerance.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 24 2011-07-01 2011-07-01 false Phytophthora palmivora; exemption from... From Tolerances § 180.1057 Phytophthora palmivora; exemption from requirement of tolerance. Phytophthora palmivora is exempted from the requirement of a tolerance in or on the raw agricultural...

  16. 78 FR 58993 - Notice of Request for Extension of Approval of an Information Collection; Phytophthora Ramorum...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-09-25

    ... Collection; Phytophthora Ramorum; Quarantine and Regulations AGENCY: Animal and Plant Health Inspection... Phytophthora ramorum. DATES: We will consider all comments that we receive on or before November 25, 2013... of regulated articles to prevent the spread of Phytophthora ramorum, contact Dr. Prakash K....

  17. 40 CFR 180.1057 - Phytophthora palmivora; exemption from requirement of tolerance.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Phytophthora palmivora; exemption from... From Tolerances § 180.1057 Phytophthora palmivora; exemption from requirement of tolerance. Phytophthora palmivora is exempted from the requirement of a tolerance in or on the raw agricultural...

  18. 40 CFR 180.1057 - Phytophthora palmivora; exemption from requirement of tolerance.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 25 2012-07-01 2012-07-01 false Phytophthora palmivora; exemption from... From Tolerances § 180.1057 Phytophthora palmivora; exemption from requirement of tolerance. Phytophthora palmivora is exempted from the requirement of a tolerance in or on the raw agricultural...

  19. 40 CFR 180.1057 - Phytophthora palmivora; exemption from requirement of tolerance.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 25 2013-07-01 2013-07-01 false Phytophthora palmivora; exemption from... From Tolerances § 180.1057 Phytophthora palmivora; exemption from requirement of tolerance. Phytophthora palmivora is exempted from the requirement of a tolerance in or on the raw agricultural...

  20. Seasonal variations in active dispersal of natural populations of Triatoma infestans in rural north-western Argentina

    PubMed Central

    VAZQUEZ-PROKOPEC, G. M.; CEBALLOS, L.A.; MARCET, P. L.; CECERE, M. C.; CARDINAL, M. V.; KITRON, U.; GÜRTLER, R. E.

    2007-01-01

    The flight dispersal of Triatoma infestans Klug (Hemiptera: Reduviidae) is one of the main mechanisms determining community re-infestation after control interventions. An empirical model of flight initiation coupled with data from a longitudinal study predicted that the flight dispersal of T. infestans would peak in summer. To test this prediction, longitudinal light trap collections were conducted during 3-8 nights in March (late summer), July (winter) and November (spring) 2003, and in March 2004 in a rural community in north-west Argentina. Following each light-trapping collection date, all peridomestic sites around light traps were inspected to assess the relative abundance and nutritional status of T. infestans at each site. A total of 21 adult and five nymph T. infestans, six Triatoma guasayana Wygodzinsky & Abalos, and nine Triatoma garciabesi Carcavallo et al. were collected in 96 light-trapping nights, whereas 696 T. infestans were collected from the peridomestic sites that surrounded the light traps. The arrival of T. infestans in the light traps occurred in 64% of catch stations and peaked in the summer surveys (10-14 bugs) compared with spring and winter surveys. When winds were < 5 km/h, the arrival of adult T. infestans at the light traps was significantly associated with maximum temperature and relative humidity. This is the first field report of seasonal variations in the flight dispersal activity of T. infestans. PMID:17044877

  1. Population structure of the Chagas disease vector Triatoma infestans in an urban environment.

    PubMed

    Khatchikian, Camilo E; Foley, Erica A; Barbu, Corentin M; Hwang, Josephine; Ancca-Juárez, Jenny; Borrini-Mayori, Katty; Quıspe-Machaca, Victor R; Naquira, Cesar; Brisson, Dustin; Levy, Michael Z

    2015-02-01

    Chagas disease is a vector-borne disease endemic in Latin America. Triatoma infestans, a common vector of this disease, has recently expanded its range into rapidly developing cities of Latin America. We aim to identify the environmental features that affect the colonization and dispersal of T. infestans in an urban environment. We amplified 13 commonly used microsatellites from 180 T. infestans samples collected from a sampled transect in the city of Arequipa, Peru, in 2007 and 2011. We assessed the clustering of subpopulations and the effect of distance, sampling year, and city block location on genetic distance among pairs of insects. Despite evidence of genetic similarity, the majority of city blocks are characterized by one dominant insect genotype, suggesting the existence of barriers to dispersal. Our analyses show that streets represent an important barrier to the colonization and dispersion of T. infestans in Arequipa. The genetic data describe a T. infestans infestation history characterized by persistent local dispersal and occasional long-distance migration events that partially parallels the history of urban development.

  2. Population Structure of the Chagas Disease Vector Triatoma infestans in an Urban Environment

    PubMed Central

    Khatchikian, Camilo E.; Foley, Erica A.; Barbu, Corentin M.; Hwang, Josephine; Ancca-Juárez, Jenny; Borrini-Mayori, Katty; Quıspe-Machaca, Victor R.; Naquira, Cesar; Brisson, Dustin; Levy, Michael Z.

    2015-01-01

    Chagas disease is a vector-borne disease endemic in Latin America. Triatoma infestans, a common vector of this disease, has recently expanded its range into rapidly developing cities of Latin America. We aim to identify the environmental features that affect the colonization and dispersal of T. infestans in an urban environment. We amplified 13 commonly used microsatellites from 180 T. infestans samples collected from a sampled transect in the city of Arequipa, Peru, in 2007 and 2011. We assessed the clustering of subpopulations and the effect of distance, sampling year, and city block location on genetic distance among pairs of insects. Despite evidence of genetic similarity, the majority of city blocks are characterized by one dominant insect genotype, suggesting the existence of barriers to dispersal. Our analyses show that streets represent an important barrier to the colonization and dispersion of T. infestans in Arequipa. The genetic data describe a T. infestans infestation history characterized by persistent local dispersal and occasional long-distance migration events that partially parallels the history of urban development. PMID:25646757

  3. First Report of Widespread Wild Populations of Triatoma infestans (Reduviidae, Triatominae) in the Valleys of La Paz, Bolivia

    PubMed Central

    Buitrago, Rosio; Waleckx, Etienne; Bosseno, Marie-France; Zoveda, Faustine; Vidaurre, Pablo; Salas, Renata; Mamani, Elio; Noireau, François; Brenière, Simone Frédérique

    2010-01-01

    Wild populations of Triatoma infestans, the main vector of Chagas disease in the Southern Cone countries, may be involved in reinfestation of human dwellings, limiting the success of vector-control campaigns in Bolivia. Knowledge of the distribution of these populations remains incomplete. We report here the detection of T. infestans wild populations in large areas in the department of La Paz, Bolivia. Among 18 sylvatic areas investigated, 17 were positive with T. infestans specimens. The infection rate of captured T. infestans with Trypanosoma cruzi was 85.7% in adult specimens. These results expand the geographical distribution of wild populations of T. infestans; it may be distributed throughout the Inter-Andean Dry Forest eco-region of Bolivia. The current information allows us to propose the hypothesis that a sylvatic origin of the reinfestation is located in the valleys of La Paz. PMID:20348501

  4. Biosynthesis of Antibiotic Leucinostatins in Bio-control Fungus Purpureocillium lilacinum and Their Inhibition on Phytophthora Revealed by Genome Mining

    PubMed Central

    Li, Erfeng; Mao, Zhenchuan; Ling, Jian; Yang, Yuhong; Yin, Wen-Bing; Xie, Bingyan

    2016-01-01

    Purpureocillium lilacinum of Ophiocordycipitaceae is one of the most promising and commercialized agents for controlling plant parasitic nematodes, as well as other insects and plant pathogens. However, how the fungus functions at the molecular level remains unknown. Here, we sequenced two isolates (PLBJ-1 and PLFJ-1) of P. lilacinum from different places Beijing and Fujian. Genomic analysis showed high synteny of the two isolates, and the phylogenetic analysis indicated they were most related to the insect pathogen Tolypocladium inflatum. A comparison with other species revealed that this fungus was enriched in carbohydrate-active enzymes (CAZymes), proteases and pathogenesis related genes. Whole genome search revealed a rich repertoire of secondary metabolites (SMs) encoding genes. The non-ribosomal peptide synthetase LcsA, which is comprised of ten C-A-PCP modules, was identified as the core biosynthetic gene of lipopeptide leucinostatins, which was specific to P. lilacinum and T. ophioglossoides, as confirmed by phylogenetic analysis. Furthermore, gene expression level was analyzed when PLBJ-1 was grown in leucinostatin-inducing and non-inducing medium, and 20 genes involved in the biosynthesis of leucionostatins were identified. Disruption mutants allowed us to propose a putative biosynthetic pathway of leucinostatin A. Moreover, overexpression of the transcription factor lcsF increased the production (1.5-fold) of leucinostatins A and B compared to wild type. Bioassays explored a new bioactivity of leucinostatins and P. lilacinum: inhibiting the growth of Phytophthora infestans and P. capsici. These results contribute to our understanding of the biosynthetic mechanism of leucinostatins and may allow us to utilize P. lilacinum better as bio-control agent. PMID:27416025

  5. Three novel phytophthora species from irrigation water in Mississippi

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The genus Phytophthora includes a number of destructive plant pathogens. Here we report three new taxa recovered from irrigation systems at an ornamental crop nursery in Mississippi. Isolates of these new taxa were recovered from rhododendron leaves submerged in ponds for 7 days in 2012. Isolat...

  6. Susceptibility of highbush blueberry cultivars to Phytophthora root rot

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora cinnamomi Rands is a ubiquitous soilborne pathogen associated with root rot in many woody perennial plant species, including highbush blueberry (Vaccinium sp.). To identify genotypes with resistance to the pathogen, cultivars and advanced selections of highbush blueberry were grown in a...

  7. PATHOGENIC PHYTOPHTHORA SPECIES IN SAN JOAQUIN VALLEY IRRIGATION WATER SOURCES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Surface sources of irrigation water including the Kings River and three canals were assayed for Phytophthora spp. at six locations in the San Joaquin Valley within 30 km of Hanford, CA. Four nylon-mesh bags, each containing three firm, green pear fruits (separated by Styrofoam blocks) as bait for Ph...

  8. Phytophthora ramorum causes cryptic bole cankers in Canyon line Oak

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Unusual mortality of large canyon live oaks was observed in natural stands in San Mateo, California starting in 2007. A survey of affected stands showed that symptomatic trees were spatially associated with California bay, the primary source of Phytophthora ramorum spores in this forest type. Trunk ...

  9. Susceptibility of sprouted oak acorns to Phytophthora ramorum zoospores

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora ramorum is a recently emerged pathogen, having established in Europe and several western U.S. states, including California and Oregon. It has a wide host range and is a threat to forest ecology and the nursery industry. In California, coast live oak (Quercus agrifolia) is a major host...

  10. Can Epiphytes reduce disease symptoms caused by Phytophthora ramorum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Leaf infection of ornamental species by Phytophthora ramorum has a significant impact on the spread of this disease. Fungicides have had limited effects on controlling this disease. With increasing concerns that repeated fungicide applications will exasperate the potential for fungicide resistance...

  11. Assessing the efficacy of pre-harvest, chlorine-based sanitizers against human pathogen indicator microorganisms and Phytophthora capsici in non-recycled surface irrigation water.

    PubMed

    Lewis Ivey, M L; Miller, S A

    2013-09-01

    Many factors must be considered in order to develop and implement treatment systems to improve the microbial quality of surface water and prevent the accidental introduction of plant and human pathogens into vegetable crops. The efficacy of chlorine gas (Cl2(g)) and chlorine dioxide (ClO2) injection systems in combination with rapid sand filtration (RSF) was evaluated in killing fecal indicator microorganisms in irrigation water in a vegetable-intensive production area. The efficacy of ClO2 and Cl2(g) was variable throughout the distribution systems and coliform bacteria never dropped below levels required by the United States Environmental Protection Agency for recreational waters. Sampling date and sampling point had a significant effect on the abundance of coliforms in Cl2(g)- and ClO2-treated water. Sampling date and sampling point also had a significant effect on the abundance of generic Escherichia coli in Cl2(g) treated water but only sampling point was significant in ClO2 treated water. Although the waterborne plant pathogen Phytophthora capsici was detected in five different sources of surface irrigation water using baiting and P. capsici-specific PCR, in vitro studies indicated that ClO2 at concentrations similar to those used to treat irrigation water did not reduce mycelial growth or direct germination of P. capsici sporangia and reduced zoospore populations by less than 50%. This study concludes that injection of ClO2 and Cl2(g) into surface water prior to rapid sand filtration is inadequate in reducing fecal indicator microorganism populations and ClO2 ineffectively kills infectious propagules of P. capsici. Additional research is needed to design a system that effectively targets and significantly reduces both plant and human pathogens that are present in surface irrigation water. A model for a multiple barrier approach to treating surface water for irrigation is proposed.

  12. The Effect of Potassium Nitrate on the Reduction of Phytophthora Stem Rot Disease of Soybeans, the Growth Rate and Zoospore Release of Phytophthora Sojae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effects of potassium nitrate (KNO3) application on Phytophthora stem rot disease reduction of Glycine max (L.) Merr. cvs. Chusei-Hikarikuro and Sachiyutaka, and fungal growth and zoospore release of a Phytophthora sojae isolate were investigated under laboratory conditions. The application of 4-...

  13. Distribution of Pyrethroid Resistant Populations of Triatoma infestans in the Southern Cone of South America

    PubMed Central

    Bustamante Gomez, Marinely; Gonçalves Diotaiuti, Liléia; Gorla, David E.

    2016-01-01

    Background A number of studies published during the last 15 years showed the occurrence of insecticide resistance in Triatoma infestans populations. The different toxicological profiles and mechanisms of resistance to insecticides is due to a genetic base and environmental factors, being the insecticide selective pressure the best studied among the last factors. The studies on insecticide resistance on T. infestans did not consider the effect of environmental factors that may influence the distribution of resistance to pyrethroid insecticides. To fill this knowledge gap, the present study aims at studying the association between the spatial distribution of pyrethroid resistant populations of T. infestans and environmental variables. Methodology/Principal Findings A total of 24 articles reporting on studies that evaluated the susceptibility to pyrethroids of 222 field-collected T. infestans populations were compiled. The relationship between resistance occurrence (according to different criteria) with environmental variables was studied using a generalized linear model. The lethal dose that kills 50% of the evaluated population (LD50) showed a strong linear relationship with the corresponding resistance ratio (RR50). The statistical descriptive analysis of showed that the frequency distribution of the Log (LD50) is bimodal, suggesting the existence of two statistical groups. A significant model including 5 environmental variables shows the geographic distribution of high and low LD50 groups with a particular concentration of the highest LD50 populations over the region identified as the putative center of dispersion of T. infestans. Conclusions/Significance The occurrence of these two groups concentrated over a particular region that coincides with the area where populations of the intermediate cytogenetic group were found might reflect the spatial heterogeneity of the genetic variability of T. infestans, that seems to be the cause of the insecticide resistance in

  14. Phytophthora x pelgrandis, a new natural hybrid pathogenic to Pelargonium grandiflorum hort.

    PubMed

    Nirenberg, Helgard I; Gerlach, Wolfram F; Gräfenhan, Tom

    2009-01-01

    A new Phytophthora hybrid of Ph. cactorum (Leb. & Cohn) Schroet. and Ph. nicotianaevan Breda de Haan pathogenic to cultivars of Pelargonium grandiflorum hort. is described as Phytophthora X pelgrandis and its morphological features are documented. Morphological, physiological (e.g., temperature requirements) and molecular data (DNA sequencing, random amplified polymorphic DNA-PCR) are presented for isolates of the Phytophthora hybrid. Pathogenicity was tested on cultivars of P. grandiflorum and Nicotiana tabacum. For comparison cultures of the parental species and additional Phytophthora taxa also were examined.

  15. Potential Role of Elicitins in the Interaction between Phytophthora Species and Tobacco

    PubMed Central

    Kamoun, Sophien; Young, Mary; Förster, Helga; Coffey, Michael D.; Tyler, Brett M.

    1994-01-01

    The potential role of extracellular elicitor proteins (elicitins) from Phytophthora species as avirulence factors in the interaction between Phytophthora and tobacco was examined. A survey of 85 Phytophthora isolates representing 14 species indicated that production of elicitin is almost ubiquitous except for isolates of Phytophthora parasitica from tobacco. The production of elicitins by isolates of P. parasitica correlated without exception with low or no virulence on tobacco. Genetic analysis was conducted by using a cross between two isolates of P. parasitica, segregating for production of elicitin and virulence on tobacco. Virulence assays of the progeny on tobacco confirmed the correlation between production of elicitin and low virulence. Images PMID:16349258

  16. Genetic variability, phylogenetic relationships and gene flow in Triatoma infestans dark morphs from the Argentinean Chaco

    PubMed Central

    Piccinali, R. V.; Marcet, P. L.; Ceballos, L. A.; Kitron, U.; Gürtler, R. E.; Dotson, E. M.

    2011-01-01

    The recent discovery of sylvatic populations of Triatoma infestans outside the Andean Valleys of Bolivia prompted an evolutionary question about the putative ancestral area of origin and dispersal of the species, and an epidemiological question regarding the possible role of these sylvatic populations in the recolonization process of insecticide-treated houses. The finding of a population of sylvatic melanic T. infestans (dark morphs) in the Argentinean dry Chaco at 7 km from a peridomestic bug population of typical coloration gave us the opportunity to test both questions simultaneously by employing phylogenetic and population genetic approaches. For this purpose we analyzed sylvatic and peridomestic bugs using sequence-based mitochondrial and nuclear markers (mtCOI and ITS-1) and microsatellites. Sylvatic bugs were confirmed to be T. infestans and not hybrids, and showed high levels of genetic variability and departures from neutral expectations for mtCOI variation. New ITS-1 and mtCOI haplotypes were recorded, as well as haplotypes shared with peridomestic and/or domestic bugs from previous records. The peridomestic population was invariant for ITS-1 and mtCOI, but showed variability for microsatellites and signatures of a population bottleneck, probably due to a limited number of founders. Phylogenetic analyses were consistent with the presence of ancestral haplotypes in sylvatic bugs. According to F-statistics and assignment methods there was a significant differentiation between sylvatic and peridomestic bugs and gene flow was low and asymmetric, with more bugs moving from the peridomicile to the sylvatic environment. These results support the hypothesis of the Chaco region as the area of origin of T. infestans, and a limited role of sylvatic melanic T. infestans in peridomestic infestation in the Argentinean Chaco. PMID:21352954

  17. Occurrence of Phytophthora plurivora and other Phytophthora species in oak forests of southern Poland and their association with site conditions and the health status of trees.

    PubMed

    Jankowiak, R; Stępniewska, H; Bilański, P; Kolařík, M

    2014-11-01

    Phytophthora plurivora and other Phytophthora species are known to be serious pathogens of forest trees. Little is known, however, about the presence of P. plurivora in Polish oak forests and their role in oak decline. The aims of this study were to identify P. plurivora in healthy and declining Quercus robur stands in southern Poland and to demonstrate the relationship between different site factors and the occurrence of P. plurivora. In addition, the virulence of P. plurivora and other Phytophthora species was evaluated through inoculations using 2-year-old oak seedlings. Rhizosphere soil was investigated from 39 oak stands representing different healthy tree statuses. The morphology and DNA sequences of the internal transcribed spacer regions (ITS) of the ribosomal DNA and the mitochondrial cox1 gene were used for identifications. P. plurivora, an oak fine root pathogen, was isolated from rhizosphere soil samples in 6 out of 39 stands. Additionally, Phytophthora cambivora, Phytophthora polonica and Phytophthora rosacearum-like were also obtained from several stands. The results showed a significant association between the presence of P. plurivora and the health status of oak trees. Similar relationships were also observed for all identified Phytophthora species. In addition, there was evidence for a connection between the presence of all identified Phytophthora species and some site conditions. Phytophthora spp. occurred more frequently in declining stands and in silt loam and sandy loam soils with pH ≥ 3.66. P. plurivora and P. cambivora were the only species capable of killing whole plants, producing extensive necrosis on seedling stems.

  18. First Report of Colonies of Sylvatic Triatoma infestans (Hemiptera: Reduviidae) in the Paraguayan Chaco, Using a Trained Dog

    PubMed Central

    Rolón, Miriam; Vega, María Celeste; Román, Fabiola; Gómez, Ana; Rojas de Arias, Antonieta

    2011-01-01

    In the Gran Chaco region, control of Triatoma infestans has been limited by persistent domestic infestations despite the efforts of the Vector Control Services. In Paraguay, this region is the highest endemic area in the country, showing high levels of indoor and outdoor infestation. Although sylvatic T. infestans have been found in the Bolivian and Argentine Chaco, similar searches for sylvatic populations of this species in Paraguay had been unsuccessful over the last 20 years. Here we present a new approach to detecting sylvatic Triatominae, using a trained dog, which has successfully confirmed sylvatic populations of T. infestans and other triatomine species in Paraguay. A total of 22 specimens corresponding to dark morph forms of T. infestans were collected, and 14 were confirmed as T. infestans by the mitochondrial cytochrome B gene analysis. Through this analysis, one of which were previously reported and a second that was a new haplotype. Triatomines were captured from amongst vegetation such as dry branches and hollows trees of different species such Aspidosperma quebracho-blanco, Bulnesia sarmientoi and Stetsonia coryne. The colonies found have been small and without apparent infection with Trypanosoma cruzi. During the study, Triatoma sordida and Triatoma guasayana have also been found in ecotopes close to those of T. infestans. PMID:21572522

  19. Phytophthora database 2.0: update and future direction.

    PubMed

    Park, Bongsoo; Martin, Frank; Geiser, David M; Kim, Hye-Seon; Mansfield, Michele A; Nikolaeva, Ekaterina; Park, Sook-Young; Coffey, Michael D; Russo, Joseph; Kim, Seong H; Balci, Yilmaz; Abad, Gloria; Burgess, Treena; Grünwald, Niklaus J; Cheong, Kyeongchae; Choi, Jaeyoung; Lee, Yong-Hwan; Kang, Seogchan

    2013-12-01

    The online community resource Phytophthora database (PD) was developed to support accurate and rapid identification of Phytophthora and to help characterize and catalog the diversity and evolutionary relationships within the genus. Since its release in 2008, the sequence database has grown to cover 1 to 12 loci for ≈2,600 isolates (representing 138 described and provisional species). Sequences of multiple mitochondrial loci were added to complement nuclear loci-based phylogenetic analyses and diagnostic tool development. Key characteristics of most newly described and provisional species have been summarized. Other additions to improve the PD functionality include: (i) geographic information system tools that enable users to visualize the geographic origins of chosen isolates on a global-scale map, (ii) a tool for comparing genetic similarity between isolates via microsatellite markers to support population genetic studies, (iii) a comprehensive review of molecular diagnostics tools and relevant references, (iv) sequence alignments used to develop polymerase chain reaction-based diagnostics tools to support their utilization and new diagnostic tool development, and (v) an online community forum for sharing and preserving experience and knowledge accumulated in the global Phytophthora community. Here we present how these improvements can support users and discuss the PD's future direction.

  20. Molecular cloning, functional characterization and expression of potato (Solanum tuberosum) 1-deoxy-d-xylulose 5-phosphate synthase 1 (StDXS1) in response to Phytophthora infestans.

    PubMed

    Henriquez, Maria Antonia; Soliman, Atta; Li, Genyi; Hannoufa, Abdelali; Ayele, Belay T; Daayf, Fouad

    2016-02-01

    1-Deoxy-D-xylulose 5-phosphate synthase (DXS) catalyzes the initial step of the plastidial 2C-methyl-D-erythritol-4-phosphate (DOXP-MEP) pathway involved in isoprenoid biosynthesis. In this study, we cloned the complete cDNA of potato DXS gene that was designated StDXS1. StDXS1 cDNA encodes for 719 amino acid residues, with MW of 77.8 kDa, and is present in one copy in the potato genome. Phylogenetic analysis and protein sequence alignments assigned StDXS1 to a group with DXS homologues from closely related species and exhibited homodomain identity with known DXS proteins from other plant species. Late blight symptoms occurred in parallel with a reduction in StDXS1 transcript levels, which may be associated with the levels of isoprenoids that contribute to plant protection against pathogens. Subcellular localization indicated that StDXS1 targets the chloroplasts where isoprenoids are synthesized. Arabidopsis expressing StDXS1 showed a higher accumulation of carotenoids and chlorophyll as compared to wild type controls. Lower levels of ABA and GA were detected in the transgenic DXS lines as compared to control plants, which reflected on higher germination rates of the transgenic DXS lines. No changes were detected in JA or SA contents. Selected downstream genes in the DOXP-MEP pathway, especially GGPPS genes, were up-regulated in the transgenic lines.

  1. Mortality of container-grown blueberry plants inoculated with Phytophthora cinnamomi Rands

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We conducted four studies to evaluate the effect of Phytophthora cinnamomi isolates and inoculum delivery methods on root rot development and mortality of container-grown blueberry plants. Phytophthora cinnamomi isolates were obtained from the root zone of symptomatic blueberry plants and identifie...

  2. An overview of Australia's Phytophthora species assemblage in natural ecosystems recovered from a survey in Victoria.

    PubMed

    Dunstan, William A; Howard, Kay; StJ Hardy, Giles E; Burgess, Treena I

    2016-06-01

    Although Phytophthora species cause serious diseases worldwide, until recently the main focus on disease in natural ecosystems in southern Australia has been on the distribution and impact of P. cinnamomi. However, new Phytophthora pathogens have emerged from natural ecosystems, and there is a need to better understand the diversity and distribution of these species in our natural forests, woodlands and heathlands. From a survey along a 70 km pipeline easement in Victoria, Phytophthora species were isolated from 249 rhizosphere samples and 25 bait bags deployed in 21 stream, river, or wetland locations. Of the 186 Phytophthora isolates recovered, 130 were identified to species based on ITS sequence data. Ninety-five isolates corresponded to 13 described Phytophthora species while additionally 35 isolates were identified as Clade 6 hybrids. Phytophthora cinnamomi was the most common species isolated (31 %), followed by P. elongata (6 %), both species were only recovered from soil. Samples from sites with the highest soil moisture at the time of sampling had the highest yield of isolates. Consistent with other studies throughout the world, Clade 6 species and their hybrids dominated water samples, although many of these species were also recovered less frequently from soil samples. Many of the species recovered in this study have not previously been reported from eastern Australia, reinforcing that Phytophthora species are widespread, abundant and diverse in natural ecosystems. We have probably been underestimating Phytophthora diversity in Australia.

  3. Amino terminal region of Phytophthora sojae cel12 endoglucanase confers tissue collapse function in Nicotiana

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora encodes an unusually large number of glycosyl hydrolases (GH), with many large gene families resulting from duplication events. There are ten copies of GH 12 (cel12) present in Phytophthora sojae. This is the only pathogen endoglucanase family to which plants produce an inhibitory pr...

  4. Fruit age and development of Phytophthora fruit rot on resistant and susceptible watermelon lines

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora fruit rot caused by Phytophthora capsici is an emerging disease in most watermelon producing regions of Southeast U.S. and has resulted in severe losses to watermelon growers especially in GA, SC, and NC. We recently released four germplasm lines (USVL203-PFR, USVL020-PFR, USVL782-PFR,...

  5. Phytophthora megakarya, a causal agent of black pod rot in Africa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In most parts of the world where Theobroma cacao is grown, Phytophthora palmivora is the major concern for causing black pod rot (BPR). Phytophthora megakarya, on the other hand, occurs only in Africa, but represents a major threat to cacao production, the countries of West Africa being the largest ...

  6. Sensitivity of Phytophthora capsici isolates from the southeast US to Fluopicolide and Cyazofamid.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The plant pathogen Phytophthora capsici is rapidly becoming an important limiting factor in vegetable production in the southeastern United States particularly on cucurbits and peppers. The diseases caused by P. capsici are known by various names such as Phytophthora blight and crown rot on peppers,...

  7. Resistance in watermelon rootstocks to crown rot caused by Phytophthora capsici

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora crown and fruit rot caused by Phytophthora capsici is becoming an important and emerging disease of watermelon (Citrullus lanatus) in south eastern United States. In recent years, the practice of grafting seedless watermelons (triploids) onto rootstocks belonging to other Cucurbitaceae...

  8. Evaluation of Commercial Watermelon Rootstocks for Tolerance to Phytophthora Blight and Watermelon Vine Decline

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora blight and fruit rot caused by Phytophthora capsici, and watermelon vine decline (WVD) caused by Squash vein yellowing virus (SqVYV), are two important and emerging diseases of watermelons (Citrullus lanatus). Recently, the practice of grafting seedless watermelons (triploids) onto roo...

  9. Effects of hydrostatic pressure, agitation and CO2 stress on Phytophthora nicotianae zoospore survival

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora nicotianae Breda de Haan was used as a model pathogen to investigate the effects of hydrostatic pressure, agitation, and aeration with CO2 or breathable air on the survival of Phytophthora zoospores in water. Injecting CO2 into 2 liters of zoospore-infested water for 5 min at 110.4 ml ...

  10. Novel quantitative trait loci for partial resistance to Phytophthora sojae in soybean PI 398841

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora root and stem rot caused by Phytophthora sojae Kaufmann and Gerdmann is one of the most severe soybean [Glycine max (L.) Merr] diseases in the US. Partial resistance is as effective in managing this disease as single-gene (Rps) mediated resistance and is more durable. The objective of t...

  11. Diversity of Phytophthora Species from Declining Mediterranean Maquis Vegetation, including Two New Species, Phytophthora crassamura and P. ornamentata sp. nov.

    PubMed

    Scanu, Bruno; Linaldeddu, Benedetto T; Deidda, Antonio; Jung, Thomas

    2015-01-01

    The Mediterranean basin is recognized as a global biodiversity hotspot accounting for more than 25,000 plant species that represent almost 10% of the world's vascular flora. In particular, the maquis vegetation on Mediterranean islands and archipelagos constitutes an important resource of the Mediterranean plant diversity due to its high rate of endemism. Since 2009, a severe and widespread dieback and mortality of Quercus ilex trees and several other plant species of the Mediterranean maquis has been observed in the National Park of La Maddalena archipelago (northeast Sardinia, Italy). Infected plants showed severe decline symptoms and a significant reduction of natural regeneration. First studies revealed the involvement of the highly invasive wide-host range pathogen Phytophthora cinnamomi and several fungal pathogens. Subsequent detailed research led to a better understanding of these epidemics showing that multiple Phytophthora spp. were involved, some of them unknown to science. In total, nine Phytophthora species were isolated from rhizosphere soil samples collected from around symptomatic trees and shrubs including Asparagus albus, Cistus sp., Juniperus phoenicea, J. oxycedrus, Pistacia lentiscus and Rhamnus alaternus. Based on morphological characters, growth-temperature relations and sequence analysis of the ITS and cox1 gene regions, the isolates were identified as Phytophthora asparagi, P. bilorbang, P. cinnamomi, P. cryptogea, P. gonapodyides, P. melonis, P. syringae and two new Clade 6 taxa which are here described as P. crassamura sp. nov. and P. ornamentata sp. nov. Pathogenicity tests supported their possible involvement in the severe decline that is currently threatening the Mediterranean maquis vegetation in the La Maddalena archipelago. PMID:26649428

  12. Diversity of Phytophthora Species from Declining Mediterranean Maquis Vegetation, including Two New Species, Phytophthora crassamura and P. ornamentata sp. nov.

    PubMed Central

    Scanu, Bruno; Linaldeddu, Benedetto T.; Deidda, Antonio; Jung, Thomas

    2015-01-01

    The Mediterranean basin is recognized as a global biodiversity hotspot accounting for more than 25,000 plant species that represent almost 10% of the world’s vascular flora. In particular, the maquis vegetation on Mediterranean islands and archipelagos constitutes an important resource of the Mediterranean plant diversity due to its high rate of endemism. Since 2009, a severe and widespread dieback and mortality of Quercus ilex trees and several other plant species of the Mediterranean maquis has been observed in the National Park of La Maddalena archipelago (northeast Sardinia, Italy). Infected plants showed severe decline symptoms and a significant reduction of natural regeneration. First studies revealed the involvement of the highly invasive wide-host range pathogen Phytophthora cinnamomi and several fungal pathogens. Subsequent detailed research led to a better understanding of these epidemics showing that multiple Phytophthora spp. were involved, some of them unknown to science. In total, nine Phytophthora species were isolated from rhizosphere soil samples collected from around symptomatic trees and shrubs including Asparagus albus, Cistus sp., Juniperus phoenicea, J. oxycedrus, Pistacia lentiscus and Rhamnus alaternus. Based on morphological characters, growth-temperature relations and sequence analysis of the ITS and cox1 gene regions, the isolates were identified as Phytophthora asparagi, P. bilorbang, P. cinnamomi, P. cryptogea, P. gonapodyides, P. melonis, P. syringae and two new Clade 6 taxa which are here described as P. crassamura sp. nov. and P. ornamentata sp. nov. Pathogenicity tests supported their possible involvement in the severe decline that is currently threatening the Mediterranean maquis vegetation in the La Maddalena archipelago. PMID:26649428

  13. Small homologous blocks in phytophthora genomes do not point to an ancient whole-genome duplication.

    PubMed

    van Hooff, Jolien J E; Snel, Berend; Seidl, Michael F

    2014-05-01

    Genomes of the plant-pathogenic genus Phytophthora are characterized by small duplicated blocks consisting of two consecutive genes (2HOM blocks) and by an elevated abundance of similarly aged gene duplicates. Both properties, in particular the presence of 2HOM blocks, have been attributed to a whole-genome duplication (WGD) at the last common ancestor of Phytophthora. However, large intraspecies synteny-compelling evidence for a WGD-has not been detected. Here, we revisited the WGD hypothesis by deducing the age of 2HOM blocks. Two independent timing methods reveal that the majority of 2HOM blocks arose after divergence of the Phytophthora lineages. In addition, a large proportion of the 2HOM block copies colocalize on the same scaffold. Therefore, the presence of 2HOM blocks does not support a WGD at the last common ancestor of Phytophthora. Thus, genome evolution of Phytophthora is likely driven by alternative mechanisms, such as bursts of transposon activity.

  14. Temporal differences in blood meal detection from the midguts of Triatoma infestans.

    PubMed

    Pinto, Jesus; Roellig, Dawn M; Gilman, Robert H; Calderón, Maritza; Bartra, Carlos; Salazar, Renzo; Bern, Caryn; Ancca-Juárez, Jenny; Levy, Michael; Náquira, Cesar; Cama, Vitaliano

    2012-01-01

    We used genus/species specific PCRs to determine the temporal persistence of host DNA in Triatoma infestans experimentally fed on blood from six common vertebrate species: humans, domestic dogs, guinea pigs, chickens, mice, and pigs. Twenty third or fourth instar nymphs per animal group were allowed to feed to engorgement, followed by fasting-maintenance in the insectary. At 7, 14, 21, or 28 days post-feeding, the midgut contents from five triatomines per group were tested with the respective PCR assay. DNA from all vertebrate species was detected in at least four of five study nymphs at seven and 14 days post-feeding. DNA of humans, domestic dogs, guinea pigs, pigs, and chickens were more successfully detected (80-100%) through day 21, and less successfully (20-100%) at day 28. Findings demonstrate that species-specific PCRs can consistently identify feeding sources of T. infestans within two weeks, a biologically relevant time interval.

  15. Phenotypic plasticity in response to food source in Triatoma infestans (Klug, 1834) (Hemiptera, Reduviidae: Triatominae).

    PubMed

    Nattero, Julieta; Malerba, Romina; Rodríguez, Claudia S; Crocco, Liliana

    2013-10-01

    In the Gran Chaco region of Argentina, Bolivia, and Paraguay, vector transmission of Trypanosoma cruzi, the etiological agent of Chagas disease, is still a severe problem because, among other causes, houses are reinfested with Triatoma infestans, the main vector of T. cruzi in southern South America. A better understanding of adaptation and evolution of T. infestans populations may contribute to the selection of appropriate vector control strategies in this region. Phenotypic plasticity is essential to understand development and maintenance of morphological variation. An experimental phenotypic plasticity study was conducted to assess if blood meal source induced head shape and size variation during development in T. infestans. Eighteen full-sib families were assigned to one of two food sources (pigeon and guinea pig) to examine the effect of food source on head shape and size in all nymph instars and adults. Data were analyzed using geometric morphometric tools and phenotypic plasticity analyses. Significant differences in head shape and size were observed between adults fed on different food sources. Allometric effects at the adult stage were observed. Head size showed significant food source × family interaction for fifth-instar nymphs and adults. For head size, significant differences between food sources were observed at stages and in ontogenetic trajectory. Phenotypic plasticity expression was found for head shape and size in adults; indeed, bugs fed on guinea pigs exhibited greater changes in head shape and larger heads than those fed on pigeon. Full-sib families exhibited different patterns of phenotypic expression in response to food source. Food source × family interaction may indicate that the observed variation in phenotypic plasticity may contribute to changes in head morphometry. These results may contribute to the selection of an appropriate control strategy for T. infestans in the Gran Chaco region, since they provide evidences of morphological

  16. Spatial Structuring of Triatoma infestans (Hemiptera, Reduviidae) Populations from Northwestern Argentina Using Wing Geometric Morphometry

    PubMed Central

    SCHACHTER-BROIDE, JUDITH; DUJARDIN, JEAN-PIERRE; KITRON, URIEL; GÜRTLER, RICARDO E.

    2005-01-01

    Wing geometric morphometry was used to study the spatial structuring of populations of Triatoma infestans from different villages, ecotopes, and sites within a village in northwestern Argentina. A total of 308 male and 197 female wings of T. infestans collected from peridomestic and domestic ecotopes in March 2000 was analyzed. On average, female bugs had a significantly larger wing size than males. Triatomines collected from domiciles or structures associated with chickens had larger wings than bugs collected from goat or pig corrals. The wing size of bugs did not differ significantly between villages. Discriminant analyses of wing shape showed significant divergence between villages, ecotopes, and individual collection sites. The study of metric variation of males between sites belonging to the same ecotope also revealed significant heterogeneity. Indeed, within the same section of the village the difference between two goat corrals was sometimes greater than that between neighboring goat and pig corrals. Thus, morphometric heterogeneity within villages may be the result not only of ecotope and host associations, but also of physical isolation between subunits. The strong structuring of T. infestans populations in the study area indicates that recolonization could be traced back to a small geographic source. PMID:15311455

  17. Changes in nuclear phenotype frequencies following sequential cold shocks in Triatoma infestans (Hemiptera, Reduviidae).

    PubMed

    Campos, Silvana G P; Rodrigues, Vera Lúcia C C; Mello, Maria Luiza S

    2002-09-01

    The nuclear phenotypes of Malpighian tubule cells in fifth instar nymphs of Triatoma infestans, one of the most important vectors of Chagas disease, were studied following sequential shocks at 0 degrees C, separated by intervals of 8 h and 24 h at 30 degrees C, under conditions of moderate fasting and full nourishment. The insects pertained to colonies reared in the laboratory and originated from domestic specimens collected in the Brazilian states of São Paulo (north) and Minas Gerais (south). Since nuclear phenotypes in this species are affected by single cold shocks, it was expected that these phenotypes could also be changed by sequential shocks. Nuclear phenotypes indicative of mechanisms of cell survival (nuclear fusion and heterochromatin decondensation) and cell death (apoptosis and necrosis) were observed concomitantly in all the conditions tested. Nuclear fusion and heterochromatin decondensation were not found relevant for the presumed acquisition of the cold-hardening response in T. infestans. The decreased frequency of apoptosis and necrosis following sequential cold shocks including under fasting conditions, indicated that tolerance to sequential cold shocks occurred in T. infestans of the mentioned origin.

  18. Fine-scale genetic structure in populations of the Chagas' disease vector Triatoma infestans (Hemiptera, Reduvidae).

    PubMed

    Pérez de Rosas, Alicia R; Segura, Elsa L; Fusco, Octavio; Guiñazú, Adolfo L Bareiro; García, Beatriz A

    2013-03-01

    Fine scale patterns of genetic structure and dispersal in Triatoma infestans populations from Argentina was analysed. A total of 314 insects from 22 domestic and peridomestic sites from the locality of San Martín (Capayán department, Catamarca province) were typed for 10 polymorphic microsatellite loci. The results confirm subdivision of T. infestans populations with restricted dispersal among sampling sites and suggest inbreeding and/or stratification within the different domestic and peridomestic structures. Spatial correlation analysis showed that the scale of structuring is approximately of 400 m, indicating that active dispersal would occur within this distance range. It was detected difference in scale of structuring among sexes, with females dispersing over greater distances than males. This study suggests that insecticide treatment and surveillance should be extended within a radius of 400 m around the infested area, which would help to reduce the probability of reinfestation by covering an area of active dispersal. The inferences made from fine-scale spatial genetic structure analyses of T. infestans populations has demonstrated to be important for community-wide control programs, providing a complementary approach to help improve vector control strategies.

  19. Experimental control of Triatoma infestans in poor rural villages of Bolivia through community participation

    PubMed Central

    Lardeux, Frédéric; Depickère, Stéphanie; Aliaga, Claudia; Chavez, Tamara; Zambrana, Lilian

    2015-01-01

    Background Triatoma infestans is the main vector of Chagas disease in the southern cone countries. Present control strategies based on indoor and outdoor residual insecticide spraying are not sufficient to control disease transmission, particularly in Bolivia. Techniques based on the management of the human environment may be good alternatives or supplements. Methods Social and entomological surveys were carried out in four villages of Bolivia situated in the dry inter-Andean Valleys and the Chaco region. Risk factors for house infestation by T. infestans were identified, and an eco-health intervention based on education and community participation was carried out to reduce the risks of house infestation. It consisted of implementing simple and low cost vector control techniques such as coating of mud walls, cleaning activities and removal of poultry that enter rooms to lay eggs. Results The eco-health intervention significantly reduced the number of infested bedrooms, the mean abundance of T. infestans in bedrooms and beds, especially in the Chaco region. Mud wall coating was well accepted and could be proposed as a supplementary tool to the National Program of Chagas Disease Control to enhance the effects of insecticide sprayings. Conclusions Even if cleaning activities were still neglected, community participation proved to be effective in reducing house infestation. PMID:25604766

  20. The expansion of Phytophthora clade 8b: three new species associated with winter grown vegetable crops.

    PubMed

    Bertier, L; Brouwer, H; de Cock, A W A M; Cooke, D E L; Olsson, C H B; Höfte, M

    2013-12-01

    Despite its association with important agricultural crops, Phytophthora clade 8b is a poorly studied group of species. The clade currently consists of three officially described species (Phytophthora porri, P. brassicae and P. primulae) that are host-specific pathogens of leek, cabbages and Primula spp., respectively. However, over the past few decades, several other clade 8b-like Phytophthoras have been found on a variety of different host plants that were all grown at low temperatures in winter seasons. In this study, a collection of 30 of these isolates was subjected to a phylogenetic study using two loci (the rDNA ITS region and the mitochondrial cox1 gene). This analysis revealed a clear clustering of isolates according to their host plants. To verify whether these isolates belong to separate species, a detailed morphological study was conducted. On the basis of genetic and morphological differences and host specificity, we now present the official description of three new species in clade 8b: Phytophthora cichorii sp. nov., P. dauci sp. nov. and P. lactucae sp. nov. Two other groups of isolates (Phytophthora taxon castitis and Phytophthora taxon parsley) might also represent new species but the data available at this time are insufficient for an official description. This brings Phytophthora clade 8b to a group of six species that are all host-specific, slow-growing and specifically infect herbaceous crops at low temperatures.

  1. Antifungal Effect of Plant Essential Oils on Controlling Phytophthora Species.

    PubMed

    Amini, Jahanshir; Farhang, Vahid; Javadi, Taimoor; Nazemi, Javad

    2016-02-01

    In this study, antifungal activity of essential oils of Cymbopogon citratus and Ocimum basilicum and two fungicides Mancozeb and Metalaxyl-Mancozeb in six different concentrations were investigated for controlling three species of Phytophthora, including P. capsici, P. drechsleri and P. melonis on pepper, cucumber and melon under in vitro and greenhouse conditions, respectively. Under the in vitro condition, the median effective concen- tration (EC50) values (ppm) of plant essential oils and fungicides were measured. In greenhouse, soil infested with Phytophthora species was treated by adding 50 ml of essential oils and fungicides (100 ppm). Disease severity was determined after 28 days. Among two tested plant essential oils, C. citratus had the lowest EC50 values for inhibition of the mycelial growth of P. capsici (31.473), P. melonis (33.097) and P. drechsleri (69.112), respectively. The mean EC50 values for Metalaxyl-Mancozeb on these pathogens were 20.87, 20.06 and 17.70, respectively. Chemical analysis of plant essential oils by GC-MS showed that, among 42 compounds identified from C. citratus, two compounds β-geranial (α-citral) (39.16%) and z-citral (30.95%) were the most abundant. Under the greenhouse condition, Metalaxyl-Mancozeb caused the greatest reduction in disease severity, 84.2%, 86.8% and 92.1% on melon, cucumber, and pepper, respectively. The C. citratus essential oil reduced disease severity from 47.4% to 60.5% compared to the untreated control (p≤0.05). Essential oils of O. basilicum had the lowest effects on the pathogens under in vitro and greenhouse conditions. These results show that essential oils may contribute to the development of new antifungal agents to protect the crops from Phytophthora diseases. PMID:26889111

  2. Host adaptation and speciation through hybridization and polyploidy in Phytophthora.

    PubMed

    Bertier, Lien; Leus, Leen; D'hondt, Liesbet; de Cock, Arthur W A M; Höfte, Monica

    2013-01-01

    It is becoming increasingly evident that interspecific hybridization is a common event in phytophthora evolution. Yet, the fundamental processes underlying interspecific hybridization and the consequences for its ecological fitness and distribution are not well understood. We studied hybridization events in phytophthora clade 8b. This is a cold-tolerant group of plant pathogenic oomycetes in which six host-specific species have been described that mostly attack winter-grown vegetables. Hybrid characterization was done by sequencing and cloning of two nuclear (ITS and Ypt1) and two mitochondrial loci (Cox1 and Nadh1) combined with DNA content estimation using flow cytometry. Three different mtDNA haplotypes were recovered among the presumed hybrid isolates, dividing the hybrids into three types, with different parental species involved. In the nuclear genes, additivity, i.e. the presence of two alleles coming from different parents, was detected. Hybrid isolates showed large variations in DNA content, which was positively correlated with the additivity in nuclear loci, indicating allopolyploid hybridization followed by a process of diploidization. Moreover, indications of homeologous recombination were found in the hybrids by cloning ITS products. The hybrid isolates have been isolated from a range of hosts that have not been reported previously for clade 8b species, indicating that they have novel pathogenic potential. Next to this, DNA content measurements of the non-hybrid clade 8b species suggest that polyploidy is a common feature of this clade. We hypothesize that interspecific hybridization and polyploidy are two linked phenomena in phytophthora, and that these processes might play an important and ongoing role in the evolution of this genus.

  3. Antifungal Effect of Plant Essential Oils on Controlling Phytophthora Species.

    PubMed

    Amini, Jahanshir; Farhang, Vahid; Javadi, Taimoor; Nazemi, Javad

    2016-02-01

    In this study, antifungal activity of essential oils of Cymbopogon citratus and Ocimum basilicum and two fungicides Mancozeb and Metalaxyl-Mancozeb in six different concentrations were investigated for controlling three species of Phytophthora, including P. capsici, P. drechsleri and P. melonis on pepper, cucumber and melon under in vitro and greenhouse conditions, respectively. Under the in vitro condition, the median effective concen- tration (EC50) values (ppm) of plant essential oils and fungicides were measured. In greenhouse, soil infested with Phytophthora species was treated by adding 50 ml of essential oils and fungicides (100 ppm). Disease severity was determined after 28 days. Among two tested plant essential oils, C. citratus had the lowest EC50 values for inhibition of the mycelial growth of P. capsici (31.473), P. melonis (33.097) and P. drechsleri (69.112), respectively. The mean EC50 values for Metalaxyl-Mancozeb on these pathogens were 20.87, 20.06 and 17.70, respectively. Chemical analysis of plant essential oils by GC-MS showed that, among 42 compounds identified from C. citratus, two compounds β-geranial (α-citral) (39.16%) and z-citral (30.95%) were the most abundant. Under the greenhouse condition, Metalaxyl-Mancozeb caused the greatest reduction in disease severity, 84.2%, 86.8% and 92.1% on melon, cucumber, and pepper, respectively. The C. citratus essential oil reduced disease severity from 47.4% to 60.5% compared to the untreated control (p≤0.05). Essential oils of O. basilicum had the lowest effects on the pathogens under in vitro and greenhouse conditions. These results show that essential oils may contribute to the development of new antifungal agents to protect the crops from Phytophthora diseases.

  4. Host Adaptation and Speciation through Hybridization and Polyploidy in Phytophthora

    PubMed Central

    Bertier, Lien; Leus, Leen; D’hondt, Liesbet; de Cock, Arthur W. A. M.; Höfte, Monica

    2013-01-01

    It is becoming increasingly evident that interspecific hybridization is a common event in phytophthora evolution. Yet, the fundamental processes underlying interspecific hybridization and the consequences for its ecological fitness and distribution are not well understood. We studied hybridization events in phytophthora clade 8b. This is a cold-tolerant group of plant pathogenic oomycetes in which six host-specific species have been described that mostly attack winter-grown vegetables. Hybrid characterization was done by sequencing and cloning of two nuclear (ITS and Ypt1) and two mitochondrial loci (Cox1 and Nadh1) combined with DNA content estimation using flow cytometry. Three different mtDNA haplotypes were recovered among the presumed hybrid isolates, dividing the hybrids into three types, with different parental species involved. In the nuclear genes, additivity, i.e. the presence of two alleles coming from different parents, was detected. Hybrid isolates showed large variations in DNA content, which was positively correlated with the additivity in nuclear loci, indicating allopolyploid hybridization followed by a process of diploidization. Moreover, indications of homeologous recombination were found in the hybrids by cloning ITS products. The hybrid isolates have been isolated from a range of hosts that have not been reported previously for clade 8b species, indicating that they have novel pathogenic potential. Next to this, DNA content measurements of the non-hybrid clade 8b species suggest that polyploidy is a common feature of this clade. We hypothesize that interspecific hybridization and polyploidy are two linked phenomena in phytophthora, and that these processes might play an important and ongoing role in the evolution of this genus. PMID:24386473

  5. Antifungal Effect of Plant Essential Oils on Controlling Phytophthora Species

    PubMed Central

    Amini, Jahanshir; Farhang, Vahid; Javadi, Taimoor; Nazemi, Javad

    2016-01-01

    In this study, antifungal activity of essential oils of Cymbopogon citratus and Ocimum basilicum and two fungicides Mancozeb and Metalaxyl-Mancozeb in six different concentrations were investigated for controlling three species of Phytophthora, including P. capsici, P. drechsleri and P. melonis on pepper, cucumber and melon under in vitro and greenhouse conditions, respectively. Under the in vitro condition, the median effective concen- tration (EC50) values (ppm) of plant essential oils and fungicides were measured. In greenhouse, soil infested with Phytophthora species was treated by adding 50 ml of essential oils and fungicides (100 ppm). Disease severity was determined after 28 days. Among two tested plant essential oils, C. citratus had the lowest EC50 values for inhibition of the mycelial growth of P. capsici (31.473), P. melonis (33.097) and P. drechsleri (69.112), respectively. The mean EC50 values for Metalaxyl-Mancozeb on these pathogens were 20.87, 20.06 and 17.70, respectively. Chemical analysis of plant essential oils by GC-MS showed that, among 42 compounds identified from C. citratus, two compounds β-geranial (α-citral) (39.16%) and z-citral (30.95%) were the most abundant. Under the greenhouse condition, Metalaxyl-Mancozeb caused the greatest reduction in disease severity, 84.2%, 86.8% and 92.1% on melon, cucumber, and pepper, respectively. The C. citratus essential oil reduced disease severity from 47.4% to 60.5% compared to the untreated control (p≤0.05). Essential oils of O. basilicum had the lowest effects on the pathogens under in vitro and greenhouse conditions. These results show that essential oils may contribute to the development of new antifungal agents to protect the crops from Phytophthora diseases. PMID:26889111

  6. Elemental variations in the germinating fungus Phytophthora palmivora

    NASA Astrophysics Data System (ADS)

    Mazzolini, A. P.; Grant, B. R.; Sealock, R. M.; Legge, G. J. F.

    1991-03-01

    We have measured the elemental variations between zoospores and germinating cystospores of the fungus Phytophthora palmivora, using a scanning proton microprobe. Averaged over a number of individual cells, our results indicate that the level of Ca is much lower in germinating cystospores than in zoospores. The levels of S, Cl, and Zn also appear to be lower, and the level of K appears to be higher. The spatial distribution of elements within the germinating cystospore is very similar for P, S, Cl, K, Mn, Fe, and Cu, but significantly different for Ca and Zn.

  7. Elicitins from Phytophthora and basic resistance in tobacco.

    PubMed Central

    Yu, L M

    1995-01-01

    Elicitins are a family of small proteins secreted by species of Phytophthora. They are thought to be major determinants of the resistance response of tobacco against these oomycetes, since purified elicitins, alone and at low concentrations, can induce vigorous defense responses in tobacco (i.e., hypersensitive cell death and resistance against subsequent pathogen attack), and in vitro elicitin production by Phytophthora isolates is strongly negatively correlated with their pathogenicity on tobacco plants. A number of elicitins have been purified and their amino acid sequences have been determined and found to be conserved. A three-dimensional structure for elicitin is emerging from nuclear magnetic resonance studies. Two structural classes, alpha and beta, are distinguished by their biological effects when applied to decapitated stems or petioles; the beta class causes more necrosis on leaves and provides better subsequent protection against pathogen attack. However, both these classes of elicitins will similarly cause necrosis when each is, instead, directly infiltrated into tobacco leaf panels. Effects of elicitins on tobacco cells include rapid electrolyte leakage, changes in protein phosphorylation and amounts of active oxygen species, and later production of ethylene and capsidiol. The sites of initial interaction with tobacco cells are unknown, but the interaction appears to induce general defense-related responses. PMID:7753775

  8. Rapid isothermal detection of Phytophthora species on plant samples using recombinase polymerase amplification

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recently several isothermal amplification techniques have been developed that are extremely tolerant towards inhibitors present in many plant extracts. Recombinase polymerase amplification (RPA) assays for the genus Phytophthora have been developed which provide a simple and rapid method to macerate...

  9. Life History Traits and Demographic Parameters of Triatoma infestans (Hemiptera: Reduviidae) Fed on Human Blood.

    PubMed

    Medone, Paula; Balsalobre, Agustin; Rabinovich, Jorge E; Marti, Gerardo A; Menu, Frédéric

    2015-11-01

    Triatoma infestans (Klug, 1834) (Hemiptera: Reduviidae), the main vector of Chagas disease in South America, feeds primarily on humans, but ethical reasons preclude carrying out demographical studies using people. Thus, most laboratory studies of T. infestans are conducted using bird or mammal live hosts that may result in different demographic parameters from those obtained on human blood. Therefore, it is of interest to determine whether the use of an artificial feeder with human blood would be operational to rear triatomines and estimate population growth rates. We estimated life history traits and demographic parameters using an artificial feeder with human blood and compared them with those obtained on live hens. Both groups of T. infestans were kept under constant conditions [28 ± 1°C, 40 ± 5% relative humidity, a photoperiod of 12:12 (L:D) h] and fed weekly. On the basis of age-specific survival and age-specific fecundity, we calculated the intrinsic rate of natural increase (r), the finite rate of population growth (λ), the net reproductive rate (Ro), and the mean generation time (Tg). Our results show differences in life history traits between blood sources, resulting in smaller population growth rates on human blood than on live hens. Although demographic growth rate was smaller on human blood than on hens, it still remains positive, so the benefit/cost ratio of this feeding method seems relatively attractive. We discuss possibility of using the artificial feeder with human blood for both ecological and behavioral studies. PMID:26373893

  10. Comparative Trial of Effectiveness of Pyrethroid Insecticides Against Peridomestic Populations of Triatoma infestans in Northwestern Argentina

    PubMed Central

    CECERE, MARÍA CARLA; VÁZQUEZ-PROKOPEC, GONZALO M.; CEBALLOS, LEONARDO A.; GUREVITZ, JUAN M.; ZÁRATE, JOAQUÍN E.; ZAIDENBERG, MARIO; KITRON, URIEL; GÜRTLER, RICARDO E.

    2007-01-01

    The effects of different pyrethroid insecticides, formulations, and doses on peridomestic populations of Triatoma infestans (Klug) were evaluated in 128 houses with 148 identified infested peridomestic sites in northwestern Argentina between October 2003 and March 2005. Four treatments were randomly assigned within each community: two doses of 5% suspension concentrate β-cypermethrin in water applied with manual compression sprayers, the standard dose (S) at 50 mg and a double dose (2S) at 100 mg active ingredient (AI)/m2; and two emulsifiable concentrates diluted in diesel fuel and applied with power sprayers, 25% cypermethrin (100 mg [AI]m2) (CF) and 10% permethrin (170 mg [AI]m2) (DF). Infestation was assessed by timed manual collections with a dislodging agent at baseline, 5, 12, and 17 mo postspraying, and the sites found to be reinfested at 5 mo postspraying were selectively resprayed. Only 2S eliminated T. infestans from all peridomestic sites up to 12 mo postspraying, and it was significantly more effective than all other treatments. At 5 mo postspraying, more sites treated with CF or DF rather than S had bug colonies that probably represented residual foci, which they also failed in eliminating after a second spray. At 17 mo postspraying, the prevalence of reinfested peridomestic sites was 5% for 2S, 29% for S, 43% for CF, and 54% for DF. The application of suspension concentrate pyrethroids in dose twice as large as that currently in use in the attack phase produces a greater initial impact and may eliminate peridomestic populations of T. infestans. PMID:17017227

  11. Life History Traits and Demographic Parameters of Triatoma infestans (Hemiptera: Reduviidae) Fed on Human Blood.

    PubMed

    Medone, Paula; Balsalobre, Agustin; Rabinovich, Jorge E; Marti, Gerardo A; Menu, Frédéric

    2015-11-01

    Triatoma infestans (Klug, 1834) (Hemiptera: Reduviidae), the main vector of Chagas disease in South America, feeds primarily on humans, but ethical reasons preclude carrying out demographical studies using people. Thus, most laboratory studies of T. infestans are conducted using bird or mammal live hosts that may result in different demographic parameters from those obtained on human blood. Therefore, it is of interest to determine whether the use of an artificial feeder with human blood would be operational to rear triatomines and estimate population growth rates. We estimated life history traits and demographic parameters using an artificial feeder with human blood and compared them with those obtained on live hens. Both groups of T. infestans were kept under constant conditions [28 ± 1°C, 40 ± 5% relative humidity, a photoperiod of 12:12 (L:D) h] and fed weekly. On the basis of age-specific survival and age-specific fecundity, we calculated the intrinsic rate of natural increase (r), the finite rate of population growth (λ), the net reproductive rate (Ro), and the mean generation time (Tg). Our results show differences in life history traits between blood sources, resulting in smaller population growth rates on human blood than on live hens. Although demographic growth rate was smaller on human blood than on hens, it still remains positive, so the benefit/cost ratio of this feeding method seems relatively attractive. We discuss possibility of using the artificial feeder with human blood for both ecological and behavioral studies.

  12. Tigutcystatin, a cysteine protease inhibitor from Triatoma infestans midgut expressed in response to Trypanosoma cruzi

    SciTech Connect

    Buarque, Diego S.; Spindola, Leticia M.N.; Martins, Rafael M.; Braz, Gloria R.C.; Tanaka, Aparecida S.

    2011-09-23

    Highlights: {yields} Tigutcystatin inhibits Trypanosoma cruzi cysteine proteases with high specificity. {yields} Tigutcystatin expression is up-regulated in response to T. cruzi infection. {yields} It is the first cysteine proteases inhibitor characterized from a triatomine insect. -- Abstract: The insect Triatoma infestans is a vector of Trypanosoma cruzi, the etiological agent of Chagas disease. A cDNA library was constructed from T. infestans anterior midgut, and 244 clones were sequenced. Among the EST sequences, an open reading frame (ORF) with homology to a cystatin type 2 precursor was identified. Then, a 288-bp cDNA fragment encoding mature cystatin (lacking signal peptide) named Tigutcystatin was cloned fused to a N-terminal His tag in pET-14b vector, and the protein expressed in Escherichia coli strain Rosetta gami. Tigutcystatin purified and cleaved by thrombin to remove His tag presented molecular mass of 11 kDa and 10,137 Da by SDS-PAGE and MALDI-TOF mass spectrometry, respectively. Purified Tigutcystatin was shown to be a tight inhibitor towards cruzain, a T. cruzi cathepsin L-like enzyme (K{sub i} = 3.29 nM) and human cathepsin L (K{sub i} = 3.78 nM). Tissue specific expression analysis showed that Tigutcystatin was mostly expressed in anterior midgut, although amplification in small intestine was also detected by semi quantitative RT-PCR. qReal time PCR confirmed that Tigutcystatin mRNA is significantly up-regulated in anterior midgut when T. infestans is infected with T. cruzi. Together, these results indicate that Tigutcystatin may be involved in modulation of T. cruzi in intestinal tract by inhibiting parasite cysteine proteases, which represent the virulence factors of this protozoan.

  13. Cross-species Global Proteomics Reveals Conserved and Unique Processes in Phytophthora sojae and Phytophthora ramorum*S⃞

    PubMed Central

    Savidor, Alon; Donahoo, Ryan S.; Hurtado-Gonzales, Oscar; Land, Miriam L.; Shah, Manesh B.; Lamour, Kurt H.; McDonald, W. Hayes

    2008-01-01

    Phytophthora ramorum and Phytophthora sojae are destructive plant pathogens. P. sojae has a narrow host range, whereas P. ramorum has a wide host range. A global proteomics comparison of the vegetative (mycelium) and infective (germinating cyst) life stages of P. sojae and P. ramorum was conducted to identify candidate proteins involved in host range, early infection, and vegetative growth. Sixty-two candidates for early infection, 26 candidates for vegetative growth, and numerous proteins that may be involved in defining host specificity were identified. In addition, common life stage proteomic trends between the organisms were observed. In mycelia, proteins involved in transport and metabolism of amino acids, carbohydrates, and other small molecules were up-regulated. In the germinating cysts, up-regulated proteins associated with lipid transport and metabolism, cytoskeleton, and protein synthesis were observed. It appears that the germinating cyst catabolizes lipid reserves through the β-oxidation pathway to drive the extensive protein synthesis necessary to produce the germ tube and initiate infection. Once inside the host, the pathogen switches to vegetative growth in which energy is derived from glycolysis and utilized for synthesis of amino acids and other molecules that assist survival in the plant tissue. PMID:18316789

  14. Pyrosequencing of environmental soil samples reveals biodiversity of the Phytophthora resident community in chestnut forests.

    PubMed

    Vannini, Andrea; Bruni, Natalia; Tomassini, Alessia; Franceschini, Selma; Vettraino, Anna Maria

    2013-09-01

    Pyrosequencing analysis was performed on soils from Italian chestnut groves to evaluate the diversity of the resident Phytophthora community. Sequences analysed with a custom database discriminated 15 pathogenic Phytophthoras including species common to chestnut soils, while a total of nine species were detected with baiting. The two sites studied differed in Phytophthora diversity and the presence of specific taxa responded to specific ecological traits of the sites. Furthermore, some species not previously recorded were represented by a discrete number of reads; among these species, Phytophthora ramorum was detected at both sites. Pyrosequencing was demonstrated to be a very sensitive technique to describe the Phytophthora community in soil and was able to detect species not easy to be isolated from soil with standard baiting techniques. In particular, pyrosequencing is an highly efficient tool for investigating the colonization of new environments by alien species, and for ecological and adaptive studies coupled with biological detection methods. This study represents the first application of pyrosequencing for describing Phytophthoras in environmental soil samples.

  15. Niche invasion, competition and coexistence amongst wild and domestic Bolivian populations of Chagas vector Triatoma infestans (Hemiptera, Reduviidae, Triatominae).

    PubMed

    Lardeux, Frédéric

    2013-04-01

    The model developed for human parasites by Bottomley et al. (2007) has been adapted to the dynamics of triatomines to better understand the processes of niche invasion, competition among species and coexistence. In Bolivia, both wild and domestic populations of Triatoma infestans exist. Their ecological niches are normally separated and the two populations do not interbreed, behaving as two distinct species. However, it has been suggested that the two populations may compete, highlighting therefore the potential risk of wild populations invading human dwellings. The model revealed the importance of the basic reproduction rates R0 of triatomine colonies for the risk of invasion. This depends not only on life traits such as survival and fecundity, but also on (1) the density-dependence phenomenon that limits triatomine establishment, (2) on house exposure to infection and (3) on the correlation between house susceptibility to domestic T. infestans and house susceptibility to wild T. infestans. Competition and coexistence amongst the two groups of T. infestans may occur under particular conditions, but are very unlikely.

  16. Antibody responses of domestic animals to salivary antigens of Triatoma infestans as biomarkers for low-level infestation of triatomines

    PubMed Central

    Schwarz, Alexandra; Sternberg, Jeremy M.; Johnston, Valerie; Medrano-Mercado, Nora; Anderson, Jennifer M.; Hume, Jen C.C.; Valenzuela, Jesus G.; Schaub, Günter A.; Billingsley, Peter F.

    2009-01-01

    Hematophagous arthropods such as Triatoma infestans, the vector of Trypanosoma cruzi, elicit host-immune responses during feeding. Characterization of antibody responses to salivary antigens offers the potential to develop immunologically based monitoring techniques for exposure to re-emergent triatomine bug populations in peridomestic animals. IgG-antibody responses to the salivary antigens of T. infestans have been detected in chickens as soon as 2 days after the first exposure to five adult bugs. Chickens and guinea pigs regularly exposed to this number of triatomines showed a significantly lower anti-saliva antibody titre than animals exposed to 25 adults and fifth instars of four different T. infestans strains originating from Bolivia and from Northern Chile. Highly immunogenic salivary antigens of 14 and 21 kDa were recognised by all chicken sera and of 79 kDa by all guinea pig sera. Cross-reactivity studies using saliva or salivary gland extracts from different hematophagous species, e.g. different triatomines, bed bugs, mosquitoes, sand flies and ticks, as well as chicken sera exposed to triatomines and mosquitoes, demonstrated that the 14 and 21 kDa salivary antigens were only found in triatomines. Sera from peridomestic chickens and guinea pigs in sites of known T. infestans challenge in Bolivia also recognised the 14 and 21 kDa antigens. These represent promising epidemiological markers for the detection of small numbers of feeding bugs and hence may be a new tool for vector surveillance in Chagas disease control programs. PMID:19248784

  17. Distribution and characterization of nitric oxide synthase in the nervous system of Triatoma infestans (Insecta: Heteroptera)

    PubMed Central

    Coronel, María F.; Nowicki, Susana; Nighorn, Alan J.; Villar, Marcelo J.

    2007-01-01

    The biochemical characterization of nitric oxide synthase (NOS) and its distribution in the central nervous system (CNS) were studied in the heteropteran bug Triatoma infestans. NOS-like immunoreactivity was found in the brain, subesophageal ganglion, and thoracic ganglia by using immunocytochemistry. In the protocerebrum, NOS-immunoreactive (IR) somata were detected in the anterior, lateral, and posterior soma rinds. In the optic lobe, numerous immunostained somata were observed at the level of the first optic chiasma, around the lobula, and in the proximal optic lobe. In the deutocerebrum, NOS-IR perikarya were mainly observed in the lateral soma rind, surrounding the sensory glomeruli, and a few cell bodies were seen in association with the antennal mechanosensory and motor neuropil. No immunostaining could be detected in the antennal nerve. The subesophageal and prothoracic ganglia contained scattered immunostained cell bodies. NOS-IR somata were present in all the neuromeres of the posterior ganglion. Western blotting showed that a universal NOS antiserum recognized a band at 134 kDa, in agreement with the expected molecular weight of the protein. Analysis of the kinetics of nitric oxide production revealed a fully active enzyme in tissue samples of the CNS of T. infestans. PMID:17235602

  18. Molecular Population Genetics and Phylogeography of the Chagas Disease Vector Triatoma infestans in South America

    PubMed Central

    Piccinali, R. V.; Marcet, P. L.; Noireau, F; Kitron, U; GÜrtler, R. E.; Dotson, E. M.

    2009-01-01

    Knowledge of the genetic variability, population structure, and evolutionary history of Triatoma infestans may be useful for developing rational vector control strategies. A 661-bp fragment of the mitochondrial gene cytochrome oxidase I (COI) was sequenced and analyzed in bugs from Argentina, Uruguay, Peru, and Bolivia, including peridomestic, domestic, Andean, and Chaco sylvatic bugs. A total of 48 polymorphic sites among 37 haplotypes were described. Nucleotide variation fluctuated among samples, with the highest nucleotide diversity observed in seven Argentinean provinces. Within this group, some populations showed patterns of variability compatible with population expansions and/or fine-scale population structure, whereas others suggested population bottlenecks and/or population admixture processes. A maximum parsimony analysis of the haplotypes showed the presence of a Bolivian/Peruvian and an Argentinean/Uruguayan clade. Bolivian sequences were further divided in Chaco sylvatic and Andean domestic and sylvatic. Two different nested clades were found within the Argentinean/Uruguayan cluster. Analysis of molecular variance (AMOVA) and KST* analysis supported a strong population structure in Argentina, where genetic differentiation was correlated with geographic distance. Departures from neutrality expectations and a nested cladistic analysis suggest a recent population expansion of T. infestans in Argentina, followed by restricted gene flow and patterns of isolation by distance. This expansion could have taken place as a two-wave process, as was shown by the phylogenetic analysis and signatures of population admixture in the southernmost Argentinean populations. PMID:19645282

  19. Inheritance of resistance to pyrethroids in Triatoma infestans, the main Chagas disease vector in South America.

    PubMed

    Cardozo, R M; Panzera, F; Gentile, A G; Segura, M A; Pérez, R; Díaz, R A; Basombrío, M A

    2010-12-01

    An outbreak of pyrethroid resistance was recently detected in Triatoma infestans from northern Argentina. To analyze the inheritance of the resistant phenotype, we carried out experimental crosses between resistant (R) and susceptible (S) strains captured in Argentina during 2005. The R strain was collected from sprayed houses in the north of the province of Salta while the S strain was collected in the province of Chaco. Both strains were bred in the laboratory for reciprocal crosses (F1), intercrosses (F2) and backcrosses (BC). The descendents were tested by a standard insecticide resistance bioassay. Resistance ratios were 1 for S strain, 103.36 for R strain and 18.34 for F1. The regression lines of F1 generations (R×S and S×R) showed no significant differences and were closer to that of the R parents, indicating that inheritance of deltamethrin resistance in T. infestans is autosomal and incompletely dominant (D=0.20). Chi-square analysis from responses of intercross and backcross progenies rejected the hypothesis of a single gene being responsible for resistance. The minimum number of independent segregation genes was three, as calculated with Lande's method. The genetic basis here described for the resistant phenotype indicate that, under pyrethroid selective pressure, the resistant genotypes could be easily spread to susceptible insects from resistant individuals, posing a major threat to vectorial control of Chagas disease.

  20. Characterisation of Phytophthora capsici isolates from black pepper in Vietnam.

    PubMed

    Truong, Nguyen V; Liew, Edward C Y; Burgess, Lester W

    2010-01-01

    Phytophthora foot rot of black pepper caused by Phytophthora capsici is a major disease of black pepper (Piper nigrum) throughout Vietnam. To understand the population structure of P. capsici, a large collection of P. capsici isolates from black pepper was studied on the basis of mating type, random amplified microsatellites (RAMS) and repetitive extragenic palindromic (REP) fingerprinting. Two mating types A1 and A2 were detected in four provinces in two climatic regions, with A1:A2 ratios ranging from 1:3 to 1:5. In several instances A1 and A2 mating types were found to co-exist in the same farm or black pepper pole, suggesting the potential for sexual reproduction of P. capsici in the field in Vietnam although its contribution to disease epidemics is uncertain. RAMS and REP DNA fingerprinting analysis of 118 isolates of P. capsici from black pepper showed that the population was genetically more diverse where two mating types were found, although the overall genetic diversity was low with most of the isolates belonging to one clonal group. The implication of these findings is discussed. The low diversity among isolates suggests that the P. capsici population may have originated from a single source. There was no genetic differentiation of isolates from different climatic regions. In addition to the large clonal group, several isolates with unique RAMS/REP phenotypes were also detected. Most of these unique phenotypes belonged to the minority A1 mating type. This may have significant implications for a gradual increase in overall genetic diversity. PMID:20960972

  1. Compost suppressiveness against Phytophthora capsicion pepper in potting trials.

    PubMed

    Pugliese, M; Marenco, M; Gullino, M L; Garibaldi, A

    2013-01-01

    Suppression of soil-borne plant diseases with composts has been widely studied. Composts suppressive to soil-borne pathogens have been detected in various cropping systems. Vegetable plants are generally propagated in pots, allowing the use of suppressive substrates to control zoospore-producing pathogens, like Phytophthora sp. The objective of the present work was to assess compost suppressiveness against Phytophthora capsici on pepper (cv. Corno di Toro). A municipal compost showing a good suppressive activity in previous trials on vegetable crops was used. Compost was mixed at 10, 20, 40, 60, 80 and 100% (v/v) with a commercial peat substrate, used as control. Substrates have been inoculated at 0.25, 0.5 and 1 g/l with wheat and hemp kernels infested with P. capsici and after one week 10 plants were transplanted for each treatment in 4 trays of 10 liters volume capacity and placed in greenhouse at 20 degrees C. Diseased plants were assessed weekly after transplanting and above-ground biomass of plants was assessed at the end of the trials. Compost applied at 80% significantly controlled the disease at high inoculum density (1 g/l), while at lower inoculums density, 0.25 and 0.5 g/l, reduced compost applications, 40% and 60% respectively, were sufficient to significantly control the disease. The application of compost at 20%, in absence of the pathogen, increased the biomass of pepper plants compared to control. The use of compost based substrates can be a suitable strategy for controlling soil-borne diseases on pepper, but results depends on application rates.

  2. Development of Phytophthora fruit rot resistant watermelon germplasm lines: USVL489-PFR, USVL782-PFR, USVL203-PFR and USVL020-PFR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora capsici, distributed worldwide, is an aggressive pathogen with a broad host range, infecting solanaceous, leguminaceous, and cucurbitaceous crops. Phytophthora fruit rot of watermelon (Citrullus lanatus) caused by Phytophthora capsici was first reported in the U.S. in 1940. Since then...

  3. Kdr mutations in Triatoma infestans from the Gran Chaco are distributed in two differentiated foci: Implications for pyrethroid resistance management.

    PubMed

    Sierra, Ivana; Capriotti, Natalia; Fronza, Georgina; Mougabure-Cueto, Gastón; Ons, Sheila

    2016-06-01

    Point mutations in the voltage-gated sodium channel, the primary target of pyrethroid insecticides, have been associated with the resistance in Triatoma infestans, an important vector of Chagas' disease. Hence, the sustainability of vector control programs requires the implementation of resistance management strategies. We determined the sensitivity of the molecular assays previously designed for early resistance detection to be used in pooled samples from a wide area of the endemic region, and validated them for their routine use in control campaigns for the monitoring of insecticide resistance in T. infestans. Consequently, we used these methods to examine the distribution of resistance-associated mutations in the sodium channel gene in populations of T. infestans from the Argentinean and Bolivian Gran Chaco. The PASA and REA assays tested proved sensitive enough to detect kdr SNPs in pooled samples, indicating these assays are suitable for routine screening in insecticide resistance surveillance. Two geographically differentiated foci were detected in T. infestans populations from the Argentinean and Bolivian Gran Chaco, with populations on the Bolivian-Argentinean border carrying L1014F mutation, and those from the Argentinean Chaco carrying L925I mutation. In all highly resistant populations analyzed, one of both kdr mutations was present, and toxicological assays determined that all pyrethroid resistant populations analyzed herein were sensitive to fenitrothion. The principal cause of pyrethroid resistance in T. infestans from the Gran Chaco ecoregion is kdr mutations in the sodium channel. Different levels of resistance occur in different populations carrying identical mutation, suggesting the existence of contributory mechanisms. PMID:26992297

  4. Hidden Sylvatic Foci of the Main Vector of Chagas Disease Triatoma infestans: Threats to the Vector Elimination Campaign?

    PubMed Central

    Schachter-Broide, Judith; Dujardin, Jean-Pierre; Dotson, Ellen M.; Kitron, Uriel; Gürtler, Ricardo E.

    2011-01-01

    Background Establishing the sources of reinfestation after residual insecticide spraying is crucial for vector elimination programs. Triatoma infestans, traditionally considered to be limited to domestic or peridomestic (abbreviated as D/PD) habitats throughout most of its range, is the target of an elimination program that has achieved limited success in the Gran Chaco region in South America. Methodology/Principal Findings During a two-year period we conducted semi-annual searches for triatomine bugs in every D/PD site and surrounding sylvatic habitats after full-coverage spraying of pyrethroid insecticides of all houses in a well-defined rural area in northwestern Argentina. We found six low-density sylvatic foci with 24 T. infestans in fallen or standing trees located 110–2,300 m from the nearest house or infested D/PD site detected after insecticide spraying, when house infestations were rare. Analysis of two mitochondrial gene fragments of 20 sylvatic specimens confirmed their species identity as T. infestans and showed that their composite haplotypes were the same as or closely related to D/PD haplotypes. Population studies with 10 polymorphic microsatellite loci and wing geometric morphometry consistently indicated the occurrence of unrestricted gene flow between local D/PD and sylvatic populations. Mitochondrial DNA and microsatellite sibship analyses in the most abundant sylvatic colony revealed descendents from five different females. Spatial analysis showed a significant association between two sylvatic foci and the nearest D/PD bug population found before insecticide spraying. Conclusions Our study shows that, despite of its high degree of domesticity, T. infestans has sylvatic colonies with normal chromatic characters (not melanic morphs) highly connected to D/PD conspecifics in the Argentinean Chaco. Sylvatic habitats may provide a transient or permanent refuge after control interventions, and function as sources for D/PD reinfestation. The

  5. A country bug in the city: urban infestation by the Chagas disease vector Triatoma infestans in Arequipa, Peru

    PubMed Central

    2013-01-01

    Background Interruption of vector-borne transmission of Trypanosoma cruzi remains an unrealized objective in many Latin American countries. The task of vector control is complicated by the emergence of vector insects in urban areas. Methods Utilizing data from a large-scale vector control program in Arequipa, Peru, we explored the spatial patterns of infestation by Triatoma infestans in an urban and peri-urban landscape. Multilevel logistic regression was utilized to assess the associations between household infestation and household- and locality-level socio-environmental measures. Results Of 37,229 households inspected for infestation, 6,982 (18.8%; 95% CI: 18.4 – 19.2%) were infested by T. infestans. Eighty clusters of infestation were identified, ranging in area from 0.1 to 68.7 hectares and containing as few as one and as many as 1,139 infested households. Spatial dependence between infested households was significant at distances up to 2,000 meters. Household T. infestans infestation was associated with household- and locality-level factors, including housing density, elevation, land surface temperature, and locality type. Conclusions High levels of T. infestans infestation, characterized by spatial heterogeneity, were found across extensive urban and peri-urban areas prior to vector control. Several environmental and social factors, which may directly or indirectly influence the biology and behavior of T. infestans, were associated with infestation. Spatial clustering of infestation in the urban context may both challenge and inform surveillance and control of vector reemergence after insecticide intervention. PMID:24171704

  6. Reaction of Phytophthora fruit rot resistant germplasm lines to a broad range of Phytophthora capsici isolates from across the United States of America

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora fruit rot limits watermelon production in most states in the Southeastern US (FL, GA, SC, NC and VA). It has also become a serious problem in recent years in northern states (IN, MD, DE). About 50% of the US watermelons are grown in the southeastern states where environmental conditions...

  7. Genome-wide association mapping of partial resistance to Phytophthora sojae in soybean plant introductions from the Republic of Korea

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora root and stem rot is one of the most yield-limiting diseases of soybean [Glycine max (L.) Merr], caused by the oomycete Phytophthora sojae. Partial resistance is controlled by several genes and, compared to single gene (Rps gene) resistance to P. sojae, places less selection pressure on...

  8. Fungicide rotation schemes for managing Phytophthora fruit rot of watermelon across Southeastern United States (NC, SC, and GA)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora capsici has been documented as a pathogen on a wide variety of vegetable crops in the family Solanaceae, Cucurbitaceae, Fabaceae, and plants belonging to 23 other families. Phytophthora fruit rot of watermelons caused by P. capsici is particularly severe in southeastern U.S where optima...

  9. Effect of cultural practices and fungicide treatments on the severity of Phytophthora root rot of blueberries grown in Mississippi

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Effect of cultural practices and fungicide treatments on the severity of Phytophthora root rot of blueberries grown in Mississippi Melinda Miller-Butler and Barbara J. Smith ABSTRACT. Phytophthora root rot is an important disease of blueberries especially when grown in areas with poor drainage. Re...

  10. Tracing the Path of Pathogens: An Ongoing Study Evaluates the Systems Approach for Managing Phytophthora Diseases in Nurseries

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Almost every nursery grower is familiar with the plant pathogen Phytophthora (from the Greek, meaning “plant destroyer”). Phytophthora species cause some of the most damaging nursery crop diseases nationwide, including root rot, dieback, leaf blight and shoot blight. These pathogens do more than ca...

  11. Identification of Phytophthora species baited and isolated from forest soil and streams in northwestern Yunnan province, China

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora species were surveyed by collecting soil samples and placing bait leaves in selected streams during June - October in the years 2005, 2006 and 2010 at three sites in oak forests in Diqing Tibetan Autonomous Prefecture of NW Yunnan province, China. Seventy-three isolates of Phytophthora ...

  12. Molecular response to the pathogen Phytophthora sojae among ten soybean near isogenic lines revealed by comparative transcriptomics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora root and stem rot (PRR) of soybean, caused by Phytophthora sojae, is effectively controlled by Rps genes in soybean. Rps genes are race-specific, yet the mechanism of resistance, as well as susceptibility, remains largely unclear. Taking advantage of RNA-seq technology, we sequenced the...

  13. Mapping of quantitative trait loci associated with partial resistance to phytophthora sojae and flooding tolerance in soybean

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora root rot (PRR) caused by Phytophthora sojae Kaufm. & Gerd. and flooding can limit growth and productivity, of soybean [Glycine max (L.) Merr.], especially on poorly drained soils. The primary objective of this research project was to map quantitative trait loci (QTL) associated with f...

  14. Molecular mapping and characterization of two genes conferring resistance to Phytophthora sojae in a soybean landrace PI 567139B

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora root and stem rot (PRR), caused by the soil-borne oomycete pathogen Phytophthora sojae, is one of the most destructive diseases of soybean. PRR can be effectively controlled by race-specific genes conferring resistance to P. sojae (Rps). However, the Rps genes are usually non-durable, a...

  15. Pathogenicity, fungicide resistance, and genetic variability of Phytophthora rubi isolates from raspberry (Rubus idaeus) in the Western United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Root rot of raspberry (Rubus idaeus), thought to be primarily caused by Phytophthora rubi, is an economically important disease in the western United States. The objectives of this study were to determine which Phytophthora species are involved in root rot, examine the efficacy of different isolatio...

  16. Sylvatic foci of the Chagas disease vector Triatoma infestans in Chile: description of a new focus and challenges for control programs.

    PubMed

    Bacigalupo, Antonella; Torres-Pérez, Fernando; Segovia, Verónica; García, Alejandro; Correa, Juana P; Moreno, Lucila; Arroyo, Patricio; Cattan, Pedro E

    2010-08-01

    Triatoma infestans is one of the main domestic vectors of Chagas disease. Reports of wild habitat occurrences have recently increased. In Chile, after a successful elimination campaign of T. infestans domestic infestation, a sylvatic focus was reported in bromeliads in the metropolitan region. Here, we report a new focus of sylvatic T. infestans inhabiting rock piles in the Valparaíso region in central Chile. All T. infestans captured were nymphal instars living among the stones, which were inhabited by several mammal species, along with the sylvatic triatomine vector Mepraia spinolai. We found a prevalence of infection with Trypanosoma cruzi of 36.54% in T. infestans, similar to the previous report for sylvatic specimens from bromeliads. Sylvatic populations of T. infestans should be studied at different geographic scales to elucidate their role in the maintenance of the sylvatic transmission cycle of T. cruzi and their possible role in threatening the domestic elimination of this vector. This information should be used to re-design the control programs in Chile to avoid the re-establishment of the domestic cycle.

  17. Soil bacteria as sources of virulence signal providers promoting plant infection by Phytophthora pathogens.

    PubMed

    Kong, Ping; Hong, Chuanxue

    2016-01-01

    Phytophthora species are known as "plant destroyers" capable of initiating single zoospore infection in the presence of a quorum of chemical signals from the same or closely related species of oomycetes. Since the natural oomycete population is too low to reach a quorum necessary to initiate a disease epidemic, creation of the quorum is reliant on alternate sources. Here, we show that a soil bacterial isolate, Bacillus megaterium Sb5, promotes plant infection by Phytophthora species. In the presence of Sb5 exudates, colonization of rhododendron leaf discs by 12 Phytophthora species/isolates was significantly enhanced, single zoospores of P. nicotianae infected annual vinca and P. sojae race 25 successfully attacked a non-host plant, Nicotiana benthamiana as well as resistant soybean cultivars with RPS1a or RPS3a. Sb5 exudates, most notably the fractions larger than 3 kDa, promoted plant infection by improving zoospore swimming, germination and plant attachment. Sb5 exudates also stimulated infection hypha growth and upregulated effector gene expression. These results suggest that environmental bacteria are important sources of virulence signal providers that promote plant infection by Phytophthora species, advancing our understanding of biotic factors in the environmental component of the Phytophthora disease triangle and of communal infection of plant pathogens. PMID:27616267

  18. Metabarcoding Analysis of Phytophthora Diversity Using Genus-Specific Primers and 454 Pyrosequencing.

    PubMed

    Prigigallo, Maria I; Abdelfattah, Ahmed; Cacciola, Santa O; Faedda, Roberto; Sanzani, Simona M; Cooke, David E L; Schena, L

    2016-03-01

    A metabarcoding method based on genus-specific primers and 454 pyrosequencing was utilized to investigate the genetic diversity of Phytophthora spp. in soil and root samples of potted plants, from eight nurseries. Pyrosequencing enabled the detection of 25 Phytophthora phylotypes distributed in seven different clades and provided a much higher resolution than a corresponding cloning/Sanger sequencing approach. Eleven of these phylotypes, including P. cactorum, P. citricola s.str., P. palmivora, P. palmivora-like, P. megasperma or P. gonapodyides, P. ramorum, and five putative new Phytophthora species phylogenetically related to clades 1, 2, 4, 6, and 7 were detected only with the 454 pyrosequencing approach. We also found an additional 18 novel records of a phylotype in a particular nursery that were not detected with cloning/Sanger sequencing. Several aspects confirmed the reliability of the method: (i) many identical sequence types were identified independently in different nurseries, (ii) most sequence types identified with 454 pyrosequencing were identical to those from the cloning/Sanger sequencing approach and/or perfectly matched GenBank deposited sequences, and (iii) the divergence noted between sequence types of putative new Phytophthora species and all other detected sequences was sufficient to rule out sequencing errors. The proposed method represents a powerful tool to study Phytophthora diversity providing that particular attention is paid to the analysis of 454 pyrosequencing raw read sequences and to the identification of sequence types.

  19. PCR amplification of species-specific DNA sequences can distinguish among Phytophthora species.

    PubMed Central

    Ersek, T; Schoelz, J E; English, J T

    1994-01-01

    We used PCR to differentiate species in the genus Phytophthora, which contains a group of devastating plant pathogenic fungi. We focused on Phytophthora parasitica, a species that can infect solanaceous plants such as tomato, and on Phytophthora citrophthora, which is primarily a citrus pathogen. Oligonucleotide primers were derived from sequences of a 1,300-bp P. parasitica-specific DNA segment and of an 800-bp P. citrophthora-specific segment. Under optimal conditions, the primers developed for P. parasitica specifically amplified a 1,000-bp sequence of DNA from isolates of P. parasitica. Primers for P. citrophthora similarly and specifically amplified a 650-bp sequence of DNA from isolates of P. citrophthora. Detectable amplification of these specific DNA sequences required picogram quantities of chromosomal DNA. Neither pair of primers amplified these sequences with DNAs from other species of Phytophthora or from the related genus Pythium. DNAs from P. parasitica and P. citrophthora growing in infected tomato stem tissue were amplified as distinctly as DNAs from axenic cultures of each fungal species. This is the first report on PCR-driven amplification with Phytophthora species-specific primers. Images PMID:8074533

  20. Multiple Phytophthora species associated with a single riparian ecosystem in South Africa.

    PubMed

    Nagel, Jan H; Slippers, Bernard; Wingfield, Michael J; Gryzenhout, Marieka

    2015-01-01

    The diversity of Phytophthora spp. in rivers and riparian ecosystems has received considerable international attention, although little such research has been conducted in South Africa. This study determined the diversity of Phytophthora spp. within a single river in Gauteng province of South Africa. Samples were collected over 1 y including biweekly river baiting with Rhododendron indicum leaves. Phytophthora isolates were identified with phylogenetic analyses of sequences for the internal transcribed spacer (ITS) region of the ribosomal DNA and the mitochondrial cytochrome oxidase c subunit I (coxI) gene. Eight Phytophthora spp. were identified, including a new taxon, P. taxon Sisulu-river, and two hybrid species from Cooke's ITS clade 6. Of these, species from Clade 6 were the most abundant, including P. chlamydospora and P. lacustris. Species residing in Clade 2 also were encountered, including P. multivora, P. plurivora and P. citrophthora. The detection of eight species in this investigation of Phytophthora diversity in a single riparian river ecosystem in northern South Africa adds to the known diversity of this genus in South Africa and globally.

  1. Soil bacteria as sources of virulence signal providers promoting plant infection by Phytophthora pathogens

    PubMed Central

    Kong, Ping; Hong, Chuanxue

    2016-01-01

    Phytophthora species are known as “plant destroyers” capable of initiating single zoospore infection in the presence of a quorum of chemical signals from the same or closely related species of oomycetes. Since the natural oomycete population is too low to reach a quorum necessary to initiate a disease epidemic, creation of the quorum is reliant on alternate sources. Here, we show that a soil bacterial isolate, Bacillus megaterium Sb5, promotes plant infection by Phytophthora species. In the presence of Sb5 exudates, colonization of rhododendron leaf discs by 12 Phytophthora species/isolates was significantly enhanced, single zoospores of P. nicotianae infected annual vinca and P. sojae race 25 successfully attacked a non-host plant, Nicotiana benthamiana as well as resistant soybean cultivars with RPS1a or RPS3a. Sb5 exudates, most notably the fractions larger than 3 kDa, promoted plant infection by improving zoospore swimming, germination and plant attachment. Sb5 exudates also stimulated infection hypha growth and upregulated effector gene expression. These results suggest that environmental bacteria are important sources of virulence signal providers that promote plant infection by Phytophthora species, advancing our understanding of biotic factors in the environmental component of the Phytophthora disease triangle and of communal infection of plant pathogens. PMID:27616267

  2. Phytophthora community structure analyses in Oregon nurseries inform systems approaches to disease management.

    PubMed

    Parke, Jennifer L; Knaus, Brian J; Fieland, Valerie J; Lewis, Carrie; Grünwald, Niklaus J

    2014-10-01

    Nursery plants are important vectors for plant pathogens. Understanding what pathogens occur in nurseries in different production stages can be useful to the development of integrated systems approaches. Four horticultural nurseries in Oregon were sampled every 2 months for 4 years to determine the identity and community structure of Phytophthora spp. associated with different sources and stages in the nursery production cycle. Plants, potting media, used containers, water, greenhouse soil, and container yard substrates were systematically sampled from propagation to the field. From 674 Phytophthora isolates recovered, 28 different species or taxa were identified. The most commonly isolated species from plants were Phytophthora plurivora (33%), P. cinnamomi (26%), P. syringae (19%), and P. citrophthora (11%). From soil and gravel substrates, P. plurivora accounted for 25% of the isolates, with P. taxon Pgchlamydo, P. cryptogea, and P. cinnamomi accounting for 18, 17, and 15%, respectively. Five species (P. plurivora, P. syringae, P. taxon Pgchlamydo, P. gonapodyides, and P. cryptogea) were found in all nurseries. The greatest diversity of taxa occurred in irrigation water reservoirs (20 taxa), with the majority of isolates belonging to internal transcribed spacer clade 6, typically including aquatic opportunists. Nurseries differed in composition of Phytophthora communities across years, seasons, and source within the nursery. These findings suggest likely contamination hazards and target critical control points for management of Phytophthora disease using a systems approach.

  3. Metabarcoding Analysis of Phytophthora Diversity Using Genus-Specific Primers and 454 Pyrosequencing.

    PubMed

    Prigigallo, Maria I; Abdelfattah, Ahmed; Cacciola, Santa O; Faedda, Roberto; Sanzani, Simona M; Cooke, David E L; Schena, L

    2016-03-01

    A metabarcoding method based on genus-specific primers and 454 pyrosequencing was utilized to investigate the genetic diversity of Phytophthora spp. in soil and root samples of potted plants, from eight nurseries. Pyrosequencing enabled the detection of 25 Phytophthora phylotypes distributed in seven different clades and provided a much higher resolution than a corresponding cloning/Sanger sequencing approach. Eleven of these phylotypes, including P. cactorum, P. citricola s.str., P. palmivora, P. palmivora-like, P. megasperma or P. gonapodyides, P. ramorum, and five putative new Phytophthora species phylogenetically related to clades 1, 2, 4, 6, and 7 were detected only with the 454 pyrosequencing approach. We also found an additional 18 novel records of a phylotype in a particular nursery that were not detected with cloning/Sanger sequencing. Several aspects confirmed the reliability of the method: (i) many identical sequence types were identified independently in different nurseries, (ii) most sequence types identified with 454 pyrosequencing were identical to those from the cloning/Sanger sequencing approach and/or perfectly matched GenBank deposited sequences, and (iii) the divergence noted between sequence types of putative new Phytophthora species and all other detected sequences was sufficient to rule out sequencing errors. The proposed method represents a powerful tool to study Phytophthora diversity providing that particular attention is paid to the analysis of 454 pyrosequencing raw read sequences and to the identification of sequence types. PMID:26574783

  4. [Survival and occurrence of molting in Triatoma infestans Klug (Hemiptera, Reduviidae) after temperature shock].

    PubMed

    Rodrigues, V L; Mello, M L; Ferraz Filho, A N; Dantas, M M

    1991-12-01

    Survival and molting occurrence were studied in specimens of Triatoma infestans over 30 days after temperature shocks. Hyperthermal and hypothermal shocks could be found to affect both survival and molting incidence as a function of temperature and period of the development phase and sex of the specimens. Considering the various test conditions, the shock at 0 degree C for 12 h was found to elicit the most deleterious effect, whereas shocks at 40 degrees C and 0 degree C even for 1 h are interpreted as affecting the hormonal balance which controls molting. Cases of a rise in post-shock survival are suggested to have been favored by heat-shock protein action.

  5. Putative Panmixia in Restricted Populations of Trypanosoma cruzi Isolated from Wild Triatoma infestans in Bolivia

    PubMed Central

    Barnabe, Christian; Buitrago, Rosio; Bremond, Philippe; Aliaga, Claudia; Salas, Renata; Vidaurre, Pablo; Herrera, Claudia; Cerqueira, Frédérique; Bosseno, Marie-France; Waleckx, Etienne; Breniere, Simone Frédérique

    2013-01-01

    Trypanosoma cruzi, the causative agent of Chagas disease, is subdivided into six discrete typing units (DTUs; TcI–TcVI) of which TcI is ubiquitous and genetically highly variable. While clonality is the dominant mode of propagation, recombinant events play a significant evolutive role. Recently, foci of wild Triatoma infestans have been described in Bolivia, mainly infected by TcI. Hence, for the first time, we evaluated the level of genetic exchange within TcI natural potentially panmictic populations (single DTU, host, area and sampling time). Seventy-nine TcI stocks from wild T. infestans, belonging to six populations were characterized at eight microsatellite loci. For each population, Hardy-Weinberg equilibrium (HWE), linkage disequilibrium (LD), and presence of repeated multilocus genotypes (MLG) were analyzed by using a total of seven statistics, to test the null hypothesis of panmixia (H0). For three populations, none of the seven statistics allowed to rejecting H0; for another one the low size did not allow us to conclude, and for the two others the tests have given contradictory results. Interestingly, apparent panmixia was only observed in very restricted areas, and was not observed when grouping populations distant of only two kilometers or more. Nevertheless it is worth stressing that for the statistic tests of "HWE", in order to minimize the type I error (i. e. incorrect rejection of a true H0), we used the Bonferroni correction (BC) known to considerably increase the type II error ( i. e. failure to reject a false H0). For the other tests (LD and MLG), we did not use BC and the risk of type II error in these cases was acceptable. Thus, these results should be considered as a good indicator of the existence of panmixia in wild environment but this must be confirmed on larger samples to reduce the risk of type II error. PMID:24312410

  6. Spatial Heterogeneity and Risk Maps of Community Infestation by Triatoma infestans in Rural Northwestern Argentina

    PubMed Central

    Vazquez-Prokopec, Gonzalo M.; Spillmann, Cynthia; Zaidenberg, Mario; Gürtler, Ricardo E.; Kitron, Uriel

    2012-01-01

    Background Fifty years of residual insecticide spraying to control Triatoma infestans in the Gran Chaco region of northern Argentina, Paraguay and Bolivia shows that vertically coordinated interventions aiming at full coverage have limited effects and are unsustainable. We quantified the spatial distribution of T. infestans domestic infestation at the district level, identified environmental factors associated with high infestation and then explored the usefulness of risk maps for the spatial stratification of interventions. Methods and Findings We performed spatial analyses of house infestation data collected by the National Chagas Service in Moreno Department, northern Argentina (1999–2002). Clusters of high domestic infestation occurred in the southwestern extreme of the district. A multi-model selection approach showed that domestic infestation clustered in areas of low elevation, with few farmlands, high density of rural houses, high mean maximum land surface temperature, large NDVI, and high percentage of degraded and deforested lands. The best model classified 98.4% of the communities in the training dataset (sensitivity, 93.3%; specificity, 95.4%). The risk map evidenced that the high-risk area only encompassed 16% of the district. By building a network-based transportation model we assessed the operational costs of spatially contiguous and spatially targeted interventions. Targeting clusters of high infestation would have reached ∼80% of all communities slated for full-coverage insecticide spraying, reducing in half the total time and economic cost incurred by a spatially contiguous strategy. Conclusions and Significance In disperse rural areas where control programs can accomplish limited coverage, consideration of infestation hot spots can contribute to the design and execution of cost-effective interventions against Chagas disease vectors. If field validated, targeted vertical control in high risk areas and horizontal control in medium to low risk

  7. [Effect of pentachloronitrobenzene (PCNB) on the ultrastructure of Mucor mucedo and Phytophthora cactorum].

    PubMed

    Casperson, G; Lyr, H

    1982-01-01

    The effect of PCNB in various concentrations on the ultrastructure of Mucor mucedo and phytophthora cactorum was analyzed after an incubation period of 2 hours. The most striking effect in both fungi was a diffuse lysis of the internal structure of the mitochondria which differs markedly from the lysis induced by etridiazol (terrazol). Moreover an enlargement of the perinuclear space and an increased formation of vacuoles was observed. In Mucor mucedo, but not in Phytophthora cactorum a pathological thickening of the cell wall was observed. Although after 2 hours incubation with PCNB Phytophthora gave similar ultrastructural reactions in the mitochondria as Mucor, in growth experiments on agar dishes this species was 5-10 times less sensitive to PCNB compared to Mucor.

  8. Phytophthora species recovered from the Connecticut River Valley in Massachusetts, USA.

    PubMed

    Brazee, Nicholas J; Wick, Robert L; Hulvey, Jonathan P

    2016-01-01

    Little is currently known about the assemblage of Phytophthora species in northeastern North America, representing a gap in our understanding of species incidence. Therefore, Phytophthora species were surveyed at 20 sites in Massachusetts, with 16 occurring in the Connecticut River Valley. Many of the sampled waterways were adjacent to active agricultural lands, yet were buffered by mature floodplain forests composed of Acer, Platanus, Populus and Ulmus. Isolates were recovered with three types of baits (rhododendron leaves, pear, green pepper) in 2013 and water filtration in 2014. Overall, 457 isolates of Phytophthora were recovered and based on morphological characters and rDNA internal transcribed spacer (ITS), β-tubulin (β-tub) and cytochrome oxidase c subunit I (cox1) sequences, 18 taxa were identified, including three new species: P. taxon intercalaris, P. taxon caryae and P. taxon pocumtuck. In addition, 49 isolates representing five species of Phytopythium also were identified. Water filtration captured a greater number of taxa (18) compared to leaf and fruit baits (12). Of the three bait types rhododendron leaves yielded the greatest number of isolates and taxa, followed by pear and green pepper, respectively. Despite the proximity to agricultural lands, none of the Phytophthora species baited are considered serious pathogens of vegetable crops in the region. However, many of the recovered species are known woody plant pathogens, including four species in the P. citricola s.l. complex that were identified: P. plurivora, P. citricola III, P. pini and a putative novel species, referred to here as P. taxon caryae. An additional novel species, P. taxon pocumtuck, is a close relative of P. borealis based on cox1 sequences. The results illustrate a high level of Phytophthora species richness in the Connecticut River Valley and that major rivers can serve as a source of inoculum for pathogenic Phytophthora species in the northeast.

  9. Phytophthora species recovered from the Connecticut River Valley in Massachusetts, USA.

    PubMed

    Brazee, Nicholas J; Wick, Robert L; Hulvey, Jonathan P

    2016-01-01

    Little is currently known about the assemblage of Phytophthora species in northeastern North America, representing a gap in our understanding of species incidence. Therefore, Phytophthora species were surveyed at 20 sites in Massachusetts, with 16 occurring in the Connecticut River Valley. Many of the sampled waterways were adjacent to active agricultural lands, yet were buffered by mature floodplain forests composed of Acer, Platanus, Populus and Ulmus. Isolates were recovered with three types of baits (rhododendron leaves, pear, green pepper) in 2013 and water filtration in 2014. Overall, 457 isolates of Phytophthora were recovered and based on morphological characters and rDNA internal transcribed spacer (ITS), β-tubulin (β-tub) and cytochrome oxidase c subunit I (cox1) sequences, 18 taxa were identified, including three new species: P. taxon intercalaris, P. taxon caryae and P. taxon pocumtuck. In addition, 49 isolates representing five species of Phytopythium also were identified. Water filtration captured a greater number of taxa (18) compared to leaf and fruit baits (12). Of the three bait types rhododendron leaves yielded the greatest number of isolates and taxa, followed by pear and green pepper, respectively. Despite the proximity to agricultural lands, none of the Phytophthora species baited are considered serious pathogens of vegetable crops in the region. However, many of the recovered species are known woody plant pathogens, including four species in the P. citricola s.l. complex that were identified: P. plurivora, P. citricola III, P. pini and a putative novel species, referred to here as P. taxon caryae. An additional novel species, P. taxon pocumtuck, is a close relative of P. borealis based on cox1 sequences. The results illustrate a high level of Phytophthora species richness in the Connecticut River Valley and that major rivers can serve as a source of inoculum for pathogenic Phytophthora species in the northeast. PMID:26553775

  10. Phytophthora austrocedri Elicitates Changes in Diterpene Profile of Austrocedrus chilensis.

    PubMed

    Olate, Verónica Rachel; Vélez, María Laura; Greslebin, Alina; Schmeda-Hirschmann, Guillermo

    2015-08-18

    The populations of the Andean Cupressaceae Austrocedrus chilensis have been severely affected by a disease caused by the phytopathogenic fungus Phytophthora austrocedri. A study was undertaken to disclose changes in the resin composition of P. austrocedri-infected individuals, including naturally infected and artificially inoculated trees, compared with healthy A. chilensis trees. GC-MS and (1)H-NMR studies showed a clear differentiation among healthy and infected resins, with the diterpene isopimara-8(9),15-dien-19-ol as a relevant constituent in resins from infected trees. The effect of resin fractions from P. austrocedri infected trees on the pathogen was assessed by measuring the mycelial growth in agar plates. The most active fractions from resin obtained from infected trees inhibited fungal growth by nearly 50% at 1 mg/dish (35.37 µg/cm(2)). The main constituent in the active fractions were 18-hydroxymanool and the aldehyde torulosal. Both compounds are oxidation products of manool and can be a chemical response of the tree to the pathogen or be formed from the pathogen as a biotransformation product of manool by microbial oxidation. While the diterpene profiles from A. chilensis tree resins can easily differentiate healthy and P. austrocedri infected individuals, the possible conversion of manool to the antifungal derivatives 4 and 6 by the microorganism remains to be established.

  11. Detection, Diversity, and Population Dynamics of Waterborne Phytophthora ramorum Populations.

    PubMed

    Eyre, C A; Garbelotto, M

    2015-01-01

    Sudden oak death, the tree disease caused by Phytophthora ramorum, has significant environmental and economic impacts on natural forests on the U.S. west coast, plantations in the United Kingdom, and in the worldwide nursery trade. Stream baiting is vital for monitoring and early detection of the pathogen in high-risk areas and is performed routinely; however, little is known about the nature of water-borne P. ramorum populations. Two drainages in an infested California forest were monitored intensively using stream-baiting for 2 years between 2009 and 2011. Pathogen presence was determined both by isolation and polymerase chain reaction (PCR) from symptomatic bait leaves. Isolates were analyzed using simple sequence repeats to study population dynamics and genetic structure through time. Isolation was successful primarily only during spring conditions, while PCR extended the period of pathogen detection to most of the year. Water populations were extremely diverse, and changed between seasons and years. A few abundant genotypes dominated the water during conditions considered optimal for aerial populations, and matched those dominant in aerial populations. Temporal patterns of genotypic diversification and evenness were identical among aerial, soil, and water populations, indicating that all three substrates are part of the same epidemiological cycle, strongly influenced by rainfall and sporulation on leaves. However, there was structuring between substrates, likely arising due to reduced selection pressure in the water. Additionally, water populations showed wholesale mixing of genotypes without the evident spatial autocorrelation present in leaf and soil populations.

  12. Strain Specific Factors Control Effector Gene Silencing in Phytophthora sojae.

    PubMed

    Shrestha, Sirjana Devi; Chapman, Patrick; Zhang, Yun; Gijzen, Mark

    2016-01-01

    The Phytophthora sojae avirulence gene Avr3a encodes an effector that is capable of triggering immunity on soybean plants carrying the resistance gene Rps3a. P. sojae strains that express Avr3a are avirulent to Rps3a plants, while strains that do not are virulent. To study the inheritance of Avr3a expression and virulence towards Rps3a, genetic crosses and self-fertilizations were performed. A cross between P. sojae strains ACR10 X P7076 causes transgenerational gene silencing of Avr3a allele, and this effect is meiotically stable up to the F5 generation. However, test-crosses of F1 progeny (ACR10 X P7076) with strain P6497 result in the release of silencing of Avr3a. Expression of Avr3a in the progeny is variable and correlates with the phenotypic penetrance of the avirulence trait. The F1 progeny from a direct cross of P6497 X ACR10 segregate for inheritance for Avr3a expression, a result that could not be explained by parental imprinting or heterozygosity. Analysis of small RNA arising from the Avr3a gene sequence in the parental strains and hybrid progeny suggests that the presence of small RNA is necessary but not sufficient for gene silencing. Overall, we conclude that inheritance of the Avr3a gene silenced phenotype relies on factors that are variable among P. sojae strains.

  13. Phytophthora austrocedri Elicitates Changes in Diterpene Profile of Austrocedrus chilensis.

    PubMed

    Olate, Verónica Rachel; Vélez, María Laura; Greslebin, Alina; Schmeda-Hirschmann, Guillermo

    2015-01-01

    The populations of the Andean Cupressaceae Austrocedrus chilensis have been severely affected by a disease caused by the phytopathogenic fungus Phytophthora austrocedri. A study was undertaken to disclose changes in the resin composition of P. austrocedri-infected individuals, including naturally infected and artificially inoculated trees, compared with healthy A. chilensis trees. GC-MS and (1)H-NMR studies showed a clear differentiation among healthy and infected resins, with the diterpene isopimara-8(9),15-dien-19-ol as a relevant constituent in resins from infected trees. The effect of resin fractions from P. austrocedri infected trees on the pathogen was assessed by measuring the mycelial growth in agar plates. The most active fractions from resin obtained from infected trees inhibited fungal growth by nearly 50% at 1 mg/dish (35.37 µg/cm(2)). The main constituent in the active fractions were 18-hydroxymanool and the aldehyde torulosal. Both compounds are oxidation products of manool and can be a chemical response of the tree to the pathogen or be formed from the pathogen as a biotransformation product of manool by microbial oxidation. While the diterpene profiles from A. chilensis tree resins can easily differentiate healthy and P. austrocedri infected individuals, the possible conversion of manool to the antifungal derivatives 4 and 6 by the microorganism remains to be established. PMID:26295220

  14. Identification of a mastigoneme protein from Phytophthora nicotianae.

    PubMed

    Blackman, Leila M; Arikawa, Mikihiko; Yamada, Shuhei; Suzaki, Toshinobu; Hardham, Adrienne R

    2011-01-01

    Tripartite tubular hairs (mastigonemes) on the anterior flagellum of protists in the stramenopile taxon are responsible for reversing the thrust of flagellar beat and for cell motility. Immunoprecipitation experiments using antibodies directed towards mastigonemes on the flagella of zoospores ofPhytophthora nicotianaehave facilitated the cloning of a gene encoding a mastigoneme shaft protein in this Oomycete. Expression of the gene, designatedPnMas2, is up-regulated during asexual sporulation, a period during which many zoospore components are synthesized. Analysis of the sequence of the PnMas2 protein has revealed that, like other stramenopile mastigoneme proteins, PnMas2 has an N-terminal secretion signal and contains four cysteine-rich epidermal growth factor (EGF)-like domains. Evidence from non-denaturing gels indicates that PnMas2 forms large oligomeric complexes, most likely through disulphide bridging. Bioinformatic analysis has revealed thatPhytophthoraspecies typically contain three or four putative mastigoneme proteins containing the four EGF-like domains. These proteins are similar in sequence to mastigoneme proteins in other stramenopile protists including the algaeOchromonas danica,Aureococcus anophagefferensandScytosiphon lomentariaand the diatomsThalassiosira pseudonana and T. weissflogii.

  15. Strain Specific Factors Control Effector Gene Silencing in Phytophthora sojae.

    PubMed

    Shrestha, Sirjana Devi; Chapman, Patrick; Zhang, Yun; Gijzen, Mark

    2016-01-01

    The Phytophthora sojae avirulence gene Avr3a encodes an effector that is capable of triggering immunity on soybean plants carrying the resistance gene Rps3a. P. sojae strains that express Avr3a are avirulent to Rps3a plants, while strains that do not are virulent. To study the inheritance of Avr3a expression and virulence towards Rps3a, genetic crosses and self-fertilizations were performed. A cross between P. sojae strains ACR10 X P7076 causes transgenerational gene silencing of Avr3a allele, and this effect is meiotically stable up to the F5 generation. However, test-crosses of F1 progeny (ACR10 X P7076) with strain P6497 result in the release of silencing of Avr3a. Expression of Avr3a in the progeny is variable and correlates with the phenotypic penetrance of the avirulence trait. The F1 progeny from a direct cross of P6497 X ACR10 segregate for inheritance for Avr3a expression, a result that could not be explained by parental imprinting or heterozygosity. Analysis of small RNA arising from the Avr3a gene sequence in the parental strains and hybrid progeny suggests that the presence of small RNA is necessary but not sufficient for gene silencing. Overall, we conclude that inheritance of the Avr3a gene silenced phenotype relies on factors that are variable among P. sojae strains. PMID:26930612

  16. The role of water vapour in the orientation behaviour of the blood-sucking bug Triatoma infestans (Hemiptera, Reduviidae).

    PubMed

    Barrozo, R B; Manrique, G; Lazzari, C R

    2003-04-01

    The behavioural response to water vapour of the haematophagous bug Triatoma infestans was analysed. Dry or humid discrete sources at different temperatures were used as stimuli for insects walking on a locomotion compensator. Humidity significantly increased the tendency of these bugs to orientate towards thermal sources. Furthermore, humid sources at room temperature were attractive to T. infestans, but this effect was limited to short-range distances. On the other hand, dynamic sources, i.e. airstreams carrying different water vapour contents did not affect the spontaneous anemotactic behaviour of this species, neither in sign (positive) nor in intensity. The anemotactic behaviour was also not influenced by the physiological water balance state of the bugs. Results are discussed in relation to the cues released by living hosts of triatomine bugs and in relation to their responses to air-currents.

  17. Susceptibility and resistance to deltamethrin of wild and domestic populations of Triatoma infestans (Reduviidae: Triatominae) in Bolivia: new discoveries.

    PubMed

    Depickère, Stéphanie; Buitrago, Rosio; Siñani, Edda; Baune, Marianne; Monje, Marcelo; Lopez, Ronald; Waleckx, Etienne; Chavez, Tamara; Brenière, Simone Frédérique

    2012-12-01

    Bolivia is a high-endemic country for Chagas disease, for which the principal vector is Triatoma infestans (Triatominae). This is a mainly domestic species that is also found in the wild environment. Recently, an increasing number of studies have shown the importance of Triatominae resistance to insecticides, especially in Bolivia. Data regarding the susceptibility/resistance of wild and domestic populations of T. infestans to deltamethrin are presented. For the first time, domestic populations of the department of Santa Cruz were tested, showing low resistance. Although most of the wild populations were found to be susceptible to deltamethrin, three populations from three departments showed a mortality rate of less than 100%. This result is emphasised here.

  18. Biological characteristics and mating type distribution of Phytophthora capsici from China.

    PubMed

    Du, Y; Gong, Z-H; Liu, G-Z; Chai, G-X; Li, C

    2014-01-01

    Phytophthora capsici from seven provinces of China were investigated for their mating type, hyphal growth, zoospore production, and virulence. All of the morphological characteristics and the results of polymerase chain reaction confirmed that these isolates were indeed Phytophthora capsici. The test of mating type showed that the mating types of 19 representative isolates from China varied. The hyphal growth and the amount of zoospores produced from these isolates differed and there was no evident relationship between them, which indicated the existence of genetic diversity among the isolates in China. Also, the isolates that were more virulent on the pepper cultivars that we checked produced more zoospores than other isolates. PMID:24535866

  19. Selective Insecticide Applications Directed Against Triatoma infestans (Hemiptera: Reduviidae) Affected a Nontarget Secondary Vector of Chagas Disease, Triatoma garciabesi.

    PubMed

    Rodríguez-Planes, L I; Vazquez-Prokopec, G M; Cecere, M C; Canale, D M; Gürtler, R E

    2016-01-01

    The control of nondomiciliated triatomine species adapted to peridomestic habitats represents a challenge because they are connected to sylvatic colonies, and pyrethroid insecticides have limited effects outdoors. The effects of residual insecticide spraying have rarely been assessed on secondary triatomines. Triatoma garciabesi (Carcavallo, Martinez, Cichero, Prosen & Ronderos, 1967) is a nontarget vector that inhabits the dry western Chaco region, and a member of the Triatoma sordida Stål 1859 complex. Little is known on the capacity of T. garciabesi to invade and establish viable domestic or peridomestic colonies, and on its response to residual insecticide sprays directed against Triatoma infestans Klug 1834. The presence and abundance of triatomines were assessed by timed manual collections annually or biannually (spring and fall) during 10 yr after a community-wide insecticide spraying campaign and selective insecticide sprays directed against T. infestans in a rural village of northwestern Argentina. T. garciabesi mainly occupied peridomestic habitats associated with chickens, and was unable to colonize human sleeping quarters. Trees with chickens occurred in nearly all houses and were infested in >25% of the occasions. The abundance of bugs at house-compound level was best explained by a generalized estimating equation model that included selective insecticide sprays during the previous semester (negative effects), chicken abundance (positive effects), seasonality, and their interactions. Our results suggest that insecticide applications targeting T. infestans affected the abundance of T. garciabesi, and reduced the likelihood of future infestation. PMID:26490000

  20. Selective Insecticide Applications Directed Against Triatoma infestans (Hemiptera: Reduviidae) Affected a Nontarget Secondary Vector of Chagas Disease, Triatoma garciabesi.

    PubMed

    Rodríguez-Planes, L I; Vazquez-Prokopec, G M; Cecere, M C; Canale, D M; Gürtler, R E

    2016-01-01

    The control of nondomiciliated triatomine species adapted to peridomestic habitats represents a challenge because they are connected to sylvatic colonies, and pyrethroid insecticides have limited effects outdoors. The effects of residual insecticide spraying have rarely been assessed on secondary triatomines. Triatoma garciabesi (Carcavallo, Martinez, Cichero, Prosen & Ronderos, 1967) is a nontarget vector that inhabits the dry western Chaco region, and a member of the Triatoma sordida Stål 1859 complex. Little is known on the capacity of T. garciabesi to invade and establish viable domestic or peridomestic colonies, and on its response to residual insecticide sprays directed against Triatoma infestans Klug 1834. The presence and abundance of triatomines were assessed by timed manual collections annually or biannually (spring and fall) during 10 yr after a community-wide insecticide spraying campaign and selective insecticide sprays directed against T. infestans in a rural village of northwestern Argentina. T. garciabesi mainly occupied peridomestic habitats associated with chickens, and was unable to colonize human sleeping quarters. Trees with chickens occurred in nearly all houses and were infested in >25% of the occasions. The abundance of bugs at house-compound level was best explained by a generalized estimating equation model that included selective insecticide sprays during the previous semester (negative effects), chicken abundance (positive effects), seasonality, and their interactions. Our results suggest that insecticide applications targeting T. infestans affected the abundance of T. garciabesi, and reduced the likelihood of future infestation.

  1. The peri-urban interface and house infestation with Triatoma infestans in the Argentine Chaco: an underreported process?

    PubMed Central

    Provecho, Yael M; Gaspe, M Sol; del Pilar Fernández, M; Enriquez, Gustavo F; Weinberg, Diego; Gürtler, Ricardo E

    2014-01-01

    Peri-urban infestations with triatomine bugs, their sources and their dynamics have rarely been investigated. Here, we corroborated the reported occurrence of Triatoma infestans in a peri-urban area and in neighbouring rural houses in Pampa del Indio, in the Argentine Chaco, and identified its putative sources using spatial analysis and demographic questionnaires. Peri-urban householders reported that 10% of their premises had triatomines, whereas T. infestans was collected by timed manual searches or community-based surveillance in only nine (3%) houses. Trypanosoma cruzi-infected T. infestans and Triatoma sordida were collected indoors only in peri-urban houses and were infected with TcV and TcI, respectively. The triatomines fed on chickens, cats and humans. Peri-urban infestations were most frequent in a squatter settlement and particularly within the recently built mud houses of rural immigrants, with large-sized households, more dogs and cats and more crowding. Several of the observed infestations were most likely associated with passive bug transport from other sources and with active bug dispersal from neighbouring foci. Thus, the households in the squatter settlement were at a greater risk of bug invasion and colonisation. In sum, the incipient process of domestic colonisation and transmission, along with persistent rural-to-urban migratory flows and unplanned urbanisation, indicate the need for active vector surveillance and control actions at the peri-urban interface of the Gran Chaco. PMID:25410997

  2. The saliva proteome of the blood-feeding insect Triatoma infestans is rich in platelet-aggregation inhibitors

    NASA Astrophysics Data System (ADS)

    Charneau, Sébastien; Junqueira, Magno; Costa, Camila M.; Pires, Daniele L.; Fernandes, Ellen S.; Bussacos, Ana C.; Sousa, Marcelo V.; Ricart, Carlos André O.; Shevchenko, Andrej; Teixeira, Antonio R. L.

    2007-12-01

    The saliva of the bloodsucking bug Triatoma infestans vector of Chagas disease contains an anti-hemostatic molecular cocktail that prevents coagulation, vasoconstriction and platelet aggregation in a vertebrate prey. In order to characterize T. infestans saliva proteome, we separated the secreted saliva by two-dimensional gel electrophoresis (2-DE). More than 200 salivary proteins were detected on the 2-DE map, mainly in the alkaline region. By nanoLC-MS/MS analysis using a LTQ-Orbitrap equipment followed by a combination of conventional and sequence-similarity searches, we identified 58 main protein spots. Most of such proteins possess potential blood-feeding associated functions, particularly anti-platelet aggregation proteins belonging to lipocalin and apyrase families. The saliva protein composition indicates a highly specific molecular mechanism of early response to platelet aggregation. This first proteome analysis of the T. infestans secreted saliva provides a basis for a better understanding of this fluid protein composition highly directed to counterpart hemostasis of the prey, thus promoting the bug's blood-feeding.

  3. Antagonistic interaction between Trichoderma asperellum and Phytophthora capsici in vitro*

    PubMed Central

    Jiang, Heng; Zhang, Liang; Zhang, Jing-ze; Ojaghian, Mohammad Reza; Hyde, Kevin D.

    2016-01-01

    Phytophthora capsici is a phytopathogen that causes a destructive pepper blight that is extremely difficult to control. Using a fungicide application against the disease is costly and relatively ineffective and there is also a huge environmental concern about the use of such chemicals. The genus Trichoderma has been known to have a potential biocontrol issue. In this paper we investigate the mechanism for causing the infection of T. asperellum against P. capsici. Trichoderma sp. (isolate CGMCC 6422) was developed to have a strong antagonistic action against hyphae of P. capsici through screening tests. The strain was identified as T. asperellum through using a combination of morphological characteristics and molecular data. T. asperellum was able to collapse the mycelium of the colonies of the pathogen through dual culture tests by breaking down the pathogenic hyphae into fragments. The scanning electron microscope showed that the hyphae of T. asperellum surrounded and penetrated the pathogens hyphae, resulting in hyphal collapse. The results show that seven days after inoculation, the hyphae of the pathogen were completely degraded in a dual culture. T. asperellum was also able to enter the P. capsici oospores through using oogonia and then developed hyphae and produced conidia, leading to the disintegration of the oogonia and oospores. Seven days after inoculation, an average 10.8% of the oospores were infected, but at this stage, the structures of oospores were still intact. Subsequently, the number of infected oospores increased and the oospores started to collapse. Forty-two days after inoculation, almost all the oospores were infected, with 9.3% of the structures of the oospores being intact and 90.7% of the oospores having collapsed.

  4. Genetic characterization of Phytophthora nicotianae by the analysis of polymorphic regions of the mitochondrial DNA.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A new method based on the analysis of mitochondrial intergenic regions characterized by intraspecific variation in DNA sequences was developed and applied to the study of the plant pathogen Phytophthora nicotianae. Two regions flanked by genes trny and rns and trnw and cox2 were identified by compa...

  5. Diverse evolutionary trajectories for small RNA biogenesis genes in the oomycete genus Phytophthora

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Gene regulation by small RNA pathways is ubiquitous among eukaryotes, but little is known about small RNA pathways in the Stramenopile kingdom. Phytophthora, a genus of filamentous oomycetes, contains many devastating plant pathogens, causing multibillion-dollar damage to crops, ornamental plants, ...

  6. Evaluation of watermelon varieties for tolerance to powdery mildew and Phytophthora fruit rot, 2014

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This experiment was conducted at the U.S. Vegetable Laboratory farm in Charleston, SC. The soil was Yonges loamy fine sand. This study was undertaken to determine the performance of seeded and seedless commercial watermelon varieties for tolerance to powdery mildew (PM) and Phytophthora fruit rot as...

  7. Phytophthora effector targets a novel component of small RNA pathway in plants to promote infection.

    PubMed

    Qiao, Yongli; Shi, Jinxia; Zhai, Yi; Hou, Yingnan; Ma, Wenbo

    2015-05-01

    A broad range of parasites rely on the functions of effector proteins to subvert host immune response and facilitate disease development. The notorious Phytophthora pathogens evolved effectors with RNA silencing suppression activity to promote infection in plant hosts. Here we report that the Phytophthora Suppressor of RNA Silencing 1 (PSR1) can bind to an evolutionarily conserved nuclear protein containing the aspartate-glutamate-alanine-histidine-box RNA helicase domain in plants. This protein, designated PSR1-Interacting Protein 1 (PINP1), regulates the accumulation of both microRNAs and endogenous small interfering RNAs in Arabidopsis. A null mutation of PINP1 causes embryonic lethality, and silencing of PINP1 leads to developmental defects and hypersusceptibility to Phytophthora infection. These phenotypes are reminiscent of transgenic plants expressing PSR1, supporting PINP1 as a direct virulence target of PSR1. We further demonstrate that the localization of the Dicer-like 1 protein complex is impaired in the nucleus of PINP1-silenced or PSR1-expressing cells, indicating that PINP1 may facilitate small RNA processing by affecting the assembly of dicing complexes. A similar function of PINP1 homologous genes in development and immunity was also observed in Nicotiana benthamiana. These findings highlight PINP1 as a previously unidentified component of RNA silencing that regulates distinct classes of small RNAs in plants. Importantly, Phytophthora has evolved effectors to target PINP1 in order to promote infection.

  8. The Phytophthora species assemblage and diversity in riparian alder ecosystems of western Oregon, USA.

    PubMed

    Sims, Laura Lee; Sutton, Wendy; Reeser, Paul; Hansen, Everett M

    2015-01-01

    Phytophthora species were systematically sampled, isolated, identified and compared for presence in streams, soil and roots of alder (Alnus species) dominated riparian ecosystems in western Oregon. We describe the species assemblage and evaluate Phytophthora diversity associated with alder. We recovered 1250 isolates of 20 Phytophthora species. Only three species were recovered from all substrates (streams, soil, alder roots): P. gonapodyides, the informally described "P. taxon Pgchlamydo", and P. siskiyouensis. P. alni ssp. uniformis along with five other species not previously recovered in Oregon forests are included in the assemblage: P.citricola s.l., P. gregata, P. gallica, P. nicotianae and P. parsiana. Phytophthora species diversity was greatest in downstream riparian locations. There was no significant difference in species diversity comparing soil and unwashed roots (the rhizosphere) to stream water. There was a difference between the predominating species from the rhizosphere compared to stream water. The most numerous species was the informally described "P. taxon Oaksoil", which was mainly recovered from, and most predominant in, stream water. The most common species from riparian forest soils and alder root systems was P. gonapodyides.

  9. Evaluation of fungicide rotations for management of Phytophthora fruit rot of watermelon, 2015

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The experiment was conducted at the U.S. Vegetable Laboratory farm in Charleston, SC. The soil was Yonges loamy fine sand. For the past 6 years, the field has been infested with Phytophthora capsici. The experimental design was a randomized complete block with four replications. Five-week-old seedli...

  10. Roles of small RNAs in soybean defense against Phytophthora sojae infection.

    PubMed

    Wong, James; Gao, Lei; Yang, Yang; Zhai, Jixian; Arikit, Siwaret; Yu, Yu; Duan, Shuyi; Chan, Vicky; Xiong, Qin; Yan, Jun; Li, Shengben; Liu, Renyi; Wang, Yuanchao; Tang, Guiliang; Meyers, Blake C; Chen, Xuemei; Ma, Wenbo

    2014-09-01

    The genus Phytophthora consists of many notorious pathogens of crops and forestry trees. At present, battling Phytophthora diseases is challenging due to a lack of understanding of their pathogenesis. We investigated the role of small RNAs in regulating soybean defense in response to infection by Phytophthora sojae, the second most destructive pathogen of soybean. Small RNAs, including microRNAs (miRNAs) and small interfering RNAs (siRNAs), are universal regulators that repress target gene expression in eukaryotes. We identified known and novel small RNAs that differentially accumulated during P. sojae infection in soybean roots. Among them, miR393 and miR166 were induced by heat-inactivated P. sojae hyphae, indicating that they may be involved in soybean basal defense. Indeed, knocking down the level of mature miR393 led to enhanced susceptibility of soybean to P. sojae; furthermore, the expression of isoflavonoid biosynthetic genes was drastically reduced in miR393 knockdown roots. These data suggest that miR393 promotes soybean defense against P. sojae. In addition to miRNAs, P. sojae infection also resulted in increased accumulation of phased siRNAs (phasiRNAs) that are predominantly generated from canonical resistance genes encoding nucleotide binding-leucine rich repeat proteins and genes encoding pentatricopeptide repeat-containing proteins. This work identifies specific miRNAs and phasiRNAs that regulate defense-associated genes in soybean during Phytophthora infection.

  11. Differences in virulence and sporulation of Phytophthora kernoviae isolates originating from two distinct geographical regions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora kernoviae has only been isolated from the United Kingdom (U.K.) and New Zealand. To understand what differences may exist between isolates from these two distinct geographical regions, virulence studies on three host plants and sporulation on host leaves were conducted on select isolat...

  12. Survival of Phytophthora ramorum hyphae following exposure to temperature extremes and various humidities

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We examined the impact of short-term exposure to high and low temperatures and a range of relative humidities on survival of Phytophthora ramorum hyphae. Spore-free hyphal colonies were grown on dialysis squares atop V8 medium. Relative humidity ranged from 41 – 93% at 20 C and 43 – 86% at 28 C. ...

  13. Sensitivity of isolates of phytophthora capsici from the eastern United States to fluopicolide

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fluopicolide, a pyridinylmethyl-benzamide fungicide, was registered in the United States in 2008 to control diseases caused by Oomycete pathogens, such as Phytophthora capsici, on cucurbit and solanaceous vegetables. The main objective of this study was to determine baseline sensitivity to fluopico...

  14. Baseline Sensitivity to Fluopicolide in Phytophthora Capsici Isolates from the Eastern United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fluopicolide was registered in 2007 to control diseases caused by Oomycete pathogens such as Phytophthora capsici on cucurbits and peppers. In this study, 69 isolates of P. capsici from Michigan (24 isolates), South Carolina (17), Georgia (14), Florida (11), and North Carolina (3) recovered from wa...

  15. Biological Control of Phytophthora palmivora Causing Root Rot of Pomelo Using Chaetomium spp.

    PubMed Central

    Wattanachai, Pongnak; Kasem, Soytong; Poaim, Supatta

    2015-01-01

    Phytophthora diseases have become a major impediment in the citrus production in Thailand. In this study, an isolate of Phytophthora denominated as PHY02 was proven to be causal pathogen of root rot of Pomelo (Citrus maxima) in Thailand. The isolate PHY02 was morphologically characterized and identified as Phytophthora palmivora based on molecular analysis of an internal transcribed spacer rDNA sequence. This work also presents in vitro evaluations of the capacities of Chaetomium spp. to control the P. palmivora PHY02. As antagonists, Chaetomium globosum CG05, Chaetomium cupreum CC3003, Chaetomium lucknowense CL01 inhibited 50~61% mycelial growth, degraded mycelia and reduced 92~99% sporangial production of P. palmivora PHY02 in bi-culture test after 30 days. Fungal metabolites from Chaetomium spp. were tested against PHY02. Results showed that, methanol extract of C. globosum CG05 expressed strongest inhibitory effects on mycelial growth and sporangium formation of P. palmivora PHY02 with effective dose ED50 values of 26.5 µg/mL and 2.3 µg/mL, respectively. It is interesting that C. lucknowense is reported for the first time as an effective antagonist against a species of Phytophthora. PMID:25892917

  16. Evaluation of Cultural Practices and Fungicides for Managing Phytophthora Fruit Rot of Watermelon, 2008

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The experiment was conducted to evaluate the efficacy of new fungicides to manage Phytophthora fruit rot of watermelon. The soil at the experimental field in Charleston, SC, was Yonges loamy fine sand. The field was sprayed with Roundup Pro (1 pt/A) and Strategy (2 pt/A) after bedding but prior to...

  17. Effect of phytophthora capsici crown rot on watermelon rootstocks and grafts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Crown and fruit rot caused by Phytophthora capsici is becoming an important and emerging disease of watermelon in the southeastern United States. In recent years, the practice of grafting seedless watermelons onto rootstocks belonging to other Cucurbitaceae genera is also gaining acceptance in our l...

  18. Virulence, sporulation, and elicitin production in three clonal lineages of Phytophthora ramorum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora ramorum populations are clonal and consist of three lineages. Recent studies have shown that the clonal lineages may have varying degrees of aggressiveness on some host species, such as Quercus rubra. In this study, we examined virulence, sporulation and elicitin production of five P. ...

  19. Interactions of Phytophthora capsici with Resistant and Susceptible Pepper Roots and Stems.

    PubMed

    Dunn, Amara R; Smart, Christine D

    2015-10-01

    Using host resistance is an important strategy for managing pepper root and crown rot caused by Phytophthora capsici. An isolate of P. capsici constitutively expressing a gene for green fluorescent protein was used to investigate pathogen interactions with roots, crowns, and stems of Phytophthora-susceptible bell pepper 'Red Knight', Phytophthora-resistant bell pepper 'Paladin', and Phytophthora-resistant landrace Criollos de Morelos 334 (CM-334). In this study, the same number of zoospores attached to and germinated on roots of all cultivars 30 and 120 min postinoculation (pi), respectively. At 3 days pi, significantly more secondary roots had necrotic lesions on Red Knight than on Paladin and CM-334 plants. By 4 days pi, necrotic lesions had formed on the taproot of Red Knight but not Paladin or CM-334 plants. Although hyphae were visible in the crowns and stems of all Red Knight plants observed at 4 days pi, hyphae were observed in crowns of only a few Paladin and in no CM-334 plants, and never in stems of either resistant cultivar at 4 days pi. These results improve our understanding of how P. capsici infects plants and may contribute to the use of resistant pepper cultivars for disease management and the development of new cultivars.

  20. Marine Phytophthora species can hamper conservation and restoration of vegetated coastal ecosystems.

    PubMed

    Govers, Laura L; Man In 't Veld, Willem A; Meffert, Johan P; Bouma, Tjeerd J; van Rijswick, Patricia C J; Heusinkveld, Jannes H T; Orth, Robert J; van Katwijk, Marieke M; van der Heide, Tjisse

    2016-08-31

    Phytophthora species are potent pathogens that can devastate terrestrial plants, causing billions of dollars of damage yearly to agricultural crops and harming fragile ecosystems worldwide. Yet, virtually nothing is known about the distribution and pathogenicity of their marine relatives. This is surprising, as marine plants form vital habitats in coastal zones worldwide (i.e. mangrove forests, salt marshes, seagrass beds), and disease may be an important bottleneck for the conservation and restoration of these rapidly declining ecosystems. We are the first to report on widespread infection of Phytophthora and Halophytophthora species on a common seagrass species, Zostera marina (eelgrass), across the northern Atlantic and Mediterranean. In addition, we tested the effects of Halophytophthora sp. Zostera and Phytophthora gemini on Z. marina seed germination in a full-factorial laboratory experiment under various environmental conditions. Results suggest that Phytophthora species are widespread as we found these oomycetes in eelgrass beds in six countries across the North Atlantic and Mediterranean. Infection by Halophytophthora sp. Zostera, P. gemini, or both, strongly affected sexual reproduction by reducing seed germination sixfold. Our findings have important implications for seagrass ecology, because these putative pathogens probably negatively affect ecosystem functioning, as well as current restoration and conservation efforts. PMID:27559058

  1. Lineage, Temperature, and Host Species Have Interacting Effects on Lesion Development in Phytophthora Ramorum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    There are four recognized clonal lineages of the pathogen Phytophthora ramorum. The two major lineages present in North America are NA1 and NA2. With a few exceptions, NA1 is found in natural forest ecosystems and nurseries, and NA2 is generally restricted to nurseries. Isolates from the NA1 and NA2...

  2. Marine Phytophthora species can hamper conservation and restoration of vegetated coastal ecosystems.

    PubMed

    Govers, Laura L; Man In 't Veld, Willem A; Meffert, Johan P; Bouma, Tjeerd J; van Rijswick, Patricia C J; Heusinkveld, Jannes H T; Orth, Robert J; van Katwijk, Marieke M; van der Heide, Tjisse

    2016-08-31

    Phytophthora species are potent pathogens that can devastate terrestrial plants, causing billions of dollars of damage yearly to agricultural crops and harming fragile ecosystems worldwide. Yet, virtually nothing is known about the distribution and pathogenicity of their marine relatives. This is surprising, as marine plants form vital habitats in coastal zones worldwide (i.e. mangrove forests, salt marshes, seagrass beds), and disease may be an important bottleneck for the conservation and restoration of these rapidly declining ecosystems. We are the first to report on widespread infection of Phytophthora and Halophytophthora species on a common seagrass species, Zostera marina (eelgrass), across the northern Atlantic and Mediterranean. In addition, we tested the effects of Halophytophthora sp. Zostera and Phytophthora gemini on Z. marina seed germination in a full-factorial laboratory experiment under various environmental conditions. Results suggest that Phytophthora species are widespread as we found these oomycetes in eelgrass beds in six countries across the North Atlantic and Mediterranean. Infection by Halophytophthora sp. Zostera, P. gemini, or both, strongly affected sexual reproduction by reducing seed germination sixfold. Our findings have important implications for seagrass ecology, because these putative pathogens probably negatively affect ecosystem functioning, as well as current restoration and conservation efforts.

  3. Draft genome sequences of seven isolates of Phytophthora ramorum EU2 from Northern Ireland.

    PubMed

    Mata Saez, Lourdes de la; McCracken, Alistair R; Cooke, Louise R; O'Neill, Paul; Grant, Murray; Studholme, David J

    2015-12-01

    Here we present draft-quality genome sequence assemblies for the oomycete Phytophthora ramorum genetic lineage EU2. We sequenced genomes of seven isolates collected in Northern Ireland between 2010 and 2012. Multiple genome sequences from P. ramorum EU2 will be valuable for identifying genetic variation within the clonal lineage that can be useful for tracking its spread. PMID:26697370

  4. Bioassay conditions for infection of Pinus radiata seedlings with Phytophthora pinifolia zoospores

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora pinifolia is known to cause a devastating disease on Monterey pines in Chile. Although this pathogen is not yet present in the U.S., there is reason for concern. The main source of Monterey pine genetic material is found in California and there is potential for other important tree sp...

  5. Evolution of an experimental population of Phytophthora capsici in the field

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Populations of the vegetable pathogen Phytophthora capsici are often highly diverse, with limited gene flow between fields. To investigate the structure of a newly established, experimental population, an uninfested research field was inoculated with two single zoospore isolates of P. capsici in Sep...

  6. Whole plant inoculations of Viburnum species and cultivars testing for susceptibility to Phytophthora ramorum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora ramorum is known to cause Ramorum blight on Viburnum species, which are commonly grown as ornamentals. This study evaluated 25 different species or cultivars for their susceptibility to P. ramorum. Whole plants were inoculated with a zoospore suspension of an NA1 isolate of P. ramorum...

  7. Root susceptibility and inoculum production from roots of eastern oak species to Phytophthora ramorum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Little is known about root susceptibility of eastern tree species to Phytophthora ramorum. In this study, we examined root susceptibility and inoculum production from roots. Oak radicles of several eastern oak species were exposed to zoospore suspensions of 1, 10, 100, or 1000 zoospores per ml at ...

  8. Root susceptibility and inoculum production from roots of Eastern United States oak species to Phytophthora ramorum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Little is known about root susceptibility of eastern U.S. tree species to Phytophthora ramorum. In this study, we examined root susceptibility and inoculum production from roots. Sprouted acorns of Q. rubra, Q. palustrus, Q. coccinia, Q. alba, Q. michauxii and Q. prinus were exposed to motile zoos...

  9. DEVELOPMENT OF AN IMPROVED PCR-BASED TECHNIQUE FOR DETECTION OF PHYTOPHTHORA CACTORUM IN STRAWBERRY PLANTS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Specific and rapid plant pathogen detection methods can aid in strawberry disease management decisions. PCR-based diagnostics for Phytophthora cactorum and other strawberry pathogens are hindered by PCR inhibitors and lack of species-specific PCR primers. We developed a DNA extraction and purificati...

  10. Antifungal activity of extracts and select compounds in heartwood of seven western conifers toward Phytophthora ramorum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Individual compounds and ethyl acetate extracts from heartwood of seven conifer species were tested for fungicidal activity against Phytophthora ramorum. Extracts from incense and western red cedar exhibited the strongest activity (EC50 589 and 646 ppm, respectively), yellow-cedar, western juniper,...

  11. Identification of quantitative trait loci conditioning partial resistance to Phytophthora sojae in soybean PI 407861A

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Improving resistance for Phytophthora root and stem rot is an important goal in soybean [Glycine max (L.) Merr.] breeding. Partial resistance can be as effective in managing this disease as single-gene (Rps) mediated resistance and is more durable. The objective of this study was to identify QTL con...

  12. Genetic mapping and characterization of two novel Phytophthora resistance genes from soybean landrace PI567139B

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora root and stem rot (PRR) disease, caused by P. sojae, is a widespread soybean disease resulting in an annual yield loss of $1~2 billion worldwide. To control the disease, breeders primarily employ race-specific resistant genes which are named Rps genes which have been identified to be lo...

  13. Characterisation and detection of Pythium and Phytophthora species associated with grapevines in South Africa.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Replant and decline diseases of grapevines not only cause quantitative and qualitative yield losses, but also results in extra costs when vineyards have to be replanted. This study investigated the role of Pythium and Phytophthora in the decline syndrome in South Africa by determining the (i) speci...

  14. The effect of salinity on the growth, sporulation and infection of Phytophthora ramorum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora ramorum, a threat to Eastern U.S. forests, has been found in waterways outside the boundaries of infested ornamental nurseries. Very little is known about what factors are conducive to its survival and sporulation in water. This study examined the effect of salt on growth, sporulation,...

  15. Interactions of Phytophthora capsici with Resistant and Susceptible Pepper Roots and Stems.

    PubMed

    Dunn, Amara R; Smart, Christine D

    2015-10-01

    Using host resistance is an important strategy for managing pepper root and crown rot caused by Phytophthora capsici. An isolate of P. capsici constitutively expressing a gene for green fluorescent protein was used to investigate pathogen interactions with roots, crowns, and stems of Phytophthora-susceptible bell pepper 'Red Knight', Phytophthora-resistant bell pepper 'Paladin', and Phytophthora-resistant landrace Criollos de Morelos 334 (CM-334). In this study, the same number of zoospores attached to and germinated on roots of all cultivars 30 and 120 min postinoculation (pi), respectively. At 3 days pi, significantly more secondary roots had necrotic lesions on Red Knight than on Paladin and CM-334 plants. By 4 days pi, necrotic lesions had formed on the taproot of Red Knight but not Paladin or CM-334 plants. Although hyphae were visible in the crowns and stems of all Red Knight plants observed at 4 days pi, hyphae were observed in crowns of only a few Paladin and in no CM-334 plants, and never in stems of either resistant cultivar at 4 days pi. These results improve our understanding of how P. capsici infects plants and may contribute to the use of resistant pepper cultivars for disease management and the development of new cultivars. PMID:26010399

  16. SPATIO-TEMPORAL ANALYSIS OF REINFESTATION BY TRIATOMA INFESTANS (HEMIPTERA: REDUVIIDAE) FOLLOWING INSECTICIDE SPRAYING IN A RURAL COMMUNITY IN NORTHWESTERN ARGENTINA

    PubMed Central

    CECERE, MARÍA C.; VAZQUEZ-PROKOPEC, GONZALO M.; GÜRTLER, RICARDO E.; KITRON, URIEL

    2005-01-01

    The spatio-temporal reinfestation patterns by Triatoma infestans following a blanket insecticide spraying in the rural community of Amamá in northwestern Argentina were analyzed using a geographic information system, satellite imagery, and spatial statistics. Domestic and peridomestic reinfestation by triatomine bugs was monitored from 1993 to 1997. Triatoma infestans was detected at least once in 75% of 2,110 sites evaluated. The prevalence of sites positive at least once for T. infestans during the study period increased sharply from 1993–1995 (0.6–2.9%) to November 1997 (32%). The initial source of T. infestans was a pig corral in southern Amamá one year post-spraying. Subsequent infestations were clustered around this initial focus at a distance of approximately 400 meters starting in 1995. In 1996, clustering was maximized in sites within the same or in neighboring compounds at distances of 25–175 meters. An effective control program on the community level will be based on the spraying of actual epicenters and sites within 450 meters of these epicenters to prevent the propagation of T. infestans. PMID:15642975

  17. Development of Rapid Isothermal Amplification Assays for Detection of Phytophthora spp. in Plant Tissue.

    PubMed

    Miles, Timothy D; Martin, Frank N; Coffey, Michael D

    2015-02-01

    Several isothermal amplification techniques recently have been developed that are tolerant of inhibitors present in many plant extracts, which can reduce the need for obtaining purified DNA for running diagnostic assays. One such commercially available technique that has similarities with real-time polymerase chain reaction (PCR) for designing primers and a labeled probe is recombinase polymerase amplification (RPA). This technology was used to develop two simple and rapid approaches for detection of Phytophthora spp.: one genus-specific assay multiplexed with a plant internal control and the other species-specific assays for Phytophthora ramorum and P. kernoviae. All assays were tested for sensitivity (ranging from 3 ng to 1 fg of DNA) and specificity using DNA extracted from more than 136 Phytophthora taxa, 21 Pythium spp., 1 Phytopythium sp., and a wide range of plant species. The lower limit of linear detection using purified DNA was 200 to 300 fg of DNA in all pathogen RPA assays. Six different extraction buffers were tested for use during plant tissue maceration and the assays were validated in the field by collecting 222 symptomatic plant samples from over 50 different hosts. Only 56 samples were culture positive for Phytophthora spp. whereas 91 were positive using the Phytophthora genus-specific RPA test and a TaqMan real-time PCR assay. A technique for the generation of sequencing templates from positive RPA amplifications to confirm species identification was also developed. These RPA assays have added benefits over traditional technologies because they are rapid (results can be obtained in as little as 15 min), do not require DNA extraction or extensive training to complete, use less expensive portable equipment than PCR-based assays, and are significantly more specific than current immunologically based methods. This should provide a rapid, field-deployable capability for pathogen detection that will facilitate point-of-sample collection processing

  18. Phytophthora capsici - Loss of Heterozygosity (LOH): A Widespread Mechanism for Rapid Adaptation ( 7th Annual SFAF Meeting, 2012)

    ScienceCinema

    Mudge, Joanne [NCGR

    2016-07-12

    Joanne Mudge on "Phytophthora capsici - Loss of Heterozygosity (LOH): A Widespread Mechanism for Rapid Mutation" at the 2012 Sequencing, Finishing, Analysis in the Future Meeting held June 5-7, 2012 in Santa Fe, New Mexico.

  19. A Phytophthora sojae effector suppresses endoplasmic reticulum stress-mediated immunity by stabilizing plant Binding immunoglobulin Proteins

    PubMed Central

    Jing, Maofeng; Guo, Baodian; Li, Haiyang; Yang, Bo; Wang, Haonan; Kong, Guanghui; Zhao, Yao; Xu, Huawei; Wang, Yan; Ye, Wenwu; Dong, Suomeng; Qiao, Yongli; Tyler, Brett M.; Ma, Wenbo; Wang, Yuanchao

    2016-01-01

    Phytophthora pathogens secrete an array of specific effector proteins to manipulate host innate immunity to promote pathogen colonization. However, little is known about the host targets of effectors and the specific mechanisms by which effectors increase susceptibility. Here we report that the soybean pathogen Phytophthora sojae uses an essential effector PsAvh262 to stabilize endoplasmic reticulum (ER)-luminal binding immunoglobulin proteins (BiPs), which act as negative regulators of plant resistance to Phytophthora. By stabilizing BiPs, PsAvh262 suppresses ER stress-triggered cell death and facilitates Phytophthora infection. The direct targeting of ER stress regulators may represent a common mechanism of host manipulation by microbes. PMID:27256489

  20. Phytophthora capsici - Loss of Heterozygosity (LOH): A Widespread Mechanism for Rapid Adaptation ( 7th Annual SFAF Meeting, 2012)

    SciTech Connect

    Mudge, Joanne

    2012-06-01

    Joanne Mudge on "Phytophthora capsici - Loss of Heterozygosity (LOH): A Widespread Mechanism for Rapid Mutation" at the 2012 Sequencing, Finishing, Analysis in the Future Meeting held June 5-7, 2012 in Santa Fe, New Mexico.

  1. The fine structure of the ocelli of Triatoma infestans (Hemiptera: Reduviidae).

    PubMed

    Insausti, T C; Lazzari, C R

    2002-12-01

    The morphology and fine structure of the ocelli of Triatoma infestans have been analyzed by means of light and electron microscopy. The two dorsal ocelli of this species are located behind the compound eyes, looking dorsally and frontally. Externally, the ocelli are marked by the corneal lenses virtually spherical in form and limited internally by a cuticular apodeme. The lens focuses the incoming rays beyond the retina. A single layer of corneagen cells lies below the cuticular lens. The corneagen cells and photoreceptors are arranged in a cup-like fashion beneath the cuticular lens. A distal retinal zone comprises the rhabdoms, which are laterally connected in an hexagonal meshwork. A middle retinal zone comprises the receptor cell segment free of rhabdom, and a proximal zone their axons. In the middle zone, the oviform nuclei and spheroids are located. Screening pigment granules are present within the retinal cell. Spherical mitochondria are homogeneously distributed in the cytoplasm of the cell body. In the axonal zone, mitochondria are found in the peripheral region. Axons from receptor cells extend into the ocellar neuropile at the base of the ocelli, to synapse with second order neurons. The large axons of second order neurons are bundled by glial cells. The ocellar plexus exhibits a high diversity of synaptic unions (i.e. axo-dendritic, axo-axonic, dendro-axonic, and dendro-dendritic).

  2. Residual infestation and recolonization during urban Triatoma infestans Bug Control Campaign, Peru.

    PubMed

    Barbu, Corentin M; Buttenheim, Alison M; Pumahuanca, Maria-Luz Hancco; Calderón, Javier E Quintanilla; Salazar, Renzo; Carrión, Malwina; Rospigliossi, Andy Catacora; Chavez, Fernando S Malaga; Alvarez, Karina Oppe; Cornejo del Carpio, Juan; Náquira, César; Levy, Michael Z

    2014-12-01

    Chagas disease vector control campaigns are being conducted in Latin America, but little is known about medium-term or long-term effectiveness of these efforts, especially in urban areas. After analyzing entomologic data for 56,491 households during the treatment phase of a Triatoma infestans bug control campaign in Arequipa, Peru, during 2003-2011, we estimated that 97.1% of residual infestations are attributable to untreated households. Multivariate models for the surveillance phase of the campaign obtained during 2009-2012 confirm that nonparticipation in the initial treatment phase is a major risk factor (odds ratio [OR] 21.5, 95% CI 3.35-138). Infestation during surveillance also increased over time (OR 1.55, 95% CI 1.15-2.09 per year). In addition, we observed a negative interaction between nonparticipation and time (OR 0.73, 95% CI 0.53-0.99), suggesting that recolonization by vectors progressively dilutes risk associated with nonparticipation. Although the treatment phase was effective, recolonization in untreated households threatens the long-term success of vector control.

  3. Residual Infestation and Recolonization during Urban Triatoma infestans Bug Control Campaign, Peru1

    PubMed Central

    Buttenheim, Alison M.; Pumahuanca, Maria-Luz Hancco; Calderón, Javier E. Quintanilla; Salazar, Renzo; Carrión, Malwina; Rospigliossi, Andy Catacora; Chavez, Fernando S. Malaga; Alvarez, Karina Oppe; Cornejo del Carpio, Juan; Náquira, César; Levy, Michael Z.

    2014-01-01

    Chagas disease vector control campaigns are being conducted in Latin America, but little is known about medium-term or long-term effectiveness of these efforts, especially in urban areas. After analyzing entomologic data for 56,491 households during the treatment phase of a Triatoma infestans bug control campaign in Arequipa, Peru, during 2003–2011, we estimated that 97.1% of residual infestations are attributable to untreated households. Multivariate models for the surveillance phase of the campaign obtained during 2009–2012 confirm that nonparticipation in the initial treatment phase is a major risk factor (odds ratio [OR] 21.5, 95% CI 3.35–138). Infestation during surveillance also increased over time (OR 1.55, 95% CI 1.15–2.09 per year). In addition, we observed a negative interaction between nonparticipation and time (OR 0.73, 95% CI 0.53–0.99), suggesting that recolonization by vectors progressively dilutes risk associated with nonparticipation. Although the treatment phase was effective, recolonization in untreated households threatens the long-term success of vector control. PMID:25423045

  4. Phytophthora bisheria sp. nov., a new species identified in isolates from the Rosaceous raspberry, rose and strawberry in three continents.

    PubMed

    Abad, Z Gloria; Abad, Jorge A; Coffey, Michael D; Oudemans, Peter V; Man in 't Veld, Willem A; de Gruyter, Hans; Cunnington, James; Louws, Frank J

    2008-01-01

    A homothallic semipapillate slow growing Phytophthora species associated with root rot of strawberries from greenhouse-grown plants in North Carolina, USA, root rot of roses in the Netherlands, and root rot of raspberry in Knoxfield, Australia, was identified. The main character of this organism is the production of paragynous antheridia with broad attachment to the oogonial wall. The morphology of the pathogen does not match that of any of the more than 85 described Phytophthora species. Phylogenetic analysis based on sequences of the internal transcribed spacer rDNA region (ITS1-5.8S-ITS2) of this taxon and those from other Phytophthora species from GenBank supports the conclusion that this organism is an unreported new species. In the phylogenetic tree with other reported Phytophthora species at the GenBank, the new species is more closely related to others in ITS clade 2 comprising semipapillate taxa including P. botryosa, P. citrophthora, P. colocasiae, P. meadii, P. citricola, P. inflata, P.tropicalis, P. capsici, Phytophthora sp. 'glovera' and P. multivesiculata. The most closely related species is P. multivesiculata isolated from Cymbidium orchid in the Netherlands. In this paper we describe the morphological characteristics and the phylogenetic relationships that support the description of this taxon as a new species Phytophthora bisheria sp. nov.

  5. Sequencing of the Litchi Downy Blight Pathogen Reveals It Is a Phytophthora Species With Downy Mildew-Like Characteristics.

    PubMed

    Ye, Wenwu; Wang, Yang; Shen, Danyu; Li, Delong; Pu, Tianhuizi; Jiang, Zide; Zhang, Zhengguang; Zheng, Xiaobo; Tyler, Brett M; Wang, Yuanchao

    2016-07-01

    On the basis of its downy mildew-like morphology, the litchi downy blight pathogen was previously named Peronophythora litchii. Recently, however, it was proposed to transfer this pathogen to Phytophthora clade 4. To better characterize this unusual oomycete species and important fruit pathogen, we obtained the genome sequence of Phytophthora litchii and compared it to those from other oomycete species. P. litchii has a small genome with tightly spaced genes. On the basis of a multilocus phylogenetic analysis, the placement of P. litchii in the genus Phytophthora is strongly supported. Effector proteins predicted included 245 RxLR, 30 necrosis-and-ethylene-inducing protein-like, and 14 crinkler proteins. The typical motifs, phylogenies, and activities of these effectors were typical for a Phytophthora species. However, like the genome features of the analyzed downy mildews, P. litchii exhibited a streamlined genome with a relatively small number of genes in both core and species-specific protein families. The low GC content and slight codon preferences of P. litchii sequences were similar to those of the analyzed downy mildews and a subset of Phytophthora species. Taken together, these observations suggest that P. litchii is a Phytophthora pathogen that is in the process of acquiring downy mildew-like genomic and morphological features. Thus P. litchii may provide a novel model for investigating morphological development and genomic adaptation in oomycete pathogens.

  6. Sequencing of the Litchi Downy Blight Pathogen Reveals It Is a Phytophthora Species With Downy Mildew-Like Characteristics.

    PubMed

    Ye, Wenwu; Wang, Yang; Shen, Danyu; Li, Delong; Pu, Tianhuizi; Jiang, Zide; Zhang, Zhengguang; Zheng, Xiaobo; Tyler, Brett M; Wang, Yuanchao

    2016-07-01

    On the basis of its downy mildew-like morphology, the litchi downy blight pathogen was previously named Peronophythora litchii. Recently, however, it was proposed to transfer this pathogen to Phytophthora clade 4. To better characterize this unusual oomycete species and important fruit pathogen, we obtained the genome sequence of Phytophthora litchii and compared it to those from other oomycete species. P. litchii has a small genome with tightly spaced genes. On the basis of a multilocus phylogenetic analysis, the placement of P. litchii in the genus Phytophthora is strongly supported. Effector proteins predicted included 245 RxLR, 30 necrosis-and-ethylene-inducing protein-like, and 14 crinkler proteins. The typical motifs, phylogenies, and activities of these effectors were typical for a Phytophthora species. However, like the genome features of the analyzed downy mildews, P. litchii exhibited a streamlined genome with a relatively small number of genes in both core and species-specific protein families. The low GC content and slight codon preferences of P. litchii sequences were similar to those of the analyzed downy mildews and a subset of Phytophthora species. Taken together, these observations suggest that P. litchii is a Phytophthora pathogen that is in the process of acquiring downy mildew-like genomic and morphological features. Thus P. litchii may provide a novel model for investigating morphological development and genomic adaptation in oomycete pathogens. PMID:27183038

  7. Ectopic expression of Dahlia merckii defensin DmAMP1 improves papaya resistance to Phytophthora palmivora by reducing pathogen vigor.

    PubMed

    Zhu, Yun J; Agbayani, Ricelle; Moore, Paul H

    2007-06-01

    Phytophthora spp., some of the more important casual agents of plant diseases, are responsible for heavy economic losses worldwide. Plant defensins have been introduced as transgenes into a range of species to increase host resistance to pathogens to which they were originally susceptible. However, the effectiveness and mechanism of interaction of the defensins with Phytophthora spp. have not been clearly characterized in planta. In this study, we expressed the Dahlia merckii defensin, DmAMP1, in papaya (Carica papaya L.), a plant highly susceptible to a root, stem, and fruit rot disease caused by Phytophthora palmivora. Extracts of total leaf proteins from transformed plants inhibited growth of Phytophthora in vitro and discs cut from the leaves of transformed plants inhibited growth of Phytophthora in a bioassay. Results from our greenhouse inoculation experiments demonstrate that expressing the DmAMP1 gene in papaya plants increased resistance against P. palmivora and that this increased resistance was associated with reduced hyphae growth of P. palmivora at the infection sites. The inhibitory effects of DmAMP1 expression in papaya suggest this approach has good potential to impart transgenic resistance against Phytophthora in papaya. PMID:17216480

  8. Wild Populations of Triatoma infestans Are Highly Connected to Intra-Peridomestic Conspecific Populations in the Bolivian Andes

    PubMed Central

    Brenière, Simone Frédérique; Salas, Renata; Buitrago, Rosio; Brémond, Philippe; Sosa, Victor; Bosseno, Marie-France; Waleckx, Etienne; Depickère, Stéphanie; Barnabé, Christian

    2013-01-01

    Triatoma infestans, the major vector of Chagas disease south of the Amazon in South America, has a large distribution of wild populations, contrary to what has previously been stated. These populations have been suspected of being the source of reinfestation of human habitats and could impede the full success of vector control campaigns. This study examined gene flow between intra-peridomestic populations and wild populations collected in the surround areas in three Andean localities in Bolivia. The populations were defined according to temporal, ecological, and spatial criteria. After DNA extraction from the legs of each insect, the samples were analyzed using seven microsatellite markers. First, the analysis of molecular variance (AMOVA) detected an absence of differentiation between wild and intra-peridomestic populations, although strong structuring was observed between the populations within each environment. Then for some populations, the Bayesian method of assignment to inferred populations showed very similar assignment patterns of the members of wild or intra-peridomestic populations in each locality. Finally, the detection of the first-generation migrants within the different populations provided evidence of insect displacement from the wild to the intra-peridomestic environment. This result indicates that, after control campaigns in the Andes, controlling this new paradigm of vector transmission risk stemming from the invasion of human habitats by wild populations of T. infestans requires long-term maintenance of public monitoring to keep the risk at a minimal level. Since wild populations of T. infestans have also been detected elsewhere in Argentina, Paraguay, and Chile, there is an urgent need to take these populations into account in future monitoring of Chagas disease transmission. PMID:24278320

  9. Examining Mechanisms of Pyrethroid Resistance in Eggs of Two Populations of the Chagas' Disease Vector Triatoma infestans (Hemiptera: Reduviidae).

    PubMed

    Roca-Acevedo, G; Picollo, M I; Capriotti, N; Sierra, I; Santo-Orihuela, P L

    2015-09-01

    Chagas disease is a zoonosis transmitted to man by blood-sucking triatomine bugs found in the Americas. Triatoma infestans (Klug, 1834) is the main vector of Chagas' disease in Argentina. The control of this illness relies heavily on vector control through the use of insecticide. However, resistance to pyrethroid insecticides associated with ineffective field treatments has been increasingly reported in T. infestans from Argentina and Bolivia. There are few reports on the expression and causes of resistance in eggs of resistant populations, and even fewer studies on insecticide resistance throughout embryonic development. In this study, we explore the biochemical and molecular mechanisms potentially associated with the deltamethrin resistance assessed in the developing eggs of the Argentinean (Campo Largo) and Bolivian (Entre Ríos) T. infestans populations.We found measurable activity of monooxigenases and pyrethroid esterases throughout embryonic development. The pyrethroid esterase activity grew steadily throughout development in all the studied populations and was highest in eggs 12 d old. Mean enzyme activity increased from 13.6 to 16.3 and 22.2 picomol 7-hydroxycoumarin/min (7-OHC) in eggs of 4-, 7-, and 12 d old from the susceptible reference bug colony. Mean activity of resistant populations increased from 16.0 to 25.9 picomol 7-OHC/min in eggs of 4- to 12 d old in Entre Ríos population, and from 15.9 to 28.9 picomol 7-OHC/min in Campo Largo population. Molecular analysis of susceptible and resistant developing eggs detected L1014F mutation in both resistant populations, but no L925I mutation was found in any of the studied populations.Higher esterase activity and L1014F presence justify the resistance to pyrethroid throughout developing eggs of both studied T. infestans populations. The description of resistance profiles including resistance mechanisms involved will allow a rational design of campaigns for the control of Chagas disease transmission

  10. [Proposal for a predator for the destruction of Triatoma infestans: Tarentola mauritanica].

    PubMed

    Castello, J A; Gil Rivas, M J

    1980-01-01

    The task of Chagas disease control is overwhelming. One of the measures is the destruction of the bugs which act as vectors. In this paper the authors suggest trying this with a predator, a voracious insectivorous lizard, a kind of gecko: the Tarentola mauritanica or Moorish Gecko. These geckos were transported fortuitously to Argentina in cargoes of cork from their natural habitat, the Mediterranean basin. They have adapted easily to their new environment. Their ability to eat the kissing bug (Triatoma infestans) was tested in the observations reported herein. Four adult geckos were caught in an area of the city of Buenos Aires, in the neighbourhood of a corkstop factory. These geckos were placed in a modified glass acuarium tank with fourteen ninphs and adult kissing bugs. The bugs were rapidly eaten by the geckos (Fig. 3). The advantages of this type of biological control are stressed. The lizard and the bugs share the same habitat and both are nocturnal. Both make themselves quite at home in human habitations. The lizards -as the birds- are refractory to Tripanosomiasis and so they cannot become reservoirs. This kind of gecko seems also to be resistant to the toxic action of Gammexane. Therefore the authors think it is worthwhile to try this possibility of bug control in the field. It will be necessary to favour the reproduction of gecko and promote an educational campaign to prevent their destruction by man emphasizing their usefulness and their domesticity. Though costs were not estimated, the authors believe that it would be cheaper to provide lizards to dwellers in the poor homes of the rural areas ("ranchos") than expensive campaigns with potentially dangerous insecticides.

  11. Enhanced Biological Control of Phytophthora Blight of Pepper by Biosurfactant-Producing Pseudomonas

    PubMed Central

    Özyilmaz, Ümit; Benlioglu, Kemal

    2013-01-01

    Pseudomonas isolates from different crop plants were screened for in vitro growth inhibition of Phytophthora capsici and production of biosurfactant. Two in vivo experiments were performed to determine the efficacy of selected Pseudomonas strains against Phytophthora blight of pepper by comparing two fungicide treatments [acibenzolar-S-methyl (ASM) and ASM + mefenoxam]. Bacterial isolates were applied by soil drenching (1 × 109 cells/ml), ASM (0.1 μg a.i./ml) and ASM + mefenoxam (0.2 mg product/ml) were applied by foliar spraying, and P. capsici inoculum was incorporated into the pot soil three days after treatments. In the first experiment, four Pseudomonas strains resulted in significant reduction from 48.4 to 61.3% in Phytophthora blight severity. In the second experiment, bacterial treatments combining with olive oil (5 mL per plant) significantly enhanced biological control activity, resulting in a reduction of disease level ranging from 56.8 to 81.1%. ASM + mefenoxam was the most effective treatment while ASM alone was less effective in both bioassays. These results indicate that our Pseudomonas fluorescens strains (6L10, 6ba6 and 3ss9) that have biosurfactant-producing abilities are effective against P. capsici on pepper, and enhanced disease suppression could be achieved when they were used in combination with olive oil. PMID:25288970

  12. Effect of plant sterols and tannins on Phytophthora ramorum growth and sporulation.

    PubMed

    Stong, Rachel A; Kolodny, Eli; Kelsey, Rick G; González-Hernández, M P; Vivanco, Jorge M; Manter, Daniel K

    2013-06-01

    Elicitin-mediated acquisition of plant sterols is required for growth and sporulation of Phytophthora spp. This study examined the interactions between elicitins, sterols, and tannins. Ground leaf tissue, sterols, and tannin-enriched extracts were obtained from three different plant species (California bay laurel, California black oak, and Oregon white oak) in order to evaluate the effect of differing sterol/tannin contents on Phytophthora ramorum growth. For all three species, high levels of foliage inhibited P. ramorum growth and sporulation, with a steeper concentration dependence for the two oak samples. Phytophthora ramorum growth and sporulation were inhibited by either phytosterols or tannin-enriched extracts. High levels of sterols diminished elicitin gene expression in P. ramorum; whereas the tannin-enriched extract decreased the amount of 'functional' or ELISA-detectable elicitin, but not gene expression. Across all treatment combinations, P. ramorum growth and sporulation correlated strongly with the amount of ELISA-detectable elicitin (R (2) = 0.791 and 0.961, respectively).

  13. Identification and Characterisation CRN Effectors in Phytophthora capsici Shows Modularity and Functional Diversity

    PubMed Central

    Stam, Remco; Jupe, Julietta; Howden, Andrew J. M.; Morris, Jenny A.; Boevink, Petra C.; Hedley, Pete E.; Huitema, Edgar

    2013-01-01

    Phytophthora species secrete a large array of effectors during infection of their host plants. The Crinkler (CRN) gene family encodes a ubiquitous but understudied class of effectors with possible but as of yet unknown roles in infection. To appreciate CRN effector function in Phytophthora, we devised a simple Crn gene identification and annotation pipeline to improve effector prediction rates. We predicted 84 full-length CRN coding genes and assessed CRN effector domain diversity in sequenced Oomycete genomes. These analyses revealed evidence of CRN domain innovation in Phytophthora and expansion in the Peronosporales. We performed gene expression analyses to validate and define two classes of CRN effectors, each possibly contributing to infection at different stages. CRN localisation studies revealed that P. capsici CRN effector domains target the nucleus and accumulate in specific sub-nuclear compartments. Phenotypic analyses showed that few CRN domains induce necrosis when expressed in planta and that one cell death inducing effector, enhances P. capsici virulence on Nicotiana benthamiana. These results suggest that the CRN protein family form an important class of intracellular effectors that target the host nucleus during infection. These results combined with domain expansion in hemi-biotrophic and necrotrophic pathogens, suggests specific contributions to pathogen lifestyles. This work will bolster CRN identification efforts in other sequenced oomycete species and set the stage for future functional studies towards understanding CRN effector functions. PMID:23536880

  14. A novel elicitor protein from Phytophthora parasitica induces plant basal immunity and systemic acquired resistance.

    PubMed

    Chang, Yi-Hsuan; Yan, Hao-Zhi; Liou, Ruey-Fen

    2015-02-01

    The interaction between Phytophthora pathogens and host plants involves the exchange of complex molecular signals from both sides. Recent studies of Phytophthora have led to the identification of various apoplastic elicitors known to trigger plant immunity. Here, we provide evidence that the protein encoded by OPEL of Phytophthora parasitica is a novel elicitor. Homologues of OPEL were identified only in oomycetes, but not in fungi and other organisms. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) revealed that OPEL is expressed throughout the development of P. parasitica and is especially highly induced after plant infection. Infiltration of OPEL recombinant protein from Escherichia coli into leaves of Nicotiana tabacum (cv. Samsun NN) resulted in cell death, callose deposition, the production of reactive oxygen species and induced expression of pathogen-associated molecular pattern (PAMP)-triggered immunity markers and salicylic acid-responsive defence genes. Moreover, the infiltration conferred systemic resistance against a broad spectrum of pathogens, including Tobacco mosaic virus, the bacteria wilt pathogen Ralstonia solanacearum and P. parasitica. In addition to the signal peptide, OPEL contains three conserved domains: a thaumatin-like domain, a glycine-rich protein domain and a glycosyl hydrolase (GH) domain. Intriguingly, mutation of a putative laminarinase active site motif in the predicted GH domain abolished its elicitor activity, which suggests enzymatic activity of OPEL in triggering the defence response.

  15. Cross-species global proteomics reveals conserved and unique processes in Phytophthora sojae and P. ramorum

    SciTech Connect

    Savidor, Alon; Donahoo, Ryan S; Hurtado-Gonzales, Oscar; Land, Miriam L; Shah, Manesh B; Lamour, Kurt H; McDonald, W Hayes

    2008-08-01

    Phytophthora ramorum and Phytophthora sojae are destructive plant pathogens. Phytophthora sojae has a narrow host range whereas P. ramorum has a wide host range. A global proteomic comparison of the vegetative (mycelium) and infective (germinating-cyst) life-stages of P. sojae and P. ramorum was conducted to identify candidate proteins involved in host range, early infection and vegetative growth. Sixty-two candidates for early infection, 26 candidates for vegetative growth, and numerous proteins that may be involved in defining host specificity were identified. In addition, common life stage proteomic trends between the organisms were observed. In mycelia, proteins involved in transport and metabolism of amino acids, carbohydrates and other small molecules were up-regulated. In the germinating cysts, up-regulated proteins associated with lipid transport and metabolism, cytoskeleton and protein synthesis were observed. It appears that the germinating cyst catabolizes lipid reserves through the -oxidation pathway to drive the extensive protein synthesis necessary to produce the germ tube and initiate infection. Once inside the host, the pathogen switches to vegetative growth, where energy is derived from glycolysis and utilized for synthesis of amino acids and other molecules that assist survival in the plant tissue.

  16. Mutations Affecting Hyphal Colonization and Pyoverdine Production in Pseudomonads Antagonistic toward Phytophthora parasitica.

    PubMed

    Yang, C H; Menge, J A; Cooksey, D A

    1994-02-01

    In previous studies, Pseudomonas putida 06909 and Pseudomonas fluorescens 09906 suppressed populations of Phytophthora parasitica in the citrus rhizosphere, suggesting that these bacteria may be useful in biological control of citrus root rot. In this study we investigated the mechanisms of antagonism between the bacteria and the fungus. Both bacteria colonized Phytophthora hyphae and inhibited the fungus on agar media. A hyphal column assay was developed to measure the colonization of bacteria on fungal hyphae and to enrich for colonization-deficient mutants. In this way we identified Tn5 mutants of each pseudomonad that were not able to colonize the hyphae and inhibit fungal growth in vitro. Colonization-deficient mutants were nonmotile and lacked flagella. Survival of nonmotile mutants in a citrus soil was similar to survival of a random Tn5 mutant over a 52-day period. Additional screening of random Tn5 mutants of both pseudomonads for loss of fungal inhibition in vitro yielded two distinct types of mutants. Mutants of the first type were deficient in production of pyoverdines and in inhibition of the fungus in vitro, although they still colonized fungal hyphae. Mutants of the second type lacked flagella and were not able to colonize the hyphae or inhibit fungal growth. No role was found for antibiotic production by the two bacteria in the inhibition of the fungus. Our results suggest that both hyphal colonization and pyoverdine production are important in the inhibition of Phytophthora parasitica by P. fluorescens and P. putida in vitro. PMID:16349177

  17. Effect of plant sterols and tannins on Phytophthora ramorum growth and sporulation.

    PubMed

    Stong, Rachel A; Kolodny, Eli; Kelsey, Rick G; González-Hernández, M P; Vivanco, Jorge M; Manter, Daniel K

    2013-06-01

    Elicitin-mediated acquisition of plant sterols is required for growth and sporulation of Phytophthora spp. This study examined the interactions between elicitins, sterols, and tannins. Ground leaf tissue, sterols, and tannin-enriched extracts were obtained from three different plant species (California bay laurel, California black oak, and Oregon white oak) in order to evaluate the effect of differing sterol/tannin contents on Phytophthora ramorum growth. For all three species, high levels of foliage inhibited P. ramorum growth and sporulation, with a steeper concentration dependence for the two oak samples. Phytophthora ramorum growth and sporulation were inhibited by either phytosterols or tannin-enriched extracts. High levels of sterols diminished elicitin gene expression in P. ramorum; whereas the tannin-enriched extract decreased the amount of 'functional' or ELISA-detectable elicitin, but not gene expression. Across all treatment combinations, P. ramorum growth and sporulation correlated strongly with the amount of ELISA-detectable elicitin (R (2) = 0.791 and 0.961, respectively). PMID:23689874

  18. Genetic Diversity, Population Structure, and Resistance to Phytophthora capsici of a Worldwide Collection of Eggplant Germplasm

    PubMed Central

    Naegele, Rachel P.; Boyle, Samantha; Quesada-Ocampo, Lina M.; Hausbeck, Mary K.

    2014-01-01

    Eggplant (Solanum melongena L.) is an important solanaceous crop with high phenotypic diversity and moderate genotypic diversity. Ninety-nine genotypes of eggplant germplasm (species (S. melongena, S. incanum, S. linnaeanum and S. gilo), landraces and heirloom cultivars) from 32 countries and five continents were evaluated for genetic diversity, population structure, fruit shape, and disease resistance to Phytophthora fruit rot. Fruits from each line were measured for fruit shape and evaluated for resistance to two Phytophthora capsici isolates seven days post inoculation. Only one accession (PI 413784) was completely resistant to both isolates evaluated. Partial resistance to Phytophthora fruit rot was found in accessions from all four eggplant species evaluated in this study. Genetic diversity and population structure were assessed using 22 polymorphic simple sequence repeats (SSRs). The polymorphism information content (PIC) for the population was moderate (0.49) in the population. Genetic analyses using the program STRUCTURE indicated the existence of four genetic clusters within the eggplant collection. Population structure was detected when eggplant lines were grouped by species, continent of origin, country of origin, fruit shape and disease resistance. PMID:24819601

  19. Genetic diversity, population structure, and resistance to Phytophthora capsici of a worldwide collection of eggplant germplasm.

    PubMed

    Naegele, Rachel P; Boyle, Samantha; Quesada-Ocampo, Lina M; Hausbeck, Mary K

    2014-01-01

    Eggplant (Solanum melongena L.) is an important solanaceous crop with high phenotypic diversity and moderate genotypic diversity. Ninety-nine genotypes of eggplant germplasm (species (S. melongena, S. incanum, S. linnaeanum and S. gilo), landraces and heirloom cultivars) from 32 countries and five continents were evaluated for genetic diversity, population structure, fruit shape, and disease resistance to Phytophthora fruit rot. Fruits from each line were measured for fruit shape and evaluated for resistance to two Phytophthora capsici isolates seven days post inoculation. Only one accession (PI 413784) was completely resistant to both isolates evaluated. Partial resistance to Phytophthora fruit rot was found in accessions from all four eggplant species evaluated in this study. Genetic diversity and population structure were assessed using 22 polymorphic simple sequence repeats (SSRs). The polymorphism information content (PIC) for the population was moderate (0.49) in the population. Genetic analyses using the program STRUCTURE indicated the existence of four genetic clusters within the eggplant collection. Population structure was detected when eggplant lines were grouped by species, continent of origin, country of origin, fruit shape and disease resistance.

  20. Antifungal Activity and Biochemical Response of Cuminic Acid against Phytophthora capsici Leonian.

    PubMed

    Wang, Yong; Sun, Yang; Zhang, Ying; Zhang, Xing; Feng, Juntao

    2016-01-01

    Phytophthora blight of pepper caused by Phytophthora capsici Leonian is a destructive disease throughout the world. Cuminic acid, extracted from the seed of Cuminum cyminum L., belongs to the benzoic acid chemical class. In this study, the sensitivity and biochemical response of P. capsici to cuminic acid was determined. The mean EC50 (50% effective concentration) values for cuminic acid in inhibiting mycelial growth and zoospore germination of the 54 studied P. capsici isolates were 14.54 ± 5.23 μg/mL and 6.97 ± 2.82 μg/mL, respectively. After treatment with cuminic acid, mycelial morphology, sporangium formation and mycelial respiration were significantly influenced; cell membrane permeability and DNA content increased markedly, but pyruvic acid content, adenosine triphosphate (ATP) content, and ATPase activity decreased compared with the untreated control. In pot experiments, cuminic acid exhibited both protective and curative activity. Importantly, POD and PAL activity of the pepper leaves increased after being treated with cuminic acid. These indicated that cuminic acid not only showed antifungal activity, but also could improve the defense capacity of the plants. All the results suggested that cuminic acid exhibits the potential to be developed as a new phytochemical fungicide, and this information increases our understanding of the mechanism of action of cuminic acid against Phytophthora capsici. PMID:27294911

  1. Direct molecular identification of Trypanosoma cruzi Discrete Typing Units in domestic and peridomestic Triatoma infestans and Triatoma sordida from the Argentine Chaco

    PubMed Central

    MAFFEY, L.; CARDINAL, M.V.; ORDÓÑEZ-KRASNOWSKI, P.C.; LANATI, L.A.; LAURICELLA, M.A.; SCHIJMAN, A.G.; GÜRTLER, R.E.

    2013-01-01

    SUMMARY We assessed the distribution of Trypanosoma cruzi Discrete Typing Units (DTUs) in domestic and peridomestic Triatoma infestans and Triatoma sordida specimens collected in a well-defined rural area in Pampa del Indio, northeastern Argentina. Microscopically-positive bugs were randomly selected with a multi-level sampling design, and DTUs were identified using direct PCR strategies. TcVI predominated in 61% of 69 T. infestans and in 56% of 9 T. sordida. TcV was the secondary DTU in T. infestans (16%) and was found in one T. sordida specimen (11%). Three T. sordida (33%) were found infected with TcI, a DTU also identified in local Didelphis albiventris opossums. Mixed DTU infections occurred rarely (5%) and were detected both directly from the bugs’ rectal ampoule and parasite cultures. The identified DTUs and bug collection sites of T. infestans were significantly associated. Bugs infected with TcV were almost exclusively captured in domiciles whereas those with TcVI were found similarly in domiciles and peridomiciles. All mixed infections occurred in domiciles. TcV-infected bugs fed more often on humans than on dogs, whereas TcVI-infected bugs showed the reverse pattern. T. sordida is a probable sylvatic vector of TcI linked to D. albiventris, and could represent a secondary vector of TcVI and TcV in the domestic/peridomestic cycle. PMID:23036510

  2. Control of experimental Triatoma infestans populations: effect of pour-on cypermethrin applied to chickens under natural conditions in the Argentinean Chaco region.

    PubMed

    Amelotti, I; Catalá, S S; Gorla, D E

    2014-06-01

    Among peridomestic structures, chicken coops are sites of major importance for the domestic ecology of Triatoma infestans (Hemiptera: Reduviidae). The aim of this study was to evaluate in an experimental context the effects of a cypermethrin pour-on formulation applied to chickens on blood intake, moulting and mortality in T. infestans, under the natural climatic conditions of a region endemic for Chagas' disease. Experimental chicken huts were made of bricks and covered with plastic mosquito nets. Ninety fourth-instar nymphs were maintained in each hut. The study used a completely random design in which chickens in the experimental group were treated with a cypermethrin pour-on formulation. Five replicates (= huts) of the experimental and control groups were conducted. The number of live T. infestans, blood intake and moults to fifth-instar stage were recorded at 1, 5, 20, 35 and 45 days after the application of cypermethrin. Cumulative mortality was higher in nymphs exposed to treated chickens (> 71%) than in control nymphs (< 50%) (P < 0.01). Blood intake and moulting rate were lower in nymphs fed on treated chickens than in control nymphs (P < 0.05). Pour-on cypermethrin was able to cause significant mortality, although it did not eliminate the experimental population of T. infestans.

  3. Microgeographic Spatial Structuring of Triatoma infestans (Hemiptera: Reduviidae) Populations Using Wing Geometric Morphometry in the Argentine Chaco

    PubMed Central

    GASPE, M. S.; SCHACHTER-BROIDE, J.; GUREVITZ, J. M.; KITRON, U.; GÜRTLER, R. E.; DUJARDIN, J. P.

    2013-01-01

    We investigated the occurrence of spatial structuring in Triatoma infestans (Klug) (Hemiptera: Reduviidae) populations 12 yr after the last community-wide insecticide spraying campaign in rural Pampa del Indio, in the Gran Chaco of northeastern Argentina. In total, 172 male and 149 female right wings collected at 16 georeferenced sites with at least 10 individuals of the same sex were analyzed using geometric morphometry. Mean female body length and wing centroid size (CS) were significantly larger than for males. Log-transformed CS and length were significantly and positively correlated both for males and females. Males collected in domiciles had significantly smaller CS than those collected in peridomestic structures both closed (kitchens or storerooms) or open (chicken coops), in agreement with our previous results elsewhere in the dry Argentine Chaco. Female wing CS was not significantly different between ecotopes. Wing shape analyses showed the occurrence of significant geographic structuring in males and females combined and in males only. Male wings showed a strong association between Mahalanobis distance and geographic distance. In general, Mahalanobis distances were significantly different between collection sites located >4 km apart. For collection sites located <4 km apart, the greater the geographic distance the larger the difference in wing shape variables. Among females, only a partial correspondence between geographic groups and Mahalanobis distances was recorded. The strong spatial structuring found in T. infestans populations may be useful for the identification of putative reinfestation sources after vector control interventions. PMID:22679857

  4. Comparison of insecticidal paint and deltamethrin against Triatoma infestans (Hemiptera: Reduviidae) feeding and mortality in simulated natural conditions

    PubMed Central

    Maloney, Kathleen M.; Ancca-Juarez, Jenny; Salazar, Renzo; Borrini-Mayori, Katty; Niemierko, Malwina; Yukich, Joshua O.; Naquira, Cesar; Keating, Joseph A.; Levy, Michael Z.

    2013-01-01

    The Chagas disease vector Triatoma infestans is largely controlled by the household application of pyrethroid insecticides. Because effective, large-scale insecticide application is costly and necessitates numerous trained personnel, alternative control techniques are badly needed. We compared the residual effect of organophosphate-based insecticidal paint (Inesfly 5A IGR™ (I5A)) to standard deltamethrin, and a negative control, against T. infestans in a simulated natural environment. We evaluated mortality, knockdown, and ability to take a blood meal among fifth instar nymphs. I5A paint caused significantly greater mortality at time points up to 9 months compared to deltamethrin (Fisher's Exact Test, p < 0.01 in all instances). A year following application mortality among nymphs in the I5A was similar to those in the deltamethrin (χ2 = 0.76, df=1, p < 0.76). At months 0 and 1 after application, fewer nymphs exposed to deltamethrin took a blood meal compared to insects exposed to paint (Fisher's Exact Tests, p < 0.01 and p < 0.01 respectively). Insecticidal paint may provide an easily-applied means of protection against Chagas disease vectors. PMID:23701602

  5. Where do these bugs come from? Phenotypic structure of Triatoma infestans populations after control interventions in the Argentine Chaco

    PubMed Central

    Gaspe, María Sol; Provecho, Yael Mariana; Piccinali, Romina Valeria; Gürtler/, Ricardo Esteban

    2015-01-01

    House re-invasion by native triatomines after insecticide-based control campaigns represents a major threat for Chagas disease vector control. We conducted a longitudinal intervention study in a rural section (Area III, 407 houses) of Pampa del Indio, northeastern Argentina, and used wing geometric morphometry to compare pre-spray and post-spray (re-infestant bugs) Triatoma infestans populations. The community-wide spraying with pyrethroids reduced the prevalence of house infestation by T. infestans from 31.9% to < 1% during a four-year follow-up, unlike our previous studies in the neighbouring Area I. Two groups of bug collection sites differing in wing shape variables before interventions (including 221 adults from 11 domiciles) were used as a reference for assigning 44 post-spray adults. Wing shape variables from post-spray, high-density bug colonies and pre-spray groups were significantly different, suggesting that re-infestant insects had an external origin. Insects from one house differed strongly in wing shape variables from all other specimens. A further comparison between insects from both areas supported the existence of independent re-infestation processes within the same district. These results point to local heterogeneities in house re-infestation dynamics and emphasise the need to expand the geographic coverage of vector surveillance and control operations to the affected region. PMID:25946158

  6. Comparison of insecticidal paint and deltamethrin against Triatoma infestans (Hemiptera: Reduviidae) feeding and mortality in simulated natural conditions.

    PubMed

    Maloney, Kathleen M; Ancca-Juarez, Jenny; Salazar, Renzo; Borrini-Mayori, Katty; Niemierko, Malwina; Yukich, Joshua O; Naquira, Cesar; Keating, Joseph A; Levy, Michael Z

    2013-06-01

    The vector of Chagas disease, Triatoma infestans, is largely controlled by the household application of pyrethroid insecticides. Because effective, large-scale insecticide application is costly and necessitates numerous trained personnel, alternative control techniques are badly needed. We compared the residual effect of organophosphate-based insecticidal paint (Inesfly 5A IGR™ (I5A)) to standard deltamethrin, and a negative control, against T. infestans in a simulated natural environment. We evaluated mortality, knockdown, and ability to take a blood meal among 5(th) instar nymphs. I5A paint caused significantly greater mortality at time points up to nine months compared to deltamethrin (Fisher's Exact Test, p < 0.01 in all instances). A year following application, mortality among nymphs in the I5A was similar to those in the deltamethrin (χ2 = 0.76, df=1, p < 0.76). At months 0 and 1 after application, fewer nymphs exposed to deltamethrin took a blood meal compared to insects exposed to paint (Fisher's Exact Tests, p < 0.01 and p < 0.01, respectively). Insecticidal paint may provide an easily-applied means of protection against vectors of Chagas disease.

  7. Hemi-Nested PCR and RFLP Methodologies for Identifying Blood Meals of the Chagas Disease Vector, Triatoma infestans

    PubMed Central

    Roellig, Dawn M.; Gomez-Puerta, Luis A.; Mead, Daniel G.; Pinto, Jesus; Ancca-Juarez, Jenny; Calderon, Maritza; Bern, Caryn; Gilman, Robert H.; Cama, Vitaliano A.

    2013-01-01

    Trypanosoma cruzi, the etiologic agent of Chagas disease, is transmitted by hematophagous reduviid bugs within the subfamily Triatominae. These vectors take blood meals from a wide range of hosts, and their feeding behaviors have been used to investigate the ecology and epidemiology of T. cruzi. In this study we describe two PCR-based methodologies that amplify a fragment of the 16S mitochondrial rDNA, aimed to improve the identification of blood meal sources for Triatoma infestans: a.- Sequence analyses of two heminested PCRs that allow the identification of mammalian and avian species, and b.- restriction fragment length polymorphism (RFLP) analysis from the mammalian PCR to identify and differentiate multi-host blood meals. Findings from both methodologies indicate that host DNA could be detected and the host species identified in samples from laboratory reared and field collected triatomines. The implications of this study are two-fold. First, these methods can be used in areas where the fauna diversity and feeding behavior of the triatomines are unknown. Secondly, the RFLP method led to the identification of multi-host DNA from T. infestans gut contents, enhancing the information provided by this assay. These tools are important contributions for ecological and epidemiological studies of vector-borne diseases. PMID:24040328

  8. cDNA isolation and characterization of two vitellogenin genes in the Chagas' disease vector Triatoma infestans (Hemiptera, Reduviidae).

    PubMed

    Blariza, María J; Soria, Néstor W; Torres, Adrián G; Grosso, Carla G; García, Beatriz A

    2014-06-10

    Two vitellogenin genes (Vg1 and Vg2) were identified in the Chagas' disease vector Triatoma infestans. The putative coding sequence corresponding to Vg2 was found to be 5553bp long, encoding 1851 amino acids in a single open reading frame. The comparative analysis of the deduced amino acid sequences from Vg1 and Vg2 cDNA fragments of T. infestans revealed 58.94% of identity with 76.43% of homology. The phylogenetic tree based on the complete Vg amino acid sequences of hemimetabolous insects unambiguously supported two clusters, one consisting of Vg sequences from dictyopteran and the other containing Vg sequences of hemipteran. The Vg1 and Vg2 mRNAs were detected in fat bodies and ovaries of adult females with the highest levels of both Vg transcripts in the first tissue. Quantitative PCR showed low expression of Vg2 in head and muscle of adult females, while the Vg1 transcript was not present in these organs. Neither Vg1 nor Vg2 was expressed in fifth instar nymph fat bodies or in adult male fat bodies, heads, and muscles. PMID:24685521

  9. The use of genus-specific amplicon pyrosequencing to assess phytophthora species diversity using eDNA from soil and water in Northern Spain.

    PubMed

    Català, Santiago; Pérez-Sierra, Ana; Abad-Campos, Paloma

    2015-01-01

    Phytophthora is one of the most important and aggressive plant pathogenic genera in agriculture and forestry. Early detection and identification of its pathways of infection and spread are of high importance to minimize the threat they pose to natural ecosystems. eDNA was extracted from soil and water from forests and plantations in the north of Spain. Phytophthora-specific primers were adapted for use in high-throughput Sequencing (HTS). Primers were tested in a control reaction containing eight Phytophthora species and applied to water and soil eDNA samples from northern Spain. Different score coverage threshold values were tested for optimal Phytophthora species separation in a custom-curated database and in the control reaction. Clustering at 99% was the optimal criteria to separate most of the Phytophthora species. Multiple Molecular Operational Taxonomic Units (MOTUs) corresponding to 36 distinct Phytophthora species were amplified in the environmental samples. Pyrosequencing of amplicons from soil samples revealed low Phytophthora diversity (13 species) in comparison with the 35 species detected in water samples. Thirteen of the MOTUs detected in rivers and streams showed no close match to sequences in international sequence databases, revealing that eDNA pyrosequencing is a useful strategy to assess Phytophthora species diversity in natural ecosystems.

  10. Phytophthora fruit rot-resistant watermelon germplasm lines: USVL489-PFR, USVL782-PFR, USVL203-PFR, and USVL020-PFR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    USVL489-PFR, USVL782-PFR, USVL203-PFR, and USVL020-PFR are watermelon (Citrullus lanatus var. lanatus (Thunb.) Matsum. & Nakai) germplasm lines that exhibit high levels of resistance to Phytophthora fruit rot caused by the plant pathogen Phytophthora capsici. Resistance in these germplasm lines is ...

  11. The Use of Genus-Specific Amplicon Pyrosequencing to Assess Phytophthora Species Diversity Using eDNA from Soil and Water in Northern Spain

    PubMed Central

    Català, Santiago; Pérez-Sierra, Ana; Abad-Campos, Paloma

    2015-01-01

    Phytophthora is one of the most important and aggressive plant pathogenic genera in agriculture and forestry. Early detection and identification of its pathways of infection and spread are of high importance to minimize the threat they pose to natural ecosystems. eDNA was extracted from soil and water from forests and plantations in the north of Spain. Phytophthora-specific primers were adapted for use in high-throughput Sequencing (HTS). Primers were tested in a control reaction containing eight Phytophthora species and applied to water and soil eDNA samples from northern Spain. Different score coverage threshold values were tested for optimal Phytophthora species separation in a custom-curated database and in the control reaction. Clustering at 99% was the optimal criteria to separate most of the Phytophthora species. Multiple Molecular Operational Taxonomic Units (MOTUs) corresponding to 36 distinct Phytophthora species were amplified in the environmental samples. Pyrosequencing of amplicons from soil samples revealed low Phytophthora diversity (13 species) in comparison with the 35 species detected in water samples. Thirteen of the MOTUs detected in rivers and streams showed no close match to sequences in international sequence databases, revealing that eDNA pyrosequencing is a useful strategy to assess Phytophthora species diversity in natural ecosystems. PMID:25775250

  12. Effectiveness of residual spraying of peridomestic ecotopes with deltamethrin and permethrin on Triatoma infestans in rural western Argentina: a district-wide randomized trial.

    PubMed Central

    Gürtler, Ricardo E.; Canale, Delmi M.; Spillmann, Cynthia; Stariolo, Raúl; Salomón, Oscar D.; Blanco, Sonia; Segura, Elsa L.

    2004-01-01

    OBJECTIVE: To compare the effectiveness of a single residual spraying of pyrethroids on the occurrence and abundance of Triatoma infestans in peridomestic ecotopes in rural La Rioja. METHODS: A total of 667 (32.8%) peridomestic sites positive for T. infestans in May 1999 were randomly assigned to treatment within each village, sprayed in December 1999, and reinspected in December 2000. Treatments included 2.5% suspension concentrate (SC) deltamethrin in water at 25 mg active ingredient (a.i.)/m(2) applied with: (a) manual compression sprayers (standard treatment) or (b) power sprayers; (c) 1.5% emulsifiable concentrate (EC) deltamethrin at 25 mg a.i./m(2); and (d) 10% EC cis-permethrin at 170 mg a.i./m(2). EC pyrethroids were diluted in soybean oil and applied with power sprayers. All habitations were sprayed with the standard treatment. FINDINGS: The prevalence of T. infestans 1-year post-spraying was significantly lower in sites treated with SC deltamethrin applied with manual (24%) or power sprayers (31%) than in sites treated with EC deltamethrin (40%) or EC permethrin (53%). The relative odds of infestation and catch of T. infestans 1-year post-spraying significantly increased with the use of EC pyrethroids, the abundance of bugs per site before spraying, total surface, and host numbers. All insecticides had poor residual effects on wooden posts. CONCLUSION: Most of the infestations probably originated from triatomines that survived exposure to insecticides at each site. Despite the standard treatment proving to be the most effective, the current tactics and procedures fail to eliminate peridomestic populations of T. infestans in semiarid rural areas and need to be revised. PMID:15112008

  13. Biological Control of the Chagas Disease Vector Triatoma infestans with the Entomopathogenic Fungus Beauveria bassiana Combined with an Aggregation Cue: Field, Laboratory and Mathematical Modeling Assessment

    PubMed Central

    Forlani, Lucas; Pedrini, Nicolás; Girotti, Juan R.; Mijailovsky, Sergio J.; Cardozo, Rubén M.; Gentile, Alberto G.; Hernández-Suárez, Carlos M.; Rabinovich, Jorge E.; Juárez, M. Patricia

    2015-01-01

    Background Current Chagas disease vector control strategies, based on chemical insecticide spraying, are growingly threatened by the emergence of pyrethroid-resistant Triatoma infestans populations in the Gran Chaco region of South America. Methodology and findings We have already shown that the entomopathogenic fungus Beauveria bassiana has the ability to breach the insect cuticle and is effective both against pyrethroid-susceptible and pyrethroid-resistant T. infestans, in laboratory as well as field assays. It is also known that T. infestans cuticle lipids play a major role as contact aggregation pheromones. We estimated the effectiveness of pheromone-based infection boxes containing B. bassiana spores to kill indoor bugs, and its effect on the vector population dynamics. Laboratory assays were performed to estimate the effect of fungal infection on female reproductive parameters. The effect of insect exuviae as an aggregation signal in the performance of the infection boxes was estimated both in the laboratory and in the field. We developed a stage-specific matrix model of T. infestans to describe the fungal infection effects on insect population dynamics, and to analyze the performance of the biopesticide device in vector biological control. Conclusions The pheromone-containing infective box is a promising new tool against indoor populations of this Chagas disease vector, with the number of boxes per house being the main driver of the reduction of the total domestic bug population. This ecologically safe approach is the first proven alternative to chemical insecticides in the control of T. infestans. The advantageous reduction in vector population by delayed-action fungal biopesticides in a contained environment is here shown supported by mathematical modeling. PMID:25969989

  14. Immunogenic Salivary Proteins of Triatoma infestans: Development of a Recombinant Antigen for the Detection of Low-Level Infestation of Triatomines

    PubMed Central

    Schwarz, Alexandra; Helling, Stefan; Collin, Nicolas; Teixeira, Clarissa R.; Medrano-Mercado, Nora; Hume, Jen C. C.; Assumpção, Teresa C.; Marcus, Katrin; Stephan, Christian; Meyer, Helmut E.; Ribeiro, José M. C.; Billingsley, Peter F.; Valenzuela, Jesus G.; Sternberg, Jeremy M.; Schaub, Günter A.

    2009-01-01

    Background Triatomines are vectors of Trypanosoma cruzi, the etiological agent of Chagas disease in Latin America. The most effective vector, Triatoma infestans, has been controlled successfully in much of Latin America using insecticide spraying. Though rarely undertaken, surveillance programs are necessary in order to identify new infestations and estimate the intensity of triatomine bug infestations in domestic and peridomestic habitats. Since hosts exposed to triatomines develop immune responses to salivary antigens, these responses can be evaluated for their usefulness as epidemiological markers to detect infestations of T. infestans. Methodology/Principal Findings T. infestans salivary proteins were separated by 2D-gel electrophoresis and tested for their immunogenicity by Western blotting using sera from chickens and guinea pigs experimentally exposed to T. infestans. From five highly immunogenic protein spots, eight salivary proteins were identified by nano liquid chromatography-electrospray ionization-tandem mass spectrometry (nanoLC-ESI-MS/MS) and comparison to the protein sequences of the National Center for Biotechnology Information (NCBI) database and expressed sequence tags of a unidirectionally cloned salivary gland cDNA library from T. infestans combined with the NCBI yeast protein sub-database. The 14.6 kDa salivary protein [gi|149689094] was produced as recombinant protein (rTiSP14.6) in a mammalian cell expression system and recognized by all animal sera. The specificity of rTiSP14.6 was confirmed by the lack of reactivity to anti-mosquito and anti-sand fly saliva antibodies. However, rTiSP14.6 was recognized by sera from chickens exposed to four other triatomine species, Triatoma brasiliensis, T. sordida, Rhodnius prolixus, and Panstrongylus megistus and by sera of chickens from an endemic area of T. infestans and Chagas disease in Bolivia. Conclusions/Significance The recombinant rTiSP14.6 is a suitable and promising epidemiological marker for

  15. An insight into the sialome of the blood-sucking bug Triatoma infestans, a vector of Chagas' disease

    PubMed Central

    Assumpção, Teresa C. F.; Francischetti, Ivo M. B.; Andersen, John F.; Schwarz, Alexandra; Santana, Jaime M.; Ribeiro, José M. C.

    2008-01-01

    Triatoma infestans is a hemiptera, vector of Chagas’ disease, that feeds exclusively on vertebrate blood in all life stages. Hematophagous insects’ salivary glands (SG) produce potent pharmacological compounds that counteract host hemostasis, including anti-clotting, anti-platelet, and vasodilatory molecules. To obtain a further insight into the salivary biochemical and pharmacological complexity of this insect, a cDNA library from its salivary glands was randomly sequenced. Also, salivary proteins were submitted to two dimentional gel (2D-gel) electrophoresis followed by MS analysis. We present the analysis of a set of 1,534 (SG) cDNA sequences, 645 of which coded for proteins of a putative secretory nature. Most salivary proteins described as lipocalins matched peptide sequences obtained from proteomic results. PMID:18207082

  16. Temporal Variations of Wing Size and Shape of Triatoma infestans (Hemiptera: Reduviidae) Populations From Northwestern Argentina Using Geometric Morphometry

    PubMed Central

    Schachter-Broide, Judith; Gürtler, Ricardo E.; Kitron, Uriel; Dujardin, Jean-Pierre

    2009-01-01

    Wing geometric morphometry of Triatoma infestans (Klug) (Hemiptera: Reduviidae) populations in northwestern Argentina showed that individual collection sites represent the discrete unit where metric differentiation took place. Here we studied temporal variations in wing size and shape of T. infestans populations from defined capture sites on three occasions between 2000 and 2003. Bugs collected from domiciles and/or storerooms had significantly larger right-wing centroid size than bugs collected at goat and/or pig corrals by the end of summer 2000 for both sexes. Conversely, male bugs collected from domiciles and/or storerooms had significantly smaller centroid size than bugs collected from pig corrals in spring 2002. The inversion in wing centroid size between seasons was consistent between sexes. Wing shape analysis from the south-central extreme of the study village showed divergence between collection dates for both sexes. Wing shape divergence was highly significant between male bugs collected by the end of summer 2000 and those collected in spring 2002 and by the end of summer 2003. For females, wing shape divergence was marginally significant between the end of summer 2000 and spring 2002, and significant between spring 2002 and the end of summer 2003. Unlike season-related variations in wing centroid size, shape differentiation was related to the time period elapsed between sample collections and suggested genetic influences acting on shape. Simultaneous consideration of wing size and shape provided complementary information on the direction and timing of bug dispersal. Morphological studies may allow determining the degree of relatedness of different bug populations and to associate morphological heterogeneity with temporal patterns of reinfestation. PMID:19769028

  17. Intensified Surveillance and Insecticide-based Control of the Chagas Disease Vector Triatoma infestans in the Argentinean Chaco

    PubMed Central

    Gurevitz, Juan M.; Gaspe, María Sol; Enriquez, Gustavo F.; Provecho, Yael M.; Kitron, Uriel; Gürtler, Ricardo E.

    2013-01-01

    Background The elimination of Triatoma infestans, the main Chagas disease vector in the Gran Chaco region, remains elusive. We implemented an intensified control strategy based on full-coverage pyrethroid spraying, followed by frequent vector surveillance and immediate selective insecticide treatment of detected foci in a well-defined rural area in northeastern Argentina with moderate pyrethroid resistance. We assessed long-term impacts, and identified factors and procedures affecting spray effectiveness. Methods and Findings After initial control interventions, timed-manual searches were performed by skilled personnel in 4,053 sites of 353–411 houses inspected every 4–7 months over a 35-month period. Residual insecticide spraying was less effective than expected throughout the three-year period, mainly because of the occurrence of moderate pyrethroid resistance and the limited effectiveness of selective treatment of infested sites only. After initial interventions, peridomestic infestation prevalence always exceeded domestic infestation, and timed-manual searches consistently outperformed householders' bug detection, except in domiciles. Most of the infestations occurred in houses infested at baseline, and were restricted to four main ecotopes. Houses with an early persistent infestation were spatially aggregated up to a distance of 2.5 km. An Akaike-based multi-model inference approach showed that new site-level infestations increased substantially with the local availability of appropriate refugia for triatomine bugs, and with proximity to the nearest site found infested at one or two preceding surveys. Conclusions and Significance Current vector control procedures have limited effectiveness in the Gran Chaco. Selective insecticide sprays must include all sites within the infested house compound. The suppression of T. infestans in rural areas with moderate pyrethroid resistance requires increased efforts and appropriate management actions. In addition to

  18. Analysing deltamethrin susceptibility and pyrethroid esterase activity variations in sylvatic and domestic Triatoma infestans at the embryonic stage

    PubMed Central

    Santo-Orihuela, Pablo Luis; Carvajal, Guillermo; Picollo, María Inés; Vassena, Claudia Viviana

    2013-01-01

    The aim of the present work was to study the deltamethrin susceptibility of eggs from Triatoma infestans populations and the contribution of pyrethroid esterases to deltamethrin degradation. Insects were collected from sylvatic areas, including Veinte de Octubre and Kirus-Mayu (Bolivia) and from domiciliary areas, including El Palmar (Bolivia) and La Pista (Argentina). Deltamethrin susceptibility was determined by dose-response bioassays. Serial dilutions of deltamethrin (0.0005-1 mg/mL) were topically applied to 12-day-old eggs. Samples from El Palmar had the highest lethal dose ratio (LDR) value (44.90) compared to the susceptible reference strain (NFS), whereas the Veinte de Octubre samples had the lowest value (0.50). Pyrethroid esterases were evaluated using 7-coumaryl permethrate (7-CP) on individually homogenised eggs from each population and from NFS. The El Palmar and La Pista samples contained 40.11 and 36.64 pmol/min/mg protein, respectively, and these values were statistically similar to NFS (34.92 pmol/min/mg protein) and different from Kirus-Mayu and Veinte de Octubre (27.49 and 22.69 pmol/min/mg protein, respectively). The toxicological data indicate that the domestic populations were resistant to deltamethrin, but no statistical contribution of 7-CP esterases was observed. The sylvatic populations had similar LDR values to NFS, but lower 7-CP esterase activities. Moreover, this is the first study of the pyrethroid esterases on T. infestans eggs employing a specific substrate (7-CP). PMID:24402155

  19. Host-Seeking Behavior and Dispersal of Triatoma infestans, a Vector of Chagas Disease, under Semi-field Conditions

    PubMed Central

    Castillo-Neyra, Ricardo; Barbu, Corentin M.; Salazar, Renzo; Borrini, Katty; Naquira, Cesar; Levy, Michael Z.

    2015-01-01

    Chagas disease affects millions of people in Latin America. The control of this vector-borne disease focuses on halting transmission by reducing or eliminating insect vector populations. Most transmission of Trypanosoma cruzi, the causative agent of Chagas disease, involves insects living within or very close to households and feeding mostly on domestic animals. As animal hosts can be intermittently present it is important to understand how host availability can modify transmission risk to humans and to characterize the host-seeking dispersal of triatomine vectors on a very fine scale. We used a semi-field system with motion-detection cameras to characterize the dispersal of Triatoma infestans, and compare the behavior of vector populations in the constant presence of hosts (guinea pigs), and after the removal of the hosts. The emigration rate – net insect population decline in original refuge – following host removal was on average 19.7% of insects per 10 days compared to 10.2% in constant host populations (p = 0.029). However, dispersal of T. infestans occurred in both directions, towards and away from the initial location of the hosts. The majority of insects that moved towards the original location of guinea pigs remained there for 4 weeks. Oviposition and mortality were observed and analyzed in the context of insect dispersal, but only mortality was higher in the group where animal hosts were removed (p-value <0.01). We discuss different survival strategies associated with the observed behavior and its implications for vector control. Removing domestic animals in infested areas increases vector dispersal from the first day of host removal. The implications of these patterns of vector dispersal in a field setting are not yet known but could result in movement towards human rooms. PMID:25569228

  20. Evidence for homoploid speciation in Phytophthora alni supports taxonomic reclassification in this species complex.

    PubMed

    Husson, C; Aguayo, J; Revellin, C; Frey, P; Ioos, R; Marçais, B

    2015-04-01

    Alder decline has been a problem along European watercourses since the early 1990s. Hybridization was identified as the main cause of this emerging disease. Indeed, the causal agent, a soil-borne pathogen named Phytophthora alni subsp. alni (Paa) is the result of interspecific hybridization between two taxa, Phytophthora alni subsp. multiformis (Pam) and Phytophthora alni subsp. uniformis (Pau), initially identified as subspecies of Paa. The aim of this work was to characterize the ploidy level within the P. alni complex that is presently poorly understood. For that, we used two complementary approaches for a set of 31 isolates of Paa, Pam and Pau: (i) quantification of allele copy number of three single-copy nuclear genes using allele-specific real-time PCR and (ii) comparison of the genome size estimated by flow cytometry. Relative quantification of alleles of the three single-copy genes showed that the copy number of a given allele in Paa was systematically half that of its parents Pau or Pam. Moreover, DNA content estimated by flow cytometry in Paa was equal to half the sum of those in Pam and Pau. Our results therefore suggest that the hybrid Paa is an allotriploid species, containing half of the genome of each of its parents Pam and Pau, which in turn are considered to be allotetraploid and diploid, respectively. Paa thus results from a homoploid speciation process. Based on published data and on results from this study, a new formal taxonomic name is proposed for the three taxa Paa, Pam and Pau which are raised to species status and renamed P. ×alni, P. ×multiformis and P. uniformis, respectively.

  1. Oxygen stress reduces zoospore survival of Phytophthora species in a simulated aquatic system

    PubMed Central

    2014-01-01

    Background The genus Phytophthora includes a group of agriculturally important pathogens and they are commonly regarded as water molds. They produce motile zoospores that can move via water currents and on their own locomotion in aquatic environments. However, zoosporic response to dissolved oxygen, an important water quality parameter, is not known. Like other water quality parameters, dissolved oxygen concentration in irrigation reservoirs fluctuates dramatically over time. The aim of this study was to determine whether and how zoospore survival may be affected by elevated and low concentrations of dissolved oxygen in water to better understand the aquatic biology of these pathogens in irrigation reservoirs. Results Zoospores of P. megasperma, P. nicotianae, P. pini and P. tropicalis were assessed for survival in 10% Hoagland’s solution at a range of dissolved concentrations from 0.9 to 20.1 mg L-1 for up to seven exposure times from 0 to 72 h. Zoospore survival was measured by resultant colony counts per ml. Zoospores of these species survived the best in control Hoagland’s solution at dissolved oxygen concentrations of 5.3 to 5.6 mg L-1. Zoospore survival rates decreased with increasing and decreasing concentration of dissolved oxygen, depending upon Phytophthora species and exposure time. Overall, P. megasperma and P. pini are less sensitive than P. nicotianae and P. tropicalis to hyperoxia and hypoxia conditions. Conclusion Zoospores in the control solution declined over time and this natural decline process was enhanced under hyperoxia and hypoxia conditions. These findings suggest that dramatic fluctuations of dissolved oxygen in irrigation reservoirs contribute to the population decline of Phytophthora species along the water path in the same reservoirs. These findings advanced our understanding of the aquatic ecology of these pathogens in irrigation reservoirs. They also provided a basis for pathogen risk mitigation by prolonging the turnover

  2. Phylogenetic relationship of Phytophthora cryptogea Pethybr. & Laff and P. drechsleri Tucker.

    PubMed

    Mostowfizadeh-Ghalamfarsa, R; Panabieres, F; Banihashemi, Z; Cooke, D E L

    2010-04-01

    The phylogeny and taxonomy of Phytophthora cryptogea and Phytophthora drechsleri has long been a matter of controversy. To re-evaluate this, a worldwide collection of 117 isolates assigned to either P. cryptogea, P. drechsleri or their sister taxon, Phytophthora erythroseptica were assessed for morphological, physiological (pathological, cultural, temperature relations, mating) and molecular traits. Multiple gene phylogenetic analysis was performed on DNA sequences of nuclear (internal transcribed spacers (ITS), ß-tubulin, translation elongation factor 1α, elicitin) and mitochondrial (cytochrome c oxidase subunit I) genes. Congruence was observed between the different phylogenetic data sets and established that P. drechsleri and P. cryptogea are distinct species. Isolates of P. drechsleri form a monophyletic grouping with low levels of intraspecific diversity whereas P. cryptogea is more variable. Three distinct phylogenetic groups were noted within P. cryptogea with an intermediate group providing strong evidence for introgression of previously isolated lineages. This evidence suggests that P. cryptogea is an operational taxonomic unit and should remain a single species. Of all the morphological and physiological traits only growth rate at higher temperatures reliably discriminated isolates of P. drechsleri and P. cryptogea. As a homothallic taxon, P. erythroseptica, considered the cause of potato pink rot, is clearly different in mating behaviour from the other two species. Pathogenicity, however, was not a reliable characteristic as all isolates of the three species formed pink rot in potato tubers. The phylogenetic evidence suggests P. erythroseptica has evolved from P. cryptogea more recently than the split from the most recent common ancestor of all three species. However, more data and more isolates of authentic P. erythroseptica are needed to fully evaluate the taxonomic position of this species.

  3. PIXE studies of changes in mineral composition of plants infected with Phytophthora cinnamomi

    NASA Astrophysics Data System (ADS)

    Chaudhri, M. Anwar; Papper, C. S.; Weste, G.

    1981-03-01

    The mineral composition of susceptible and resistant plants from native forests infected with Phytophthora cinnamomi was compared between themselves and with the same species from disease-free areas. Root and shoot samples from different plants were carefully ashed, compressed into pellets and analysed with the thick target PIXE technique. A number of elements, Ca, Ti, Mn, Fe, Ni, Cu, Zn, Rb, Sr, Zr, Sn and Pb, were detected. Many of the elements showed variations, sometimes large, between the composition of susceptible and resistant plants, and between the same species from infected and disease-free forests.

  4. Risk of transmission of Trypanosoma cruzi by wild Triatoma infestans (Hemiptera: Reduviidae) in Bolivia supported by the detection of human blood meals.

    PubMed

    Buitrago, Nelly Lilian Rosio; Bosseno, Marie France; Waleckx, Etienne; Brémond, Philippe; Vidaurre, Pablo; Zoveda, Faustine; Brenière, Simone Frédérique

    2013-10-01

    We analyzed the food sources of Bolivian wild Triatoma infestans (the main vector of Chagas disease in this country), to assess the role of these populations in the epidemiological context of Chagas disease. Ninety-eight blood meals were identified by heteroduplex assay and sequencing. Most of them were from wild mammals but surprisingly 27 were from humans. This brings to light the occurrence of human-vector contacts at risk of Trypanosoma cruzi transmission in the wild environment by highly infected insects.

  5. Response of U.S. Bottle Gourd (Lagenaria siceraria) Plant Introductions (PI) to Crown Rot caused by Phytophthora Capsici

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora capsici can cause severe damage to cucurbit crops grown in open fields in the southeast regions of US. In recent years, there has been a growing interest in the US in grafting watermelon plants onto various cucurbit rootstocks including bottle gourds for managing soil borne diseases. ...

  6. Evaluation of molecular markers for Phytophthora ramorum detection and identification; testing for specificity using a standardized library of isolates.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A number of molecular techniques have been developed for detection of Phytophthora ramorum from infected tissue. These have been based on spacer regions (the rDNA ITS region, the spacer region between the cox I and II gene) or specific genes (beta tubulin, elicitin) and have been configured for use ...

  7. Effects of Phytophthora cinnamomi isolate, inoculum delivery method, flood, and drought on vigor, disease severity and mortality of blueberry plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Four studies evaluated the effects of cultivar, inoculum delivery method, flood, and drought on plant vigor, disease severity, and mortality of blueberry plants grown in pots in the greenhouse. Phytophthora cinnamomi isolates were obtained from the root zone of blueberry plants displaying symptoms...

  8. Real-time PCR assay to distinguish the four Phytophthora ramorum lineages using cellulose binding elicitor lectin (CBEL) locus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora ramorum is a pathogenic oomycete responsible for causing sudden oak death in the Western United States and sudden larch death in the United Kingdom. This pathogen has so far caused extensive mortality of oak and tanoak in California and of Japanese larch in the United Kingdom. Until rec...

  9. Biological control of Black Pod Disease and Seedling Blight of cacao caused by Phytophthora Species using Trichoderma from Aceh Sumatra

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The cocoa tree, Theobroma cacao L., suffers large yield losses in Aceh Indonesia to the disease black pod rot, caused by Phytophthora spp. Despite having the largest area under cacao production in Sumatra, farmers in the Aceh region have low overall production because of losses to insect pests and b...

  10. Pre- and post-harvest development of Phytophthora fruit rot on watermelons treated with fungicides in the field

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fruit rot, caused by Phytophthora capsici, is a serious disease in most watermelon producing regions in southeastern U.S., and has caused devastating loss over the past few years. In many instances, severe losses occurred after harvest during transportation. Experiments were conducted in 2010, 201...

  11. Improved Phytophthora resistance in commercial chickpea (Cicer arietinum) varieties negatively impacts symbiotic gene signalling and symbiotic potential in some varieties.

    PubMed

    Plett, Jonathan M; Plett, Krista L; Bithell, Sean L; Mitchell, Chris; Moore, Kevin; Powell, Jeff R; Anderson, Ian C

    2016-08-01

    Breeding disease-resistant varieties is one of the most effective and economical means to combat soilborne diseases in pulse crops. Commonalities between pathogenic and mutualistic microbe colonization strategies, however, raises the concern that reduced susceptibility to pathogens may simultaneously reduce colonization by beneficial microbes. We investigate here the degree of overlap in the transcriptional response of the Phytophthora medicaginis susceptible chickpea variety 'Sonali' to the early colonization stages of either Phytophthora, rhizobial bacteria or arbuscular mycorrhizal fungi. From a total of 6476 genes differentially expressed in Sonali roots during colonization by any of the microbes tested, 10.2% were regulated in a similar manner regardless of whether it was the pathogenic oomycete or a mutualistic microbe colonizing the roots. Of these genes, 49.7% were oppositely regulated under the same conditions in the moderately Phytophthora resistant chickpea variety 'PBA HatTrick'. Chickpea varieties with improved resistance to Phytophthora also displayed lower colonization by rhizobial bacteria and mycorrhizal fungi leading to an increased reliance on N and P from soil. Together, our results suggest that marker-based breeding in crops such as chickpea should be further investigated such that plant disease resistance can be tailored to a specific pathogen without affecting mutualistic plant:microbe interactions.

  12. First report of the EU1 clonal lineage of Phytophthora ramorum on tanoak in an Oregon forest

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Initially reported in California as the causal agent of sudden oak death (SOD), efforts to limit spread of Phytophthora ramorum in Oregon natural forests have concentrated on quarantine regulations and eradication of the pathogen from infested areas. P. ramorum has four clonal lineages NA1, NA2, EU1...

  13. Characterization of fungi (Fusarium and Rhizoctonia) and oomycetes (Phytophthora and Pythium) associated with apple orchards in South Africa.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Several species of fungi and oomycetes including Fusarium, Rhizoctonia, Phytophthora and Pythium have been reported as root pathogens of apple where they contribute to a phenomenon known as apple replant disease. In South Africa, very little is known about the specific species in these genera and th...

  14. Improved Phytophthora resistance in commercial chickpea (Cicer arietinum) varieties negatively impacts symbiotic gene signalling and symbiotic potential in some varieties.

    PubMed

    Plett, Jonathan M; Plett, Krista L; Bithell, Sean L; Mitchell, Chris; Moore, Kevin; Powell, Jeff R; Anderson, Ian C

    2016-08-01

    Breeding disease-resistant varieties is one of the most effective and economical means to combat soilborne diseases in pulse crops. Commonalities between pathogenic and mutualistic microbe colonization strategies, however, raises the concern that reduced susceptibility to pathogens may simultaneously reduce colonization by beneficial microbes. We investigate here the degree of overlap in the transcriptional response of the Phytophthora medicaginis susceptible chickpea variety 'Sonali' to the early colonization stages of either Phytophthora, rhizobial bacteria or arbuscular mycorrhizal fungi. From a total of 6476 genes differentially expressed in Sonali roots during colonization by any of the microbes tested, 10.2% were regulated in a similar manner regardless of whether it was the pathogenic oomycete or a mutualistic microbe colonizing the roots. Of these genes, 49.7% were oppositely regulated under the same conditions in the moderately Phytophthora resistant chickpea variety 'PBA HatTrick'. Chickpea varieties with improved resistance to Phytophthora also displayed lower colonization by rhizobial bacteria and mycorrhizal fungi leading to an increased reliance on N and P from soil. Together, our results suggest that marker-based breeding in crops such as chickpea should be further investigated such that plant disease resistance can be tailored to a specific pathogen without affecting mutualistic plant:microbe interactions. PMID:27103212

  15. Phytophthora species recovered from irrigation reservoirs in Mississippi and Alabama nurseries and pathogenicity of three new species

    Technology Transfer Automated Retrieval System (TEKTRAN)

    From a survey of containment ponds for Phytophthora spp. at one nursery each in Alabama and Mississippi, eight species and one taxon were recovered with P. gonapodyides dominant in cooler months and P. hydropathica in warmer months, accounting for 39.6% and 46.6% overall recovery, respectively. Amo...

  16. Phytophthora species recovered from irrigation reservoirs in Mississippi and Alabama nurseries and pathogenicity of three new species.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    From a survey of containment ponds for Phytophthora spp. at one nursery each in Alabama and Mississippi, eight species and one taxon were recovered with P. gonapodyides dominant in cooler months and P. hydropathica in warmer months, accounting for 39.6% and 46.6% overall recovery, respectively. Amo...

  17. Joint QTL analyses for partial resistance to Phytophthora sojae using six nested inbred populations with heterogeneous conditions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Partial resistance to Phytophthora sojae in soybean is controlled by multiple quantitative trait loci (QTL). With traditional QTL mapping approaches, power to detect these QTL, frequently of small effect, can be limited by population size. Joint linkage QTL analysis of nested recombinant inbred li...

  18. L925I Mutation in the Para-Type Sodium Channel Is Associated with Pyrethroid Resistance in Triatoma infestans from the Gran Chaco Region

    PubMed Central

    Capriotti, Natalia; Mougabure-Cueto, Gastón; Rivera-Pomar, Rolando; Ons, Sheila

    2014-01-01

    Background Chagas' disease is an important public health concern in Latin America. Despite intensive vector control efforts using pyrethroid insecticides, the elimination of Triatoma infestans has failed in the Gran Chaco, an ecoregion that extends over Argentina, Paraguay, Bolivia and Brazil. The voltage-gated sodium channel is the target site of pyrethroid insecticides. Point mutations in domain II region of the channel have been implicated in pyrethroid resistance of several insect species. Methods and Findings In the present paper, we identify L925I, a new pyrethroid resistance-conferring mutation in T. infestans. This mutation has been found only in hemipterans. In T. infestans, L925I mutation occurs in a resistant population from the Gran Chaco region and is associated with inefficiency in the control campaigns. We also describe a method to detect L925I mutation in individuals from the field. Conclusions and Significance The findings have important implications in the implementation of strategies for resistance management and in the rational design of campaigns for the control of Chagas' disease transmission. PMID:24466362

  19. Is imidacloprid an effective alternative for controlling pyrethroid-resistant populations of Triatoma infestans (Hemiptera: Reduviidae) in the Gran Chaco ecoregion?

    PubMed Central

    Carvajal, Guillermo; Picollo, María Inés; Toloza, Ariel Ceferino

    2014-01-01

    The prevention of Chagas disease is based primarily on the chemical control of Triatoma infestans (Klug) using pyrethroid insecticides. However, high resistance levels, correlated with control failures, have been detected in Argentina and Bolivia. A previous study at our laboratory found that imidacloprid could serve as an alternative to pyrethroid insecticides. We studied the delayed toxicity of imidacloprid and the influence of the blood feeding condition of the insect on the toxicity of this insecticide; we also studied the effectiveness of various commercial imidacloprid formulations against a pyrethroid-resistant T. infestans population from the Gran Chaco ecoregion. Variations in the toxic effects of imidacloprid were not observed up to 72 h after exposure and were not found to depend on the blood feeding condition of susceptible and resistant individuals. Of the three different studied formulations of imidacloprid on glass and filter paper, only the spot-on formulation was effective. This formulation was applied to pigeons at doses of 1, 5, 20 and 40 mg/bird. The nymphs that fed on pigeons treated with 20 mg or 40 mg of the formulation showed a higher mortality rate than the control group one day and seven days post-treatment (p < 0.01). A spot-on formulation of imidacloprid was effective against pyrethroid-resistant T. infestans populations at the laboratory level. PMID:25141281

  20. Microbiota Characterization of Compost Using Omics Approaches Opens New Perspectives for Phytophthora Root Rot Control.

    PubMed

    Blaya, Josefa; Marhuenda, Frutos C; Pascual, Jose A; Ros, Margarita

    2016-01-01

    Phytophthora root rot caused by Phytophthora nicotianae is an economically important disease in pepper crops. The use of suppressive composts is a low environmental impact method for its control. Although attempts have been made to reveal the relationship between microbiota and compost suppressiveness, little is known about the microorganisms associated with disease suppression. Here, an Ion Torrent platform was used to assess the microbial composition of composts made of different agro-industrial waste and with different levels of suppressiveness against P. nicotianae. Both bacterial and fungal populations responded differently depending on the chemical heterogeneity of materials used during the composting process. High proportions (67-75%) of vineyard pruning waste were used in the most suppressive composts, COM-A and COM-B. This material may have promoted the presence of higher relative abundance of Ascomycota as well as higher microbial activity, which have proved to be essential for controlling the disease. Although no unique fungi or bacteria have been detected in neither suppressive nor conducive composts, relatively high abundance of Fusarium and Zopfiella were found in compost COM-B and COM-A, respectively. To the best of our knowledge, this is the first work that studies compost metabolome. Surprisingly, composts and peat clustered together in principal component analysis of the metabolic data according to their levels of suppressiveness achieved. This study demonstrated the need for combining the information provided by different techniques, including metagenomics and metametabolomics, to better understand the ability of compost to control plant diseases.

  1. A test system to quantify inoculum in runoff from Phytophthora ramorum-infected plant roots.

    PubMed

    Shishkoff, Nina

    2011-12-01

    Foliar hosts of Phytophthora ramorum are often susceptible to root infection but the epidemiological significance of such infections is unknown. A standardized test system was developed to quantify inoculum in runoff from root-infected Viburnum tinus ?Spring Bouquet? or Rhododendron ?Cunningham's White? cuttings. Cuttings of both species gave off a maximum amount of inoculum 1 to 3 weeks after inoculation. The greatest amount of inoculum was recovered from Viburnum roots that were 48 to 70 days old at the time of inoculation, or roots incubated at 15 to 20?C rather than 25?C. Inoculum in runoff from inoculated Viburnum roots was similar for four different isolates of P. ramorum representing both the NA1 and EU1 lineages. When Rhododendron cuttings were inoculated with P. ramorum, P. citricola, or P. cactorum, inoculum of all three pathogens was recovered from runoff, with the highest amount recovered from plants inoculated with P. citricola, followed by the other two. Compared with the other two pathogens, P. ramorum colonized root tissue to a smaller extent. The epidemiology of root infection by P. ramorum is important in itself but the assay might lend itself for use in risk analysis for root infection of other plant species and evaluation of control measures, and also shed light on other root-infecting Phytophthora spp.

  2. Influence of electron beam irradiation on growth of Phytophthora cinnamomi and its control in substrates

    NASA Astrophysics Data System (ADS)

    MigdaŁ, Wojciech; Orlikowski, Leszek B.; Ptaszek, Magdalena; Gryczka, Urszula

    2012-08-01

    Very extensive production procedure, especially in plants growing under covering, require methods, which would allow quick elimination or substantial reduction of populations of specific pathogens without affecting the growth and development of the cultivated plants. Among soil-borne pathogens, the Phytophthora species are especially dangerous for horticultural plants. In this study, irradiation with electron beam was applied to control Phytophthora cinnamomi. The influence of irradiation dose on the reduction of in vitro growth and the population density of the pathogen in treated peat and its mixture with composted pine bark (1:1), as well as the health of Chamaecyparis lawsoniana and Lavandula angustifolia plants were evaluated. Application of irradiation at a dose of 1.5 kGy completely inhibited the in vitro development of P. cinnamomi. This irradiation effect was connected with the disintegration of the hyphae and spores of the species. Irradiation of peat and its mixture with composted pine bark with 10 kGy resulted in the inhibition of stem base rot development in Ch. lawsoniana. Symptoms of the disease were not observed when the substrates were treated with 15 kGy. In the case of L. angustifolia, stem root rot was not observed on cuttings transplanted to infected peat irradiated at a dose of 10 kGy. Irradiation of the horticultural substrates did not affect plant growth.

  3. Microbiota Characterization of Compost Using Omics Approaches Opens New Perspectives for Phytophthora Root Rot Control.

    PubMed

    Blaya, Josefa; Marhuenda, Frutos C; Pascual, Jose A; Ros, Margarita

    2016-01-01

    Phytophthora root rot caused by Phytophthora nicotianae is an economically important disease in pepper crops. The use of suppressive composts is a low environmental impact method for its control. Although attempts have been made to reveal the relationship between microbiota and compost suppressiveness, little is known about the microorganisms associated with disease suppression. Here, an Ion Torrent platform was used to assess the microbial composition of composts made of different agro-industrial waste and with different levels of suppressiveness against P. nicotianae. Both bacterial and fungal populations responded differently depending on the chemical heterogeneity of materials used during the composting process. High proportions (67-75%) of vineyard pruning waste were used in the most suppressive composts, COM-A and COM-B. This material may have promoted the presence of higher relative abundance of Ascomycota as well as higher microbial activity, which have proved to be essential for controlling the disease. Although no unique fungi or bacteria have been detected in neither suppressive nor conducive composts, relatively high abundance of Fusarium and Zopfiella were found in compost COM-B and COM-A, respectively. To the best of our knowledge, this is the first work that studies compost metabolome. Surprisingly, composts and peat clustered together in principal component analysis of the metabolic data according to their levels of suppressiveness achieved. This study demonstrated the need for combining the information provided by different techniques, including metagenomics and metametabolomics, to better understand the ability of compost to control plant diseases. PMID:27490955

  4. Microbiota Characterization of Compost Using Omics Approaches Opens New Perspectives for Phytophthora Root Rot Control

    PubMed Central

    Blaya, Josefa; Marhuenda, Frutos C.; Pascual, Jose A.; Ros, Margarita

    2016-01-01

    Phytophthora root rot caused by Phytophthora nicotianae is an economically important disease in pepper crops. The use of suppressive composts is a low environmental impact method for its control. Although attempts have been made to reveal the relationship between microbiota and compost suppressiveness, little is known about the microorganisms associated with disease suppression. Here, an Ion Torrent platform was used to assess the microbial composition of composts made of different agro-industrial waste and with different levels of suppressiveness against P. nicotianae. Both bacterial and fungal populations responded differently depending on the chemical heterogeneity of materials used during the composting process. High proportions (67–75%) of vineyard pruning waste were used in the most suppressive composts, COM-A and COM-B. This material may have promoted the presence of higher relative abundance of Ascomycota as well as higher microbial activity, which have proved to be essential for controlling the disease. Although no unique fungi or bacteria have been detected in neither suppressive nor conducive composts, relatively high abundance of Fusarium and Zopfiella were found in compost COM-B and COM-A, respectively. To the best of our knowledge, this is the first work that studies compost metabolome. Surprisingly, composts and peat clustered together in principal component analysis of the metabolic data according to their levels of suppressiveness achieved. This study demonstrated the need for combining the information provided by different techniques, including metagenomics and metametabolomics, to better understand the ability of compost to control plant diseases. PMID:27490955

  5. Purification and characterization of elicitor protein from Phytophthora colocasiae and basic resistance in Colocasia esculenta.

    PubMed

    Mishra, Ajay Kumar; Sharma, Kamal; Misra, Raj Shekhar

    2009-01-01

    An elicitor was identified in the fungus Phytophthora colocasiae. The molecular weight of the purified elicitor was estimated by means of gel filtration chromatography and SDS-PAGE and was estimated as 15kDa. Protease treatment severely reduced its activity, allowing the conclusion that the elicitor is proteinaceous. Infiltration of a few nanograms of this proteinaceous elicitor into taro leaves caused the formation of lesions that closely resemble hypersensitive response lesions. The elicitation of the cells was effective in the induction of the activity of lipoxygenase. Cellular damage, restricted to the infiltrated zone, occurred only several hours later, after the infiltration of the elicitor protein. After few days, systemic acquired resistance was also induced. Thus, taro plant cells that perceived the glycoprotein generated a cascade of signals acting at local, short, and long distances, and causing the coordinate expression of specific defence. The obtained results give important information regarding the plant-pathogen interactions, mainly as subsidy for taro improvement against Phytophthora leaf blight.

  6. Phytophthora capsici epidemic dispersion on commercial pepper fields in Aguascalientes, Mexico.

    PubMed

    Zapata-Vázquez, Adrián; Sánchez-Sánchez, Mario; del-Río-Robledo, Alicia; Silos-Espino, Héctor; Perales-Segovia, Catarino; Flores-Benítez, Silvia; González-Chavira, Mario Martín; Valera-Montero, Luis Lorenzo

    2012-01-01

    Chili pepper blight observed on pepper farms from north Aguascalientes was monitored for the presence of Phytophthora capsici during 2008-2010. Initially, ELISA tests were directed to plant samples from greenhouses and rustic nurseries, showing an 86% of positive samples. Later, samples of wilted plants from the farms during the first survey were tested with ELISA. The subsequent survey on soil samples included mycelia isolation and PCR amplification of a 560 bp fragment of ITS-specific DNA sequence of P. capsici. Data was analyzed according to four geographical areas defined by coordinates to ease the dispersal assessment. In general, one-third of the samples from surveyed fields contained P. capsici, inferring that this may be the pathogen responsible of the observed wilt. Nevertheless, only five sites from a total of 92 were consistently negative to P. capsici. The presence of this pathogen was detected through ELISA and confirmed through PCR. The other two-thirds of the negative samples may be attributable to Fusarium and Rhizoctonia, both isolated instead of Phytophthora in these areas. Due to these striking results, this information would be of interest for local plant protection committees and farmers to avoid further dispersal of pathogens to new lands. PMID:22629131

  7. Efficacy of Chaetomium Species as Biological Control Agents against Phytophthora nicotianae Root Rot in Citrus

    PubMed Central

    Wattanachai, Pongnak; Kasem, Soytong; Poeaim, Supattra

    2015-01-01

    Thailand is one of the largest citrus producers in Southeast Asia. Pathogenic infection by Phytophthora, however, has become one of major impediments to production. This study identified a pathogenic oomycete isolated from rotted roots of pomelo (Citrus maxima) in Thailand as Phytophthora nicotianae by the internal transcribed spacer ribosomal DNA sequence analysis. Then, we examined the in vitro and in vivo effects of Chaetomium globosum, Chaetomium lucknowense, Chaetomium cupreum and their crude extracts as biological control agents in controlling this P. nicotianae strain. Represent as antagonists in biculture test, the tested Chaetomium species inhibited mycelial growth by 50~56% and parasitized the hyphae, resulting in degradation of P. nicotianae mycelia after 30 days. The crude extracts of these Chaetomium species exhibited antifungal activities against mycelial growth of P. nicotianae, with effective doses of 2.6~101.4 µg/mL. Under greenhouse conditions, application of spores and methanol extracts of these Chaetomium species to pomelo seedlings inoculated with P. nicotianae reduced root rot by 66~71% and increased plant weight by 72~85% compared to that in the control. The method of application of antagonistic spores to control the disease was simple and economical, and it may thus be applicable for large-scale, highly effective biological control of this pathogen. PMID:26539045

  8. Suppression of phytophthora blight in bell pepper by a no-till wheat cover crop.

    PubMed

    Ristaino, J B; Parra, G; Campbell, C L

    1997-03-01

    ABSTRACT Four mechanisms of dispersal of propagules of Phytophthora capsici were investigated through modifications in cultural practices and fungicide applications in field plots of bell pepper (Capsicum annuum). Dispersal of soil inoculum was suppressed, and final incidence of disease was 2.5 to 43% when stubble from a fall-sown, no-till, wheat cover crop was present. Final disease incidence was 71 to 72% and pathogen spread occurred within and across rows when all dispersal mechanisms were operative in plots of pepper planted into bare soil. Final disease incidence was 42 to 78% with black plastic mulch when a sporulating pepper fruit placed on the surface served as the source of initial inoculum. The fungicide metalaxyl applied in the irrigation system did not suppress within-row spread of surface inoculum from a sporulating fruit on plastic, but did limit across-row spread; final disease incidence in metalaxyl-treated plots was 11.5 to 14%. Pathogen dispersal mechanisms were modified most dramatically by the no-till cropping system. Thus, simple changes in cultural practices can have dramatic effects on the development of Phytophthora epidemics. Ecologically based disease management strategies have the potential to reduce our reliance on agrichemicals in this and similar pathosystems.

  9. Phytophthora capsici Epidemic Dispersion on Commercial Pepper Fields in Aguascalientes, Mexico

    PubMed Central

    Zapata-Vázquez, Adrián; Sánchez-Sánchez, Mario; del-Río-Robledo, Alicia; Silos-Espino, Héctor; Perales-Segovia, Catarino; Flores-Benítez, Silvia; González-Chavira, Mario Martín; Valera-Montero, Luis Lorenzo

    2012-01-01

    Chili pepper blight observed on pepper farms from north Aguascalientes was monitored for the presence of Phytophthora capsici during 2008–2010. Initially, ELISA tests were directed to plant samples from greenhouses and rustic nurseries, showing an 86% of positive samples. Later, samples of wilted plants from the farms during the first survey were tested with ELISA. The subsequent survey on soil samples included mycelia isolation and PCR amplification of a 560 bp fragment of ITS-specific DNA sequence of P. capsici. Data was analyzed according to four geographical areas defined by coordinates to ease the dispersal assessment. In general, one-third of the samples from surveyed fields contained P. capsici, inferring that this may be the pathogen responsible of the observed wilt. Nevertheless, only five sites from a total of 92 were consistently negative to P. capsici. The presence of this pathogen was detected through ELISA and confirmed through PCR. The other two-thirds of the negative samples may be attributable to Fusarium and Rhizoctonia, both isolated instead of Phytophthora in these areas. Due to these striking results, this information would be of interest for local plant protection committees and farmers to avoid further dispersal of pathogens to new lands. PMID:22629131

  10. Pattern swimming of Phytophthora citricola zoospores: an example of microbial bioconvection.

    PubMed

    Ochiai, Naoyuki; Dragiila, Maria Ines; Parke, Jennifer L

    2011-03-01

    The genus Phytophthora, belonging to the class Oomycota, comprises a group of over fifty fungus-like plant pathogens in both managed and unmanaged ecosystems. A unique feature of the oomycete lifecycle is a zoosporic stage in which motile, unicellular propagules, serving as the primary agents of dispersal, are produced and released in the presence of water. In Petri dish suspensions, zoospores frequently exhibit 'pattern swimming', whereby they spontaneously form concentrated swimming masses, visible to the naked eye, even in the absence of a chemical or electrical gradient. The nature of this behaviour is unclear, but is of interest because of the potential for auto-attraction and implications for cohort recruitment during infection. Similar behaviour observed in a variety of motile bacteria, algae, and protists is attributed to 'bioconvection' that results from instability in fluid density due to the organisms' upward-swimming tendency and greater-than-water density. In this investigation, we determined that Phytophthora citricola zoospore 'pattern swimming' is unrelated to phototaxis, surface tension-driven (Marangoni) convection, or auto-attraction and that the observed convective pattern, directional swimming, and depth- and concentration dependence are consistent with bioconvection.

  11. Cloning and characterization of cDNA encoding an elicitor of Phytophthora colocasiae.

    PubMed

    Mishra, Ajay Kumar; Sharma, Kamal; Misra, Raj Shekhar

    2010-02-28

    The rapid and effective activation of disease resistance responses is essential for plant defense against pathogen attack. These responses are initiated when pathogen-derived molecules (elicitors) are recognized by the host. A cDNA encoding elicitor, the major secreted extracellular glycoprotein of Phytophthora colocasiae, a pathogen of taro (Colocasia esculenta) plants, was isolated, sequenced and characterized. The expression of the corresponding elicitor gene during the disease cycle of P. colocasiae was analyzed. Elicitor was shown to be expressed in mycelium grown in culture media, whereas it was not expressed in sporangiospores and zoospores. In planta, during infection of taro, particularly during the biotrophic stage, expression of elicitor was down-regulated compared to in vitro. The highest levels of expression of elicitor were observed in in vitro grown mycelium and in late stages of infection when profuse sporulation and leaf necrosis occur. The elicitation of the suspension-cultured taro cells was effective in the induction of the enzyme activity of l-phenylalanine-ammonia lyase, peroxidase and lipoxygenase as well as the expression of defense-related endochitinase gene. All these biological activities were exerted within a low concentration range. The glycoprotein represents a powerful tool to investigate further the signals and their transduction pathways involved in induced disease resistance. It may also be useful to engineer broad disease protection in taro plant against Phytophthora leaf blight. PMID:19230634

  12. Transport and Retention of Phytophthora capsici Zoospores in Saturated Porous Media.

    PubMed

    Jeon, Sangho; Krasnow, Charles S; Kirby, Caitlin K; Granke, Leah L; Hausbeck, Mary K; Zhang, Wei

    2016-09-01

    Phytophthora capsici is an important plant pathogen capable of infecting several major vegetable crops. Water-induced P. capsici transport is considered to be a significant contributor to disease outbreaks and subsequent crop loss. However, little is known about factors controlling P. capsici zoospore transport in porous media, thus impeding our understanding of their environmental dispersal and development of filtration techniques for contaminated irrigation water. This study investigated the transport and retention of P. capsici zoospores in saturated columns packed with iron-oxide-coated sand (IOCS) or uncoated sand in Na(+) or Ca(2+) background solution at pH 7.7 ± 0.5 or 4.0 ± 0.3, in combination with XDLVO interaction energy calculations and microscopic visualizations. Significantly more encysted zoospores were retained in IOCS than in uncoated sand, and at pH 4.0 than at pH 7.7, which likely resulted from increased electrostatic attraction between zoospores and grain surface. At pH 7.7, up to 99% and 96% of the encysted zoospores were removed in IOCS and uncoated sand, respectively, due to a combination of strong surface attachment, pore straining, and adhesive interactions. Motile biflagellate zoospores were more readily transported than encysted zoospores, thus posing a greater dispersal and infection risk. This study has broad implications in environmental transport of Phytophthora zoospores in natural soils as well as in cost-effective engineered filtration systems. PMID:27517718

  13. Domestic Animal Hosts Strongly Influence Human-Feeding Rates of the Chagas Disease Vector Triatoma infestans in Argentina

    PubMed Central

    Gürtler, Ricardo E.; Cecere, María C.; Vázquez-Prokopec, Gonzalo M.; Ceballos, Leonardo A.; Gurevitz, Juan M.; Fernández, María del Pilar; Kitron, Uriel; Cohen, Joel E.

    2014-01-01

    Background The host species composition in a household and their relative availability affect the host-feeding choices of blood-sucking insects and parasite transmission risks. We investigated four hypotheses regarding factors that affect blood-feeding rates, proportion of human-fed bugs (human blood index), and daily human-feeding rates of Triatoma infestans, the main vector of Chagas disease. Methods A cross-sectional survey collected triatomines in human sleeping quarters (domiciles) of 49 of 270 rural houses in northwestern Argentina. We developed an improved way of estimating the human-feeding rate of domestic T. infestans populations. We fitted generalized linear mixed-effects models to a global model with six explanatory variables (chicken blood index, dog blood index, bug stage, numbers of human residents, bug abundance, and maximum temperature during the night preceding bug catch) and three response variables (daily blood-feeding rate, human blood index, and daily human-feeding rate). Coefficients were estimated via multimodel inference with model averaging. Findings Median blood-feeding intervals per late-stage bug were 4.1 days, with large variations among households. The main bloodmeal sources were humans (68%), chickens (22%), and dogs (9%). Blood-feeding rates decreased with increases in the chicken blood index. Both the human blood index and daily human-feeding rate decreased substantially with increasing proportions of chicken- or dog-fed bugs, or the presence of chickens indoors. Improved calculations estimated the mean daily human-feeding rate per late-stage bug at 0.231 (95% confidence interval, 0.157–0.305). Conclusions and Significance Based on the changing availability of chickens in domiciles during spring-summer and the much larger infectivity of dogs compared with humans, we infer that the net effects of chickens in the presence of transmission-competent hosts may be more adequately described by zoopotentiation than by zooprophylaxis

  14. Label-free detection of Phytophthora ramorum using surface-enhanced Raman spectroscopy.

    PubMed

    Yüksel, Sezin; Schwenkbier, Lydia; Pollok, Sibyll; Weber, Karina; Cialla-May, Dana; Popp, Jürgen

    2015-11-01

    In this study, we report on a novel approach for the label-free and species-specific detection of the plant pathogen Phytophthora ramorum from real samples using surface enhanced Raman scattering (SERS). In this context, we consider the entire analysis chain including sample preparation, DNA isolation, amplification and hybridization on SERS substrate-immobilized adenine-free capture probes. Thus, the SERS-based detection of target DNA is verified by the strong spectral feature of adenine which indicates the presence of hybridized target DNA. This property was realized by replacing adenine moieties in the species-specific capture probes with 2-aminopurine. In the case of the matching capture and target sequence, the characteristic adenine peak serves as an indicator for specific DNA hybridization. Altogether, this is the first assay demonstrating the detection of a plant pathogen from an infected plant material by label-free SERS employing DNA hybridization on planar SERS substrates consisting of silver nanoparticles.

  15. Tobacco Rotated with Rapeseed for Soil-Borne Phytophthora Pathogen Biocontrol: Mediated by Rapeseed Root Exudates.

    PubMed

    Fang, Yuting; Zhang, Limeng; Jiao, Yongge; Liao, Jingjing; Luo, Lifen; Ji, Sigui; Li, Jiangzhou; Dai, Kuai; Zhu, Shusheng; Yang, Min

    2016-01-01

    Black shank, caused by Phytophthora parasitica var. nicotianae, is a widespread and destructive disease of tobacco. Crop rotation is essential in controlling black shank. Here, we confirmed that rotating black shank-infested fields with rapeseed (Brassica napus) suppressed the incidence this disease. Further study demonstrated that rapeseed roots have a strong ability to attract zoospores and subsequently stop the swimming of zoospores into cystospores. Then, rapeseed roots secrete a series of antimicrobial compounds, including 2-butenoic acid, benzothiazole, 2-(methylthio)benzothiazole, 1-(4-ethylphenyl)-ethanone, and 4-methoxyindole, to inhibit the cystospore germination and mycelial growth of P. parasitica var. nicotianae. Thus, rapeseed rotated with tobacco suppresses tobacco black shank disease through the chemical weapons secreted by rapeseed roots. PMID:27379037

  16. Chemotactic Preferences and Strain Variation in the Response of Phytophthora sojae Zoospores to Host Isoflavones

    PubMed Central

    Tyler, B. M.; Wu, M.; Wang, J.; Cheung, W.; Morris, P. F.

    1996-01-01

    The zoospores of Phytophthora sojae are chemotactically attracted to the isoflavones genistein and daidzein that are released by soybean roots. In this study we have examined the response of P. sojae zoospores to a wide range of compounds having some structural similarity to genistein and daidzein, including isoflavones, flavones, chalcones, stilbenes, benzoins, benzoates, benzophenones, acetophenones, and coumarins. Of 59 compounds examined, 43 elicited some response. A comparison of the chemotactic responses elicited by the various compounds revealed a primary role for the phenolic 4(prm1)- and 7-hydroxyl groups on the isoflavone structure. A few compounds acted as repellents, notably methylated flavones with a hydrophobic B ring. The chemotactic response to many of the analogs was markedly different among different strains of P. sojae. PMID:16535375

  17. Tobacco Rotated with Rapeseed for Soil-Borne Phytophthora Pathogen Biocontrol: Mediated by Rapeseed Root Exudates

    PubMed Central

    Fang, Yuting; Zhang, Limeng; Jiao, Yongge; Liao, Jingjing; Luo, Lifen; Ji, Sigui; Li, Jiangzhou; Dai, Kuai; Zhu, Shusheng; Yang, Min

    2016-01-01

    Black shank, caused by Phytophthora parasitica var. nicotianae, is a widespread and destructive disease of tobacco. Crop rotation is essential in controlling black shank. Here, we confirmed that rotating black shank-infested fields with rapeseed (Brassica napus) suppressed the incidence this disease. Further study demonstrated that rapeseed roots have a strong ability to attract zoospores and subsequently stop the swimming of zoospores into cystospores. Then, rapeseed roots secrete a series of antimicrobial compounds, including 2-butenoic acid, benzothiazole, 2-(methylthio)benzothiazole, 1-(4-ethylphenyl)-ethanone, and 4-methoxyindole, to inhibit the cystospore germination and mycelial growth of P. parasitica var. nicotianae. Thus, rapeseed rotated with tobacco suppresses tobacco black shank disease through the chemical weapons secreted by rapeseed roots. PMID:27379037

  18. Acetylome analysis reveals the involvement of lysine acetylation in diverse biological processes in Phytophthora sojae

    PubMed Central

    Li, Delong; Lv, Binna; Tan, Lingling; Yang, Qianqian; Liang, Wenxing

    2016-01-01

    Lysine acetylation is a dynamic and highly conserved post-translational modification that plays an important regulatory role in almost every aspects of cell metabolism in both eukaryotes and prokaryotes. Phytophthora sojae is one of the most important plant pathogens due to its huge economic impact. However, to date, little is known about the functions of lysine acetylation in this Phytopthora. Here, we conducted a lysine acetylome in P. sojae. Overall, 2197 lysine acetylation sites in 1150 proteins were identified. The modified proteins are involved in diverse biological processes and are localized to multiple cellular compartments. Importantly, 7 proteins involved in the pathogenicity or the secretion pathway of P. sojae were found to be acetylated. These data provide the first comprehensive view of the acetylome of P. sojae and serve as an important resource for functional analysis of lysine acetylation in plant pathogens. PMID:27412925

  19. Toxicological and biochemical analysis of the susceptibility of sylvatic Triatoma infestans from the Andean Valley of Bolivia to organophosphate insecticide.

    PubMed

    Santo-Orihuela, Pablo Luis; Carvajal, Guillermo; Picollo, María Inés; Vassena, Claudia Viviana

    2013-09-01

    To increase our knowledge of the natural susceptibility of Triatoma infestans to an organophosphate insecticide, we performed toxicological and biochemical studies on three sylvatic populations from Bolivia and two populations from domestic dwellings from Bolivia and Argentina. Fifty-per-cent lethal doses (LD50) were determined based on the topical application of fenitrothion on first instar nymphs and mortality was assessed at 24 h. Both type of populations exhibited LD50ratios significantly higher than 1 with a range of the values (1.42-2.47); the maximum value were found in a sylvatic (-S) population, Veinte de Octubre-S. Samples were biochemically analysed using a glutathione S-transferase activity assay. The highest significant activity was obtained for Veinte de Octubre-S and the lowest activity was obtained for the reference population (102.69 and 54.23 pmol per minute per mg of protein respectively). Two out of the three sylvatic populations (Veinte de Octubre-S and Kirus Mayu-S) exhibited significantly higher glutathione S-transferase activity than that of the reference population. Based on this analysis of the natural susceptibility of this organism to organophosphate insecticides, continental and focal surveys of organophosphate susceptibility should be conducted to evaluate the evolution and distribution of this phenomenon.

  20. Distribution and pathogenicity of Trypanosoma cruzi isolated from peridomestic populations of Triatoma infestans and Triatoma guasayana from rural western Argentina

    PubMed Central

    Lauricella, Marta A; Stariolo, Raúl L; Riarte, Adelina R; Segura, Elsa L; Gürtler, Ricardo E

    2011-01-01

    We assessed the distribution of Trypanosoma cruzi infection in peridomestic triatomines collected manually at a district-wide scale in rural villages around Olta, western Argentina, and typed the isolated strains according to their pathogenicity to laboratory mice. Of 1623 triatomines examined, only 14 (0.9%) were infected with T. cruzi based on microscopical examination of feces. The prevalence of T. cruzi infection was 0.8% in Triatoma infestans, 2.3% in T. guasayana, and nil in T. garciabesi, T. platensis, and T. eratyrusiformis. Local transmission occurred in kitchens, store-rooms and goat corrals or nearby, though at very low levels. T. cruzi was detected by at least one parasitological method in 11 (79%) of 14 microscope-positive bugs. Hemoculture was the most sensitive method (67%) followed by culture of organ homogenates, histopathology or xenodiagnosis of inoculated suckling mice (55-58%), and culture of microscope-positive bug feces (46%). The evidence suggests that most of the isolated T. cruzi strains would be myotropic type III. Our study establishes for the first time that peridomestic, microscope-positive T. guasayana nymphs were actually infected with T. cruzi, and may be implicated as a putative secondary vector of T. cruzi in domestic or peridomestic sites. PMID:16021298

  1. Geographical structuring of Trypanosoma cruzi populations from Chilean Triatoma infestans triatomines and their genetic relationship with other Latino American counterparts

    PubMed Central

    Venegas, J; Rojas, T; DÍaz, F; Miranda, S; Jercic, M I; González, C; Coñoepán, W; Pichuantes, S; RodrÍguez, J; Gajardo, M; Sánchez, G

    2011-01-01

    In order to obtain more information about the population structure of Chilean Trypanosoma cruzi, and their genetic relationship with other Latino American counterparts, we performed the study of T. cruzi samples detected in the midgut content of Triatoma infestans insects from three endemic regions of Chile. The genetic characteristics of these samples were analysed using microsatellite markers and PCR conditions that allow the detection of predominant T. cruzi clones directly in triatomine midgut content. Population genetic analyses using the Fisher’s exact method, analysis of molecular variance (AMOVA) and the determination of FST showed that the northern T. cruzi population sample was genetically differentiated from the two southern population counterparts. Further analysis showed that the cause of this genetic differentiation was the asymmetrical distribution of TcIII T. cruzi predominant clones. Considering all triatomines from the three regions, the most frequent predominant lineages were TcIII (38%), followed by TcI (34%) and hybrid (8%). No TcII lineage was observed along the predominant T. cruzi clones. The best phylogenetic reconstruction using the shared allelic genetic distance was concordant with the population genetic analysis and tree topology previously described studying foreign samples. The correlation studies showed that the lineage TcIII from the III region was genetically differentiated from the other two, and this differentiation was correlated with geographical distance including Chilean and mainly Brazilian samples. It will be interesting to investigate whether this geographical structure may be related with different clinical manifestation of Chagas disease. PMID:22325822

  2. Probability of transmission of Chagas disease by Triatoma infestans (Hemiptera: Reduviidae) in an endemic area of Santiago del Estero, Argentina.

    PubMed Central

    Rabinovich, J. E.; Wisnivesky-Colli, C.; Solarz, N. D.; Gürtler, R. E.

    1990-01-01

    The daily probability (P) of transmission of Trypanosoma cruzi to a noninfected human host by an infected Triatoma infestans bug was estimated using field data from a 2-year longitudinal study carried out in a rural settlement of 20 households in Amamá, Santiago del Estero, Argentina. The following information was used for this purpose: the bug density and the proportion of infected bugs; the bug biting rate and the distribution of bites between humans and animals; the age-specific seropositivity to T. cruzi of the human population; and the actual number of new cases of human infection. The 2-year accumulated number of infective contacts per house estimated using a binomial model shows a statistically significant logistic correlation with the observed proportion of new cases per house. An average house where new cases of human infection were registered in the 2-year period had a P value of 0.0012, while an average general house (i.e., with and without new cases) had a P value of 0.0009. The observed range of P is discussed in terms of the chain of factors that affects the individual human risk of acquiring the infection and the possible entomological sampling errors. PMID:2127382

  3. Toxicological and biochemical analysis of the susceptibility of sylvatic Triatoma infestans from the Andean Valley of Bolivia to organophosphate insecticide

    PubMed Central

    Santo-Orihuela, Pablo Luis; Carvajal, Guillermo; Picollo, María Inés; Vassena, Claudia Viviana

    2013-01-01

    To increase our knowledge of the natural susceptibility of Triatoma infestans to an organophosphate insecticide, we performed toxicological and biochemical studies on three sylvatic populations from Bolivia and two populations from domestic dwellings from Bolivia and Argentina. Fifty-per-cent lethal doses (LD50) were determined based on the topical application of fenitrothion on first instar nymphs and mortality was assessed at 24 h. Both type of populations exhibited LD50ratios significantly higher than 1 with a range of the values (1.42-2.47); the maximum value were found in a sylvatic (-S) population, Veinte de Octubre-S. Samples were biochemically analysed using a glutathione S-transferase activity assay. The highest significant activity was obtained for Veinte de Octubre-S and the lowest activity was obtained for the reference population (102.69 and 54.23 pmol per minute per mg of protein respectively). Two out of the three sylvatic populations (Veinte de Octubre-S and Kirus Mayu-S) exhibited significantly higher glutathione S-transferase activity than that of the reference population. Based on this analysis of the natural susceptibility of this organism to organophosphate insecticides, continental and focal surveys of organophosphate susceptibility should be conducted to evaluate the evolution and distribution of this phenomenon. PMID:24037203

  4. Characteristic of the Pepper CaRGA2 Gene in Defense Responses against Phytophthora capsici Leonian

    PubMed Central

    Zhang, Ying-Li; Jia, Qing-Li; Li, Da-Wei; Wang, Jun-E; Yin, Yan-Xu; Gong, Zhen-Hui

    2013-01-01

    The most significant threat to pepper production worldwide is the Phytophthora blight, which is caused by the oomycete pathogen, Phytophthora capsici Leonian. In an effort to help control this disease, we isolated and characterized a P. capsici resistance gene, CaRGA2, from a high resistant pepper (C. annuum CM334) and analyzed its function by the method of real-time PCR and virus-induced gene silencing (VIGS). The CaRGA2 has a full-length cDNA of 3,018 bp with 2,874 bp open reading frame (ORF) and encodes a 957-aa protein. The protein has a predicted molecular weight of 108.6 kDa, and the isoelectric point is 8.106. Quantitative real-time PCR indicated that CaRGA2 expression was rapidly induced by P. capsici. The gene expression pattern was different between the resistant and susceptible cultivars. CaRGA2 was quickly expressed in the resistant cultivar, CM334, and reached to a peak at 24 h after inoculation with P. capsici, five-fold higher than that of susceptible cultivar. Our results suggest that CaRGA2 has a distinct pattern of expression and plays a critical role in P. capsici stress tolerance. When the CaRGA2 gene was silenced via VIGS, the resistance level was clearly suppressed, an observation that was supported by semi-quantitative RT-PCR and detached leave inoculation. VIGS analysis revealed their importance in the surveillance to P. capsici in pepper. Our results support the idea that the CaRGA2 gene may show their response in resistance against P. capsici. These analyses will aid in an effort towards breeding for broad and durable resistance in economically important pepper cultivars. PMID:23698759

  5. Two cytoplasmic effectors of Phytophthora sojae regulate plant cell death via interactions with plant catalases.

    PubMed

    Zhang, Meixiang; Li, Qi; Liu, Tingli; Liu, Li; Shen, Danyu; Zhu, Ye; Liu, Peihan; Zhou, Jian-Min; Dou, Daolong

    2015-01-01

    Plant pathogenic oomycetes, such as Phytophthora sojae, secrete an arsenal of host cytoplasmic effectors to promote infection. We have shown previously that P. sojae PsCRN63 (for crinkling- and necrosis-inducing proteins) induces programmed cell death (PCD) while PsCRN115 blocks PCD in planta; however, they are jointly required for full pathogenesis. Here, we find that PsCRN63 alone or PsCRN63 and PsCRN115 together might suppress the immune responses of Nicotiana benthamiana and demonstrate that these two cytoplasmic effectors interact with catalases from N. benthamiana and soybean (Glycine max). Transient expression of PsCRN63 increases hydrogen peroxide (H(2)O(2)) accumulation, whereas PsCRN115 suppresses this process. Transient overexpression of NbCAT1 (for N. benthamiana CATALASE1) or GmCAT1 specifically alleviates PsCRN63-induced PCD. Suppression of the PsCRN63-induced PCD by PsCRN115 is compromised when catalases are silenced in N. benthamiana. Interestingly, the NbCAT1 is recruited into the plant nucleus in the presence of PsCRN63 or PsCRN115; NbCAT1 and GmCAT1 are destabilized when PsCRN63 is coexpressed, and PsCRN115 inhibits the processes. Thus, PsCRN63/115 manipulates plant PCD through interfering with catalases and perturbing H(2)O(2) homeostasis. Furthermore, silencing of catalase genes enhances susceptibility to Phytophthora capsici, indicating that catalases are essential for plant resistance. Taken together, we suggest that P. sojae secretes these two effectors to regulate plant PCD and H(2)O(2) homeostasis through direct interaction with catalases and, therefore, overcome host immune responses.

  6. NPP1, a Phytophthora-associated trigger of plant defense in parsley and Arabidopsis.

    PubMed

    Fellbrich, Guido; Romanski, Annette; Varet, Anne; Blume, Beatrix; Brunner, Frédéric; Engelhardt, Stefan; Felix, Georg; Kemmerling, Birgit; Krzymowska, Magdalena; Nürnberger, Thorsten

    2002-11-01

    Activation of non-cultivar-specific plant defense against attempted microbial infection is mediated through the recognition of pathogen-derived elicitors. Previously, we have identified a peptide fragment (Pep-13) within a 42-kDa cell wall transglutaminase from various Phytophthora species that triggers a multifacetted defense response in parsley cells. Many of these oomycete species have now been shown to possess another cell wall protein (24 kDa), that evoked the same pattern of responses in parsley as Pep-13. Unlike Pep-13, necrosis-inducing Phytophthora protein 1 (NPP1) purified from P. parasitica also induced hypersensitive cell death-like lesions in parsley. NPP1 structural homologs were found in oomycetes, fungi, and bacteria, but not in plants. Structure-activity relationship studies revealed the intact protein as well as two cysteine residues to be essential for elicitor activity. NPP1-mediated activation of pathogen defense in parsley does not employ the Pep-13 receptor. However, early induced cellular responses implicated in elicitor signal transmission (increased levels of cytoplasmic calcium, production of reactive oxygen species, MAP kinase activation) were stimulated by either elicitor, suggesting the existence of converging signaling pathways in parsley. Infiltration of NPP1 into leaves of Arabidopsis thaliana Col-0 plants resulted in transcript accumulation of pathogenesis-related (PR) genes, production of ROS and ethylene, callose apposition, and HR-like cell death. NPP1-mediated induction of the PR1 gene is salicylic acid-dependent, and, unlike the P. syringae pv. tomato DC3000(avrRpm1)-induced PR1 gene expression, requires both functional NDR1 and PAD4. In summary, Arabidopsis plants infiltrated with NPP1 constitute an experimental system that is amenable to forward genetic approaches aiming at the dissection of signaling pathways implicated in the activation of non-cultivar-specific plant defense.

  7. Diverse Evolutionary Trajectories for Small RNA Biogenesis Genes in the Oomycete Genus Phytophthora

    PubMed Central

    Bollmann, Stephanie R.; Fang, Yufeng; Press, Caroline M.; Tyler, Brett M.; Grünwald, Niklaus J.

    2016-01-01

    Gene regulation by small RNA pathways is ubiquitous among eukaryotes, but little is known about small RNA pathways in the Stramenopile kingdom. Phytophthora, a genus of filamentous oomycetes, contains many devastating plant pathogens, causing multibillion-dollar damage to crops, ornamental plants, and natural environments. The genomes of several oomycetes including Phytophthora species such as the soybean pathogen P. sojae, have been sequenced, allowing evolutionary analysis of small RNA-processing enzymes. This study examined the evolutionary origins of the oomycete small RNA-related genes Dicer-like (DCL), and RNA-dependent RNA polymerase (RDR) through broad phylogenetic analyses of the key domains. Two Dicer gene homologs, DCL1 and DCL2, and one RDR homolog were cloned and analyzed from P. sojae. Gene expression analysis revealed only minor changes in transcript levels among different life stages. Oomycete DCL1 homologs clustered with animal and plant Dicer homologs in evolutionary trees, whereas oomycete DCL2 homologs clustered basally to the tree along with Drosha homologs. Phylogenetic analysis of the RDR homologs confirmed a previous study that suggested the last common eukaryote ancestor possessed three RDR homologs, which were selectively retained or lost in later lineages. Our analysis clarifies the position of some Unikont and Chromalveolate RDR lineages within the tree, including oomycete homologs. Finally, we analyzed alterations in the domain structure of oomycete Dicer and RDR homologs, specifically focusing on the proposed domain transfer of the DEAD-box helicase domain from Dicer to RDR. Implications of the oomycete domain structure are discussed, and possible roles of the two oomycete Dicer homologs are proposed. PMID:27014308

  8. Levels of Polyamines and Kinetic Characterization of Their Uptake in the Soybean Pathogen Phytophthora sojae

    PubMed Central

    Chibucos, M. Constantine; Morris, Paul F.

    2006-01-01

    Polyamines are ubiquitous biologically active aliphatic cations that are at least transiently available in the soil from decaying organic matter. Our objectives in this study were to characterize polyamine uptake kinetics in Phytophthora sojae zoospores and to quantify endogenous polyamines in hyphae, zoospores, and soybean roots. Zoospores contained 10 times more free putrescine than spermidine, while hyphae contained only 4 times as much free putrescine as spermidine. Zoospores contained no conjugated putrescine, but conjugated spermidine was present. Hyphae contained both conjugated putrescine and spermidine at levels comparable to the hyphal free putrescine and spermidine levels. In soybean roots, cadaverine was the most abundant polyamine, but only putrescine efflux was detected. The selective efflux of putrescine suggests that the regulation of polyamine availability is part of the overall plant strategy to influence microbial growth in the rhizosphere. In zoospores, uptake experiments with [1,4-14C]putrescine and [1,4-14C]spermidine confirmed the existence of high-affinity polyamine transport for both polyamines. Putrescine uptake was reduced by high levels of exogenous spermidine, but spermidine uptake was not reduced by exogenous putrescine. These observations suggest that P. sojae zoospores express at least two high-affinity polyamine transporters, one that is spermidine specific and a second that is putrescine specific or putrescine preferential. Disruption of polyamine uptake or metabolism has major effects on a wide range of cellular activities in other organisms and has been proposed as a potential control strategy for Phytophthora. Inhibition of polyamine uptake may be a means of reducing the fitness of the zoospore along with subsequent developmental stages that precede infection. PMID:16672477

  9. Modeling climate impact on an emerging disease, the Phytophthora alni-induced alder decline.

    PubMed

    Aguayo, Jaime; Elegbede, Fabrice; Husson, Claude; Saintonge, François-Xavier; Marçais, Benoît

    2014-10-01

    Alder decline caused by Phytophthora alni is one of the most important emerging diseases in natural ecosystems in Europe, where it has threatened riparian ecosystems for the past 20 years. Environmental factors, such as mean site temperature and soil characteristics, play an important role in the occurrence of the disease. The objective of the present work was to model and forecast the effect of environment on the severity of alder Phytophthora outbreaks, and to determine whether recent climate change might explain the disease emergence. Two alder sites networks in NE and SW France were surveyed to assess the crown health of trees; the oomycete soil inoculum was also monitored in the NE network. The main factors explaining the temporal annual variation in alder crown decline or crown recovery were the mean previous winter and previous summer temperatures. Both low winter temperatures and high summer temperatures were unfavorable to the disease. Cold winters promoted tree recovery because of poor survival of the pathogen, while hot summer temperature limited the incidence of tree decline. An SIS model explaining the dynamics of the P. alni-induced alder decline was developed using the data of the NE site network and validated using the SW site network. This model was then used to simulate the frequency of declining alder over time with historical climate data. The last 40 years' weather conditions have been generally favorable to the establishment of the disease, indicating that others factors may be implicated in its emergence. The model, however, showed that the climate of SW France was much more favorable for the disease than that of the Northeast, because it seldom limited the overwintering of the pathogen. Depending on the European area, climate change could either enhance or decrease the severity of the alder decline.

  10. Diverse Evolutionary Trajectories for Small RNA Biogenesis Genes in the Oomycete Genus Phytophthora.

    PubMed

    Bollmann, Stephanie R; Fang, Yufeng; Press, Caroline M; Tyler, Brett M; Grünwald, Niklaus J

    2016-01-01

    Gene regulation by small RNA pathways is ubiquitous among eukaryotes, but little is known about small RNA pathways in the Stramenopile kingdom. Phytophthora, a genus of filamentous oomycetes, contains many devastating plant pathogens, causing multibillion-dollar damage to crops, ornamental plants, and natural environments. The genomes of several oomycetes including Phytophthora species such as the soybean pathogen P. sojae, have been sequenced, allowing evolutionary analysis of small RNA-processing enzymes. This study examined the evolutionary origins of the oomycete small RNA-related genes Dicer-like (DCL), and RNA-dependent RNA polymerase (RDR) through broad phylogenetic analyses of the key domains. Two Dicer gene homologs, DCL1 and DCL2, and one RDR homolog were cloned and analyzed from P. sojae. Gene expression analysis revealed only minor changes in transcript levels among different life stages. Oomycete DCL1 homologs clustered with animal and plant Dicer homologs in evolutionary trees, whereas oomycete DCL2 homologs clustered basally to the tree along with Drosha homologs. Phylogenetic analysis of the RDR homologs confirmed a previous study that suggested the last common eukaryote ancestor possessed three RDR homologs, which were selectively retained or lost in later lineages. Our analysis clarifies the position of some Unikont and Chromalveolate RDR lineages within the tree, including oomycete homologs. Finally, we analyzed alterations in the domain structure of oomycete Dicer and RDR homologs, specifically focusing on the proposed domain transfer of the DEAD-box helicase domain from Dicer to RDR. Implications of the oomycete domain structure are discussed, and possible roles of the two oomycete Dicer homologs are proposed. PMID:27014308

  11. Arabidopsis Lectin Receptor Kinases LecRK-IX.1 and LecRK-IX.2 Are Functional Analogs in Regulating Phytophthora Resistance and Plant Cell Death.

    PubMed

    Wang, Yan; Cordewener, Jan H G; America, Antoine H P; Shan, Weixing; Bouwmeester, Klaas; Govers, Francine

    2015-09-01

    L-type lectin receptor kinases (LecRK) are potential immune receptors. Here, we characterized two closely-related Arabidopsis LecRK, LecRK-IX.1 and LecRK-IX.2, of which T-DNA insertion mutants showed compromised resistance to Phytophthora brassicae and Phytophthora capsici, with double mutants showing additive susceptibility. Overexpression of LecRK-IX.1 or LecRK-IX.2 in Arabidopsis and transient expression in Nicotiana benthamiana increased Phytophthora resistance but also induced cell death. Phytophthora resistance required both the lectin domain and kinase activity, but for cell death, the lectin domain was not needed. Silencing of the two closely related mitogen-activated protein kinase genes NbSIPK and NbNTF4 in N. benthamiana completely abolished LecRK-IX.1-induced cell death but not Phytophthora resistance. Liquid chromatography-mass spectrometry analysis of protein complexes coimmunoprecipitated in planta with LecRK-IX.1 or LecRK-IX.2 as bait, resulted in the identification of the N. benthamiana ABC transporter NbPDR1 as a potential interactor of both LecRK. The closest homolog of NbPDR1 in Arabidopsis is ABCG40, and coimmunoprecipitation experiments showed that ABCG40 associates with LecRK-IX.1 and LecRK-IX.2 in planta. Similar to the LecRK mutants, ABCG40 mutants showed compromised Phytophthora resistance. This study shows that LecRK-IX.1 and LecRK-IX.2 are Phytophthora resistance components that function independent of each other and independent of the cell-death phenotype. They both interact with the same ABC transporter, suggesting that they exploit similar signal transduction pathways.

  12. The Pectin Methylesterase Gene Complement of Phytophthora sojae: Structural and Functional Analyses, and the Evolutionary Relationships with Its Oomycete Homologs

    PubMed Central

    Horowitz, Brent B.; Ospina-Giraldo, Manuel D.

    2015-01-01

    Phytophthora sojae is an oomycete pathogen that causes the disease known as root and stem rot in soybean plants, frequently leading to massive economic damage. Additionally, P. sojae is increasingly being utilized as a model for phytopathogenic oomycete research. Despite the economic and scientific importance of P. sojae, the mechanism by which it penetrates the host roots is not yet fully understood. It has been found that oomycetes are not capable of penetrating the cell wall solely through mechanical force, suggesting that alternative factors facilitate breakdown of the host cell wall. Pectin methylesterases have been suggested to be important for Phytophthora pathogenicity, but no data exist on their role in the P. sojae infection process. We have scanned the newly revised version of the annotated P. sojae genome for the presence of putative pectin methylesterases genes and conducted a sequence analysis of all gene models found. We also searched for potential regulatory motifs in the promoter region of the proposed P. sojae models, and investigated the gene expression levels throughout the early course of infection on soybean plants. We found that P. sojae contains a large repertoire of pectin methylesterase-coding genes and that most of these genes display similar motifs in the promoter region, indicating the possibility of a shared regulatory mechanism. Phylogenetic analyses confirmed the evolutionary relatedness of the pectin methylesterase-coding genes within and across Phytophthora spp. In addition, the gene duplication events that led to the emergence of this gene family appear to have occurred prior to many speciation events in the genus Phytophthora. Our results also indicate that the highest levels of expression occurred in the first 24 hours post inoculation, with expression falling after this time. Our study provides evidence that pectin methylesterases may be important for the early action of the P. sojae infection process. PMID:26544849

  13. The Pectin Methylesterase Gene Complement of Phytophthora sojae: Structural and Functional Analyses, and the Evolutionary Relationships with Its Oomycete Homologs.

    PubMed

    Horowitz, Brent B; Ospina-Giraldo, Manuel D

    2015-01-01

    Phytophthora sojae is an oomycete pathogen that causes the disease known as root and stem rot in soybean plants, frequently leading to massive economic damage. Additionally, P. sojae is increasingly being utilized as a model for phytopathogenic oomycete research. Despite the economic and scientific importance of P. sojae, the mechanism by which it penetrates the host roots is not yet fully understood. It has been found that oomycetes are not capable of penetrating the cell wall solely through mechanical force, suggesting that alternative factors facilitate breakdown of the host cell wall. Pectin methylesterases have been suggested to be important for Phytophthora pathogenicity, but no data exist on their role in the P. sojae infection process. We have scanned the newly revised version of the annotated P. sojae genome for the presence of putative pectin methylesterases genes and conducted a sequence analysis of all gene models found. We also searched for potential regulatory motifs in the promoter region of the proposed P. sojae models, and investigated the gene expression levels throughout the early course of infection on soybean plants. We found that P. sojae contains a large repertoire of pectin methylesterase-coding genes and that most of these genes display similar motifs in the promoter region, indicating the possibility of a shared regulatory mechanism. Phylogenetic analyses confirmed the evolutionary relatedness of the pectin methylesterase-coding genes within and across Phytophthora spp. In addition, the gene duplication events that led to the emergence of this gene family appear to have occurred prior to many speciation events in the genus Phytophthora. Our results also indicate that the highest levels of expression occurred in the first 24 hours post inoculation, with expression falling after this time. Our study provides evidence that pectin methylesterases may be important for the early action of the P. sojae infection process.

  14. Host-feeding sources and habitats jointly affect wing developmental stability depending on sex in the major Chagas disease vector Triatoma infestans.

    PubMed

    Nattero, Julieta; Dujardin, Jean-Pierre; del Pilar Fernández, María; Gürtler, Ricardo E

    2015-12-01

    Fluctuating asymmetry (FA), a slight and random departure from bilateral symmetry that is normally distributed around a 0 mean, has been widely used to infer developmental instability. We investigated whether habitats (ecotopes) and host-feeding sources influenced wing FA of the hematophagous bug Triatoma infestans. Because bug populations occupying distinct habitats differed substantially and consistently in various aspects such as feeding rates, engorgement status and the proportion of gravid females, we predicted that bugs from more open peridomestic habitats (i.e., goat corrals) were more likely to exhibit higher FA than bugs from domiciles. We examined patterns of asymmetry and the amount of wing size and shape FA in 196 adult T. infestans collected across a gradient of habitat suitability and stability that decreased from domiciles, storerooms, kitchens, chicken coops, pig corrals, to goat corrals in a well-defined area of Figueroa, northwestern Argentina. The bugs had unmixed blood meals on human, chicken, pig and goat depending on the bug collection ecotope. We documented the occurrence of FA in wing shape for bugs fed on all host-feeding sources and in all ecotopes except for females from domiciles or fed on humans. FA indices for wing shape differed significantly among host-feeding sources, ecotopes and sexes. The patterns of wing asymmetry in females from domiciles and from goat corrals were significantly different; differences in male FA were congruent with evidence showing that they had higher mobility than females across habitats. The host-feeding sources and habitats of T. infestans affected wing developmental stability depending on sex.

  15. Identification of three cytochrome P450 genes in the Chagas' disease vector Triatoma infestans: Expression analysis in deltamethrin susceptible and resistant populations.

    PubMed

    Grosso, Carla G; Blariza, María J; Mougabure-Cueto, Gastón; Picollo, María I; García, Beatriz A

    2016-10-01

    Cytochrome P450 monooxygenases play a predominant role in the metabolism of insecticides. Many insect P450 genes have frequently been associated with detoxification processes allowing the insect to become tolerant or resistant to insecticides. The increases of expression of P450 genes at transcriptional level are often consider responsible for increasing the metabolism of insecticides and seems to be a common phenomenon in the evolution of resistance development in insects. As pyrethroid resistance has been detected in Triatoma infestans, it was of interest to analyze genes associated with resistance to insecticides such as those encoding for cytochromes P450. With this purpose, the cDNA sequences of three cytochrome P450 genes (CYP4EM7, CYP3085B1, and CYP3092A6) were identified in this species. Primers and specific Taqman probes were designed from these sequences to determine their expression by quantitative PCR. The mRNA levels of the cytochrome P450 genes identified were determined from total RNA extracted from pools of fat body collected from individuals of different resistant and susceptible strains of T. infestans, and at different interval times after the topical application of the lethal doses 50% (LD50) of deltamethrin on the ventral abdomen of insects belonging to the different populations analyzed. It was detected overexpression of the CYP4EM7 gene in the most resistant strain of T. infestans and the expression of the three cytochrome P450 genes isolated was induced by deltamethrin in the susceptible and resistant populations included in this study. These results suggest that these genes would be involved in the detoxification of deltamethrin and support the hypothesis that considers to the cytochrome P450 genes of importance in the development of pyrethroid resistance. PMID:27461853

  16. Assessing the effectiveness of Byssochlamys nivea and Scopulariopsis brumptii in pentachlorophenol removal and biological control of two Phytophthora species.

    PubMed

    Bosso, Luciano; Scelza, Rosalia; Varlese, Rosaria; Meca, Giuseppe; Testa, Antonino; Rao, Maria A; Cristinzio, Gennaro

    2016-04-01

    Bioremediation and biological-control by fungi have made tremendous strides in numerous biotechnology applications. The aim of this study was to test Byssochlamys nivea and Scopulariopsis brumptii in sensitivity and degradation to pentachlorophenol (PCP) and in biological-control of Phytophthora cinnamomi and Phytophthora cambivora. B. nivea and S. brumptii were tested in PCP sensitivity and degradation in microbiological media while the experiments of biological-control were carried out in microbiological media and soil. The fungal strains showed low PCP sensitivity at 12.5 and 25 mg PCP L(-1) although the hyphal size, fungal mat, patulin, and spore production decreased with increasing PCP concentrations. B. nivea and S. brumptii depleted completely 12.5 and 25 mg PCP L(-1) in liquid culture after 28 d of incubation at 28 °C. Electrolyte leakage assays showed that both fungi have low sensitivity to 25 mg PCP L(-1) and produced no toxic compounds for the plant. B. nivea and S. brumptii were able to inhibit the growth of the two plant pathogens in laboratory studies and reduce the mortality of chestnut plants caused by two Phytophthorae in greenhouse experiments. The two fungal strains did not produce volatile organic compounds able to reduce the growth of two plant pathogens tested. PMID:27020163

  17. Efficient disruption and replacement of an effector gene in the oomycete Phytophthora sojae using CRISPR/Cas9.

    PubMed

    Fang, Yufeng; Tyler, Brett M

    2016-01-01

    Phytophthora sojae is an oomycete pathogen of soybean. As a result of its economic importance, P. sojae has become a model for the study of oomycete genetics, physiology and pathology. The lack of efficient techniques for targeted mutagenesis and gene replacement have long hampered genetic studies of pathogenicity in Phytophthora species. Here, we describe a CRISPR/Cas9 system enabling rapid and efficient genome editing in P. sojae. Using the RXLR effector gene Avr4/6 as a target, we observed that, in the absence of a homologous template, the repair of Cas9-induced DNA double-strand breaks (DSBs) in P. sojae was mediated by non-homologous end-joining (NHEJ), primarily resulting in short indels. Most mutants were homozygous, presumably as a result of gene conversion triggered by Cas9-mediated cleavage of non-mutant alleles. When donor DNA was present, homology-directed repair (HDR) was observed, which resulted in the replacement of Avr4/6 with the NPT II gene. By testing the specific virulence of several NHEJ mutants and HDR-mediated gene replacements in soybean, we have validated the contribution of Avr4/6 to recognition by soybean R gene loci, Rps4 and Rps6, but also uncovered additional contributions to resistance by these two loci. Our results establish a powerful tool for the study of functional genomics in Phytophthora, which provides new avenues for better control of this pathogen.

  18. Touchdown nested multiplex PCR detection of Phytophthora cinnamomi and P. cambivora from French and English chestnut grove soils.

    PubMed

    Langrell, Stephen R H; Morel, Olivier; Robin, Cécile

    2011-07-01

    Soil borne Phytophthora cinnamomi and Phytophthora cambivora are considered the most pathogenic species associated with chestnut (Castanea sativa) decline in Europe. Mapping their incidence and distribution from nursery and plantation soils may offer valuable information for limiting spread. As conventional biological baiting and taxonomic confirmation is generally time consuming, labour, logistically and space intensive, we have focused on the development of a specific touchdown nested multiplex Polymerase Chain Reaction (PCR) approach for the simultaneous detection of both species direct from soil. Pre-existing and novel primers, based on Internal Transcribed Spacer (ITS) sequences, have been evaluated for their specificity and use in a multiplex capacity in various combinations. Coupled to this we have modified a mechanical lysis procedure for DNA extraction from up to 10 g of chestnut under storey soils (ranging from 0.5 to 25 μg DNA g(-1)fresh soil). Using serial dilutions and/or polyvinylpolypyrrolidone chromatography purification, both species have been successfully detected, in artificially and naturally infected soils. Levels of assay detection are comparable to other Phytophthora species where PCR based diagnostic systems have been reported. A qualitative evaluation of this approach against conventional baiting is presented.

  19. Pistachio gummosis disease caused by Phytophthora species and its control management with soil solarisation method in Iran.

    PubMed

    Saremi, H; Okhovvat, S M; Saremi, Ha

    2008-01-01

    Pistachio (Pistacia vera L.) is the most important commercial product in Iran and root rot or crown rot (Gummosis) is the most serious diseases of this crop. During 2005-2007 Infected trees of Pistachio orchards were visited and plant samples plus soils around the infected trees collected from Kerman province in Iran. Samples were transferred to laboratory and cultured on common medium and using citrus leaves pieces as baits on water-saturated soils. Different Phytophthora species were isolated and studied to be identified. Three Phytophthora species including P. megasperma, P. drechsleri and P. citrophthora were the principal cause of pistachio gummosis and root rot in Iran. However, Phytophthora pistaciae as new species was introduced as aggressive species to different Pistachio cultivars. Since chemical control was not property managed the disease, soil disinfestations by soil solarisation method was carried in Kenrman as the nearly warmer climate in studied areas to manage the pathogen. Application of this method reduced population density of the pathogen from 1300 to 200 CFU -g/soil after 6 weeks. This method was effective, non negative side and economic which can be used in all agricultural areas.

  20. Ecological and Sociodemographic Determinants of House Infestation by Triatoma infestans in Indigenous Communities of the Argentine Chaco

    PubMed Central

    Gaspe, M. Sol; Provecho, Yael M.; Cardinal, M. Victoria; del Pilar Fernández, M.; Gürtler, Ricardo E.

    2015-01-01

    Background The Gran Chaco ecoregion, a hotspot for Chagas and other neglected tropical diseases, is home to >20 indigenous peoples. Our objective was to identify the main ecological and sociodemographic determinants of house infestation and abundance of Triatoma infestans in traditional Qom populations including a Creole minority in Pampa del Indio, northeastern Argentina. Methods A cross-sectional survey determined house infestation by timed-manual searches with a dislodging aerosol in 386 inhabited houses and administered questionnaires on selected variables before full-coverage insecticide spraying and annual vector surveillance. We fitted generalized linear models to two global models of domestic infestation and bug abundance, and estimated coefficients via multimodel inference with model averaging. Principal Findings Most Qom households were larger and lived in small-sized, recently-built, precarious houses with fewer peridomestic structures, and fewer livestock and poultry than Creoles’. Qom households had lower educational level and unexpectedly high residential mobility. House infestation (31.9%) was much lower than expected from lack of recent insecticide spraying campaigns and was spatially aggregated. Nearly half of the infested houses examined had infected vectors. Qom households had higher prevalence of domestic infestation (29.2%) than Creoles’ (10.0%), although there is large uncertainty around the adjusted OR. Factors with high relative importance for domestic infestation and/or bug abundance were refuge availability, distance to the nearest infested house, domestic insecticide use, indoor presence of poultry, residential overcrowding, and household educational level. Conclusions and Significance Our study highlights the importance of sociodemographic determinants of domestic infestation such as overcrowding, education and proximity to the nearest infested house, and corroborates the role of refuge availability, domestic use of insecticides and

  1. Origins of house reinfestation with Triatoma infestans after insecticide spraying in the Argentine Chaco using wing geometric morphometry

    PubMed Central

    Gaspe, M. Sol; Gurevitz, Juan M.; Gürtler, Ricardo E.; Dujardin, Jean-Pierre

    2013-01-01

    Identifying the origins of insect vectors collected after community-wide residual insecticide applications is a relevant challenge in the Gran Chaco region where the main vector of Chagas disease Triatoma infestans usually reinfests human dwellings. Wing geometric morphometry was used to compare the right wings of 63 males and 54 females collected at 4 months post-spraying (MPS) with those from 165 males and 111 females collected before full-coverage spraying with pyrethroids in a well-defined rural area in northeastern Argentina. Male and female wing centroid size resulted significantly larger at 4 MPS than before interventions, but no significant changes in shape were detected. Metric disparity (variance of shape) varied significantly in males but not in females. Using shape variables, a relatively large fraction of post-spraying males (70%) and females (54%) could not be differentiated from those collected at the same source house or at the nearest infested house before interventions. Bugs collected at 4 and 8 MPS in a persistently infested house were mainly assigned to the source house. These results support the hypothesis of persistent bug populations that survived the insecticide application at local spatial scales, and are consistent with the occurrence of vector control failures most likely related to moderate pyrethroid resistance. Wing geometric morphometry is a useful tool for identifying sources of reinfestation, but it is limited by the spatial structure found in the reference populations. Combined with field and genetic data, this approach may contribute to the understanding of the reinfestation process and improvement of vector control strategies. PMID:23557838

  2. Key Source Habitats and Potential Dispersal of Triatoma infestans Populations in Northwestern Argentina: Implications for Vector Control

    PubMed Central

    Gürtler, Ricardo E.; Cecere, María C.; Fernández, María del Pilar; Vazquez-Prokopec, Gonzalo M.; Ceballos, Leonardo A.; Gurevitz, Juan M.; Kitron, Uriel; Cohen, Joel E.

    2014-01-01

    Background Triatoma infestans —the principal vector of the infection that causes Chagas disease— defies elimination efforts in the Gran Chaco region. This study identifies the types of human-made or -used structures that are key sources of these bugs in the initial stages of house reinfestation after an insecticide spraying campaign. Methodology and Principal Findings We measured demographic and blood-feeding parameters at two geographic scales in 11 rural communities in Figueroa, northwest Argentina. Of 1,297 sites searched in spring, 279 (21.5%) were infested. Bug abundance per site and female fecundity differed significantly among habitat types (ecotopes) and were highly aggregated. Domiciles (human sleeping quarters) had maximum infestation prevalence (38.7%), human-feeding bugs and total egg production, with submaximal values for other demographic and blood-feeding attributes. Taken collectively peridomestic sites were three times more often infested than domiciles. Chicken coops had greater bug abundance, blood-feeding rates, engorgement status, and female fecundity than pig and goat corrals. The host-feeding patterns were spatially structured yet there was strong evidence of active dispersal of late-stage bugs between ecotopes. Two flight indices predicted that female fliers were more likely to originate from kitchens and domiciles, rejecting our initial hypothesis that goat and pig corrals would dominate. Conclusions and Significance Chicken coops and domiciles were key source habitats fueling rapid house reinfestation. Focusing control efforts on ecotopes with human-fed bugs (domiciles, storerooms, goat corrals) would neither eliminate the substantial contributions to bug population growth from kitchens, chicken coops, and pig corrals nor stop dispersal of adult female bugs from kitchens. Rather, comprehensive control of the linked network of ecotopes is required to prevent feeding on humans, bug population growth, and bug dispersal simultaneously. Our

  3. Infection of Citrus Roots by Tylenchulus semipenetrans Reduces Root Infection by Phytophthora nicotianae

    PubMed Central

    El-Borai, F. E.; Duncan, L. W.; Graham, J. H.

    2002-01-01

    Bioassays and whole-plant experiments were conducted to investigate the interaction between Tylenchulus semipenetrans and Phytophthora nicotianae. Both organisms are parasites of the citrus fibrous root cortex. Nematode-infected and non-infected root segments were excised from naturally infected field roots and placed on water agar in close proximity to agar plugs of P. nicotianae and then transferred to a Phytophthora-selective medium. At 10 and 12 days, 50% fewer nematode-infected segments were infected by P. nicotianae than non-infected segments. In whole-plant experiments in glass test tubes, sour orange seedlings were inoculated with two densities (8,000 or 80,000 eggs and second-stage juveniles) of T. semipenetrans, and after establishment of infection were inoculated with two densities (9,000 and 90,000 zoospores) of P. nicotianae. In the first experiment, fungal protein was 53% to 65% lower in the roots infected by both organisms than in roots infected by the fungus only. Compared to plants infected only by P. nicotianae, shoot weights were 33% to 50% greater (P ≤ 0.05) in plants infected by both parasites, regardless of inoculum density. Fibrous and tap root weights were 5% to 23% and 19% to 34% greater (P ≤ 0.05), respectively, in nematode-fungus combination treatments compared to the fungus alone. A second experiment was conducted, where plants were infected by the fungus, the nematode, both organisms, or neither organism. The soil mixture pH for 50% of the plants was adjusted from 4.5 to 7.0 to favor nematode infection. A higher rate of nematode infection of plants growing at pH 7.0 compared to pH 4.5 resulted in greater suppression of fungal development and greater inhibition of fungal damage to the plant. Compared to plants infected only by P. nicotianae, shoot and root weights were 37% and 33% greater (P ≤ 0.05), respectively, in plants infected by both parasites. These experiments have revealed antagonism between T. semipenetrans and P

  4. Global gene expression of Poncirus trifoliata, Citrus sunki and their hybrids under infection of Phytophthora parasitica

    PubMed Central

    2011-01-01

    Background Gummosis and root rot caused by Phytophthora are among the most economically important diseases in citrus. Four F1 resistant hybrids (Pool R), and four F1 susceptible hybrids (Pool S) to P. parasitica, were selected from a cross between susceptible Citrus sunki and resistant Poncirus trifoliata cv. Rubidoux. We investigated gene expression in pools of four resistant and four susceptible hybrids in comparison with their parents 48 hours after P. parasitica inoculation. We proposed that genes differentially expressed between resistant and susceptible parents and between their resistant and susceptible hybrids provide promising candidates for identifying transcripts involved in disease resistance. A microarray containing 62,876 UniGene transcripts selected from the CitEST database and prepared by NimbleGen Systems was used for analyzing global gene expression 48 hours after infection with P. parasitica. Results Three pairs of data comparisons (P. trifoliata/C. sunki, Pool R/C. sunki and Pool R/Pool S) were performed. With a filter of false-discovery rate less than 0.05 and fold change greater than 3.0, 21 UniGene transcripts common to the three pairwise comparative were found to be up-regulated, and 3 UniGene transcripts were down-regulated. Among them, our results indicated that the selected transcripts were probably involved in the whole process of plant defense responses to pathogen attack, including transcriptional regulation, signaling, activation of defense genes participating in HR, single dominant genes (R gene) such as TIR-NBS-LRR and RPS4 and switch of defense-related metabolism pathway. Differentially expressed genes were validated by RT-qPCR in susceptible and resistant plants and between inoculated and uninoculated control plants Conclusions Twenty four UniGene transcripts were identified as candidate genes for Citrus response to P. parasitica. UniGene transcripts were likely to be involved in disease resistance, such as genes potentially

  5. Composition and anti-insect activity of essential oils from Tagetes L. species (Asteraceae, Helenieae) on Ceratitis capitata Wiedemann and Triatoma infestans Klug.

    PubMed

    López, Sandra B; López, María L; Aragón, Liliana M; Tereschuk, María L; Slanis, Alberto C; Feresin, Gabriela E; Zygadlo, Julio A; Tapia, Alejandro A

    2011-05-25

    Essential oils from four species of the genus Tagetes L. (Asteraceae, Helenieae) collected in Tucumán province, Argentina, were evaluated for their chemical composition, toxicity, and olfactory activity on Mediterranean fruit fly Ceratitis capitata Wiedemann adults and for repellent properties on Triatoma infestans (Klug) (Chagas disease vector). Yields of essential oils range from 0.2 to 0.8% (v/w). The same main constituents among Tagetes minuta L., Tagestes rupestris Cabrera, and Tagetes terniflora Kunth, (cis-trans)-ocimenes, (cis-trans)-tagetones, and (cis-trans)-ocimenones showed important differences in their relative compositions. Tagetes filifolia Lag. was characterized by the recognized phenylpropanoids methylchavicol and trans-anethole as the main components. LD(50) was ≤20 μg/insect in topical bioassays. T. rupestris was the most toxic to C. capitata females, whereas the other oils presented similar toxicities against males and females. Tagetes rupestris oil attracted both sexes of C. capitata at 5 μg, whereas T. minuta showed opposite activities between males (attractant) and females (repellent). Oils from T. minuta and T. filifolia were the most repellent to T. infestans. The results suggest that compositions of essential oils influence their insecticidal and olfactory properties. The essential oils from Tagetes species show an important potential as infochemical agents on insects' behaviors. This study highlights the chemical variability of essential oils as a source of variation of anti-insect properties. PMID:21469658

  6. Overexpression of Soybean Isoflavone Reductase (GmIFR) Enhances Resistance to Phytophthora sojae in Soybean

    PubMed Central

    Cheng, Qun; Li, Ninghui; Dong, Lidong; Zhang, Dayong; Fan, Sujie; Jiang, Liangyu; Wang, Xin; Xu, Pengfei; Zhang, Shuzhen

    2015-01-01

    Isoflavone reductase (IFR) is an enzyme involved in the biosynthetic pathway of isoflavonoid phytoalexin in plants. IFRs are unique to the plant kingdom and are considered to have crucial roles in plant response to various biotic and abiotic environmental stresses. Here, we report the characterization of a novel member of the soybean isoflavone reductase gene family GmIFR. Overexpression of GmIFR transgenic soybean exhibited enhanced resistance to Phytophthora sojae. Following stress treatments, GmIFR was significantly induced by P. sojae, ethephon (ET), abscisic acid (placeCityABA), salicylic acid (SA). It is located in the cytoplasm when transiently expressed in soybean protoplasts. The daidzein levels reduced greatly for the seeds of transgenic plants, while the relative content of glyceollins in transgenic plants was significantly higher than that of non-transgenic plants. Furthermore, we found that the relative expression levels of reactive oxygen species (ROS) of transgenic soybean plants were significantly lower than those of non-transgenic plants after incubation with P. sojae, suggesting an important role of GmIFR might function as an antioxidant to reduce ROS in soybean. The enzyme activity assay suggested that GmIFR has isoflavone reductase activity. PMID:26635848

  7. Overexpression of Soybean Isoflavone Reductase (GmIFR) Enhances Resistance to Phytophthora sojae in Soybean.

    PubMed

    Cheng, Qun; Li, Ninghui; Dong, Lidong; Zhang, Dayong; Fan, Sujie; Jiang, Liangyu; Wang, Xin; Xu, Pengfei; Zhang, Shuzhen

    2015-01-01

    Isoflavone reductase (IFR) is an enzyme involved in the biosynthetic pathway of isoflavonoid phytoalexin in plants. IFRs are unique to the plant kingdom and are considered to have crucial roles in plant response to various biotic and abiotic environmental stresses. Here, we report the characterization of a novel member of the soybean isoflavone reductase gene family GmIFR. Overexpression of GmIFR transgenic soybean exhibited enhanced resistance to Phytophthora sojae. Following stress treatments, GmIFR was significantly induced by P. sojae, ethephon (ET), abscisic acid (placeCityABA), salicylic acid (SA). It is located in the cytoplasm when transiently expressed in soybean protoplasts. The daidzein levels reduced greatly for the seeds of transgenic plants, while the relative content of glyceollins in transgenic plants was significantly higher than that of non-transgenic plants. Furthermore, we found that the relative expression levels of reactive oxygen species (ROS) of transgenic soybean plants were significantly lower than those of non-transgenic plants after incubation with P. sojae, suggesting an important role of GmIFR might function as an antioxidant to reduce ROS in soybean. The enzyme activity assay suggested that GmIFR has isoflavone reductase activity.

  8. Genetic diversity of Phytophthora capsici isolates from pepper and pumpkin in Argentina.

    PubMed

    Gobena, Daniel; Roig, Julián; Galmarini, Claudio; Hulvey, Jon; Lamour, Kurt

    2012-01-01

    Phytophthora capsici is a soilborne oomycete plant pathogen that limits pepper production worldwide. The population structure varies significantly depending on the location (e.g. Peru vs. USA) and little is known about the diversity of P. capsici in Argentina. Our objective was to assess the diversity of P. capsici in Argentina at key pepper production areas. Forty isolates were recovered 2006-2009 from pepper and one isolate from pumpkin at 11 locations. Isolates were assessed for mating type, mefenoxam sensitivity and multilocus single nucleotide polymorphism (SNP) genotype profiles. Ten isolates with identical SNP profiles also were genotyped with amplified fragment length polymorphism (AFLP) markers. All 41 isolates had the A1 mating type and were sensitive to mefenoxam. Genotypic analysis using eight polymorphic SNP markers indicated 87% of the isolates had the same multilocus genotype, which is fixed for heterozygosity at seven of the eight SNP sites. AFLP analyses confirmed these findings, and overall it appears that clonal reproduction drives the population structure of P. capsici in Argentina. The implications for breeding resistant peppers and overall disease management are discussed. PMID:21933926

  9. Loss of heterozygosity drives clonal diversity of Phytophthora capsici in China.

    PubMed

    Hu, Jian; Diao, Yongzhao; Zhou, Yuxin; Lin, Dong; Bi, Yang; Pang, Zhili; Trout Fryxell, Rebecca; Liu, Xili; Lamour, Kurt

    2013-01-01

    Phytophthora capsici causes significant loss to pepper (Capsicum annum) in China and our goal was to develop single nucleotide polymorphism (SNP) markers for P. capsici and characterize genetic diversity nationwide. Eighteen isolates of P. capsici from locations worldwide were re-sequenced and candidate nuclear and mitochondrial SNPs identified. From 2006 to 2012, 276 isolates of P. capsici were recovered from 136 locations in 27 provinces and genotyped using 45 nuclear and 2 mitochondrial SNPs. There were two main mitochondrial haplotypes and 95 multi-locus genotypes (MLGs) identified. Genetic diversity was geographically structured with a high level of genotypic diversity in the north and on Hainan Island in the south, suggesting outcrossing contributes to diversity in these areas. The remaining areas of China are dominated by four clonal lineages that share mitochondrial haplotypes, are almost exclusively the A1 or A2 mating type and appear to exhibit extensive diversity based on loss of heterozygosity (LOH). Analysis of SNPs directly from infected peppers confirmed LOH in field populations. One clonal lineage is dominant throughout much of the country. The overall implications for long-lived genetically diverse clonal lineages amidst a widely dispersed sexual population are discussed. PMID:24349339

  10. RNA-Seq Reveals Infection-Related Gene Expression Changes in Phytophthora capsici

    PubMed Central

    Chen, Xiao-Ren; Xing, Yu-Ping; Li, Yan-Peng; Tong, Yun-Hui; Xu, Jing-You

    2013-01-01

    Phytophthora capsici is a soilborne plant pathogen capable of infecting a wide range of plants, including many solanaceous crops. However, genetic resistance and fungicides often fail to manage P. capsici due to limited knowledge on the molecular biology and basis of P. capsici pathogenicity. To begin to rectify this situation, Illumina RNA-Seq was used to perform massively parallel sequencing of three cDNA samples derived from P. capsici mycelia (MY), zoospores (ZO) and germinating cysts with germ tubes (GC). Over 11 million reads were generated for each cDNA library analyzed. After read mapping to the gene models of P. capsici reference genome, 13,901, 14,633 and 14,695 putative genes were identified from the reads of the MY, ZO and GC libraries, respectively. Comparative analysis between two of samples showed major differences between the expressed gene content of MY, ZO and GC stages. A large number of genes associated with specific stages and pathogenicity were identified, including 98 predicted effector genes. The transcriptional levels of 19 effector genes during the developmental and host infection stages of P. capsici were validated by RT-PCR. Ectopic expression in Nicotiana benthamiana showed that P. capsici RXLR and Crinkler effectors can suppress host cell death triggered by diverse elicitors including P. capsici elicitin and NLP effectors. This study provides a first look at the transcriptome and effector arsenal of P. capsici during the important pre-infection stages. PMID:24019970

  11. Genetic Analysis of Phytophthora nicotianae Populations from Different Hosts Using Microsatellite Markers.

    PubMed

    Biasi, Antonio; Martin, Frank N; Cacciola, Santa O; di San Lio, Gaetano Magnano; Grünwald, Niklaus J; Schena, Leonardo

    2016-09-01

    In all, 231 isolates of Phytophthora nicotianae representing 14 populations from different host genera, including agricultural crops (Citrus, Nicotiana, and Lycopersicon), potted ornamental species in nurseries (Lavandula, Convolvulus, Myrtus, Correa, and Ruta), and other plant genera were characterized using simple-sequence repeat markers. In total, 99 multilocus genotypes (MLG) were identified, revealing a strong association between genetic grouping and host of recovery, with most MLG being associated with a single host genus. Significant differences in the structure of populations were revealed but clonality prevailed in all populations. Isolates from Citrus were found to be genetically related regardless of their geographic origin and were characterized by high genetic uniformity and high inbreeding coefficients. Higher variability was observed for other populations and a significant geographical structuring was determined for isolates from Nicotiana. Detected differences were related to the propagation and cultivation systems of different crops. Isolates obtained from Citrus spp. are more likely to be distributed worldwide with infected plant material whereas Nicotiana and Lycopersicon spp. are propagated by seed, which would not contribute to the spread of the pathogen and result in a greater chance for geographic isolation of lineages. With regard to ornamental species in nurseries, the high genetic variation is likely the result of the admixture of diverse pathogen genotypes through the trade of infected plant material from various geographic origins, the presence of several hosts in the same nursery, and genetic recombination through sexual reproduction of this heterothallic species. PMID:27111805

  12. Race-specific molecules that protect soybeans from Phytophthora megasperma var. sojae*

    PubMed Central

    Wade, Mark; Albersheim, Peter

    1979-01-01

    Phytophthora megasperma var. sojae (A. A. Hildebrand) is a fungal stem and root rot-causing pathogen of soybeans. Glycoproteins secreted into the medium of the aseptically cultured fungus have been partially purified by (NH4)2SO4 precipitation and by column chromatography on norleucine-substituted Sepharose 4B and on DEAE-cellulose. Glycoprotein preparations from P. megasperma var. sojae races 1, 2, and 3 have been tested on four cultivars of soybeans. The partially purified glycoproteins from incompatible races of the pathogen (races that cannot successfully infect the plant), but not those from compatible races (races that can kill the plant), protect soybean seedlings from attack by compatible races. The seedlings are protected by introducing the glycoproteins into hypocotyl wounds of seedlings either 90 min prior to or at the time of inoculation of the wounds with mycelia of one of the pathogens. The glycoprotein preparations are poor nonspecific elicitors of phytoalexin accumulation; the glycoproteins have less than 1.0% of the elicitor activity of the glucans present in the mycelial walls of the pathogen. Images PMID:16592713

  13. Strong genetic differentiation between North American and European populations of Phytophthora alni subsp. uniformis.

    PubMed

    Aguayo, Jaime; Adams, Gerard C; Halkett, Fabien; Catal, Mursel; Husson, Claude; Nagy, Zoltán Á; Hansen, Everett M; Marçais, Benoît; Frey, Pascal

    2013-02-01

    Alder decline caused by Phytophthora alni has been one of the most important diseases of natural ecosystems in Europe during the last 20 years. The emergence of P. alni subsp. alni -the pathogen responsible for the epidemic-is linked to an interspecific hybridization event between two parental species: P. alni subsp. multiformis and P. alni subsp. uniformis. One of the parental species, P. alni subsp. uniformis, has been isolated in several European countries and, recently, in North America. The objective of this work was to assess the level of genetic diversity, the population genetic structure, and the putative reproduction mode and mating system of P. alni subsp. uniformis. Five new polymorphic microsatellite markers were used to contrast both geographical populations. The study comprised 71 isolates of P. alni subsp. uniformis collected from eight European countries and 10 locations in North America. Our results revealed strong differences between continental populations (Fst = 0.88; Rst = 0.74), with no evidence for gene flow. European isolates showed extremely low genetic diversity compared with the North American collection. Selfing appears to be the predominant mating system in both continental collections. The results suggest that the European P. alni subsp. uniformis population is most likely alien and derives from the introduction of a few individuals, whereas the North American population probably is an indigenous population. PMID:23095465

  14. Strong genetic differentiation between North American and European populations of Phytophthora alni subsp. uniformis.

    PubMed

    Aguayo, Jaime; Adams, Gerard C; Halkett, Fabien; Catal, Mursel; Husson, Claude; Nagy, Zoltán Á; Hansen, Everett M; Marçais, Benoît; Frey, Pascal

    2013-02-01

    Alder decline caused by Phytophthora alni has been one of the most important diseases of natural ecosystems in Europe during the last 20 years. The emergence of P. alni subsp. alni -the pathogen responsible for the epidemic-is linked to an interspecific hybridization event between two parental species: P. alni subsp. multiformis and P. alni subsp. uniformis. One of the parental species, P. alni subsp. uniformis, has been isolated in several European countries and, recently, in North America. The objective of this work was to assess the level of genetic diversity, the population genetic structure, and the putative reproduction mode and mating system of P. alni subsp. uniformis. Five new polymorphic microsatellite markers were used to contrast both geographical populations. The study comprised 71 isolates of P. alni subsp. uniformis collected from eight European countries and 10 locations in North America. Our results revealed strong differences between continental populations (Fst = 0.88; Rst = 0.74), with no evidence for gene flow. European isolates showed extremely low genetic diversity compared with the North American collection. Selfing appears to be the predominant mating system in both continental collections. The results suggest that the European P. alni subsp. uniformis population is most likely alien and derives from the introduction of a few individuals, whereas the North American population probably is an indigenous population.

  15. Winter Conditions Correlate with Phytophthora alni Subspecies Distribution in Southern Sweden.

    PubMed

    Redondo, Miguel A; Boberg, Johanna; Olsson, Christer H B; Oliva, Jonàs

    2015-09-01

    During the last century, the number of forest pathogen invasions has increased substantially. Environmental variables can play a crucial role in determining the establishment of invasive species. The objective of the present work was to determine the correlation between winter climatic conditions and distribution of two subspecies of the invasive forest pathogen Phytophthora alni: P. alni subspp. alni and uniformis killing black alder (Alnus glutinosa) in southern Sweden. It is known from laboratory experiments that P. alni subsp. alni is more pathogenic than P. alni subsp. uniformis, and that P. alni subsp. alni is sensitive to low temperatures and long frost periods. By studying the distribution of these two subspecies at the northern limit of the host species, we could investigate whether winter conditions can affect the geographical distribution of P. alni subsp. alni spreading northward. Sixteen major river systems of southern Sweden were systematically surveyed and isolations were performed from active cankers. The distribution of the two studied subspecies was highly correlated with winter temperature and duration of periods with heavy frost. While P. alni subsp. uniformis covered the whole range of temperatures of the host, P. alni subsp. alni was recovered in areas subjected to milder winter temperatures and shorter frost periods. Our observations suggest that winter conditions can play an important role in limiting P. alni subsp. alni establishment in cold locations, thus affecting the distribution of the different subspecies of P. alni in boreal regions.

  16. Identification and validation of polymorphic microsatellite loci for the analysis of Phytophthora nicotianae populations.

    PubMed

    Biasi, Antonio; Martin, Frank; Schena, Leonardo

    2015-03-01

    A large number of SSR loci were screened in the genomic assemblies of 14 different isolates of Phytophthora nicotianae and primers were developed for amplification of 17 markers distributed among different contigs. These loci were highly polymorphic and amplified from genetically distant isolates of the pathogen. Among these, nine were further validated using a multiplexed genotyping assay with differentially labeled primers (FAM or HEX) to allow for duplex PCR amplification. The use of reverse primers with a 5' PIG tail was important to increase the quality and reliability of the analyses. A total of 46 alleles were detected in 5 tester isolates of P. nicotianae representing the breadth of diversity in the species. Furthermore, a high incidence of heterozygosity was determined with two alleles detected in 67% of the primer/isolate combinations. Three different alleles where detected for a single locus/isolate combination, indicating variation in ploidy. These markers represent a valuable new tool for the characterization of populations of P. nicotianae.

  17. Proteomic analysis on zoxamide-induced sensitivity changes in Phytophthora cactorum.

    PubMed

    Mei, Xinyue; Yang, Min; Jiang, Bingbing; Ding, Xupo; Deng, Weiping; Dong, Yumei; Chen, Lei; Liu, Xili; Zhu, Shusheng

    2015-09-01

    Zoxamide is an important fungicide for oomycete disease management. In this study, we established the baseline sensitivity of Phytophthora cactorum to zoxamide and assessed the risk of developing resistance to zoxamide using ultraviolet irradiation and fungicide taming methods. All 73 studied isolates were sensitive to zoxamide, with effective concentrations for 50% inhibition of mycelial growth ranging from 0.04 to 0.29 mg/L and mean of 0.15 mg/L. Stable zoxamide-resistant mutants of P. cactorum were not obtained from four arbitrarily selected isolates by either treating mycelial cultures with ultraviolet irradiation or adapting mycelial cultures to the addition of increasing zoxamide concentrations. However, the sensitivity of the isolates to zoxamide could be easily reduced by successive zoxamide treatments. In addition to displaying decreased sensitivity to zoxamide, these isolates also showed decreased sensitivity to the fungicides flumorph and cymoxanil. Proteomic analysis indicated that some proteins involved in antioxidant detoxification, ATP-dependent multidrug resistance, and anti-apoptosis activity, are likely responsible for the induced decrease in the sensitivity of P. cactorum to zoxamide compared to controls. Further semi-quantitative PCR analysis demonstrated that the gene expression profiles of most of above proteins were consistent with the proteomic analysis. Based on the above results, P. cactorum shows low resistance risk to zoxamide; however, the fungicidal effect of zoxamide might be decreased due to induced resistance when this fungicide is continuously applied.

  18. Antagonism of antifungal metabolites from Streptomyces griseus H7602 against Phytophthora capsici.

    PubMed

    Nguyen, Xuan Hoa; Naing, Kyaw Wai; Lee, Young Seong; Kim, Yong Hwan; Moon, Jae Hak; Kim, Kil Yong

    2015-01-01

    In this study, evidences for antagonism were established by production of antifungal metabolites from Streptomyces griseus H7602, which were active to inhibit mycelial growth of Phytophthora capsici in the in vitro assays. Mycelial growth and zoosporangia formation of P. capsici was strongly inhibited in the medium containing the cell free culture filtrate of S. griseus H7602. Antifungal metabolites from the cell free culture filtrate of S. griseus H7602 showed substantial antagonistic effects on P. capsici. In addition, a purified antifungal compound was separated from the antifungal metabolites of S. griseus H7602 and identified to be 1H-pyrrole-2-carboxylic acid (PCA) by spectra analyses. PCA showed strong antifungal activity and was evaluated for the first time for its antagonism against P. capsici under in vitro conditions. Minimum inhibitory concentration (MIC) value of PCA was low (4 µg ml(-1)), and the mycelial growth of P. capsici was almost inhibited at concentration of 64 µg ml(-1). This study suggests that the PCA may be useful as biofungicides against P. capsici, and the prominent antagonism of antifungal metabolites from S. griseus H7602 highlights it as a candidate for biocontrol of P. capsici.

  19. Microarray profiling reveals microRNAs involving soybean resistance to Phytophthora sojae.

    PubMed

    Guo, Na; Ye, Wen-Wu; Wu, Xiao-Ling; Shen, Dan-Yu; Wang, Yuan-Chao; Xing, Han; Dou, Dao-Long

    2011-11-01

    MicroRNAs (miRNAs), a group of small noncoding RNAs, may serve as a class of post-transcriptional regulators in plant immune systems. Nevertheless, little is known about their roles in plant immune response to the oomycete pathogens. To identify miRNAs involved in the response of soybean to Phytophthora sojae, we examined expressional patterns of miRNAs upon infection by P. sojae by microarray analysis in three soybean cultivars: Williams (susceptible), Conrad (quantitative resistance), and Williams 82 (qualitative resistance). Expression of a number of miRNAs was significantly altered upon infection and (or) in the different genotypes. qRT-PCR data with some miRNAs further confirmed the microarray results. Comparative analysis of the selected miRNAs and their targeted gene expression datasets uncovered many reciprocally expressed miRNA-target pairs, which could proposed a feedback circuit between miRNA(s) and protein-coding genes. These results may serve as a basis for further in-depth studies of miRNAs involved in soybean resistance to P. sojae.

  20. Effect of a benzothiadiazole on inducing resistance of soybean to Phytophthora sojae.

    PubMed

    Han, Qingmei; Feng, Hao; Zhao, Haiyan; Huang, Lili; Wang, Xiaojie; Wang, Xiaodong; Kang, Zhensheng

    2013-04-01

    Effects of benzothidiazole (BTH), an inducer of resistance, were examined in a compatible interaction of soybean seedlings and Phytophthora sojae using electron microscopy and quantitative real-time polymerase chain reaction (qRT-PCR) techniques. Seedlings were sprayed with BTH 2 days before inoculation of hypocotyls with zoospore suspension of P. sojae. In hypocotyls treated with BTH, the infection process of P. sojae was significantly delayed, and also the structures of hyphae and haustorium-like bodies were remarkably altered. These changes included increased vacuolation, plasmolysis, degeneration of cytoplasm, and collapse of hyphae and haustorium-like bodies. Large morphological differences were detected in P. sojae-infected hypocotyl tissue treated with BTH compared with infected but non-treated control tissue. Very thick layers of wall appositions were formed in the host cells contacting with hyphae, whereas such structures were never observed in only P. sojae-infected control hypocotyls. In addition, five pathogenesis-related (PR)-genes were selected to detect their transcription changes using qRT-PCR. Expression of PR-1, PR-3a, PR-3b, PR-9, and PR-10 genes were induced in BTH-treated and P. sojae-inoculated tissue at different times and levels. The up-regulated expression of these genes as well as the morphological defense structures may contribute to disease resistance in soybean hypocotyls to P. sojae. PMID:22777214

  1. Transient fusion and selective secretion of vesicle proteins in Phytophthora nicotianae zoospores

    PubMed Central

    Zhang, Weiwei; Blackman, Leila M.

    2013-01-01

    Secretion of pathogen proteins is crucial for the establishment of disease in animals and plants. Typically, early interactions between host and pathogen trigger regulated secretion of pathogenicity factors that function in pathogen adhesion and host penetration. During the onset of plant infection by spores of the Oomycete, Phytophthora nicotianae, proteins are secreted from three types of cortical vesicles. Following induction of spore encystment, two vesicle types undergo full fusion, releasing their entire contents onto the cell surface. However, the third vesicle type, so-called large peripheral vesicles, selectively secretes a small Sushi domain-containing protein, PnCcp, while retaining a large glycoprotein, PnLpv, before moving away from the plasma membrane. Selective secretion of PnCcp is associated with its compartmentalization within the vesicle periphery. Pharmacological inhibition of dynamin function, purportedly in vesicle fission, by dynasore treatment provides evidence that selective secretion of PnCcp requires transient fusion of the large peripheral vesicles. This is the first report of selective protein secretion via transient fusion outside mammalian cells. Selective secretion is likely to be an important aspect of plant infection by this destructive pathogen. PMID:24392285

  2. Identification and analysis of Phytophthora cactorum genes up-regulated during cyst germination and strawberry infection.

    PubMed

    Chen, Xiaoren; Klemsdal, Sonja Sletner; Brurberg, May Bente

    2011-10-01

    The oomycete Phytophthora cactorum can cause economically important diseases on numerous host plants worldwide, such as crown rot on strawberry. To explore the molecular mechanisms underlying the pathogenicity of P. cactorum on strawberry, transcriptional analysis of P. cactorum during strawberry infection and cyst germination was performed by applying suppression subtractive hybridization (SSH) and effector-specific differential display (ESDD) techniques. Two SSH cDNA libraries were generated, enriched for P. cactorum genes expressed during infection or during cyst germination, respectively, and 137 unique differentially expressed genes were identified. To specifically select RxLR effector genes from P. cactorum, ESDD was performed using RxLR and EER motif-based degenerate primers. Eight RxLR effector candidate genes as well as 67 other genes were identified out of 124 selected fragments. The expression levels of 20 putatively up-regulated genes were further analyzed using real-time RT-PCR, showing that, indeed 19 of these 20 genes were up-regulated during at least one of the studied developmental stages or during strawberry crown invasion, relative to the mycelium. This study provides a first overview of P. cactorum genes that are up-regulated immediately prior to or during strawberry infection and also provides a novel method for selecting RxLR effector genes from the unsequenced genome of P. cactorum.

  3. A method to quantify infection and colonization of holm oak (Quercus ilex) roots by Phytophthora cinnamomi

    PubMed Central

    2012-01-01

    Phytophthora cinnamomi Rands. is an important root rot pathogen widely distributed in the north hemisphere, with a large host range. Among others diseases, it is known to be a principal factor in the decline of holm oak and cork oak, the most important tree species in the “dehesa” ecosystem of south-western Spain. Previously, the focus of studies on P. cinnamomi and holm oak have been on molecular tools for identification, functional responses of the host, together with other physiological and morphological host variables. However, a microscopic index to describe the degree of infection and colonization in the plant tissues has not yet been developed. A colonization or infection index would be a useful tool for studies that examine differences between individuals subjected to different treatments or to individuals belonging to different breeding accessions, together with their specific responses to the pathogen. This work presents a methodology based on the capture and digital treatment of microscopic images, using simple and accessible software, together with a range of variables that quantify the infection and colonization process. PMID:22974221

  4. Expression of resistance gene analogs in woodland strawberry (Fragaria vesca) during infection with Phytophthora cactorum.

    PubMed

    Chen, Xiao-Ren; Brurberg, May Bente; Elameen, Abdelhameed; Klemsdal, Sonja Sletner; Martinussen, Inger

    2016-10-01

    Important losses in strawberry production are often caused by the oomycete Phytophthora cactorum, the causal agent of crown rot. However, very limited studies at molecular levels exist of the mechanisms related to strawberry resistance against this pathogen. To begin to rectify this situation, a PCR-based approach (NBS profiling) was used to isolate strawberry resistance gene analogs (RGAs) with altered expression in response to P. cactorum during a time course (2, 4, 6, 24, 48, 96 and 192 h post-infection). Twenty-three distinct RGA fragments of the NB-LRR type were identified from a resistance genotype (Bukammen) of the wild species Fragaria vesca. The gene transcriptional profiles after infection showed that the response of most RGAs was quicker and stronger in the resistance genotype (Bukammen) than in the susceptible one (FDP821) during the early infection stage. The transcriptional patterns of one RGA (RGA109) were further monitored and compared during the P. cactorum infection of two pairs of resistant and susceptible genotype combinations (Bukammen/FDP821 and FDR1218/1603). The 5' end sequence was cloned, and its putative protein was characteristic of NBS-LRR R protein. Our results yielded a first insight into the strawberry RGAs responding to P. cactorum infection at molecular level.

  5. In planta selfing and oospore production of Phytophthora cinnamomi in the presence of Acacia pulchella.

    PubMed

    Jayasekera, Arunodini U; McComb, Jen A; Shearer, Bryan L; Hardy, Giles E St J

    2007-03-01

    This paper provides the first evidence of A2 type 1 and type 2 isolates of Phytophthora cinnamomi producing selfed oospores in planta in an Australian soil and in a potting mix. Oospores were observed in infected lupin (Lupinus angustifolius) roots incubated for 7d either in the substrate under potted Acacia pulchella plants, or in soils collected from under and near varieties of A. pulchella in jarrah (Eucalyptus marginata) forest. The A2 type isolates varied in their ability to produce selfed oospores and none were produced by A1 isolates. The gametangial association was amphigynous and spores were predominantly spherical with diameters from 13-40 microm. The oospores were viable but dormant. Two A2 type isolates produced small numbers of selfed oospores with amphigynous antheridia axenically in Ribeiro's liquid medium within 30 d, and one A2 type 2 isolate produced oospores after mating with an A1 strain. Evidence is presented that the presence of roots of Acacia pulchella, and particularly A. pulchella var. glaberrima and var. goadbyi, enhances the production of oospores. PMID:17350243

  6. Host Phenology and Leaf Effects on Susceptibility of California Bay Laurel to Phytophthora ramorum.

    PubMed

    Johnston, Steven F; Cohen, Michael F; Torok, Tamas; Meentemeyer, Ross K; Rank, Nathan E

    2016-01-01

    Spread of the plant pathogen Phytophthora ramorum, causal agent of the forest disease sudden oak death, is driven by a few competent hosts that support spore production from foliar lesions. The relationship between traits of a principal foliar host, California bay laurel (Umbellularia californica), and susceptibility to P. ramorum infection were investigated with multiple P. ramorum isolates and leaves collected from multiple trees in leaf-droplet assays. We examined whether susceptibility varies with season, leaf age, or inoculum position. Bay laurel susceptibility was highest during spring and summer and lowest in winter. Older leaves (>1 year) were more susceptible than younger ones (8 to 11 months). Susceptibility was greater at leaf tips and edges than the middle of the leaf. Leaf surfaces wiped with 70% ethanol were more susceptible to P. ramorum infection than untreated leaf surfaces. Our results indicate that seasonal changes in susceptibility of U. californica significantly influence P. ramorum infection levels. Thus, in addition to environmental variables such as temperature and moisture, variability in host plant susceptibility contributes to disease establishment of P. ramorum.

  7. Forest type influences transmission of Phytophthora ramorum in California oak woodlands.

    PubMed

    Davidson, Jennifer M; Patterson, Heather A; Wickland, Allison C; Fichtner, Elizabeth J; Rizzo, David M

    2011-04-01

    The transmission ecology of Phytophthora ramorum from bay laurel (Umbellularia californica) leaves was compared between mixed-evergreen and redwood forest types throughout winter and summer disease cycles in central, coastal California. In a preliminary multisite study, we found that abscission rates of infected leaves were higher at mixed-evergreen sites. In addition, final infection counts were slightly higher at mixed-evergreen sites or not significantly different than at redwood sites, in part due to competition from other foliar pathogens at redwood sites. In a subsequent, detailed study of paired sites where P. ramorum was the main foliar pathogen, summer survival of P. ramorum in bay laurel leaves was lower in mixed-evergreen forest due to lower recovery from infected attached leaves and higher abscission rates of infected leaves. Onset of inoculum production and new infections of bay laurel leaves occurred later in mixed-evergreen forest. Mean inoculum levels in rainwater and final infection counts on leaves were higher in redwood forest. Based on these two studies, lower summer survival of reservoir inoculum in bay laurel leaves in mixed-evergreen forest may result in delayed onset of both inoculum production and new infections, leading to slower disease progress in the early rainy season compared with redwood forest. Although final infection counts also will depend on other foliar pathogens and disease history, in sites where P. ramorum is the main foliar pathogen, these transmission patterns suggest higher rates of disease spread in redwood forests during rainy seasons of short or average length.

  8. Comparative Genomic Analysis among Four Representative Isolates of Phytophthora sojae Reveals Genes under Evolutionary Selection

    PubMed Central

    Ye, Wenwu; Wang, Yang; Tyler, Brett M.; Wang, Yuanchao

    2016-01-01

    Comparative genomic analysis is useful for identifying genes affected by evolutionary selection and for studying adaptive variation in gene functions. In Phytophthora sojae, a model oomycete plant pathogen, the related study is lacking. We compared sequence data among four isolates of P. sojae, which represent its four major genotypes. These isolates exhibited >99.688%, >99.864%, and >98.981% sequence identities at genome, gene, and non-gene regions, respectively. One hundred and fifty-three positive selection and 139 negative selection candidate genes were identified. Between the two categories of genes, the positive selection genes were flanked by larger intergenic regions, poorly annotated in function, and less conserved; they had relatively lower transcription levels but many genes had increased transcripts during infection. Genes coding for predicted secreted proteins, particularly effectors, were overrepresented in positive selection. Several RxLR effector genes were identified as positive selection genes, exhibiting much stronger positive selection levels. In addition, candidate genes with presence/absence polymorphism were analyzed. This study provides a landscape of genomic variation among four representative P. sojae isolates and characterized several evolutionary selection-affected gene candidates. The results suggest a relatively covert two-speed genome evolution pattern in P. sojae and will provide clues for identification of new virulence factors in the oomycete plant pathogens. PMID:27746768

  9. Selection of genetically diverse Trichoderma spp. isolates for suppression of Phytophthora capsici on bell pepper.

    PubMed

    Roberts, Daniel P; Maul, Jude E; McKenna, Laurie F; Emche, Sarah E; Meyer, Susan L F; Collins, Ronald T; Bowers, John H

    2010-10-01

    Environmentally compatible control measures are needed for suppression of Phytophthora capsici on pepper. Twenty-three isolates of Trichoderma were screened for suppression of a mixture of 4 genetically distinct isolates of this pathogen on bell pepper (Capsicum anuum) in greenhouse pot assays. Of these 23 isolates, GL12, GL13, and Th23 provided significant suppression of P. capsici in at least 2 assays. These isolates were then compared with Trichoderma virens isolates GL3 and GL21 for suppression of this disease in the presence and absence of the harpin-based natural product Messenger. Isolates GL3 and Th23 provided significant disease suppression (P ≤ 0.05) in 3 of 4 assays, while GL12, GL13, and GL21 provided significant suppression in 2 of 4 assays. There was no apparent benefit from the application of Messenger. Phylogenetic analysis of these 5 isolates (based on the ITS1 region of the nuclear rDNA cluster and tef1), and an additional 9 isolates that suppressed P. capsici in at least 1 assay, separated isolates into 2 clades, with 1 clade containing GL3, GL12, GL13, and GL21. There were also 2 more distantly related isolates, one of which was Th23. We report here the identification of genetically distinct Trichoderma isolates for potential use in disease management strategies employing isolate combinations directed at suppression of P. capsici on pepper.

  10. Standardizing the nomenclature for clonal lineages of the sudden oak death pathogen, Phytophthora ramorum.

    PubMed

    Grünwald, Niklaus J; Goss, Erica M; Ivors, Kelly; Garbelotto, Matteo; Martin, Frank N; Prospero, Simone; Hansen, Everett; Bonants, Peter J M; Hamelin, Richard C; Chastagner, Gary; Werres, Sabine; Rizzo, David M; Abad, Gloria; Beales, Paul; Bilodeau, Guillaume J; Blomquist, Cheryl L; Brasier, Clive; Brière, Stephan C; Chandelier, Anne; Davidson, Jennifer M; Denman, Sandra; Elliott, Marianne; Frankel, Susan J; Goheen, Ellen M; de Gruyter, Hans; Heungens, Kurt; James, Delano; Kanaskie, Alan; McWilliams, Michael G; Man in 't Veld, Willem; Moralejo, Eduardo; Osterbauer, Nancy K; Palm, Mary E; Parke, Jennifer L; Sierra, Ana Maria Perez; Shamoun, Simon F; Shishkoff, Nina; Tooley, Paul W; Vettraino, Anna Maria; Webber, Joan; Widmer, Timothy L

    2009-07-01

    Phytophthora ramorum, the causal agent of sudden oak death and ramorum blight, is known to exist as three distinct clonal lineages which can only be distinguished by performing molecular marker-based analyses. However, in the recent literature there exists no consensus on naming of these lineages. Here we propose a system for naming clonal lineages of P. ramorum based on a consensus established by the P. ramorum research community. Clonal lineages are named with a two letter identifier for the continent on which they were first found (e.g., NA = North America; EU = Europe) followed by a number indicating order of appearance. Clonal lineages known to date are designated NA1 (mating type: A2; distribution: North America; environment: forest and nurseries), NA2 (A2; North America; nurseries), and EU1 (predominantly A1, rarely A2; Europe and North America; nurseries and gardens). It is expected that novel lineages or new variants within the existing three clonal lineages could in time emerge.

  11. Influence of climatic conditions on white tip disease (Phytophthora porri) in leek (Allium porrum).

    PubMed

    De Jonghe, K; Keirsebilck, D; Martens, K; Buysens, S; Höfte, M

    2002-01-01

    In leek, one of the major vegetable crops in Belgium, Phytophthora porri causes the so-called white tip disease. During the growing seasons of 1999, 2000 and 2001 the incidence of the white tip disease and the role of environmental conditions in the appearance were investigated on several non-treated leek fields in Flanders (Belgium). The first symptoms of white tip disease on leek where recorded in July and the disease progressed until March. Lesions appeared after an incubation period of 91 to 204 DD (degree days above -3 degrees C) (t0) and were diagnostic at 120 DD. The obtained data confirmed a disease increase corresponding with an amount of rainfall of more than 20 l/m2 in 4 days in the period t = t0-92 to t = t0-154 DD. A good correlation was found between the daily disease increase on one hand and the leaf wetness, relative humidity and temperature (negative correlation) on the other hand. Daily disease increase only weakly correlated with rainfall. Based on these results recommendations can be made (for further studies) to develop a model, combining several of the climatic factors, to predict infection periods with high risk on disease increase in the production of leek. PMID:12701432

  12. Host Phenology and Leaf Effects on Susceptibility of California Bay Laurel to Phytophthora ramorum.

    PubMed

    Johnston, Steven F; Cohen, Michael F; Torok, Tamas; Meentemeyer, Ross K; Rank, Nathan E

    2016-01-01

    Spread of the plant pathogen Phytophthora ramorum, causal agent of the forest disease sudden oak death, is driven by a few competent hosts that support spore production from foliar lesions. The relationship between traits of a principal foliar host, California bay laurel (Umbellularia californica), and susceptibility to P. ramorum infection were investigated with multiple P. ramorum isolates and leaves collected from multiple trees in leaf-droplet assays. We examined whether susceptibility varies with season, leaf age, or inoculum position. Bay laurel susceptibility was highest during spring and summer and lowest in winter. Older leaves (>1 year) were more susceptible than younger ones (8 to 11 months). Susceptibility was greater at leaf tips and edges than the middle of the leaf. Leaf surfaces wiped with 70% ethanol were more susceptible to P. ramorum infection than untreated leaf surfaces. Our results indicate that seasonal changes in susceptibility of U. californica significantly influence P. ramorum infection levels. Thus, in addition to environmental variables such as temperature and moisture, variability in host plant susceptibility contributes to disease establishment of P. ramorum. PMID:26439707

  13. A Phytophthora sojae cytoplasmic effector mediates disease resistance and abiotic stress tolerance in Nicotiana benthamiana.

    PubMed

    Zhang, Meixiang; Ahmed Rajput, Nasir; Shen, Danyu; Sun, Peng; Zeng, Wentao; Liu, Tingli; Juma Mafurah, Joseph; Dou, Daolong

    2015-06-03

    Each oomycete pathogen encodes a large number of effectors. Some effectors can be used in crop disease resistance breeding, such as to accelerate R gene cloning and utilisation. Since cytoplasmic effectors may cause acute physiological changes in host cells at very low concentrations, we assume that some of these effectors can serve as functional genes for transgenic plants. Here, we generated transgenic Nicotiana benthamiana plants that express a Phytophthora sojae CRN (crinkling and necrosis) effector, PsCRN115. We showed that its expression did not significantly affect the growth and development of N. benthamiana, but significantly improved disease resistance and tolerance to salt and drought stresses. Furthermore, we found that expression of heat-shock-protein and cytochrome-P450 encoding genes were unregulated in PsCRN115-transgenic N. benthamiana based on digital gene expression profiling analyses, suggesting the increased plant defence may be achieved by upregulation of these stress-related genes in transgenic plants. Thus, PsCRN115 may be used to improve plant tolerance to biotic and abiotic stresses.

  14. Influence of climatic conditions on white tip disease (Phytophthora porri) in leek (Allium porrum).

    PubMed

    De Jonghe, K; Keirsebilck, D; Martens, K; Buysens, S; Höfte, M

    2002-01-01

    In leek, one of the major vegetable crops in Belgium, Phytophthora porri causes the so-called white tip disease. During the growing seasons of 1999, 2000 and 2001 the incidence of the white tip disease and the role of environmental conditions in the appearance were investigated on several non-treated leek fields in Flanders (Belgium). The first symptoms of white tip disease on leek where recorded in July and the disease progressed until March. Lesions appeared after an incubation period of 91 to 204 DD (degree days above -3 degrees C) (t0) and were diagnostic at 120 DD. The obtained data confirmed a disease increase corresponding with an amount of rainfall of more than 20 l/m2 in 4 days in the period t = t0-92 to t = t0-154 DD. A good correlation was found between the daily disease increase on one hand and the leaf wetness, relative humidity and temperature (negative correlation) on the other hand. Daily disease increase only weakly correlated with rainfall. Based on these results recommendations can be made (for further studies) to develop a model, combining several of the climatic factors, to predict infection periods with high risk on disease increase in the production of leek.

  15. Population History and Pathways of Spread of the Plant Pathogen Phytophthora plurivora

    PubMed Central

    Schoebel, Corine N.; Stewart, Jane; Gruenwald, Niklaus J.; Rigling, Daniel; Prospero, Simone

    2014-01-01

    Human activity has been shown to considerably affect the spread of dangerous pests and pathogens worldwide. Therefore, strict regulations of international trade exist for particularly harmful pathogenic organisms. Phytophthora plurivora, which is not subject to regulations, is a plant pathogen frequently found on a broad range of host species, both in natural and artificial environments. It is supposed to be native to Europe while resident populations are also present in the US. We characterized a hierarchical sample of isolates from Europe and the US and conducted coalescent-, migration, and population genetic analysis of sequence and microsatellite data, to determine the pathways of spread and the demographic history of this pathogen. We found P. plurivora populations to be moderately diverse but not geographically structured. High levels of gene flow were observed within Europe and unidirectional from Europe to the US. Coalescent analyses revealed a signal of a recent expansion of the global P. plurivora population. Our study shows that P. plurivora has most likely been spread around the world by nursery trade of diseased plant material. In particular, P. plurivora was introduced into the US from Europe. International trade has allowed the pathogen to colonize new environments and/or hosts, resulting in population growth. PMID:24427303

  16. Expression of resistance gene analogs in woodland strawberry (Fragaria vesca) during infection with Phytophthora cactorum.

    PubMed

    Chen, Xiao-Ren; Brurberg, May Bente; Elameen, Abdelhameed; Klemsdal, Sonja Sletner; Martinussen, Inger

    2016-10-01

    Important losses in strawberry production are often caused by the oomycete Phytophthora cactorum, the causal agent of crown rot. However, very limited studies at molecular levels exist of the mechanisms related to strawberry resistance against this pathogen. To begin to rectify this situation, a PCR-based approach (NBS profiling) was used to isolate strawberry resistance gene analogs (RGAs) with altered expression in response to P. cactorum during a time course (2, 4, 6, 24, 48, 96 and 192 h post-infection). Twenty-three distinct RGA fragments of the NB-LRR type were identified from a resistance genotype (Bukammen) of the wild species Fragaria vesca. The gene transcriptional profiles after infection showed that the response of most RGAs was quicker and stronger in the resistance genotype (Bukammen) than in the susceptible one (FDP821) during the early infection stage. The transcriptional patterns of one RGA (RGA109) were further monitored and compared during the P. cactorum infection of two pairs of resistant and susceptible genotype combinations (Bukammen/FDP821 and FDR1218/1603). The 5' end sequence was cloned, and its putative protein was characteristic of NBS-LRR R protein. Our results yielded a first insight into the strawberry RGAs responding to P. cactorum infection at molecular level. PMID:27447867

  17. Phytophthora parasitica biofilm formation: installation and organization of microcolonies on the surface of a host plant.

    PubMed

    Galiana, Eric; Fourré, Sandra; Engler, Gilbert

    2008-08-01

    Zoospores of the oomycete Phytophthora parasitica establish microbial spheroid microcolonies and biofilms on the surface of wounded leaves of their host, Nicotiana tabacum. The formation of microcolonies involves the movement of some zoospores towards attractants from wound sites, followed by their irreversible adsorption and the formation of a cluster of cells. These cells drive the migration of a second wave of zoospores (several hundreds cells) by setting up an external chemotactic gradient leading to massive zoospore encystment and cyst-orientated germination. Zoospores that are still swimming at this stage circulate within the nascent biofilm by opening channels. Concomitantly, the cell population secretes various substances to elaborate an extracellular mucilage. Embedded within the extracellular matrix, biofilm cells are organized into a structured community as coacervates. The granular surface is composed of individual cysts, located on the outside of the microcolony. Hyphae from these cysts plunge downwards towards the dense core formed by the founder cells. This report is the first to show the installation and organization of a biofilm formed by eukaryotic cells on plant surfaces. The P. parasitica microcolonies constitute heterogeneous microenvironments for the embedded and circulating cells. They may affect plant-pathogen interactions by serving as reservoirs for pathogenic microorganisms, as protecting niche against host defences or as structures for infecting populations.

  18. Transient fusion and selective secretion of vesicle proteins in Phytophthora nicotianae zoospores.

    PubMed

    Zhang, Weiwei; Blackman, Leila M; Hardham, Adrienne R

    2013-01-01

    Secretion of pathogen proteins is crucial for the establishment of disease in animals and plants. Typically, early interactions between host and pathogen trigger regulated secretion of pathogenicity factors that function in pathogen adhesion and host penetration. During the onset of plant infection by spores of the Oomycete, Phytophthora nicotianae, proteins are secreted from three types of cortical vesicles. Following induction of spore encystment, two vesicle types undergo full fusion, releasing their entire contents onto the cell surface. However, the third vesicle type, so-called large peripheral vesicles, selectively secretes a small Sushi domain-containing protein, PnCcp, while retaining a large glycoprotein, PnLpv, before moving away from the plasma membrane. Selective secretion of PnCcp is associated with its compartmentalization within the vesicle periphery. Pharmacological inhibition of dynamin function, purportedly in vesicle fission, by dynasore treatment provides evidence that selective secretion of PnCcp requires transient fusion of the large peripheral vesicles. This is the first report of selective protein secretion via transient fusion outside mammalian cells. Selective secretion is likely to be an important aspect of plant infection by this destructive pathogen. PMID:24392285

  19. A method to quantify infection and colonization of holm oak (Quercus ilex) roots by Phytophthora cinnamomi.

    PubMed

    Ruiz-Gómez, Francisco J; Sánchez-Cuesta, Rafael; Navarro-Cerrillo, Rafael M; Pérez-de-Luque, Alejandro

    2012-01-01

    Phytophthora cinnamomi Rands. is an important root rot pathogen widely distributed in the north hemisphere, with a large host range. Among others diseases, it is known to be a principal factor in the decline of holm oak and cork oak, the most important tree species in the "dehesa" ecosystem of south-western Spain. Previously, the focus of studies on P. cinnamomi and holm oak have been on molecular tools for identification, functional responses of the host, together with other physiological and morphological host variables. However, a microscopic index to describe the degree of infection and colonization in the plant tissues has not yet been developed. A colonization or infection index would be a useful tool for studies that examine differences between individuals subjected to different treatments or to individuals belonging to different breeding accessions, together with their specific responses to the pathogen. This work presents a methodology based on the capture and digital treatment of microscopic images, using simple and accessible software, together with a range of variables that quantify the infection and colonization process.

  20. Screening of bacterial antagonists for biological control of Phytophthora blight of pepper.

    PubMed

    Rajkumar, M; Lee, Wang Hyu; Lee, Kui Jae

    2005-01-01

    The aim of this study was to assess the potential of bacterial antagonists to control Phytophthora blight of pepper caused by P. capsici using different screening methods. Among a collection of fluorescent pseudomonas isolated from the rhizosphere of pepper, twelve isolates were initially selected based on dual culture assay on potato dextrose agar and corn meal agar. Further, these twelve isolates were screened for the reduction of disease severity caused by P. capsici using detached leaves and seedling assay. Most of the antagonists showed varying levels of antagonism against P. capsici in both detached leaves and seedlings assay. In addition, few isolates increased shoot and root length of pepper in seedling assays. Among them, isolate PS119 showing highest ability to reduce the disease severity in the in vitro seedling assay was found to be the most efficient antagonists against P. capsici in the in vivo biological control tests. These results indicate that the in vitro seedling assay can be used as a rapid and more accurate technique for the selection of promising biocontrol agents against P. capsici. PMID:15678563