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Sample records for infect rice citrus

  1. Citrus Functional Genomics and Molecular Modeling in Relation to Citrus sinensis (Sweet Orange) Infection with Xylella fastidiosa (Citrus Variegated Chlorosis).

    PubMed

    Dwivedi, Upendra N; Tiwari, Sameeksha; Prasanna, Pragya; Awasthi, Manika; Singh, Swati; Pandey, Veda P

    2016-08-01

    Citrus are among the economically most important fruit tree crops in the world. Citrus variegated chlorosis (CVC), caused by Xylella fastidiosa infection, is a serious disease limiting citrus production at a global scale. With availability of citrus genomic resources, it is now possible to compare citrus expressed sequence tag (EST) data sets and identify single-nucleotide polymorphisms (SNPs) within and among different citrus cultivars that can be exploited for citrus resistance to infections, citrus breeding, among others. We report here, for the first time, SNPs in the EST data sets of X. fastidiosa-infected Citrus sinensis (sweet orange) and their functional annotation that revealed the involvement of eight C. sinensis candidate genes in CVC pathogenesis. Among these genes were xyloglucan endotransglycosylase, myo-inositol-1-phosphate synthase, and peroxidase were found to be involved in plant cell wall metabolism. These have been further investigated by molecular modeling for their role in CVC infection and defense. Molecular docking analyses of the wild and the mutant (SNP containing) types of the selected three enzymes with their respective substrates revealed a significant decrease in the binding affinity of substrates for the mutant enzymes, thus suggesting a decrease in the catalytic efficiency of these enzymes during infection, thereby facilitating a favorable condition for infection by the pathogen. These findings offer novel agrigenomics insights in developing future molecular targets and strategies for citrus fruit cultivation in ways that are resistant to X. fastidiosa infection, and by extension, with greater harvesting efficiency and economic value.

  2. Pathogen infection drives patterns of nutrient resorption in citrus plants

    PubMed Central

    Cao, Jirong; Cheng, Chunzhen; Yang, Junjie; Wang, Qibing

    2015-01-01

    Nutrient resorption processes in the plants infected by pathogen remain poorly understood. Huanglongbing (HLB) is a destructive disease of citrus. HLB-pathogen ‘Candidatus Liberibacter asiaticus’ grows specifically in the phloem of hosts and may cause problems in the plant vascular system after infection. Therefore, it brings a great concern about the phloem nutrient transport and nutrient intra-cycling in HLB-affected plants. We investigated the effects of ‘Ca. L. asiaticus’ infection on nitrogen (N) and phosphorus (P) concentrations and resorption in different citrus species (i.e. Citrus reticulata, Citrus limon and Citrus maxima). HLB-pathogen infection had distinctive impacts on nutrient resorption in different species. P resorption efficiency substantially decreased in infected C. reticulata plants relative to the healthy plants in summer, which may account for the marked decrease in the average fruit yield. P resorption was more efficient in infected C. limon plants than in the healthy plants. However, for C. maxima plants, HLB had no significant effects on N:P ratio in live leaves and resorption efficiency as well as on fruit yield. Keeping efficient internal nutrient cycling can be a strategy of citrus species being tolerant to HLB. PMID:26419510

  3. Pathogen infection drives patterns of nutrient resorption in citrus plants.

    PubMed

    Cao, Jirong; Cheng, Chunzhen; Yang, Junjie; Wang, Qibing

    2015-09-30

    Nutrient resorption processes in the plants infected by pathogen remain poorly understood. Huanglongbing (HLB) is a destructive disease of citrus. HLB-pathogen 'Candidatus Liberibacter asiaticus' grows specifically in the phloem of hosts and may cause problems in the plant vascular system after infection. Therefore, it brings a great concern about the phloem nutrient transport and nutrient intra-cycling in HLB-affected plants. We investigated the effects of 'Ca. L. asiaticus' infection on nitrogen (N) and phosphorus (P) concentrations and resorption in different citrus species (i.e. Citrus reticulata, Citrus limon and Citrus maxima). HLB-pathogen infection had distinctive impacts on nutrient resorption in different species. P resorption efficiency substantially decreased in infected C. reticulata plants relative to the healthy plants in summer, which may account for the marked decrease in the average fruit yield. P resorption was more efficient in infected C. limon plants than in the healthy plants. However, for C. maxima plants, HLB had no significant effects on N:P ratio in live leaves and resorption efficiency as well as on fruit yield. Keeping efficient internal nutrient cycling can be a strategy of citrus species being tolerant to HLB.

  4. Identification of large masses of citrus fruit and rice fields in eastern Spain

    NASA Technical Reports Server (NTRS)

    Desagredo, F. L.; Salinas, F. G.

    1973-01-01

    ERTS-1 imagery has been successfully used for the identification of large areas of citrus groves and rice fields in the Valencia region of Eastern Spain. Results are encouraging and will facilitate the elaboration of a land use map with a fair degree of definition once methods prove to be fully operational.

  5. Spectral difference analysis and airborne imaging classification for citrus greening infected trees

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus greening, also called Huanglongbing (HLB), became a devastating disease spread through citrus groves in Florida, since it was first found in 2005. Multispectral (MS) and hyperspectral (HS) airborne images of citrus groves in Florida were acquired to detect citrus greening infected trees in 20...

  6. Infectivity of Two Biotypes of the Citrus Nematode on Citrus and on Some Other Hosts

    PubMed Central

    Baines, R. C.; Miyakawa, T.; Cameron, J. W.; Small, R. H.

    1969-01-01

    The infectivity and development of two biotypes of citrus nematode (Tylenchulus semipenetrans) were compared on highly resistant Poncirus trifoliata selection 'Pomeroy,' moderately susceptible 'Troyer' citrange, and highly susceptible sweet orange selection 'Homosassa' small seedlings in a glasshouse. Biotype-1 was more infective on the above hosts and developed faster on sweet orange and on 'Troyer' citrange than Biotype-2. The differences in infectivity were interpreted to reflect differences in the ability of the nematodes to penetrate the epidermis and hypodermis and become established in host roots. Poncirus selections 'Pomeroy,' 'Webber-Fawcett,' and 'Rubidoux' seedlings were highly resistant to the citrus nematode in California, but seedlings of 'Pomeroy' and 'Rubidoux' were only moderately resistant in Japan. These differences in degree of infection may indicate different biotypes of the nematode. Host range tests with California Biotype-1 indicate that it differs from those occurring in Israel. PMID:19325669

  7. Characterization of a virus infecting Citrus volkameriana with citrus leprosis-like symptoms.

    PubMed

    Melzer, Michael J; Sether, Diane M; Borth, Wayne B; Hu, John S

    2012-01-01

    A Citrus volkameriana tree displaying symptoms similar to citrus leprosis on its leaves and bark was found in Hawaii. Citrus leprosis virus C (CiLV-C)-specific detection assays, however, were negative for all tissues tested. Short, bacilliform virus-like particles were observed by transmission electron microscopy in the cytoplasm of symptomatic leaves but not in healthy controls. Double-stranded (ds) RNAs ≈8 and 3 kbp in size were present in symptomatic leaf tissue but not in healthy controls. Excluding poly(A) tails, the largest molecule, RNA1, was 8,354 bp in length. The ≈3 kbp dsRNA band was found to be composed of two distinct molecules, RNA2 and RNA3, which were 3,169 and 3,113 bp, respectively. Phylogenetic analyses indicated that the RNA-dependent RNA polymerase (RdRp) domain located in RNA1 was most closely related to the RdRp domain of CiLV-C. A reverse-transcription polymerase chain reaction assay developed for the detection of this virus was used to screen nearby citrus trees as well as Hibiscus arnottianus plants with symptoms of hibiscus green spot, a disease associated with infection by Hibiscus green spot virus (HGSV). All nearby citrus trees tested negative with the assay; however, symptomatic H. arnottianus plants were positive. All three RNAs were present in symptomatic H. arnottianus and were >98% identical to the RNAs isolated from C. volkameriana. We contend that the virus described in this study is HGSV, and propose that it be the type member of a new virus genus, Higrevirus.

  8. Morphological, molecular and virulence characterization of three Lencanicillium species infecting Asian citrus psyllids in Huangyan citrus groves.

    PubMed

    Lu, Lianming; Cheng, Baoping; Du, Danchao; Hu, Xiurong; Peng, Aitian; Pu, Zhanxu; Zhang, Xiaoya; Huang, Zhendong; Chen, Guoqing

    2015-02-01

    Citrus greening or Huanglongbing (HLB) is caused by the infection of Candidatus Liberibacter spp. in citrus plants. Since Asian citrus psyllid is the primary vector of this bacterial pathogen, the spread of HLB can be mitigated by suppressing Asian citrus psyllid populations in citrus groves using entomopathogens. To expand the current data on entomopathogens infecting Asian citrus psyllids, we isolated and characterized three different entomopathogens. Strains ZJLSP07, ZJLA08, and ZJLP09 infected the Asian citrus psyllid, Diaphorina citri Kuwayama, in Huangyan citrus groves. Based on molecular and morphological analyses, two were identified as Lecanicillium attenuatum and Lecanicillium psalliotae, and the third was recognized as an unidentified species of the genus, Lecanicillium. The corrected mortalities caused by strains ZJLSP07, ZJLA08 were 100% at 7days post-inoculation, while by ZJLP09 complete mortality occurred at 6days after inoculation, with 1.0×10(8)conidia/ml at 25°C and a relative humidity of 90% in the laboratory. Under the same condition, the corrected mortalities caused by strains ZJLSP07, ZJLA08 and ZJLP09 were 100%, 92.55% and 100%, respectively at 9days post-inoculation in the greenhouse. Our findings also revealed that these fungal strains infected D. citri using hyphae that penetrated deep into the insect tissues. Further, all three strains secreted the enzymes proteinases, chitinases and lipases with a potential to destroy insect tissues. Interestingly, strain ZJLP09 had an earlier invasion time and the highest levels of enzyme activities when compared to the other two strains. These findings have expanded the existing pool of entomopathogenic fungi that infect D. citri and can be potentially used for the management of D. citri populations.

  9. Early detection of CLas infections in citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    “Early” detection of CLas infection is essential to minimize the risk of Huanglongbing (HLB) epidemics in areas where the pathogen has been recently introduced. Any delay in confirmation of CLas infection results in delays of regulatory and management actions, and increased spread of the pathogen ev...

  10. Near-Infrared Spectroscopy for discrimination of huanglongbing-infected citrus leaves from uninfected leaves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus greening, also called Huanglongbing (HLB) or yellow dragon disease, is one of the more serious diseases of citrus and is a threat to the U.S. industry. An infected tree produces fruit that is unsuitable for sale as fresh fruit or for juice. The only definitive method of diagnosis of trees su...

  11. Digital Gene Expression Analysis of Ponkan Mandarin (Citrus reticulata Blanco) in Response to Asia Citrus Psyllid-Vectored Huanglongbing Infection.

    PubMed

    Zhong, Yun; Cheng, Chunzhen; Jiang, Bo; Jiang, Nonghui; Zhang, Yongyan; Hu, Minlun; Zhong, Guangyan

    2016-07-02

    Citrus Huanglongbing (HLB), the most destructive citrus disease, can be transmitted by psyllids and diseased budwoods. Although the final symptoms of the two main HLB transmission ways were similar and hard to distinguish, the host responses might be different. In this study, the global gene changes in leaves of ponkan (Citrus reticulata) mandarin trees following psyllid-transmission of HLB were analyzed at the early symptomatic stage (13 weeks post inoculation, wpi) and late symptomatic stage (26 wpi) using digital gene expression (DGE) profiling. At 13 wpi, 2452 genes were downregulated while only 604 genes were upregulated in HLB infected ponkan leaves but no pathway enrichment was identified. Gene function analysis showed impairment in defense at the early stage of infection. At late stage of 26 wpi, however, differentially expressed genes (DEGs) involved in carbohydrate metabolism, plant defense, hormone signaling, secondary metabolism, transcription regulation were overwhelmingly upregulated, indicating that the defense reactions were eventually activated. The results indicated that HLB bacterial infection significantly influenced ponkan gene expression, and a delayed response of the host to the fast growing bacteria might be responsible for its failure in fighting against the bacteria.

  12. Digital Gene Expression Analysis of Ponkan Mandarin (Citrus reticulata Blanco) in Response to Asia Citrus Psyllid-Vectored Huanglongbing Infection

    PubMed Central

    Zhong, Yun; Cheng, Chunzhen; Jiang, Bo; Jiang, Nonghui; Zhang, Yongyan; Hu, Minlun; Zhong, Guangyan

    2016-01-01

    Citrus Huanglongbing (HLB), the most destructive citrus disease, can be transmitted by psyllids and diseased budwoods. Although the final symptoms of the two main HLB transmission ways were similar and hard to distinguish, the host responses might be different. In this study, the global gene changes in leaves of ponkan (Citrus reticulata) mandarin trees following psyllid-transmission of HLB were analyzed at the early symptomatic stage (13 weeks post inoculation, wpi) and late symptomatic stage (26 wpi) using digital gene expression (DGE) profiling. At 13 wpi, 2452 genes were downregulated while only 604 genes were upregulated in HLB infected ponkan leaves but no pathway enrichment was identified. Gene function analysis showed impairment in defense at the early stage of infection. At late stage of 26 wpi, however, differentially expressed genes (DEGs) involved in carbohydrate metabolism, plant defense, hormone signaling, secondary metabolism, transcription regulation were overwhelmingly upregulated, indicating that the defense reactions were eventually activated. The results indicated that HLB bacterial infection significantly influenced ponkan gene expression, and a delayed response of the host to the fast growing bacteria might be responsible for its failure in fighting against the bacteria. PMID:27384559

  13. A sensitive and reliable RT-nested PCR assay for detection of Citrus tristeza virus from naturally infected citrus plants.

    PubMed

    Adkar-Purushothama, Charith Raj; Maheshwar, P K; Sano, Teruo; Janardhana, G R

    2011-05-01

    A specific and sensitive reverse transcriptase-nested polymerase chain reaction assay (RT-nPCR) was developed for the detection of Citrus tristeza virus (CTV) from naturally infected citrus samples. Two sets of primer pairs were designed by alignment of nucleotide sequences available in GenBank database for different genotypes of CTV. RT-nPCR reaction components and thermal cycling parameters were optimized and reaction conditions were standardized. Sequencing of the PCR products from direct and nested-PCR reactions confirmed the specificity of both primer pairs. Presence of CTV specific amplicons in asymptomatic samples which were collected from diseased orchards indicated the sensitivity of the test. As RT-nPCR technique, developed in the present study, is specific and efficient in detecting CTV, this could be envisioned for diagnostic applications and surveillance.

  14. Development of a full-genome cDNA clone of Citrus leaf blotch virus and infection of citrus plants.

    PubMed

    Vives, María Carmen; Martín, Susana; Ambrós, Silvia; Renovell, Agueda; Navarro, Luis; Pina, Jose Antonio; Moreno, Pedro; Guerri, José

    2008-11-01

    Citrus leaf blotch virus (CLBV), a member of the family Flexiviridae, has a ~9-kb single-stranded, positive-sense genomic RNA encapsidated by a 41-kDa coat protein. CLBV isolates are associated with symptom production in citrus including leaf blotching of Dweet tangor and stem pitting in Etrog citron (Dweet mottle disease), and some isolates are associated with bud union crease on trifoliate rootstocks, but Koch's postulates for this virus were not fulfilled. A full-genome cDNA of CLBV isolate SRA-153, which induces bud union crease, was placed under the T7 promoter (clone T7-CLBV), or between the 35S promoter and the Nos-t terminator, with or without a ribozyme sequence downstream of the CLBV sequence (clones 35SRbz-CLBV and 35S-CLBV). RNA transcripts from T7-CLBV failed to infect Etrog citron and Nicotiana occidentalis and N. benthamiana plants, whereas agro-inoculation with binary vectors carrying 35SRbz-CLBV or 35S-CLBV, and the p19 silencing suppressor, caused systemic infection and production of normal CLBV virions. Virus accumulation was similar in citron plants directly agro-infiltrated, or mechanically inoculated with wild-type or 35SRbz-CLBV-derived virions from Nicotiana, and the three sources incited the symptoms characteristic of Dweet mottle disease, but not bud union crease. Our results show that (1) virions derived from an infectious clone show the same replication, movement and pathogenicity characteristics as the wild-type CLBV; (2) CLBV is the causal agent of Dweet mottle disease but not of the bud union crease syndrome; and (3) for the first time an RNA virus could be successfully agro-inoculated on citrus plants. This infectious clone may become a useful viral vector for citrus genomic studies.

  15. Simultaneous determination of nickel, lead, zinc, and copper in citrus leaves and rice flour by liquid chromatography with hexamethylenedithiocarbamate extraction

    SciTech Connect

    Ichinoki, S.; Yamazaki, M.

    1985-10-01

    Reversed-phase liquid chromatography followed by solvent extraction with hexamethyleneammonium hexamethylenedithiocarbamate (HMA-HMDC) was carried out to determine Ni, Pb, Zn, and Cu in standard reference citrus leaves and rice flour. These samples (250 mg) were ashed with nitric acid and perchloric acid. The metals in the ash were extracted into chloroform as HMDC chelates which were then separated on a C 18 column and monitored at 260 nm. The microgram per gram levels of Ni, Pb, Zn, and Cu in the standard biological materials were simultaneously determined within 25 min. 22 references, 3 figures, 1 table.

  16. Volatile Organic Compound (VOC) profiling of Citrus tristeza virus (CTV) infection in sweet orange citrus varietals using thermal desorption gas chromatography time of flight mass spectrometry (TD-GC/TOF-MS)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) is a plant pathogen which predominately infects economically important citrus crops such as sweet orange, clementine, lime and grapefruit varietals. Within the last 70 years, an estimated 100 million citrus trees on sour orange rootstock have been destroyed due to CTV inf...

  17. Rootstock-scion interaction affecting citrus response to CTV infection: a proteomic view.

    PubMed

    Laino, Paolo; Russo, Maria P; Guardo, Maria; Reforgiato-Recupero, Giuseppe; Valè, Giampiero; Cattivelli, Luigi; Moliterni, Vita M C

    2016-04-01

    Citrus tristeza virus (CTV) is the causal agent of various diseases with dramatic effects on citrus crops worldwide. Most Citrus species, grown on their own roots, are symptomless hosts for many CTV isolates. However, depending on different scion-rootstock combination, CTV infection should result in distinct syndromes, being 'tristeza' the more severe one, leading to a complete decline of the susceptible plants in a few weeks. Transcriptomic analyses revealed several genes involved either in defense response, or systemic acquired resistance, as well as transcription factors and components of the phosphorylation cascades, to be differentially regulated during CTV infection in Citrus aurantifolia species. To date little is known about the molecular mechanism of this host-pathogen interaction, and about the rootstock effect on citrus response to CTV infection. In this work, the response to CTV infection has been investigated in tolerant and susceptible scion-rootstock combinations by two-dimensional gel electrophoresis (2DE). A total of 125 protein spots have been found to be differently accumulated and/or phosphorylated between the two rootstock combinations. Downregulation in tolerant plants upon CTV infection was detected for proteins involved in reactive oxygen species (ROS) scavenging and defense response, suggesting a probable acclimation response able to minimize the systemic effects of virus infection. Some of these proteins resulted to be modulated also in absence of virus infection, revealing a rootstock effect on scion proteome modulation. Moreover, the phospho-modulation of proteins involved in ROS scavenging and defense response, further supports their involvement either in scion-rootstock crosstalk or in the establishment of tolerance/susceptibility to CTV infection.

  18. Effects of Oxamyl on the Citrus Nematode, Tylenchulus semipenetrans, and on Infection of Sweet Orange

    PubMed Central

    Baines, R. C.; Small, R. H.

    1976-01-01

    Foliar sprays of 4 μg/ml oxamyl on sweet orange trees in a greenhouse slightly depressed the number of Tylenchulus semipenetrans larvae obtained from roots and soil, but similar treatments were not effective in two orchards. Soil drench treatments decreased the number of citrus nematode larvae obtained from roots or soil of citrus plants grown itt a greenhouse and in orchards. Exposure to 5-10 μg/ml of oxamyl in water was lethal to only a few second-stage larvae treated 10 days, and many second-stage larvae in 2.0 μg/ml oxamyl recovered motility when transferred to fresh water. Aqueous solutions of 50 and 100 μg/ml of oxamyl were toxic to citrus nematode larvae. Additional observations indicate that oxamyl interfered with hatch of citrus nematode larvae and was nematistatic and/or protected sweet orange roots from infection. Oxamyl degraded at different rates in two soils. The number of citrus nematode larvae that infected and developed on sweet orange roots was increased by an undetermined product of the degradation of oxamyl in soil, water, and possibly within plants. This product apparently was translocated in roots. PMID:19308209

  19. Acquisition, Replication and Inoculation of Candidatus Liberibacter asiaticus following Various Acquisition Periods on Huanglongbing-Infected Citrus by Nymphs and Adults of the Asian Citrus Psyllid

    PubMed Central

    Ammar, El-Desouky; Ramos, John E.; Hall, David G.; Dawson, William O.; Shatters, Robert G.

    2016-01-01

    The Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae), is the primary vector of Candidatus Liberibacter asiaticus (Las) implicated as causative agent of citrus huanglongbing (citrus greening), currently the most serious citrus disease worldwide. Las is transmitted by D. citri in a persistent-circulative manner, but the question of replication of this bacterium in its psyllid vector has not been resolved. Thus, we studied the effects of the acquisition access period (AAP) by nymphs and adults of D. citri on Las acquisition, multiplication and inoculation/transmission. D. citri nymphs or adults (previously non-exposed to Las) were caged on Las-infected citrus plants for an AAP of 1, 7 or 14 days. These ‘Las-exposed’ psyllids were then transferred weekly to healthy citrus or orange jasmine plants, and sampled via quantitative polymerase chain reaction (qPCR) analysis 1–42 days post-first access to diseased plants (padp); all tested nymphs became adults 7–14 days padp. Our results indicate that following 1 or 7 day AAP as nymphs 49–59% of Las-exposed psyllids became Las-infected (qPCR-positive), whereas only 8–29% of the psyllids were infected following 1–14 day AAP as adults. Q-PCR analysis also indicated that Las titer in the Las-exposed psyllids (relative to that of the psyllid S20 ribosomal protein gene) was: 1) significantly higher, and increasing at a faster rate, following Las acquisition as nymphs compared to that following Las acquisition as adults; 2) higher as post-acquisition time of psyllids on healthy plants increased reaching a peak at 14–28 days padp for nymphs and 21–35 days padp for adults, with Las titer decreasing or fluctuating after that; 3) higher with longer AAP on infected plants, especially with acquisition as adults. Our results strongly suggest that Las multiplies in both nymphs and adults of D. citri but attains much higher levels in a shorter period of time post-acquisition when acquired by nymphs than when

  20. Lack of effect of feeding citrus by-products in reducing Salmonella in experimentally infected weanling pigs

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of the current research was to determine if feeding citrus by-products D’Limonene (DL) and citrus molasses (MOL) would reduce the concentration and prevalence of Salmonella in weanling pigs experimentally infected with Salmonella Typhimurium. Twenty crossbred weanling pigs (avg. BW = ...

  1. Analysis of full-length sequences of two Citrus yellow mosaic badnavirus isolates infecting Citrus jambhiri (Rough Lemon) and Citrus sinensis L. Osbeck (Sweet Orange) from a nursery in India.

    PubMed

    Anthony Johnson, A M; Borah, B K; Sai Gopal, D V R; Dasgupta, I

    2012-12-01

    Citrus yellow mosaic badna virus (CMBV), a member of the Family Caulimoviridae, Genus Badnavirus is the causative agent of mosaic disease among Citrus species in southern India. Despite its reported prevalence in several citrus species, complete information on clear functional genomics or functional information of full-length genomes from all the CMBV isolates infecting citrus species are not available in publicly accessible databases. CMBV isolates from Rough Lemon and Sweet Orange collected from a nursery were cloned and sequenced. The analysis revealed high sequence homology of the two CMBV isolates with previously reported CMBV sequences implying that they represent new variants. Based on computational analysis of the predicted secondary structures, the possible functions of some CMBV proteins have been analyzed.

  2. Infection of rice plants by rice black streaked dwarf virus improves an egg parasitoid, Anagrus nilaparvatae (Hymenoptera: Mymaridae), of rice planthoppers.

    PubMed

    Xu, Hongxing; He, Xiaochan; Zheng, Xusong; Yang, Yajun; Tian, Junce; Lu, Zhongxian

    2014-10-01

    The effects of rice plants infected by rice black streaked dwarf virus (RBSDV) on the host preference, duration of immature stages, sex ratio, and adult longevity and parasitic capacity of an egg parasitoid, Anagrus nilaparvatae Pang et Wang, of rice brown planthopper, Nilaparvata lugens Stål, were evaluated. Tests of response to plant volatiles using an olfactometer showed that A. nilaparvatae preferred rice plants harboring rice brown planthopper eggs over plants free of rice brown planthopper eggs. However, both the response to plant volatiles and the host selectivity test showed no significant differences in host preference between RBSDV-infected plants and healthy plants when both contained rice brown planthopper eggs. The developmental duration at immature stage of the male A. nilaparvatae in rice brown planthopper eggs on RBSDV-infected rice plants was significantly prolonged, and the parasitic capacity of rice brown planthopper eggs was significantly increased in comparison with the A. nilaparvatae parasite in rice brown planthopper eggs on healthy rice plants. There were no significant differences between RBSDV-infected rice plants and healthy rice plants in other ecological fitness parameters, including the developmental duration of female adults, female percentage, and adult longevity of A. nilaparvatae.

  3. Metabolomic analysis of citrus infection by 'Candidatus Liberibacter' reveals insight into pathogenicity.

    PubMed

    Slisz, Anne M; Breksa, Andrew P; Mishchuk, Darya O; McCollum, Greg; Slupsky, Carolyn M

    2012-08-03

    Huanglongbing (HLB), considered the most serious citrus disease in the world, is associated with the nonculturable bacterium 'Candidatus Liberibacter asiaticus' (Las). Infection of citrus by this pathogen leads to reduced plant vigor and productivity, ultimately resulting in death of the infected tree. It can take up to two years following initial infection before outward symptoms become apparent, making detection difficult. The existing knowledge gap in our understanding of Las and its pathogenesis leading to HLB has stymied development of treatments and methods to mitigate the pathogen's influence. To evaluate the influence of Las on fruit quality in both symptomatic and asymptomatic fruit, and gain further insight into the pathogenesis of the disease, a 1H NMR metabolomics investigation, complemented with physicochemical and analyte-specific analyses, was undertaken. Comparison of the juice obtained from oranges gathered from Las+ (symptomatic and asymptomatic) and Las- (healthy) trees revealed significant differences in the concentrations of sugars, amino and organic acids, limonin glucoside, and limonin. This study demonstrates differing metabolic profiles in the juice of oranges from Las+ and Las- and proposes how Las may be able to evade citrus defense responses.

  4. Identification and characterization of Citrus yellow vein clearing virus, a putative new member of the genus Mandarivirus infecting Citrus spp.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Yellow vein clearing virus, an uncharacterized filamentous virus, was first observed in Pakistan in 1988 and later in India in 1997 in Etrog citron (Citrus medica). Based on electron microscopic evidence of filamentous particles, the virus, provisionally named Citrus yellow vein clearing virus (CYVC...

  5. Transcriptional and microscopic analyses of citrus stem and root responses to Candidatus Liberibacter asiaticus infection.

    PubMed

    Aritua, Valente; Achor, Diann; Gmitter, Frederick G; Albrigo, Gene; Wang, Nian

    2013-01-01

    Huanglongbing (HLB) is the most destructive disease that affects citrus worldwide. The disease has been associated with Candidatus Liberibacter. HLB diseased citrus plants develop a multitude of symptoms including zinc and copper deficiencies, blotchy mottle, corky veins, stunting, and twig dieback. Ca. L. asiaticus infection also seriously affects the roots. Previous study focused on gene expression of leaves and fruit to Ca. L. asiaticus infection. In this study, we compared the gene expression levels of stems and roots of healthy plants with those in Ca. L. asiaticus infected plants using microarrays. Affymetrix microarray analysis showed a total of 988 genes were significantly altered in expression, of which 885 were in the stems, and 111 in the roots. Of these, 551 and 56 were up-regulated, while 334 and 55 were down-regulated in the stem and root samples of HLB diseased trees compared to healthy plants, respectively. Dramatic differences in the transcriptional responses were observed between citrus stems and roots to Ca. L. asiaticus infection, with only 8 genes affected in both the roots and stems. The affected genes are involved in diverse cellular functions, including carbohydrate metabolism, cell wall biogenesis, biotic and abiotic stress responses, signaling and transcriptional factors, transportation, cell organization, protein modification and degradation, development, hormone signaling, metal handling, and redox. Microscopy analysis showed the depletion of starch in the roots of the infected plants but not in healthy plants. Collapse and thickening of cell walls were observed in HLB affected roots, but not as severe as in the stems. This study provides insight into the host response of the stems and roots to Ca. L. asiaticus infection.

  6. Transcriptional and Microscopic Analyses of Citrus Stem and Root Responses to Candidatus Liberibacter asiaticus Infection

    PubMed Central

    Aritua, Valente; Achor, Diann; Gmitter, Frederick G.; Albrigo, Gene; Wang, Nian

    2013-01-01

    Huanglongbing (HLB) is the most destructive disease that affects citrus worldwide. The disease has been associated with Candidatus Liberibacter. HLB diseased citrus plants develop a multitude of symptoms including zinc and copper deficiencies, blotchy mottle, corky veins, stunting, and twig dieback. Ca. L. asiaticus infection also seriously affects the roots. Previous study focused on gene expression of leaves and fruit to Ca. L. asiaticus infection. In this study, we compared the gene expression levels of stems and roots of healthy plants with those in Ca. L. asiaticus infected plants using microarrays. Affymetrix microarray analysis showed a total of 988 genes were significantly altered in expression, of which 885 were in the stems, and 111 in the roots. Of these, 551 and 56 were up-regulated, while 334 and 55 were down-regulated in the stem and root samples of HLB diseased trees compared to healthy plants, respectively. Dramatic differences in the transcriptional responses were observed between citrus stems and roots to Ca. L. asiaticus infection, with only 8 genes affected in both the roots and stems. The affected genes are involved in diverse cellular functions, including carbohydrate metabolism, cell wall biogenesis, biotic and abiotic stress responses, signaling and transcriptional factors, transportation, cell organization, protein modification and degradation, development, hormone signaling, metal handling, and redox. Microscopy analysis showed the depletion of starch in the roots of the infected plants but not in healthy plants. Collapse and thickening of cell walls were observed in HLB affected roots, but not as severe as in the stems. This study provides insight into the host response of the stems and roots to Ca. L. asiaticus infection. PMID:24058486

  7. A transcriptomic approach highlights induction of secondary metabolism in citrus fruit in response to Penicillium digitatum infection

    PubMed Central

    2010-01-01

    Background Postharvest losses of citrus fruit due to green mold decay, caused by the fungus Penicillium digitaum, have a considerable economic impact. However, little is known about the molecular processes underlying the response of citrus fruit to P. digitatum. Results Here we describe the construction of a subtracted cDNA library enriched in citrus genes preferentially expressed in response to pathogen infection followed by cDNA macroarray hybridization to investigate gene expression during the early stages of colonization of the fruit's peel by P. digitatum. Sequence annotation of clones from the subtracted cDNA library revealed that induction of secondary and amino acid metabolisms constitutes the major response of citrus fruits to P. digitatum infection. Macroarray hybridization analysis was conducted with RNA from either control, wounded, ethylene treated or P. digitatum infected fruit. Results indicate an extensive overlap in the response triggered by the three treatments, but also demonstrated specific patterns of gene expression in response to each stimulus. Collectively our data indicate a significant presence of isoprenoid, alkaloid and phenylpropanoid biosynthetic genes in the transcriptomic response of citrus fruits to P. digitatum infection. About half of the genes that are up-regulated in response to pathogen infection are also induced by ethylene, but many examples of ethylene-independent gene regulation were also found. Two notable examples of this regulation pattern are the genes showing homology to a caffeine synthase and a berberine bridge enzyme, two proteins involved in alkaloid biosynthesis, which are among the most induced genes upon P. digitatum infection but are not responsive to ethylene. Conclusions This study provided the first global picture of the gene expression changes in citrus fruit in response to P. digitatum infection, emphasizing differences and commonalities with those triggered by wounding or exogenous ethylene treatment

  8. Infection Density Dynamics of the Citrus Greening Bacterium “Candidatus Liberibacter asiaticus” in Field Populations of the Psyllid Diaphorina citri and Its Relevance to the Efficiency of Pathogen Transmission to Citrus Plants

    PubMed Central

    Ukuda-Hosokawa, Rie; Sadoyama, Yasutsune; Kishaba, Misaki; Kuriwada, Takashi; Anbutsu, Hisashi

    2015-01-01

    Huanglongbing, or citrus greening, is a devastating disease of citrus plants recently spreading worldwide, which is caused by an uncultivable bacterial pathogen, “Candidatus Liberibacter asiaticus,” and vectored by a phloem-sucking insect, Diaphorina citri. We investigated the infection density dynamics of “Ca. Liberibacter asiaticus” in field populations of D. citri with experiments using field-collected insects to address how “Ca. Liberibacter asiaticus” infection density in the vector insect is relevant to pathogen transmission to citrus plants. Of 500 insects continuously collected from “Ca. Liberibacter asiaticus”-infected citrus trees with pathological symptoms in the spring and autumn of 2009, 497 (99.4%) were “Ca. Liberibacter asiaticus” positive. The infections were systemic across head-thorax and abdomen, ranging from 103 to 107 bacteria per insect. In spring, the infection densities were low in March, at ∼103 bacteria per insect, increasing up to 106 to 107 bacteria per insect in April and May, and decreasing to 105 to 106 bacteria per insect in late May, whereas the infection densities were constantly ∼106 to 107 bacteria per insect in autumn. Statistical analysis suggested that several factors, such as insect sex, host trees, and collection dates, may be correlated with “Ca. Liberibacter asiaticus” infection densities in field D. citri populations. Inoculation experiments with citrus seedlings using field-collected “Ca. Liberibacter asiaticus”-infected insects suggested that (i) “Ca. Liberibacter asiaticus”-transmitting insects tend to exhibit higher infection densities than do nontransmitting insects, (ii) a threshold level (∼106 bacteria per insect) of “Ca. Liberibacter asiaticus” density in D. citri is required for successful transmission to citrus plants, and (iii) D. citri attaining the threshold infection level transmits “Ca. Liberibacter asiaticus” to citrus plants in a stochastic manner. These

  9. Infection Density Dynamics of the Citrus Greening Bacterium "Candidatus Liberibacter asiaticus" in Field Populations of the Psyllid Diaphorina citri and Its Relevance to the Efficiency of Pathogen Transmission to Citrus Plants.

    PubMed

    Ukuda-Hosokawa, Rie; Sadoyama, Yasutsune; Kishaba, Misaki; Kuriwada, Takashi; Anbutsu, Hisashi; Fukatsu, Takema

    2015-06-01

    Huanglongbing, or citrus greening, is a devastating disease of citrus plants recently spreading worldwide, which is caused by an uncultivable bacterial pathogen, "Candidatus Liberibacter asiaticus," and vectored by a phloem-sucking insect, Diaphorina citri. We investigated the infection density dynamics of "Ca. Liberibacter asiaticus" in field populations of D. citri with experiments using field-collected insects to address how "Ca. Liberibacter asiaticus" infection density in the vector insect is relevant to pathogen transmission to citrus plants. Of 500 insects continuously collected from "Ca. Liberibacter asiaticus"-infected citrus trees with pathological symptoms in the spring and autumn of 2009, 497 (99.4%) were "Ca. Liberibacter asiaticus" positive. The infections were systemic across head-thorax and abdomen, ranging from 10(3) to 10(7) bacteria per insect. In spring, the infection densities were low in March, at ∼ 10(3) bacteria per insect, increasing up to 10(6) to 10(7) bacteria per insect in April and May, and decreasing to 10(5) to 10(6) bacteria per insect in late May, whereas the infection densities were constantly ∼ 10(6) to 10(7) bacteria per insect in autumn. Statistical analysis suggested that several factors, such as insect sex, host trees, and collection dates, may be correlated with "Ca. Liberibacter asiaticus" infection densities in field D. citri populations. Inoculation experiments with citrus seedlings using field-collected "Ca. Liberibacter asiaticus"-infected insects suggested that (i) "Ca. Liberibacter asiaticus"-transmitting insects tend to exhibit higher infection densities than do nontransmitting insects, (ii) a threshold level (∼ 10(6) bacteria per insect) of "Ca. Liberibacter asiaticus" density in D. citri is required for successful transmission to citrus plants, and (iii) D. citri attaining the threshold infection level transmits "Ca. Liberibacter asiaticus" to citrus plants in a stochastic manner. These findings provide

  10. Temporal interactions of plant - insect - predator after infection of bacterial pathogen on rice plants

    PubMed Central

    Sun, Ze; Liu, Zhuang; Zhou, Wen; Jin, Huanan; Liu, Hao; Zhou, Aiming; Zhang, Aijun; Wang, Man-Qun

    2016-01-01

    Pathogenic infection on plants may affect interactions of host-plants with their herbivores, as well as the herbivores with their predators. In this study, the effects of infection by pathogenic bacterium Xanthomonas oryzae pv. oryzae (Xoo), which causes a vascular disease in rice, on rice plants and consequent interactions with a rice herbivore, brown rice planthopper (BPH) Nilaparvata lugens, and its major predator, Cyrtorhinus lividipennis, were investigated. The results showed that the rice plants exhibited increased resistance to BPH only at 3 d post-inoculation of Xoo, while the Xoo infection did not affect the development and fecundity of BPH. BPH exhibited a higher preference to Xoo infected rice plants, whereas C. lividipennis preferred the Xoo infected rice plants after BPH fed, but preferred healthy rice plants without BPH fed. Volatile organic compounds emitted from Xoo rice were significantly higher than those from healthy rice plants, Xoo infection on BPH fed plants caused rice plants to emit more the herbivore-induced plant volatiles, while all of these changes correlated to the temporal dimension. These results demonstrated that Xoo infection significantly influenced the interactions of rice plants with two non-vectors, BPH and its predator, although these effects exhibited in a temporal pattern after infection. PMID:27185548

  11. Temporal interactions of plant - insect - predator after infection of bacterial pathogen on rice plants.

    PubMed

    Sun, Ze; Liu, Zhuang; Zhou, Wen; Jin, Huanan; Liu, Hao; Zhou, Aiming; Zhang, Aijun; Wang, Man-Qun

    2016-05-17

    Pathogenic infection on plants may affect interactions of host-plants with their herbivores, as well as the herbivores with their predators. In this study, the effects of infection by pathogenic bacterium Xanthomonas oryzae pv. oryzae (Xoo), which causes a vascular disease in rice, on rice plants and consequent interactions with a rice herbivore, brown rice planthopper (BPH) Nilaparvata lugens, and its major predator, Cyrtorhinus lividipennis, were investigated. The results showed that the rice plants exhibited increased resistance to BPH only at 3 d post-inoculation of Xoo, while the Xoo infection did not affect the development and fecundity of BPH. BPH exhibited a higher preference to Xoo infected rice plants, whereas C. lividipennis preferred the Xoo infected rice plants after BPH fed, but preferred healthy rice plants without BPH fed. Volatile organic compounds emitted from Xoo rice were significantly higher than those from healthy rice plants, Xoo infection on BPH fed plants caused rice plants to emit more the herbivore-induced plant volatiles, while all of these changes correlated to the temporal dimension. These results demonstrated that Xoo infection significantly influenced the interactions of rice plants with two non-vectors, BPH and its predator, although these effects exhibited in a temporal pattern after infection.

  12. A Lipoxygenase Pathway Is Activated in Rice after Infection with the Rice Blast Fungus Magnaporthe grisea.

    PubMed

    Ohta, H; Shida, K; Peng, Y L; Furusawa, I; Shishiyama, J; Aibara, S; Morita, Y

    1991-09-01

    Lipoxygenase (LOX) and lipid hydroperoxide-decomposing activity (LHDA) markedly increased in the fifth leaves of rice (Oryza sativa cv Aichiasahi) after infection with the rice blast fungus, Magnaporthe grisea. The increases in the enzyme activities were significantly higher in response to infection with an incompatible strain (race 131) compared with infection with a compatible strain (race 007) of the fungus. Using ion-exchange chromatography, we isolated three LOX activities (leaf LOX-1, -2, -3) from both uninoculated and infected leaves. The activity of leaf LOX-3, in particular, increased in the incompatible race-infected leaves. The leaf LOX-3 had a pH optimum of 5.0 and produced preferentially 13-l-hydroperoxy-9,11 (Z,E)-octadecadienoic acid (13-HPODD) from linoleic acid. 13-HPODD and 13-l-hydroxy-9,11 (Z,E)-octadecadienoic acid, one of the reaction products from 13-HPODD by LHDA, were highly inhibitory to the germination of conidia of the fungus. The present study provides correlative evidence for important roles of LOX and LHDA in the resistance response of rice against the blast fungus.

  13. Variation in flavonoid levels in Citrus benikoji Hort. ex. Tan. infected by Colletotrichum gloeosporioides.

    PubMed

    Jeong, Sung Woo; Kim, Hae Gyeong; Park, Semin; Lee, Jung Han; Kim, Yun-Hi; Kim, Gon-Sup; Jin, Jong Sung; Kwak, Youn-Sig; Huh, Moo Ryong; Lee, Ji Eun; Song, Yi; Shin, Sung Chul

    2014-04-01

    Anthracnose caused by Colletotrichum gloeosporioides is one of the most serious postharvest diseases of citrus fruit. Fruit peels infected with C. gloeosporioides and the peels of healthy fruit were analysed for flavonoids, using high performance liquid chromatography-tandem mass spectroscopy, to evaluate variations in flavonoid levels in Citrus benikoji Hort. ex. Tan. Seventeen flavonoids were characterised from the fruit peels. The flavonoids were validated using structurally related standards and quantified. Among the 17 flavonoids, the concentration of component 3 was the highest and that of component 10 was the lowest. During 8 days after inoculation, the concentration of the seven flavonoids 1-3, 5, 6, 13, and 14 increased gradually up to day 8. Flavonoid 4 was detected from day 7. The seven minor flavonoid components, 8-12, 15, and 16 increased to day 5 and then decreased. However, flavonoids level variations were not significantly different from that of the non-infected fruits during the experimental period. The monitoring suggested that the constitutively formed seven polymethoxyflavones (8-12, 15, and 16) may act as phytoanticipins in the defence mechanism against the fungus, and that hespertin 7-O-glucoside (4), produced de novo on day 6 after infection, may function as a phytoalexin.

  14. NAC transcription factor family genes are differentially expressed in rice during infections with Rice dwarf virus, Rice black-streaked dwarf virus, Rice grassy stunt virus, Rice ragged stunt virus, and Rice transitory yellowing virus

    PubMed Central

    Nuruzzaman, Mohammed; Sharoni, Akhter M.; Satoh, Kouji; Karim, Mohammad Rezaul; Harikrishna, Jennifer A.; Shimizu, Takumi; Sasaya, Takahide; Omura, Toshihiro; Haque, Mohammad A.; Hasan, Sayed M. Z.; Ahmad, Aziz; Kikuchi, Shoshi

    2015-01-01

    Expression levels of the NAC gene family were studied in rice infected with Rice dwarf virus (RDV), Rice black-streaked dwarf virus (RBSDV), Rice grassy stunt virus (RGSV), Rice ragged stunt virus (RRSV), and Rice transitory yellowing virus (RTYV). Microarray analysis showed that 75 (68%) OsNAC genes were differentially regulated during infection with RDV, RBSDV, RGSV, and RRSV compared with the control. The number of OsNAC genes up-regulated was highest during RGSV infection, while the lowest number was found during RTYV infection. These phenomena correlate with the severity of the syndromes induced by the virus infections. Most of the genes in the NAC subgroups NAC22, SND, ONAC2, ANAC34, and ONAC3 were down-regulated for all virus infections. These OsNAC genes might be related to the health stage maintenance of the host plants. Interestingly, most of the genes in the subgroups TIP and SNAC were more highly expressed during RBSDV and RGSV infections. These results suggested that OsNAC genes might be related to the responses induced by the virus infection. All of the genes assigned to the TIP subgroups were highly expressed during RGSV infection when compared with the control. For RDV infection, the number of activated genes was greatest during infection with the S-strain, followed by the D84-strain and the O-strain, with seven OsNAC genes up-regulated during infection by all three strains. The Os12g03050 and Os11g05614 genes showed higher expression during infection with four of the five viruses, and Os11g03310, Os11g03370, and Os07g37920 genes showed high expression during at least three viral infections. We identified some duplicate genes that are classified as neofunctional and subfunctional according to their expression levels in different viral infections. A number of putative cis-elements were identified, which may help to clarify the function of these key genes in network pathways. PMID:26442000

  15. Micronutrient-Fortified Rice Can Increase Hookworm Infection Risk: A Cluster Randomized Trial

    PubMed Central

    de Gier, Brechje; Campos Ponce, Maiza; Perignon, Marlene; Fiorentino, Marion; Khov, Kuong; Chamnan, Chhoun; de Boer, Michiel R.; Parker, Megan E.; Burja, Kurt; Dijkhuizen, Marjoleine A.; Berger, Jacques; Polman, Katja; Wieringa, Frank T.

    2016-01-01

    Background Fortification of staple foods is considered an effective and safe strategy to combat micronutrient deficiencies, thereby improving health. While improving micronutrient status might be expected to have positive effects on immunity, some studies have reported increases in infections or inflammation after iron supplementation. Objective To study effects of micronutrient-fortified rice on hookworm infection in Cambodian schoolchildren. Methods A double-blinded, cluster-randomized trial was conducted in 16 Cambodian primary schools partaking in the World Food Program school meal program. Three types of multi-micronutrient fortified rice were tested against placebo rice within the school meal program: UltraRice_original, UltraRice_improved and NutriRice. Four schools were randomly assigned to each study group (placebo n = 492, UltraRice_original n = 479, UltraRice_improved n = 500, NutriRice n = 506). Intestinal parasite infection was measured in fecal samples by Kato-Katz method at baseline and after three and seven months. In a subgroup (N = 330), fecal calprotectin was measured by ELISA as a marker for intestinal inflammation. Results Baseline prevalence of hookworm infection was 18.6%, but differed considerably among schools (range 0%- 48.1%).Micronutrient-fortified rice significantly increased risk of new hookworm infection. This effect was modified by baseline hookworm prevalence at the school; hookworm infection risk was increased by all three types of fortified rice in schools where baseline prevalence was high (>15%), and only by UltraRice_original in schools with low baseline prevalence. Neither hookworm infection nor fortified rice was related to fecal calprotectin. Conclusions Consumption of rice fortified with micronutrients can increase hookworm prevalence, especially in environments with high infection pressure. When considering fortification of staple foods, a careful risk-benefit analysis is warranted, taking into account severity of

  16. Processess involved in the dispersal of Xanthomonas citri pv. citri from canker-infected citrus canopies, and in the infection of citrus foliage

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker (Xanthomonas citri subsp. citri, Xcc) is now considered endemic in Florida, and epidemics result in yield loss and market penalties both in Florida and elsewhere, where the pathogen occurs and susceptible citrus is cultivated. The bacterium is dispersed in rain splash, and storms with...

  17. Wolbachia infection density in populations of the Asian citrus psyllid (Hemiptera: Liviidae).

    PubMed

    Hoffmann, M; Coy, M R; Kingdom Gibbard, H N; Pelz-Stelinski, K S

    2014-10-01

    The symbiotic relationships between bacteria of the genus Wolbachia (order Rickettsiales) and their arthropod hosts are diverse and can range from mutualism to parasitism. Whereas effects of Wolbachia on host biology are well investigated, little is known about diversity and abundance of Wolbachia in their natural hosts. The phloem-feeding Asian citrus psyllid, Diaphorina citri (Kuwayama) (Hemiptera: Liviidae), is naturally infected with Wolbachia (wDi). In the current study, we calculated the within-host density of Wolbachia in Florida D. citri populations using quantitative polymerase chain reaction for detection of the Wolbachia outer surface protein gene, wsp. Gene quantities were normalized to the D. citri wingless gene (Wg) to estimate Wolbachia abundance in individual D. citri. Using this method, significant geographic differences in Wolbachia densities were detected among Florida D. citri populations, with higher infection levels occurring in male versus female hosts.

  18. Detection and diagnosis of rice-infecting viruses

    PubMed Central

    Uehara-Ichiki, Tamaki; Shiba, Takuya; Matsukura, Keiichiro; Ueno, Takanori; Hirae, Masahiro; Sasaya, Takahide

    2013-01-01

    Rice-infecting viruses have caused serious damage to rice production in Asian, American, and African countries, where about 30 rice viruses and diseases have been reported. To control these diseases, developing accurate, quick methods to detect and diagnose the viruses in the host plants and any insect vectors of the viruses is very important. Based on an antigen–antibody reaction, serological methods such as latex agglutination reaction and enzyme-linked immunosorbent assay have advanced to detect viral particles or major proteins derived from viruses. They aid in forecasting disease and surveying disease spread and are widely used for virus detection at plant protection stations and research laboratories. From the early 2000s, based on sequence information for the target virus, several other methods such as reverse transcription-polymerase chain reaction (RT-PCR) and reverse transcription-loop-mediated isothermal amplification have been developed that are sensitive, rapid, and able to differentiate closely related viruses. Recent techniques such as real-time RT-PCR can be used to quantify the pathogen in target samples and monitor population dynamics of a virus, and metagenomic analyses using next-generation sequencing and microarrays show potential for use in the diagnosis of rice diseases. PMID:24130554

  19. Identification and Molecular Characterization of Nuclear Citrus leprosis virus, a Member of the Proposed Dichorhavirus Genus Infecting Multiple Citrus Species in Mexico.

    PubMed

    Roy, Avijit; Stone, Andrew L; Shao, Jonathan; Otero-Colina, Gabriel; Wei, Gang; Choudhary, Nandlal; Achor, Diann; Levy, Laurene; Nakhla, Mark K; Hartung, John S; Schneider, William L; Brlansky, Ronald H

    2015-04-01

    Citrus leprosis is one of the most destructive diseases of Citrus spp. and is associated with two unrelated virus groups that produce particles primarily in either the cytoplasm or nucleus of infected plant cells. Symptoms of leprosis, including chlorotic spots surrounded by yellow haloes on leaves and necrotic spots on twigs and fruit, were observed on leprosis-affected mandarin and navel sweet orange trees in the state of Querétaro, Mexico. Serological and molecular assays showed that the cytoplasmic types of Citrus leprosis virus (CiLV-C) often associated with leprosis symptomatic tissues were absent. However, using transmission electron microscopy, bullet-shaped rhabdovirus-like virions were observed in the nuclei and cytoplasm of the citrus leprosis-infected leaf tissues. An analysis of small RNA populations from symptomatic tissue was carried out to determine the genome sequence of the rhabdovirus-like particles observed in the citrus leprosis samples. The complete genome sequence showed that the nuclear type of CiLV (CiLV-N) present in the samples consisted of two negative-sense RNAs: 6,268-nucleotide (nt)-long RNA1 and 5,847-nt-long RNA2, excluding the poly(A) tails. CiLV-N had a genome organization identical to that of Orchid fleck virus (OFV), with the exception of shorter 5' untranslated regions in RNA1 (53 versus 205 nt) and RNA2 (34 versus 182 nt). Phylogenetic trees constructed with the amino acid sequences of the nucleocapsid (N) and glycoproteins (G) and the RNA polymerase (L protein) showed that CiLV-N clusters with OFV. Furthermore, phylogenetic analyses of N protein established CiLV-N as a member of the proposed genus Dichorhavirus. Reverse-transcription polymerase chain reaction primers for the detection of CiLV-N were designed based on the sequence of the N gene and the assay was optimized and tested to detect the presence of CiLV-N in both diseased and symptom-free plants.

  20. Acquisition, replication and inoculation of Candidatus Liberibacter asiaticus following various acquisition periods on Huanglongbing-infected citrus by nymphs and adults of the Asian citrus psyllid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae), is the primary vector of Candidatus Liberibacter asiaticus (Las) implicated as causative agent of citrus huanglongbing (citrus greening), currently the most serious citrus disease worldwide. Las is transmitted by D. citri in a persist...

  1. Predisposition of citrus foliage to infection with Xanthomonas citri subsp. citri

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker (caused by Xanthomonas citri subsp. citri, Xcc) is a serious disease of susceptible citrus in Florida and other citrus-growing areas of the world. The effect of leaf preconditioning as a route for entry of the bacteria is poorly characterized. A series of experiments were designed to i...

  2. First Report of Dodder Transmission of Huanglongbing from Naturally Infected Murraya paniculata to Citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB) or “greening” disease of citrus is caused by phloem-limited, uncultured bacteria in the genus “Candidatus Liberibacter”. HLB is one of the most destructive diseases of citrus worldwide and is considered so dangerous to a U.S. citrus production that the USDA has listed “Ca. Liberi...

  3. Killing Two Birds with One Stone: Natural Rice Rhizospheric Microbes Reduce Arsenic Uptake and Blast Infections in Rice.

    PubMed

    Lakshmanan, Venkatachalam; Cottone, Jonathon; Bais, Harsh P

    2016-01-01

    Our recent work has shown that a rice thizospheric natural isolate, a Pantoea sp (hereafter EA106) attenuates Arsenic (As) uptake in rice. In parallel, yet another natural rice rhizospheric isolate, a Pseudomonas chlororaphis (hereafter EA105), was shown to inhibit rice blast pathogen Magnaporthe oryzae. Considering the above, we envisaged to evaluate the importance of mixed stress regime in rice plants subjected to both As toxicity and blast infections. Plants subjected to As regime showed increased susceptibility to blast infections compared to As-untreated plants. Rice blast pathogen M. oryzae showed significant resistance against As toxicity compared to other non-host fungal pathogens. Interestingly, plants treated with EA106 showed reduced susceptibility against blast infections in plants pre-treated with As. This data also corresponded with lower As uptake in plants primed with EA106. In addition, we also evaluated the expression of defense related genes in host plants subjected to As treatment. The data showed that plants primed with EA106 upregulated defense-related genes with or without As treatment. The data shows the first evidence of how rice plants cope with mixed stress regimes. Our work highlights the importance of natural association of plant microbiome which determines the efficacy of benign microbes to promote the development of beneficial traits in plants.

  4. Killing Two Birds with One Stone: Natural Rice Rhizospheric Microbes Reduce Arsenic Uptake and Blast Infections in Rice

    PubMed Central

    Lakshmanan, Venkatachalam; Cottone, Jonathon; Bais, Harsh P.

    2016-01-01

    Our recent work has shown that a rice thizospheric natural isolate, a Pantoea sp (hereafter EA106) attenuates Arsenic (As) uptake in rice. In parallel, yet another natural rice rhizospheric isolate, a Pseudomonas chlororaphis (hereafter EA105), was shown to inhibit rice blast pathogen Magnaporthe oryzae. Considering the above, we envisaged to evaluate the importance of mixed stress regime in rice plants subjected to both As toxicity and blast infections. Plants subjected to As regime showed increased susceptibility to blast infections compared to As-untreated plants. Rice blast pathogen M. oryzae showed significant resistance against As toxicity compared to other non-host fungal pathogens. Interestingly, plants treated with EA106 showed reduced susceptibility against blast infections in plants pre-treated with As. This data also corresponded with lower As uptake in plants primed with EA106. In addition, we also evaluated the expression of defense related genes in host plants subjected to As treatment. The data showed that plants primed with EA106 upregulated defense-related genes with or without As treatment. The data shows the first evidence of how rice plants cope with mixed stress regimes. Our work highlights the importance of natural association of plant microbiome which determines the efficacy of benign microbes to promote the development of beneficial traits in plants. PMID:27790229

  5. Biological Effects of Weak Electromagnetic Field on Healthy and Infected Lime (Citrus aurantifolia) Trees with Phytoplasma

    PubMed Central

    Abdollahi, Fatemeh; Niknam, Vahid; Ghanati, Faezeh; Masroor, Faribors; Noorbakhsh, Seyyed Nasr

    2012-01-01

    Exposure to electromagnetic fields (EMF) has become an issue of concern for a great many people and is an active area of research. Phytoplasmas, also known as mycoplasma-like organisms, are wall-less prokaryotes that are pathogens of many plant species throughout the world. Effects of electromagnetic fields on the changes of lipid peroxidation, content of H2O2, proline, protein, and carbohydrates were investigated in leaves of two-year-old trees of lime (Citrus aurantifolia) infected by the Candidatus Phytoplasma aurantifoliae. The healthy and infected plants were discontinuously exposed to a 10 KHz quadratic EMF with maximum power of 9 W for 5 days, each 5 h, at 25°C. Fresh and dry weight of leaves, content of MDA, proline, and protein increased in both healthy and infected plants under electromagnetic fields, compared with those of the control plants. Electromagnetic fields decreased hydrogen peroxide and carbohydrates content in both healthy and infected plants compared to those of the controls. PMID:22649313

  6. Rice Stripe Mosaic Virus, a Novel Cytorhabdovirus Infecting Rice via Leafhopper Transmission

    PubMed Central

    Yang, Xin; Huang, Jilei; Liu, Chuanhe; Chen, Biao; Zhang, Tong; Zhou, Guohui

    2017-01-01

    A new rice viral disease exhibiting distinct symptoms—yellow stripes, mosaic and twisted tips on leaves—was found in China. Electron microscopy of infected leaf cells revealed the presence of bacilliform virions and electron-translucent granular-fibrillar viroplasm in the cytoplasm. The enveloped viral particles were 300 to 375 nm long and 45 to 55 nm wide. The leafhopper Recilia dorsalis was able to transmit the virus to rice seedlings, which subsequently exhibited symptoms similar to those observed in fields. The complete genome of the virus was obtained by small-RNA deep sequencing and reverse transcription-PCR product sequencing. The anti-genome contains seven open reading frames (ORFs). The deduced amino acids of ORF1, ORF5, and ORF7 are, respectively, homologous to the nucleocapsid protein (N), glycoprotein (G), and large polymerase protein (L) of known rhabdoviruses. The predicted product of ORF2 is identified as a phosphoprotein (P) based on its multiple potential phosphorylation sites and 12.6 to 21.0% amino acid (aa) identities with the P proteins of plant rhabdoviruses. The product of ORF4 is presumed to be the viral matrix (M) protein for it shares 10.3 to 14.3% aa identities with those of other rhabdoviruses. The above five products were confirmed as the viral structural proteins by SDS-PAGE and aa sequencing analyses of purified virus preparation. ORF3 and ORF6 are considered to encode two nonstructural proteins with unknown functions. Phylogenetic analysis based on protein N, G, and L amino acid sequences indicated that the isolated virus, which we have tentatively named Rice stripe mosaic virus (RSMV), is a new species in the genus Cytorhabdovirus. To our knowledge, RSMV is the only cytorhabdovirus naturally infecting rice and the first reported leafhopper-transmitted cytorhabdovirus. Our surveys of rice fields indicate that RSMV occurs frequently in Guangdong Province, China. Although the disease incidence is low at present, it might become

  7. Temporal interactions of plant - insect - predator after infection of bacterial pathogen on rice plant

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pathogenic infection on plants may affect interactions of the host-plants with their herbivores, as well as the herbivores with their predators. In this study, the effects of infection by pathogenic bacterium Xanthomonas oryzae pv. oryzae (Xoo), which causes a vascular disease in rice, in rice plant...

  8. Study on citrus response to huanglongbing highlights a down-regulation of defense-related proteins in lemon plants upon 'Ca. Liberibacter asiaticus' infection.

    PubMed

    Nwugo, Chika C; Duan, Yongping; Lin, Hong

    2013-01-01

    Citrus huanglongbing (HLB) is a highly destructive disease of citrus presumably caused by 'Candidatus Liberibacterasiaticus' (Las), a gram-negative, insect-transmitted, phloem-limited α-proteobacterium. Although almost all citrus plants are susceptible to HLB, reports have shown reduced susceptibility to Las infection in lemon (Citrus limon) plants. The aim of this study is to identify intra-species specific molecular mechanisms associated with Las-induced responses in lemon plants. To achieve this, comparative 2-DE and mass spectrometry, in addition to Inductively Coupled Plasma Spectroscopy (ICPS) analyses, were applied to investigate differences in protein accumulation and the concentrations of cationic elements in leaves of healthy and Las-infected lemon plants. Results showed a differential accumulation of 27 proteins, including an increase in accumulation of starch synthase but decrease in the production of photosynthesis-related proteins in Las-infected lemon plants compared to healthy plants. Furthermore, there was a 6% increase (P > 0.05) in K concentration in leaves of lemon plants upon Las infection, which support results from previous studies and might represent a common response pattern of citrus plants to Las infection. Interestingly, contrary to reports from prior studies, this study showed a general reduction in the production of defense-related pathogen-response proteins but a 128% increase in Zn concentration in lemon plants in response to Las infection. Taken together, this study sheds light on general and intra-species specific responses associated with the response of citrus plants to Las.

  9. Chemical characterization of orange juice from trees infected with citrus greening (Huanglongbing).

    PubMed

    Dagulo, Lilibeth; Danyluk, Michelle D; Spann, Timothy M; Valim, M Filomena; Goodrich-Schneider, Renée; Sims, Charles; Rouseff, Russell

    2010-03-01

    The effects due to Candidatus Liberibacter infection, commonly called citrus greening or Huanglongbing (HLB), on volatile and nonvolatile components of orange juices, OJ, were examined using GC-MS and high-performance liquid chromatography (HPLC). HLB symptomatic, asymptomatic, and control "Hamlin" and "Valencia" oranges were harvested from December to May during the 2007 to 2008 harvest season. Brix/acid levels in control and asymptomatic juices were similar but symptomatic juices were as much as 62% lower than control juices. No bitter flavanone neohesperidosides were detected and polymethoxyflavone concentrations were well below bitter taste thresholds. Limonin concentrations were significantly higher (91% to 425%) in symptomatic juice compared to control but still below juice bitterness taste thresholds. Juice terpenes, such as gamma-terpinene and alpha-terpinolene, were as much as 1320% and 62% higher in symptomatic juice than control. Average ethyl butanoate concentrations were 45% lower and average linalool was 356% higher in symptomatic Valencia OJ compared to control. Symptomatic Valencia OJ had on average only 40% the total esters, 48% the total aldehydes, and 33% as much total sesquiterpenes as control juice. Total volatiles between control and symptomatic juices were similar due to elevated levels of alcohols and terpenes in symptomatic juice. There were no consistent differences between asymptomatic and control juices. The chemical composition of juice from HLB/greening symptomatic fruit appears to mimic that of juice from less mature fruit. The reported off-flavor associated with symptomatic juices probably stem from lower concentrations of sugars, higher concentrations of acid as all known citrus bitter compounds were either below taste thresholds or absent.

  10. Expression profiling of common and specific defense responses of rice to Magnaporthe oryzae infection using deep sequencing technologies

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rice blast caused by Magnaporthe oryzae is a serious disease in rice production. Wild type Nipponbare and transgenic rice plants (carrying the Pi9 blast resistance gene) were challenged with the rice blast strain KJ201 to identify the early, mid and late host responses to M. oryzae infection at the ...

  11. Entomopathogenic fungi infecting the Asian citrus psyllid, Diaphorina citri (Homoptera: Psyllidae), in Florida

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Asian citrus psyllid, Diaphorina citri is an invasive pest that vectors citrus greening disease. In 2005-2006 mycosed psyllids displaying two phenotypes were collected in central Florida. The major pathogen, identified by morphological and genetic analyses, was a novel isolate related to Hirsute...

  12. Transcriptome comparison and gene coexpression network analysis provide a systems view of citrus response to ‘Candidatus Liberibacter asiaticus’ infection

    PubMed Central

    2013-01-01

    Background Huanglongbing (HLB) is arguably the most destructive disease for the citrus industry. HLB is caused by infection of the bacterium, Candidatus Liberibacter spp. Several citrus GeneChip studies have revealed thousands of genes that are up- or down-regulated by infection with Ca. Liberibacter asiaticus. However, whether and how these host genes act to protect against HLB remains poorly understood. Results As a first step towards a mechanistic view of citrus in response to the HLB bacterial infection, we performed a comparative transcriptome analysis and found that a total of 21 Probesets are commonly up-regulated by the HLB bacterial infection. In addition, a number of genes are likely regulated specifically at early, late or very late stages of the infection. Furthermore, using Pearson correlation coefficient-based gene coexpression analysis, we constructed a citrus HLB response network consisting of 3,507 Probesets and 56,287 interactions. Genes involved in carbohydrate and nitrogen metabolic processes, transport, defense, signaling and hormone response were overrepresented in the HLB response network and the subnetworks for these processes were constructed. Analysis of the defense and hormone response subnetworks indicates that hormone response is interconnected with defense response. In addition, mapping the commonly up-regulated HLB responsive genes into the HLB response network resulted in a core subnetwork where transport plays a key role in the citrus response to the HLB bacterial infection. Moreover, analysis of a phloem protein subnetwork indicates a role for this protein and zinc transporters or zinc-binding proteins in the citrus HLB defense response. Conclusion Through integrating transcriptome comparison and gene coexpression network analysis, we have provided for the first time a systems view of citrus in response to the Ca. Liberibacter spp. infection causing HLB. PMID:23324561

  13. Callose deposition in the phloem plasmodesmata and inhibition of phloem transport in citrus leaves infected with "Candidatus Liberibacter asiaticus".

    PubMed

    Koh, Eun-Ji; Zhou, Lijuan; Williams, Donna S; Park, Jiyoung; Ding, Ningyuan; Duan, Yong-Ping; Kang, Byung-Ho

    2012-07-01

    Huanglongbing (HLB) is a destructive disease of citrus trees caused by phloem-limited bacteria, Candidatus Liberibacter spp. One of the early microscopic manifestations of HLB is excessive starch accumulation in leaf chloroplasts. We hypothesize that the causative bacteria in the phloem may intervene photoassimilate export, causing the starch to over-accumulate. We examined citrus leaf phloem cells by microscopy methods to characterize plant responses to Liberibacter infection and the contribution of these responses to the pathogenicity of HLB. Plasmodesmata pore units (PPUs) connecting companion cells and sieve elements were stained with a callose-specific dye in the Liberibacter-infected leaf phloem cells; callose accumulated around PPUs before starch began to accumulate in the chloroplasts. When examined by transmission electron microscopy, PPUs with abnormally large callose deposits were more abundant in the Liberibacter-infected samples than in the uninfected samples. We demonstrated an impairment of symplastic dye movement into the vascular tissue and delayed photoassimilate export in the Liberibacter-infected leaves. Liberibacter infection was also linked to callose deposition in the sieve plates, which effectively reduced the sizes of sieve pores. Our results indicate that Liberibacter infection is accompanied by callose deposition in PPUs and sieve pores of the sieve tubes and suggest that the phloem plugging by callose inhibits phloem transport, contributing to the development of HLB symptoms.

  14. The genome sequence of an isolate of Indian citrus ringspot virus infecting the sweet orange in India.

    PubMed

    K, Prabha; Baranwal, V K

    2012-11-01

    Whole-genome sequencing of an isolate of Mandarivirus infecting the sweet orange [Citrus sinensis (L) Blanco] in the western part of India (Pune) was done. The single-stranded positive-sense RNA genome of Indian citrus ringspot virus (ICRSV) Pune has 7,560 nucleotides (nt), excluding a poly(A) tail, comprised of 27.98% (2,115 nt) A, 32.12% (2,428 nt) C, 19.68% (1,488 nt) G, and 20.22% (1,529 nt) T residues. The genome, organized into six open reading frames (ORFs), shares 97.7% sequence identity with the complete genome of the ICRSV K1 isolate (AF406744.1) infecting the kinnow (Citrus reticulate Blanco, a hybrid between King and Willow mandarins) in north India. The ICRSV Pune genome formed a complex secondary structure with a large number of unpaired cytosine-rich regions, and recombination analysis highlighted potential recombination in the ICRSV genome.

  15. RL-SAGE and microarray analysis of the rice defense transcriptome after Rhizoctonia solani infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sheath blight caused by the fungal pathogen Rhizoctonia solani is an emerging problem in rice production worldwide. To elucidate the molecular basis of rice defense to the pathogen, RNA isolated from R. solani-infected leaves of Jasmine 85 was used for both RL-SAGE library construction and microarra...

  16. Possible Contribution of Blast Spores to the Oxidative Burst in the Infection Droplet on Rice Leaf

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The infection-induced overproduction of reactive oxygen species (ROS) in resistant plants is usually ascribed to the host. Here we tested the possible contribution of the parasite, the rice blast fungus to ROS production. Droplets of spore suspensions or water were kept on rice leaves or on plastic....

  17. The effectiveness of grapefruit (Citrus paradisi) seeds in treating urinary tract infections.

    PubMed

    Oyelami, O A; Agbakwuru, E A; Adeyemi, L A; Adedeji, G B

    2005-04-01

    Three middle-aged males and one female were diagnosed as having urinary tract infections (UTIs) between 2001 and 2003 in the Wesley Guild Hospital, Ilesa, a unit of Obafemi Awolowo University Teaching Hospitals Complex, Ile Ife, Osun State, Nigeria. Of the 4 patients, only the female was asymptomatic. The 3 males had Pseudomonas aeruginosa, Klebsiella species, and Staphylococcus aureus, respectively, in their urine samples, while the female had Escherichia coli. All 4 patients were treated with grapefruit seeds (Citrus paradisi) orally for 2 weeks and they all responded satisfactorily to the treatment except the man with P. aeruginosa isolate. However, the initial profuse growth of Pseudomonas isolate in the patient that was resistant to gentamicin, tarivid, and augmentin later subsided to mild growth with reversal of the antibiotic resistance pattern after 2 weeks' treatment with grapefruit seeds. These preliminary data thus suggest an antibacterial characteristic of dried or fresh grapefruit seeds (C. paradisi) when taken at a dosage of 5 to 6 seeds every 8 hours, that is comparable to that of proven antibacterial drugs.

  18. Global Gene Expression of Rice after Infections with Rice Blast and Sheath blight Pathogens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rice (Oryza sativa) production worldwide has been challenged by increased new virulent pathogens. Over the years, genetic diversity needed for fighting diseases has been decreasing in cultivated rice around the globe. This presents a real challenge for rice crop protection. In an effort to develo...

  19. Spectral sensitivity of the Asian citrus psyllid, Diaphorina citri

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Asian Citrus psyllid, Diaphorina citri, as a vector of the bacteria causing citrus greening, is considered one of the most important citrus pests globally. Movement of infected psyllids onto uninfected young citrus remains a key concern for the maintenance of citrus production. Attraction of d...

  20. Exacerbation of citrus canker by citrus leafminer, Phyllocnistis citrella in Florida

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker (caused by Xanthomonas citri subsp. citri, Xcc) is an important bacterial disease of citrus that is spread naturally by rain and wind. Damage to citrus leaves by the citrus leafminer (CLM) , Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae), has been shown to promote infect...

  1. Processes involved in the dispersal of Xanthomonas citri pv. citri from canker-infectd citrus canopies, and in the infection of citrus foliage

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker (Xanthomonas citri subsp. citri, Xcc) is now considered endemic in Florida, and epidemics result in yield loss and market penalties both in Florida, and elsewhere where the pathogen occurs, and susceptible citrus is cultivated. The bacterium is dispersed in rain splash, and storms wit...

  2. Deciphering the Bacterial Microbiome of Citrus Plants in Response to ‘Candidatus Liberibacter asiaticus’-Infection and Antibiotic Treatments

    PubMed Central

    Zhang, Muqing; Powell, Charles A.; Benyon, Lesley S.; Zhou, Hui; Duan, Yongping

    2013-01-01

    The bacterial microbiomes of citrus plants were characterized in response to ‘Candidatus Liberibacter asiaticus’ (Las)-infection and treatments with ampicillin (Amp) and gentamicin (Gm) by Phylochip-based metagenomics. The results revealed that 7,407 of over 50,000 known Operational Taxonomic Units (OTUs) in 53 phyla were detected in citrus leaf midribs using the PhyloChip™ G3 array, of which five phyla were dominant, Proteobacteria (38.7%), Firmicutes (29.0%), Actinobacteria (16.1%), Bacteroidetes (6.2%) and Cyanobacteria (2.3%). The OTU62806, representing ‘Candidatus Liberibacter’, was present with a high titer in the plants graft-inoculated with Las-infected scions treated with Gm at 100 mg/L and in the water-treated control (CK1). However, the Las bacterium was not detected in the plants graft-inoculated with Las-infected scions treated with Amp at 1.0 g/L or in plants graft-inoculated with Las-free scions (CK2). The PhyloChip array demonstrated that more OTUs, at a higher abundance, were detected in the Gm-treated plants than in the other treatment and the controls. Pairwise comparisons indicated that 23 OTUs from the Achromobacter spp. and 12 OTUs from the Methylobacterium spp. were more abundant in CK2 and CK1, respectively. Ten abundant OTUs from the Stenotrophomonas spp. were detected only in the Amp-treatment. These results provide new insights into microbial communities that may be associated with the progression of citrus huanglongbing (HLB) and the potential effects of antibiotics on the disease and microbial ecology. PMID:24250784

  3. Infection of Ustilaginoidea virens intercepts rice seed formation but activates grain-filling-related genes.

    PubMed

    Fan, Jing; Guo, Xiao-Yi; Li, Liang; Huang, Fu; Sun, Wen-Xian; Li, Yan; Huang, Yan-Yan; Xu, Yong-Ju; Shi, Jun; Lei, Yang; Zheng, Ai-Ping; Wang, Wen-Ming

    2015-06-01

    Rice false smut has become an increasingly serious disease in rice (Oryza sativa L.) production worldwide. The typical feature of this disease is that the fungal pathogen Ustilaginoidea virens (Uv) specifically infects rice flower and forms false smut ball, the ustiloxin-containing ball-like fungal colony, of which the size is usually several times larger than that of a mature rice seed. However, the underlying mechanisms of Uv-rice interaction are poorly understood. Here, we applied time-course microscopic and transcriptional approaches to investigate rice responses to Uv infection. The results demonstrated that the flower-opening process and expression of associated transcription factors, including ARF6 and ARF8, were inhibited in Uv-infected spikelets. The ovaries in infected spikelets were interrupted in fertilization and thus were unable to set seeds. However, a number of grain-filling-related genes, including seed storage protein genes, starch anabolism genes and endosperm-specific transcription factors (RISBZ1 and RPBF), were highly transcribed as if the ovaries were fertilized. In addition, critical defense-related genes like NPR1 and PR1 were downregulated by Uv infection. Our data imply that Uv may hijack host nutrient reservoir by activation of the grain-filling network because of growth and formation of false smut balls.

  4. Physical Changes in Satsuma Mandarin Leaf after Infection of Elsinoë fawcettii Causing Citrus Scab Disease

    PubMed Central

    Paudyal, Dilli Prasad; Hyun, Jae-Wook

    2015-01-01

    Citrus scab disease is one of the destructive diseases that reduce the value of fruit for the fresh market. We analyzed the process of symptom development after infection with scab pathogen Elsinoë fawcettii in the susceptible satsuma mandarin leaves to observe the structural modification against pathogen. The cuticle and epidermal cells along with 3–5 layers of mesophyll tissue were degraded 1–2 days post inoculation. Surrounding peripheral cells of degraded tissues grew rapidly and then enveloped the necrotic area along with the growing conidia. Cross sections through the lesion revealed hyphal colonization in epidermis and mesophyll tissues. In response to the pathogen colonization, host cell walls were lignified, inner cells were rapidly compartmentalized and a semi-circular boundary was formed that separated the infected region from the non-infected region, and finally prevented the intercellular pathogen spread. PMID:26674386

  5. Incidence of Candidatus Liberibacter asiaticus infection in abandoned citrus occurring in proximity to commercially managed groves.

    PubMed

    Tiwari, Siddharth; Lewis-Rosenblum, Hannah; Pelz-Stelinski, Kirsten; Stelinski, Lukasz L

    2010-12-01

    Huanglongbing is one of the most devastating diseases of citrus (Citrus spp.). One management tactic against huanglongbing is aggressive management of the vector, the Asian citrus psyllid (Diaphorina citri Kuwayama), with insecticide applications. However, D. citri in abandoned groves are not controlled and therefore pose a risk of reinfestation for nearby commercial citrus. These abandoned groves could serve as a reservoir for the vector, as well as a source of the presumed causal agent for huanglongbing in Florida, Candidatus Liberibacter asiaticus (Las). The current study was conducted to determine the degree to which Las is present in abandoned Florida citrus groves and to compare relative inoculum levels in nearby managed and abandoned groves during times of the year when D. citri are abundant (June, July, and August). In addition, the movement of Las by dispersing D. citri adults from inner and edge rows of abandoned grove plots to the corresponding rows of managed plots was quantified during the same 3 mo. The results of the current study confirmed the presence of Las in both D. citri and plant tissue in abandoned groves at statistically equivalent levels to those in nearby managed groves. The mean number of D. citri adults dispersing from abandoned to managed grove plots ranged from 7.25 +/- 1.70 to 70.25 +/- 21.25 per 4-d intervals. Of those, the mean number of dispersing D. citri adults that were carrying the Las pathogen ranged from 1.00 +/- 0.58 to 1.50 +/- 0.50. Our results indicate that abandoned citrus groves are a significant source of Ca. Las and that dispersing D. citri move this pathogen into nearby managed groves.

  6. Supplementation of moist and dehydrated citrus pulp in the diets of sheep artificially and naturally infected with gastrointestinal nematodes on the parasitological parameters and performance.

    PubMed

    Nordi, E C P; Costa, R L D; David, C M G; Parren, G A E; Freitas, A C B; Lameirinha, L P; Katiki, L M; Bueno, M S; Quirino, C R; Gama, P E; Bizzo, H R; Chagas, A C S

    2014-10-15

    The inclusion of industrial byproducts such as citrus pulp in the composition of animal diets has been widely recommended due to sustainability aspects and their high level of carbohydrates. Limonene is found in citrus pulp and has been described elsewhere as a major compound of citrus essential oils with excellent anthelmintic activity. The objective of this study was to evaluate the parasitological parameters of lambs artificially infected (Experiment 1) with Haemonchus contortus and naturally infected (Experiment 2) by gastrointestinal nematodes, fed diets with dehydrated citrus pulp or silage of moist orange pulp. Both experiments had three treatments (C: control, DP: diet+dehydrated citrus pulp, and MP: diet+silage of moist orange pulp). The diets were isoproteic (11% crude protein) and the concentrate was corrected every 14 days according to animal weight. Parasitological parameters were evaluated for both experiments each 14 days (body weight, body condition; fecal egg counts-FEC, egg hatch assay-EHA, coproculture, and packed cell volume-PCV). Analysis of variance (GLM of the SAS software) was performed with repeated measures in time, and the means were compared by the Tukey test. Gas chromatography with mass spectrometry was used to detect constituents of dry or moist citrus pulp. Dehydrated citrus pulp had 0.02% essential oil (major compounds were 85.9% limonene and 7.6% valencene). Moist orange pulp contained 1.5% essential oil (major compounds were 65.5% limonene and 31.2% alpha- and gamma-terpineol). In both experiments, the weight gain among the treatments was similar (p>0.05) demonstrating that both moist and dehydrated orange pulp can be used to replace corn kernels to feed infected lambs. The supplementation with orange pulp did not decrease natural or artificial infections of gastrointestinal nematodes according to the FEC results (p>0.05). However, PCV increased from animals fed dehydrated and moist pulp in natural infection (Experiment 2, p<0

  7. Diaphorina citri Induces Huanglongbing-Infected Citrus Plant Volatiles to Repel and Reduce the Performance of Propylaea japonica

    PubMed Central

    Lin, Yongwen; Lin, Sheng; Akutse, Komivi S.; Hussain, Mubasher; Wang, Liande

    2016-01-01

    Transmission of plant pathogens through insect vectors is a complex biological process involving interactions between the host plants, insects, and pathogens. Simultaneous impact of the insect damage and pathogenic bacteria in infected host plants induce volatiles that modify not only the behavior of its insect vector but also of their natural enemies, such as parasitoid wasps. Therefore, it is essential to understand how insects such as the predator ladybird beetle responds to volatiles emitted from a host plant and how the disease transmission alters the interactions between predators, vector, pathogens, and plants. In this study, we investigated the response of Propylaea japonica to volatiles from citrus plants damaged by Diaphorina citri and Candidatus Liberibacter asiaticus through olfactometer bioassays. Synthetic chemical blends were also used to determine the active compounds in the plant volatile. The results showed that volatiles emitted by healthy plants attracted more P. japonica than other treatments, due to the presence of high quantities of D-limonene and beta-ocimene, and the lack of methyl salicylate. When using synthetic chemicals in the olfactory tests, we found that D-limonene attracted P. japonica while methyl salicylate repelled the predator. However, beta-ocimene attracted the insects at lower concentrations but repelled them at higher concentrations. These results indicate that P. japonica could not efficiently search for its host by using volatile cues emitted from psyllids- and Las bacteria-infected citrus plants. PMID:28083006

  8. Relevance of the transcription factor PdSte12 in Penicillium digitatum conidiation and virulence during citrus fruit infection.

    PubMed

    Vilanova, Laura; Teixidó, Neus; Torres, Rosario; Usall, Josep; Viñas, Inmaculada; Sánchez-Torres, Paloma

    2016-10-17

    Green mould, resulting from Penicillium digitatum, is the most important postharvest disease of citrus. In a previous study, the PdSte12 transcription factor gene was identified, and disruption mutants were obtained. In the present study, the ΔPdSte12 mutants generated through gene replacement showed significantly reduced virulence during citrus fruit infection. Virulence was affected not only in mature fruit but also in immature fruit, and disease severity was markedly reduced when the oranges were stored at 20 or 4°C. In addition, the ΔPdSte12 mutants were defective in asexual reproduction, producing few conidia. The conidiophores of these mutants had longer metulae with fewer branches at the tip of the hyphae. Gene expression analysis revealed that PdSte12 might act as a negative regulator of several transporter-encoding genes and a positive regulator of two sterol demethylases, all of which are involved in fungicide resistance and fungal virulence. Moreover, PdSte12 exhibited the negative regulation of another transcription factor PdMut3, putatively involved in fungal pathogenesis but with no effect on the MAPK SLT2 P. digitatum orthologue belonging to different transcription pathways relevant to cell integrity. These results indicate the PdSte12 transcription factor is functionally conserved in P. digitatum for infection and asexual reproduction, similar to other Ste12 fungal plant pathogens.

  9. XA21-specific induction of stress-related genes following Xanthomonas infection of detached rice leaves

    PubMed Central

    Liu, Furong; Chen, Huamin; Wei, Tong; Nguyen, Yen P.; Shaker, Isaac W.F.

    2016-01-01

    The rice XA21 receptor kinase confers robust resistance to the bacterial pathogen Xanthomonas oryzaepv. oryzae (Xoo). We developed a detached leaf infection assay to quickly and reliably measure activation of the XA21-mediated immune response using genetic markers. We used RNA sequencing of elf18 treated EFR:XA21:GFP plants to identify candidate genes that could serve as markers for XA21 activation. From this analysis, we identified eight genes that are up-regulated in both in elf18 treated EFR:XA21:GFP rice leaves and Xoo infected XA21 rice leaves. These results provide a rapid and reliable method to assess bacterial-rice interactions. PMID:27703843

  10. Effect of chemical compounds on the ‘Cadidatus Liberibacter asiaticus’ infected pomelo (Citrus maxima)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB), also known as citrus greening, is one of the most destructive diseases affecting Rutaceae plants in many parts of the world. HLB is associated with three species of ‘Candidatus Liberibacter’ with ‘Ca. L. asiaticus’ (Las), being the most widely distributed in Thailand and Asia. T...

  11. Transcriptional response of susceptible and tolerant citrus to infection with Candidatus Liberibacter asiaticus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Candidatus Liberibacter asiaticus (Las), a non-culturable phloem-limited bacterium, is the suspected causal agent of Huanglongbing (HLB) in Florida. HLB is one of the most devastating diseases of citrus and no resistant cultivars have been identified to date, though tolerance has been observed in t...

  12. Current understanding on Villosiclava virens, a unique flower-infecting fungus causing rice false smut disease.

    PubMed

    Fan, Jing; Yang, Juan; Wang, Yu-Qiu; Li, Guo-Bang; Li, Yan; Huang, Fu; Wang, Wen-Ming

    2016-12-01

    Villosiclava virens (Vv) is an ascomycete fungal pathogen that causes false smut disease in rice. Recent reports have revealed some interesting aspects of the enigmatic pathogen to address the question of why it specifically infects rice flowers and converts a grain into a false smut ball. Comparative and functional genomics have suggested specific adaptation of Vv in the colonization of rice flowers. Anatomical studies have disclosed that Vv specifically infects rice stamen filaments before heading and intercepts seed formation. In addition, Vv can occupy the whole inner space of a spikelet embracing all floral organs and activate the rice grain-filling network, presumably for nutrient acquisition to support the development of the false smut ball. This profile provides a general overview of the rice false smut pathogen, and summarizes advances in the Vv life cycle, genomics and genetics, and the molecular Vv-rice interaction. Current understandings of the Vv-rice pathosystem indicate that it is a unique and interesting system which can enrich the study of plant-pathogen interactions. Taxonomy: Ustilaginoidea virens is the anamorph form of the pathogen (Kingdom Fungi; Phylum Ascomycota; Class Ascomycetes; Subclass Incertae sedis; Order Incertae sedis; Family Incertae sedis; Genus Ustilaginoidea). The teleomorph form is Villosiclava virens (Kingdom Fungi; Phylum Ascomycota; Class Ascomycetes; Subclass Sordariomycetes; Order Hypocreales; Family Clavicipitaceae; Genus Villosiclava). Disease symptoms: The only visible symptom is the replacement of rice grains by ball-shaped fungal mycelia, namely false smut balls. When maturing, the false smut ball is covered with powdery chlamydospores, and the colour changes to yellowish, yellowish orange, green, olive green and, finally, to greenish black. Sclerotia are often formed on the false smut balls in autumn. Identification and detection: Vv conidia are round to elliptical, measuring 3-5 μm in diameter. Chlamydospores are

  13. Effect of rice cultivation on the prevalence and infection rates of Schistosoma intermediate host.

    PubMed

    el-Hawy, A M; Negm, I A; el-Alamy, M A; Agina, A A

    1993-12-01

    In this study, it was aimed to explore the effect of rice cultivation on the prevalence, density and infection rates on the snail intermediate hosts of human schistosomiasis. A village was chosen and examine for the main water canal supplying the rice field by making three successive scoops at 10.20, 30, 40 and 50 meters up stream the site of the pump supplying the rice field. The same was done in the large irrigation canal supplying 50 feddans of rice up to 100 meters (i.e. 10 sampling sites). Also the drains in between the rice fields were examined at 10 meters distance throughout the drain. The hunted snails were sorted, the Biomphalaria alexanderina and Bulinus truncatus were separated, their age was determined then crushed to look for immature stages under the microscope. Also the vegetations coming in the scoops were examined. The results showed that rice cultivation and irrigation is a suitable site for the prevalence and intensity of snails. Both species of snails were found in the main canal, irrigation canal and drains. All Bulinus truncatus snails were free from infections. The number of infected Biomphalaria alexanderina snails was significantly higher in the irrigation canal and drains than main canal which may be attributed to pollution of the drain water and irrigation canal by micturition and defecation or to the use of water in ablution.

  14. Secondary metabolite profiles of leaves of healthy and huanglongbing-infected orange (Citrus sinensis L.) seedlings measured by HPLC-fluorescence detection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Leaves of greenhouse-grown ‘Hamlin’ and ‘Valencia’ orange (Citrus sinensis L.) seedlings were analyzed by high performance liquid chromatography in a study of the progression of changes in secondary metabolite profiles resulting from infection by Candidatus Liberibacter asiaticus (CLas), the Huanglo...

  15. Proteomics analysis reveals novel host molecular mechanisms associated with thermotherapy of ‘Ca. Liberibacter asiaticus’-infected citrus plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus huanglongbing (HLB) is the most devastating disease of citrus plants, and longer-term measures for controlling the disease via breeding or genetic engineering are hampered by the fact that all cultivated citrus species are susceptible to the disease. However, the degree of HLB susceptibility ...

  16. Rice Xa21 primed genes and pathways that are critical for combating bacterial blight infection.

    PubMed

    Peng, Hai; Chen, Zheng; Fang, Zhiwei; Zhou, Junfei; Xia, Zhihui; Gao, Lifen; Chen, Lihong; Li, Lili; Li, Tiantian; Zhai, Wenxue; Zhang, Weixiong

    2015-07-17

    Rice bacterial blight (BB) is a devastating rice disease. The Xa21 gene confers a broad and persistent resistance against BB. We introduced Xa21 into Oryza sativa L ssp indica (rice 9311), through multi-generation backcrossing, and generated a nearly isogenic, blight-resistant 9311/Xa21 rice. Using next-generation sequencing, we profiled the transcriptomes of both varieties before and within four days after infection of bacterium Xanthomonas oryzae pv. oryzae. The identified differentially expressed (DE) genes and signaling pathways revealed insights into the functions of Xa21. Surprisingly, before infection 1,889 genes on 135 of the 316 signaling pathways were DE between the 9311/Xa21 and 9311 plants. These Xa21-mediated basal pathways included mainly those related to the basic material and energy metabolisms and many related to phytohormones such as cytokinin, suggesting that Xa21 triggered redistribution of energy, phytohormones and resources among essential cellular activities before invasion. Counter-intuitively, after infection, the DE genes between the two plants were only one third of that before the infection; other than a few stress-related pathways, the affected pathways after infection constituted a small subset of the Xa21-mediated basal pathways. These results suggested that Xa21 primed critically important genes and signaling pathways, enhancing its resistance against bacterial infection.

  17. Plant defence suppression is mediated by a fungal sirtuin during rice infection by Magnaporthe oryzae.

    PubMed

    Fernandez, Jessie; Marroquin-Guzman, Margarita; Nandakumar, Renu; Shijo, Sara; Cornwell, Kathryn M; Li, Gang; Wilson, Richard A

    2014-10-01

    Crop destruction by the hemibiotrophic rice pathogen Magnaporthe oryzae requires plant defence suppression to facilitate extensive biotrophic growth in host cells before the onset of necrosis. How this is achieved at the genetic level is not well understood. Here, we report that a M. oryzae sirtuin, MoSir2, plays an essential role in rice defence suppression and colonization by controlling superoxide dismutase (SOD) gene expression. Loss of MoSir2 function in Δsir2 strains did not affect appressorial function, but biotrophic growth in rice cells was attenuated. Compared to wild type, Δsir2 strains failed to neutralize plant-derived reactive oxygen species (ROS) and elicited robust defence responses in rice epidermal cells that included elevated pathogenesis-related gene expression and granular depositions. Deletion of a SOD-encoding gene under MoSir2 control generated Δsod1 deletion strains that mimicked Δsir2 for impaired rice defence suppression, confirming SOD activity as a downstream output of MoSir2. In addition, comparative protein acetylation studies and forward genetic analyses identified a JmjC domain-containing protein as a likely target of MoSir2, and a Δsir2 Δjmjc double mutant was restored for MoSOD1 expression and defence suppression in rice epidermal cells. Together, this work reveals MoSir2 and MoJmjC as novel regulators of early rice cell infection.

  18. Rice-based oral antibody fragment prophylaxis and therapy against rotavirus infection

    PubMed Central

    Tokuhara, Daisuke; ρlvarez, Beatriz; Mejima, Mio; Hiroiwa, Tomoko; Takahashi, Yuko; Kurokawa, Shiho; Kuroda, Masaharu; Oyama, Masaaki; Kozuka-Hata, Hiroko; Nochi, Tomonori; Sagara, Hiroshi; Aladin, Farah; Marcotte, Harold; Frenken, Leon G.J.; Iturriza-Gómara, Miren; Kiyono, Hiroshi; Hammarström, Lennart; Yuki, Yoshikazu

    2013-01-01

    Rotavirus-induced diarrhea is a life-threatening disease in immunocompromised individuals and in children in developing countries. We have developed a system for prophylaxis and therapy against rotavirus disease using transgenic rice expressing the neutralizing variable domain of a rotavirus-specific llama heavy-chain antibody fragment (MucoRice-ARP1). MucoRice-ARP1 was produced at high levels in rice seeds using an overexpression system and RNAi technology to suppress the production of major rice endogenous storage proteins. Orally administered MucoRice-ARP1 markedly decreased the viral load in immunocompetent and immunodeficient mice. The antibody retained in vitro neutralizing activity after long-term storage (>1 yr) and boiling and conferred protection in mice even after heat treatment at 94°C for 30 minutes. High-yield, water-soluble, and purification-free MucoRice-ARP1 thus forms the basis for orally administered prophylaxis and therapy against rotavirus infections. PMID:23925294

  19. Southern rice black-streaked dwarf virus alters insect vectors' host orientation preferences to enhance spread and increase rice ragged stunt virus co-infection.

    PubMed

    Wang, Han; Xu, Donglin; Pu, Lingling; Zhou, Guohui

    2014-02-01

    In recent years, Southern rice black-streaked dwarf virus (SRBSDV), a tentative species in the genus Fijivirus (family Reoviridae), has spread rapidly and caused serious rice losses in eastern and southeastern Asia. With this virus spread, Rice ragged stunt virus (RRSV, genus Oryzavirus, family Reoviridae) became more common in southern China, usually in co-infection with the former. SRBSDV and RRSV are transmitted by two different species of planthoppers, white-backed planthopper (WBPH, Sogatella furcifera) and brown planthopper (BPH, Nilaparvata lugens), respectively, in a persistent, circulative, propagative manner. In this study, using a Y-shape olfactometer-based device, we tested the host preference of three types of macropterous WBPH adults for healthy or SRBSDV-infected rice plants. The results showed that virus-free WBPHs significantly preferred infected rice plants to healthy plants, whereas both the viruliferous and nonviruliferous WBPHs preferred healthy plants to infected plants. In additional tests, we found that the BPHs significantly preferred healthy plants when they were virus free, whereas RRSV-carrying BPHs preferred SRBSDV-infected rice plants. From these findings, we propose that plant viruses may alter host selection preference of vectors to enhance their spread and that of insects vectoring another virus to result in co-infection with more than one virus.

  20. Transcriptome Analysis of Early Responsive Genes in Rice during Magnaporthe oryzae Infection

    PubMed Central

    Wang, Yiming; Kwon, Soon Jae; Wu, Jingni; Choi, Jaeyoung; Lee, Yong-Hwan; Agrawal, Ganesh Kumar; Tamogami, Shigeru; Rakwal, Randeep; Park, Sang-Ryeol; Kim, Beom-Gi; Jung, Ki-Hong; Kang, Kyu Young; Kim, Sang Gon; Kim, Sun Tae

    2014-01-01

    Rice blast disease caused by Magnaporthe oryzae is one of the most serious diseases of cultivated rice (Oryza sativa L.) in most rice-growing regions of the world. In order to investigate early response genes in rice, we utilized the transcriptome analysis approach using a 300 K tilling microarray to rice leaves infected with compatible and incompatible M. oryzae strains. Prior to the microarray experiment, total RNA was validated by measuring the differential expression of rice defense-related marker genes (chitinase 2, barwin, PBZ1, and PR-10) by RT-PCR, and phytoalexins (sakuranetin and momilactone A) with HPLC. Microarray analysis revealed that 231 genes were up-regulated (>2 fold change, p < 0.05) in the incompatible interaction compared to the compatible one. Highly expressed genes were functionally characterized into metabolic processes and oxidation-reduction categories. The oxidative stress response was induced in both early and later infection stages. Biotic stress overview from MapMan analysis revealed that the phytohormone ethylene as well as signaling molecules jasmonic acid and salicylic acid is important for defense gene regulation. WRKY and Myb transcription factors were also involved in signal transduction processes. Additionally, receptor-like kinases were more likely associated with the defense response, and their expression patterns were validated by RT-PCR. Our results suggest that candidate genes, including receptor-like protein kinases, may play a key role in disease resistance against M. oryzae attack. PMID:25506299

  1. Transcriptome Analysis of Early Responsive Genes in Rice during Magnaporthe oryzae Infection.

    PubMed

    Wang, Yiming; Kwon, Soon Jae; Wu, Jingni; Choi, Jaeyoung; Lee, Yong-Hwan; Agrawal, Ganesh Kumar; Tamogami, Shigeru; Rakwal, Randeep; Park, Sang-Ryeol; Kim, Beom-Gi; Jung, Ki-Hong; Kang, Kyu Young; Kim, Sang Gon; Kim, Sun Tae

    2014-12-01

    Rice blast disease caused by Magnaporthe oryzae is one of the most serious diseases of cultivated rice (Oryza sativa L.) in most rice-growing regions of the world. In order to investigate early response genes in rice, we utilized the transcriptome analysis approach using a 300 K tilling microarray to rice leaves infected with compatible and incompatible M. oryzae strains. Prior to the microarray experiment, total RNA was validated by measuring the differential expression of rice defense-related marker genes (chitinase 2, barwin, PBZ1, and PR-10) by RT-PCR, and phytoalexins (sakuranetin and momilactone A) with HPLC. Microarray analysis revealed that 231 genes were up-regulated (>2 fold change, p < 0.05) in the incompatible interaction compared to the compatible one. Highly expressed genes were functionally characterized into metabolic processes and oxidation-reduction categories. The oxidative stress response was induced in both early and later infection stages. Biotic stress overview from MapMan analysis revealed that the phytohormone ethylene as well as signaling molecules jasmonic acid and salicylic acid is important for defense gene regulation. WRKY and Myb transcription factors were also involved in signal transduction processes. Additionally, receptor-like kinases were more likely associated with the defense response, and their expression patterns were validated by RT-PCR. Our results suggest that candidate genes, including receptor-like protein kinases, may play a key role in disease resistance against M. oryzae attack.

  2. Systemic Suppression of the Shoot Metabolism upon Rice Root Nematode Infection

    PubMed Central

    Kyndt, Tina; Denil, Simon; Bauters, Lander; Van Criekinge, Wim; De Meyer, Tim

    2014-01-01

    Hirschmanniella oryzae is the most common plant-parasitic nematode in flooded rice cultivation systems. These migratory animals penetrate the plant roots and feed on the root cells, creating large cavities, extensive root necrosis and rotting. The objective of this study was to investigate the systemic response of the rice plant upon root infection by this nematode. RNA sequencing was applied on the above-ground parts of the rice plants at 3 and 7 days post inoculation. The data revealed significant modifications in the primary metabolism of the plant shoot, with a general suppression of for instance chlorophyll biosynthesis, the brassinosteroid pathway, and amino acid production. In the secondary metabolism, we detected a repression of the isoprenoid and shikimate pathways. These molecular changes can have dramatic consequences for the growth and yield of the rice plants, and could potentially change their susceptibility to above-ground pathogens and pests. PMID:25216177

  3. Systemic suppression of the shoot metabolism upon rice root nematode infection.

    PubMed

    Kyndt, Tina; Denil, Simon; Bauters, Lander; Van Criekinge, Wim; De Meyer, Tim

    2014-01-01

    Hirschmanniella oryzae is the most common plant-parasitic nematode in flooded rice cultivation systems. These migratory animals penetrate the plant roots and feed on the root cells, creating large cavities, extensive root necrosis and rotting. The objective of this study was to investigate the systemic response of the rice plant upon root infection by this nematode. RNA sequencing was applied on the above-ground parts of the rice plants at 3 and 7 days post inoculation. The data revealed significant modifications in the primary metabolism of the plant shoot, with a general suppression of for instance chlorophyll biosynthesis, the brassinosteroid pathway, and amino acid production. In the secondary metabolism, we detected a repression of the isoprenoid and shikimate pathways. These molecular changes can have dramatic consequences for the growth and yield of the rice plants, and could potentially change their susceptibility to above-ground pathogens and pests.

  4. Metabolomic study of two rice lines infected by Rhizoctonia solani in negative ion mode by CE/TOF-MS.

    PubMed

    Suharti, Woro Sri; Nose, Akihiro; Zheng, Shao-Hui

    2016-11-01

    Rhizoctonia solani is a fungal pathogen that causes sheath blight disease in rice plants. In this study, metabolomic analysis using CE/TOF-MS in negative ion mode was used to investigate the resistance response of resistant and susceptible rice lines (32R and 29S, respectively) due to R. solani infection. Two rice lines showed different responses to the infection of R. solani. In 32R, R. solani infection induced significant increases in adenosine diphosphate (ADP), glyceric acid, mucic acid and jasmonic acid. In 29S, inosine monophosphate (IMP) was involved in the plant response to R. solani infection. Phenol compounds showed an increase as a response of the rice lines to R. solani infection. The study suggests that R. solani infection effects in 32R are associated with the induction of plant metabolic processes such as respiration, photorespiration, pectin synthesis, and lignin accumulation. In 29S, the R. solani infection is suggested to correlate with nitrogen metabolism.

  5. Rice-Infecting Pseudomonas Genomes Are Highly Accessorized and Harbor Multiple Putative Virulence Mechanisms to Cause Sheath Brown Rot

    PubMed Central

    Quibod, Ian Lorenzo; Grande, Genelou; Oreiro, Eula Gems; Borja, Frances Nikki; Dossa, Gerbert Sylvestre; Mauleon, Ramil; Cruz, Casiana Vera; Oliva, Ricardo

    2015-01-01

    Sheath rot complex and seed discoloration in rice involve a number of pathogenic bacteria that cannot be associated with distinctive symptoms. These pathogens can easily travel on asymptomatic seeds and therefore represent a threat to rice cropping systems. Among the rice-infecting Pseudomonas, P. fuscovaginae has been associated with sheath brown rot disease in several rice growing areas around the world. The appearance of a similar Pseudomonas population, which here we named P. fuscovaginae-like, represents a perfect opportunity to understand common genomic features that can explain the infection mechanism in rice. We showed that the novel population is indeed closely related to P. fuscovaginae. A comparative genomics approach on eight rice-infecting Pseudomonas revealed heterogeneous genomes and a high number of strain-specific genes. The genomes of P. fuscovaginae-like harbor four secretion systems (Type I, II, III, and VI) and other important pathogenicity machinery that could probably facilitate rice colonization. We identified 123 core secreted proteins, most of which have strong signatures of positive selection suggesting functional adaptation. Transcript accumulation of putative pathogenicity-related genes during rice colonization revealed a concerted virulence mechanism. The study suggests that rice-infecting Pseudomonas causing sheath brown rot are intrinsically diverse and maintain a variable set of metabolic capabilities as a potential strategy to occupy a range of environments. PMID:26422147

  6. Rice-Infecting Pseudomonas Genomes Are Highly Accessorized and Harbor Multiple Putative Virulence Mechanisms to Cause Sheath Brown Rot.

    PubMed

    Quibod, Ian Lorenzo; Grande, Genelou; Oreiro, Eula Gems; Borja, Frances Nikki; Dossa, Gerbert Sylvestre; Mauleon, Ramil; Cruz, Casiana Vera; Oliva, Ricardo

    2015-01-01

    Sheath rot complex and seed discoloration in rice involve a number of pathogenic bacteria that cannot be associated with distinctive symptoms. These pathogens can easily travel on asymptomatic seeds and therefore represent a threat to rice cropping systems. Among the rice-infecting Pseudomonas, P. fuscovaginae has been associated with sheath brown rot disease in several rice growing areas around the world. The appearance of a similar Pseudomonas population, which here we named P. fuscovaginae-like, represents a perfect opportunity to understand common genomic features that can explain the infection mechanism in rice. We showed that the novel population is indeed closely related to P. fuscovaginae. A comparative genomics approach on eight rice-infecting Pseudomonas revealed heterogeneous genomes and a high number of strain-specific genes. The genomes of P. fuscovaginae-like harbor four secretion systems (Type I, II, III, and VI) and other important pathogenicity machinery that could probably facilitate rice colonization. We identified 123 core secreted proteins, most of which have strong signatures of positive selection suggesting functional adaptation. Transcript accumulation of putative pathogenicity-related genes during rice colonization revealed a concerted virulence mechanism. The study suggests that rice-infecting Pseudomonas causing sheath brown rot are intrinsically diverse and maintain a variable set of metabolic capabilities as a potential strategy to occupy a range of environments.

  7. Combined use of a new SNP-based assay and multilocus SSR markers to assess genetic diversity of Xylella fastidiosa subsp. pauca infecting citrus and coffee plants.

    PubMed

    Montes-Borrego, Miguel; Lopes, Joao R S; Jiménez-Díaz, Rafael M; Landa, Blanca B

    2015-03-01

    Two haplotypes of Xylella fastidiosa subsp. pauca (Xfp) that correlated with their host of origin were identified in a collection of 90 isolates infecting citrus and coffee plants in Brazil, based on a single-nucleotide polymorphism in the gyrB sequence. A new single-nucleotide primer extension (SNuPE) protocol was designed for rapid identification of Xfp according to the host source. The protocol proved to be robust for the prediction of the Xfp host source in blind tests using DNA from cultures of the bacterium, infected plants, and insect vectors allowed to feed on Xfp-infected citrus plants. AMOVA and STRUCTURE analyses of microsatellite data separated most Xfp populations on the basis of their host source, indicating that they were genetically distinct. The combined use of the SNaPshot protocol and three previously developed multilocus SSR markers showed that two haplotypes and distinct isolates of Xfp infect citrus and coffee in Brazil and that multiple, genetically different isolates can be present in a single orchard or infect a single tree. This combined approach will be very useful in studies of the epidemiology of Xfp-induced diseases, host specificity of bacterial genotypes, the occurrence of Xfp host jumping, vector feeding habits, etc., in economically important cultivated plants or weed host reservoirs of Xfp in Brazil and elsewhere.

  8. Herbivory by the insect diaphorina citri induces greater change in citrus plant volatile profile than does infection by the bacterium, Candidatus Liberibacter asiaticus

    PubMed Central

    Hijaz, Faraj; El-Shesheny, Ibrahim; Killiny, Nabil

    2013-01-01

    The volatile organic compound (VOC) profile in plant leaves often changes after biotic and abiotic stresses. Monitoring changes in VOCs in plant leaves could provide valuable information about multitrophic interactions. In the current study, we investigated the effect of Asian citrus psyllid (ACP) infestation, citrus greening pathogen (Candidatus Liberibacter asiaticus [CLas]) infection, and simultaneous attack by ACP and CLas on the VOC content of citrus leaves. Leaf volatiles were extracted using hexane and analyzed with gas chromatography-mass spectrometry (GC-MS). Although ACP is a phloem-sucking insect that causes minimal damage to plant tissues, the relative amount of 21 out of the 27 VOCs increased 2- to 10-fold in ACP-infested plants. The relative amount of d-limonene, β-phelandrene, citronellal, and undecanal were increased 4- to 20- fold in CLas-infected plants. A principle component analysis (PCA) and cluster analysis (CA) showed that VOC patterns of ACP-infested and CLas-infected plants were different from each other and were also different from the controls, while the VOC pattern of double-attacked plants was more like that of the controls than that of ACP-infested or CLas-infected plants. VOC amounts from leaves were compromised when plants were attacked by ACP and CLas. The results of this study showed that a simple direct extraction of citrus leaf volatiles could be successfully used to discriminate between healthy and CLas-infected plants. Information about the effects of insect and pathogen attack on the VOC content profile of plants might contribute to a better understanding of biotic stress. PMID:23857364

  9. Citrus phenylpropanoids and defence against pathogens. Part II: gene expression and metabolite accumulation in the response of fruits to Penicillium digitatum infection.

    PubMed

    Ballester, Ana-Rosa; Teresa Lafuente, M; González-Candelas, Luis

    2013-01-01

    The effect of infection of Citrus sinensis (var. Navelina) fruits with Penicillium digitatum was studied at gene expression and metabolite levels. In this study, expression of genes involved in the phenylpropanoid pathway was studied in the flavedo (outer coloured part of the peel) and albedo (inner white part) in response to pathogen infection. Results of the time-course experiment showed that maximal expression of 10 out of 17 phenylpropanoid genes analysed occurred at 48h post-inoculation, when decay symptoms started to appear, and mRNA levels either kept constant or decreased after 72h post-inoculation. To further investigate the putative involvement of the phenylpropanoid pathway in the defence of citrus fruit, changes in the metabolic profile of both tissues infected with P. digitatum was studied by means of HPLC-PDA-FD. Metabolite accumulation levels along the time course suggest that flavanones, flavones, polymethoxylated flavones and scoparone are induced in citrus fruit in response to P. digitatum infection, although with different trends depending on the tissue.

  10. Involvement of ethylene biosynthesis and perception in the susceptibility of citrus fruits to Penicillium digitatum infection and the accumulation of defence-related mRNAs.

    PubMed

    Marcos, Jose F; González-Candelas, Luis; Zacarías, Lorenzo

    2005-08-01

    Citrus fruits infected with the fungus Penicillium digitatum substantially increase the production of the plant hormone ethylene. In this study, the regulation of ethylene biosynthesis in Citrus sinensis-infected fruits and its putative involvement in an active defence response against P. digitatum infection is examined. Ethylene production is demonstrated as being the result of the co-ordinated and differential up-regulation of at least three ethylene biosynthetic genes: ACS1, ACS2, and ACO. Blocking ethylene perception by 1-MCP resulted in an increased ethylene production and ACS2 expression during infection and mechanical wounding, suggesting that this gene is negatively regulated by ethylene. ACO expression was induced by ethylene in the absence of wounding or infection, although further results indicate that its induction during the course of infection may not be primarily mediated by ethylene. Treatment with 1-MCP also increased susceptibility to Penicillium decay, showing an involvement of ethylene perception in promoting defence responses in citrus fruits. The changes in the expression of two defence-related genes up-regulated during infection were also studied: the ones coding for phenylalanine ammonia-lyase (PAL) and an acidic class II chitinase (ACR311). The onset of PAL expression after mechanical wounding or inoculation was not changed in 1-MCP-pretreated fruits, while its later increase during the course of infection was abolished. Chitinase gene induction was more related to mechanical damage and was partially repressed by ethylene. These studies indicate distinct possible regulatory mechanisms of plant fruit defence genes in the context of fungal infection and ethylene perception.

  11. Rice Dwarf Virus P2 Protein Hijacks Auxin Signaling by Directly Targeting the Rice OsIAA10 Protein, Enhancing Viral Infection and Disease Development

    PubMed Central

    Jin, Lian; Qin, Qingqing; Wang, Yu; Pu, Yingying; Liu, Lifang; Wen, Xing; Ji, Shaoyi; Wu, Jianguo; Wei, Chunhong; Li, Yi

    2016-01-01

    The phytohormone auxin plays critical roles in regulating myriads of plant growth and developmental processes. Microbe infection can disturb auxin signaling resulting in defects in these processes, but the underlying mechanisms are poorly understood. Auxin signaling begins with perception of auxin by a transient co-receptor complex consisting of an F-box transport inhibitor response 1/auxin signaling F-box (TIR1/AFB) protein and an auxin/indole-3-acetic acid (Aux/IAA) protein. Auxin binding to the co-receptor triggers ubiquitination and 26S proteasome degradation of the Aux/IAA proteins, leading to subsequent events, including expression of auxin-responsive genes. Here we report that Rice dwarf virus (RDV), a devastating pathogen of rice, causes disease symptoms including dwarfing, increased tiller number and short crown roots in infected rice as a result of reduced sensitivity to auxin signaling. The RDV capsid protein P2 binds OsIAA10, blocking the interaction between OsIAA10 and OsTIR1 and inhibiting 26S proteasome-mediated OsIAA10 degradation. Transgenic rice plants overexpressing wild-type or a dominant-negative (degradation-resistant) mutant of OsIAA10 phenocopy RDV symptoms are more susceptible to RDV infection; however, knockdown of OsIAA10 enhances the resistance of rice to RDV infection. Our findings reveal a previously unknown mechanism of viral protein reprogramming of a key step in auxin signaling initiation that enhances viral infection and pathogenesis. PMID:27606959

  12. Transcriptional reprogramming by root knot and migratory nematode infection in rice.

    PubMed

    Kyndt, Tina; Denil, Simon; Haegeman, Annelies; Trooskens, Geert; Bauters, Lander; Van Criekinge, Wim; De Meyer, Tim; Gheysen, Godelieve

    2012-11-01

    Rice is one of the most important staple crops worldwide, but its yield is compromised by different pathogens, including plant-parasitic nematodes. In this study we have characterized specific and general responses of rice (Oryza sativa) roots challenged with two endoparasitic nematodes with very different modes of action. Local transcriptional changes in rice roots upon root knot (Meloidogyne graminicola) and root rot nematode (RRN, Hirschmanniella oryzae) infection were studied at two time points (3 and 7 d after infection, dai), using mRNA-seq. Our results confirm that root knot nematodes (RKNs), which feed as sedentary endoparasites, stimulate metabolic pathways in the root, and enhance nutrient transport towards the induced root gall. The migratory RRNs, on the other hand, induce programmed cell death and oxidative stress, and obstruct the normal metabolic activity of the root. While RRN infection causes up-regulation of biotic stress-related genes early in the infection, the sedentary RKNs suppress the local defense pathways (e.g. salicylic acid and ethylene pathways). Interestingly, hormone pathways mainly involved in plant development were strongly induced (gibberellin) or repressed (cytokinin) at 3 dai. These results uncover previously unrecognized nematode-induced expression profiles related to their specific infection strategy.

  13. The Ability of the Antagonist Yeast Pichia Guilliermondii Strain Z1 to Suppress Green Mould Infection in Citrus Fruit

    PubMed Central

    Hamadi, Younes; El Guilli, Mohammed; Jijakli, M. Haissam

    2014-01-01

    In previous studies it was shown that Pichia guilliermondii strain Z1, isolated from healthy Moroccan citrus Valencia-Late oranges, was effective against Penicillium italicum. Here the effectiveness of strain Z1 was assessed against Penicillium digitatum, the causal agent of green mould, under different temperature (5-25°C) and relative humidity (RH) (45-100%) regimes for its reliable and large-scale application in packinghouse. All main effects and interactions were significant (P<0.0001). In the pathogen control, the largest lesion diameter was at an RH range between 98 and 100%, regardless of the incubation temperature. The efficacy of strain Z1 was not dependent on the environment and reduced disease incidence by >80%. Its applications as a formulated product significantly reduced the incidence of infected fruit (55%) and the percentage of infected wounds (47%) compared to the only pathogen control treatment. However, disease control with formulated product was significantly less than that obtained with thiabendazole (30%) or strain Z1 culturable cells (35%). These results highlight that strain Z1 is an effective biological control agent for control of green mould under varying environmental conditions, and control may be optimized by combining its use with other environmentally-safe post-harvest treatments or improved formulation. PMID:27800373

  14. The Ability of the Antagonist Yeast Pichia Guilliermondii Strain Z1 to Suppress Green Mould Infection in Citrus Fruit.

    PubMed

    Lahlali, Rachid; Hamadi, Younes; El Guilli, Mohammed; Jijakli, M Haissam

    2014-12-09

    In previous studies it was shown that Pichia guilliermondii strain Z1, isolated from healthy Moroccan citrus Valencia-Late oranges, was effective against Penicillium italicum. Here the effectiveness of strain Z1 was assessed against Penicillium digitatum, the causal agent of green mould, under different temperature (5-25°C) and relative humidity (RH) (45-100%) regimes for its reliable and large-scale application in packinghouse. All main effects and interactions were significant (P<0.0001). In the pathogen control, the largest lesion diameter was at an RH range between 98 and 100%, regardless of the incubation temperature. The efficacy of strain Z1 was not dependent on the environment and reduced disease incidence by >80%. Its applications as a formulated product significantly reduced the incidence of infected fruit (55%) and the percentage of infected wounds (47%) compared to the only pathogen control treatment. However, disease control with formulated product was significantly less than that obtained with thiabendazole (30%) or strain Z1 culturable cells (35%). These results highlight that strain Z1 is an effective biological control agent for control of green mould under varying environmental conditions, and control may be optimized by combining its use with other environmentally-safe post-harvest treatments or improved formulation.

  15. Citrus Genomics

    PubMed Central

    Talon, Manuel; Gmitter Jr., Fred G.

    2008-01-01

    Citrus is one of the most widespread fruit crops globally, with great economic and health value. It is among the most difficult plants to improve through traditional breeding approaches. Currently, there is risk of devastation by diseases threatening to limit production and future availability to the human population. As technologies rapidly advance in genomic science, they are quickly adapted to address the biological challenges of the citrus plant system and the world's industries. The historical developments of linkage mapping, markers and breeding, EST projects, physical mapping, an international citrus genome sequencing project, and critical functional analysis are described. Despite the challenges of working with citrus, there has been substantial progress. Citrus researchers engaged in international collaborations provide optimism about future productivity and contributions to the benefit of citrus industries worldwide and to the human population who can rely on future widespread availability of this health-promoting and aesthetically pleasing fruit crop. PMID:18509486

  16. Transcriptional responses and carbohydrate metabolism of citrus infected with Candidatus Liberibacter asiaticus, the causal agent of Huanglongbing in Florida

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB) is a devastating disease of citrus occurring in most citrus growing regions worldwide. It is caused by a phloem-limited, fastidious bacterium of the genus Candidatus Liberibacter, which is transmitted through insect vectors. HLB causes the decline of trees within few years afte...

  17. Differential stylet penetration behaviors of two Aphis gossypii biotypes in relation to host or vector infection with Citrus tristeza virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) is one of the most important citrus disease agents worldwide. The impact of CTV on American agriculture has been significant, affecting 50 million trees with economic losses of several hundred million dollars. In California, this virus is predominantly transmitted by tw...

  18. Transcriptome analysis of sweet orange trees infected with ‘Candidatus Liberibacter asiaticus’ and two strains of citrus tristeza virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB) and tristeza, are diseases of citrus caused by a member of the a-proteobacteria, ‘Candidatus Liberibacter asiaticus’ (CaLas), and Citrus tristeza virus (CTV) respectively. HLB is a devastating disease, but CTV strains vary from very severe to very mild. Both CaLas and CTV are p...

  19. Wolbachia infections in mosquitoes and their predators inhabiting rice field communities in Thailand and China.

    PubMed

    Wiwatanaratanabutr, Itsanun; Zhang, Chongxing

    2016-07-01

    Wolbachia are inherited, endocytoplasmic bacteria that infect a wide range of arthropods. Here is the first systematic report on the study of Wolbachia infection in mosquitoes and their predators from both Thailand and China. In Thailand, 632 mosquito specimens (20 spp.) and 424 insect predators (23 spp.) were collected from the rice agroecosystem, mostly from the Central region, followed by the Northeast, the North and the South and were inhabiting rice fields, wetlands and ditches. In China, 928 mosquitoes (15 spp.) and 149 insect predators (16 spp.) were collected from rice fields along the Weishan Lake in Shandong province. Specimens were classified in the orders Diptera, Coleoptera, Odonata and Hemiptera. Using wsp, ftsZ, 16S rRNA and groE gene amplifications, Wolbachia were detected in 12 mosquito spp. and 6 predator spp. from Thailand and 11 mosquito spp. and 5 predator spp. from China. The relative Wolbachia densities of these species were determined using quantitative real-time PCR. The mosquito, Aedes albopictus, and the predator, Agriocnemis femina, had the highest bacterial densities. These results imply that Wolbachia of supergroup B are distributed throughout these insects, probably via horizontal transmission in rice agroecosystems.

  20. Genome-wide identification of sweet orange (Citrus sinensis) histone modification gene families and their expression analysis during the fruit development and fruit-blue mold infection process

    PubMed Central

    Xu, Jidi; Xu, Haidan; Liu, Yuanlong; Wang, Xia; Xu, Qiang; Deng, Xiuxin

    2015-01-01

    In eukaryotes, histone acetylation and methylation have been known to be involved in regulating diverse developmental processes and plant defense. These histone modification events are controlled by a series of histone modification gene families. To date, there is no study regarding genome-wide characterization of histone modification related genes in citrus species. Based on the two recent sequenced sweet orange genome databases, a total of 136 CsHMs (Citrus sinensis histone modification genes), including 47 CsHMTs (histone methyltransferase genes), 23 CsHDMs (histone demethylase genes), 50 CsHATs (histone acetyltransferase genes), and 16 CsHDACs (histone deacetylase genes) were identified. These genes were categorized to 11 gene families. A comprehensive analysis of these 11 gene families was performed with chromosome locations, phylogenetic comparison, gene structures, and conserved domain compositions of proteins. In order to gain an insight into the potential roles of these genes in citrus fruit development, 42 CsHMs with high mRNA abundance in fruit tissues were selected to further analyze their expression profiles at six stages of fruit development. Interestingly, a numbers of genes were expressed highly in flesh of ripening fruit and some of them showed the increasing expression levels along with the fruit development. Furthermore, we analyzed the expression patterns of all 136 CsHMs response to the infection of blue mold (Penicillium digitatum), which is the most devastating pathogen in citrus post-harvest process. The results indicated that 20 of them showed the strong alterations of their expression levels during the fruit-pathogen infection. In conclusion, this study presents a comprehensive analysis of the histone modification gene families in sweet orange and further elucidates their behaviors during the fruit development and the blue mold infection responses. PMID:26300904

  1. Genome-wide identification of sweet orange (Citrus sinensis) histone modification gene families and their expression analysis during the fruit development and fruit-blue mold infection process.

    PubMed

    Xu, Jidi; Xu, Haidan; Liu, Yuanlong; Wang, Xia; Xu, Qiang; Deng, Xiuxin

    2015-01-01

    In eukaryotes, histone acetylation and methylation have been known to be involved in regulating diverse developmental processes and plant defense. These histone modification events are controlled by a series of histone modification gene families. To date, there is no study regarding genome-wide characterization of histone modification related genes in citrus species. Based on the two recent sequenced sweet orange genome databases, a total of 136 CsHMs (Citrus sinensis histone modification genes), including 47 CsHMTs (histone methyltransferase genes), 23 CsHDMs (histone demethylase genes), 50 CsHATs (histone acetyltransferase genes), and 16 CsHDACs (histone deacetylase genes) were identified. These genes were categorized to 11 gene families. A comprehensive analysis of these 11 gene families was performed with chromosome locations, phylogenetic comparison, gene structures, and conserved domain compositions of proteins. In order to gain an insight into the potential roles of these genes in citrus fruit development, 42 CsHMs with high mRNA abundance in fruit tissues were selected to further analyze their expression profiles at six stages of fruit development. Interestingly, a numbers of genes were expressed highly in flesh of ripening fruit and some of them showed the increasing expression levels along with the fruit development. Furthermore, we analyzed the expression patterns of all 136 CsHMs response to the infection of blue mold (Penicillium digitatum), which is the most devastating pathogen in citrus post-harvest process. The results indicated that 20 of them showed the strong alterations of their expression levels during the fruit-pathogen infection. In conclusion, this study presents a comprehensive analysis of the histone modification gene families in sweet orange and further elucidates their behaviors during the fruit development and the blue mold infection responses.

  2. Bacterial diversity analysis of Huanglongbing pathogen-infected citrus, using PhyloChip and 16S rRNA gene clone library sequencing

    SciTech Connect

    Shankar Sagaram, U.; DeAngelis, K.M.; Trivedi, P.; Andersen, G.L.; Lu, S.-E.; Wang, N.

    2009-03-01

    The bacterial diversity associated with citrus leaf midribs was characterized 1 from citrus groves that contained the Huanglongbing (HLB) pathogen, which has yet to be cultivated in vitro. We employed a combination of high-density phylogenetic 16S rDNA microarray and 16S rDNA clone library sequencing to determine the microbial community composition of symptomatic and asymptomatic citrus midribs. Our results revealed that citrus leaf midribs can support a diversity of microbes. PhyloChip analysis indicated that 47 orders of bacteria from 15 phyla were present in the citrus leaf midribs while 20 orders from phyla were observed with the cloning and sequencing method. PhyloChip arrays indicated that nine taxa were significantly more abundant in symptomatic midribs compared to asymptomatic midribs. Candidatus Liberibacter asiaticus (Las) was detected at a very low level in asymptomatic plants, but was over 200 times more abundant in symptomatic plants. The PhyloChip analysis was further verified by sequencing 16S rDNA clone libraries, which indicated the dominance of Las in symptomatic leaves. These data implicate Las as the pathogen responsible for HLB disease. Citrus is the most important commercial fruit crop in Florida. In recent years, citrus Huanglongbing (HLB), also called citrus greening, has severely affected Florida's citrus production and hence has drawn an enormous amount of attention. HLB is one of the most devastating diseases of citrus (6,13), characterized by blotchy mottling with green islands on leaves, as well as stunting, fruit decline, and small, lopsided fruits with poor coloration. The disease tends to be associated with a phloem-limited fastidious {alpha}-proteobacterium given a provisional Candidatus status (Candidatus Liberobacter spp. later changed to Candidatus Liberibacter spp.) in nomenclature (18,25,34). Previous studies indicate that HLB infection causes disorder in the phloem and severely impairs the translocation of assimilates in host

  3. Contribution of Ethylene Biosynthesis for Resistance to Blast Fungus Infection in Young Rice Plants1[OA

    PubMed Central

    Iwai, Takayoshi; Miyasaka, Atsushi; Seo, Shigemi; Ohashi, Yuko

    2006-01-01

    The role of ethylene (ET) in resistance to infection with blast fungus (Magnaporthe grisea) in rice (Oryza sativa) is poorly understood. To study it, we quantified ET levels after inoculation, using young rice plants at the four-leaf stage of rice cv Nipponbare (wild type) and its isogenic plant (IL7), which contains the Pi-i resistance gene to blast fungus race 003. Small necrotic lesions by hypersensitive reaction (HR) were formed at 42 to 72 h postinoculation (hpi) in resistant IL7 leaves, and whitish expanding lesions at 96 hpi in susceptible wild-type leaves. Notable was the enhanced ET emission at 48 hpi accompanied by increased 1-aminocyclopropane-1-carboxylic acid (ACC) levels and highly elevated ACC oxidase (ACO) activity in IL7 leaves, whereas only an enhanced ACC increase at 96 hpi in wild-type leaves. Among six ACC synthase (ACS) and seven ACO genes found in the rice genome, OsACS2 was transiently expressed at 48 hpi in IL7 and at 96 hpi in wild type, and OsACO7 was expressed at 48 hpi in IL7. Treatment with an inhibitor for ACS, aminooxyacetic acid, suppressed enhanced ET emission at 48 hpi in IL7, resulting in expanding lesions instead of HR lesions. Exogenously supplied ACC compromised the aminooxyacetic acid-induced breakdown of resistance in IL7, and treatment with 1-methylcyclopropene and silver thiosulfate, inhibitors of ET action, did not suppress resistance. These findings suggest the importance of ET biosynthesis and, consequently, the coproduct, cyanide, for HR-accompanied resistance to blast fungus in young rice plants and the contribution of induced OsACS2 and OsACO7 gene expression to it. PMID:17012402

  4. Plant-parasitic nematode infections in rice: molecular and cellular insights.

    PubMed

    Kyndt, Tina; Fernandez, Diana; Gheysen, Godelieve

    2014-01-01

    Being one of the major staple foods in the world, and an interesting model monocot plant, rice (Oryza sativa L.) has recently received attention from molecular nematologists studying the cellular and molecular aspects of the interaction between this crop and plant-parasitic nematodes. In this review, we highlight recent advances in this field, with a focus on the best-studied root-knot nematodes. Histological studies have revealed the cellular changes inside root-knot nematode-induced feeding sites, both in the compatible interaction with Oryza sativa and the incompatible interaction with the related species Oryza glaberrima. After comparing the published data from transcriptome analyses, mutant studies, and exogenous hormone applications, we provide a comprehensive model showing the role and interaction of plant hormone pathways in defense of this monocot crop against root nematodes, where jasmonate seems to play a key role. Finally, recent evidence indicates that effectors secreted from rice-infecting nematodes can suppress plant defense.

  5. The epidemiological significance of post-packinghouse survival of Xanthomonas citri ssp. citri for dissemination of Asiatic citrus canker via infected fruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The risk of introduction of Xanthomonas citri spp. citri (Xcc) to new, unaffected citrus producing areas is a major concern for those citrus industries attempting to remain free of citrus canker. Citrus fruit, as a potential pathway for Xcc to enter and become established in these areas, is assumed...

  6. Visible/near-infrared spectroscopy for discrimination of HLB-infected citrus leaves from healthy leaves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Researchers have used various hyperspectral systems, covering several areas of the electromagnetic spectrum to investigate all types of disease/plant interactions. The purpose of this research was to investigate using visible and near-infrared (400-1100nm) spectroscopy to differentiate HLB infected...

  7. Quantification and localization of hesperidin and rutin in Citrus sinensis grafted on C. limonia after Xylella fastidiosa infection by HPLC-UV and MALDI imaging mass spectrometry.

    PubMed

    Soares, Márcio Santos; da Silva, Danielle Fernandes; Forim, Moacir Rossi; da Silva, Maria Fátima das Graças Fernandes; Fernandes, João Batista; Vieira, Paulo Cezar; Silva, Denise Brentan; Lopes, Norberto Peporine; de Carvalho, Sérgio Alves; de Souza, Alessandra Alves; Machado, Marcos Antônio

    2015-07-01

    A high performance liquid chromatography-ultraviolet (HPLC-UV) method was developed for quantifying hesperidin and rutin levels in leaves and stems of Citrus limonia, with a good linearity over a range of 1.0-80.0 and 1.0-50.0 μg mL(-1) respectively, with r(2)>0.999 for all curves. The limits of detection (LOD) for both flavonoids were 0.6 and 0.5 μg mL(-1), respectively, with quantification (LOQ) being 2.0 and 1.0 μg mL(-1), respectively. The quantification method was applied to Citrus sinensis grafted onto C. limonia with and without CVC (citrus variegated chlorosis) symptoms after Xylella fastidiosa infection. The total content of rutin was low and practically constant in all analyses in comparison with hesperidin, which showed a significant increase in its amount in symptomatic leaves. Scanning electron microscopy studies on leaves with CVC symptoms showed vessel occlusion by biofilm, and a crystallized material was noted. Considering the difficulty in isolating these crystals for analysis, tissue sections were analyzed by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) to confirm the presence of hesperidin at the site of infection. The images constructed from MS/MS data with a specific diagnostic fragment ion (m/z 483) also showed higher ion intensities for it in infected plants than in healthy ones, mainly in the vessel regions. These data suggest that hesperidin plays a role in the plant-pathogen interaction, probably as a phytoanticipin. This method was also applied to C. sinensis and C. limonia seedlings, and comparison with the graft results showed that the rootstock had an increased hesperidin content ∼3.6 fold greater in the graft stem than in the stem of C. sinensis seedlings. Increase in hesperidin content by rootstock can be related to induced internal defense mechanisms.

  8. Phenylacetaldehyde O-methyloxime: a volatile compound produced by grapefruit leaves infected with the citrus canker pathogen, Xanthomonas axonopodis pv. citri.

    PubMed

    Zhang, Aijun; Hartung, John S

    2005-06-29

    An aldehyde oxime O-methyl ether, phenylacetaldehyde O-methyloxime, was detected using solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) in the headspace above grapefruit leaves infected with Xanthomonas axonopodis pv. citri, the causal agent of citrus bacterial canker disease (CBCD). This disease is a major phytosanitary concern, and an eradication campaign against it is currently underway in Florida. Phenylacetaldehyde O-methyloxime has been reported to be produced by other plants and fragrant flowers, but it was not observed in the headspace above uninfected grapefruit leaves, the pathogenic bacterium X. axonopodis pv. citri itself, or grapefruit leaves infected with another closely related bacterial pathogen, X. axonopodis pv. citrumelo, which causes citrus bacterial spot, a disease of no phytosanitary significance. It was also not detected from CBCD infected fruits, including orange, lemon, grapefruit, and lime. We conclude that phenylacetaldehyde O-methyloxime may potentially be used to identify CBCD infestations. However, more intensive studies will be required to fully evaluate the potential of phenylacetaldehyde O-methyloxime as a diagnostic compound for CBCD. Using SPME and GC-MS to measure phenylacetaldehyde O-methyloxime may provide an easy and feasible tool to complement current methods used to detect X. axonopodis pv. citri in environmental samples.

  9. Anti-infective potential of Citrus bergamia Risso et Poiteau (bergamot) derivatives: a systematic review.

    PubMed

    Cirmi, Santa; Bisignano, Carlo; Mandalari, Giuseppina; Navarra, Michele

    2016-09-01

    Infectious diseases remain among the leading causes of morbidity and mortality worldwide, mainly because of the increase of resistance to chemotherapeutic drugs. Nature is the major source of anti-infective drugs and could represent a font of medicines that may help overcome antibiotic resistance. Recently, the potential antimicrobial effect of certain plant extracts has attracted attention within the scientific community as alternatives to synthetic drugs. Here, we present a systematic review on the anti-infective properties of bergamot derivatives that highlight the activity of bergamot essential oil against bacteria, mycetes and larvae, as well as the anti-Helicobacter pylori effect of bergamot juice and the antimicrobial properties of extracts from bergamot peel. Findings presented herein could be used to develop novel and alternative preventive and therapeutic strategies aimed to overcome antibiotic resistance. Copyright © 2016 John Wiley & Sons, Ltd.

  10. Biological Indexing of Graft Transmissible Diseases of Citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Biological indexing for the detection of graft transmissible diseases of citrus is essential for maintaining a citrus certification program. Many of the graft transmissible diseases of citrus are harbored as latent infections in the scions, but when propagated on a susceptible rootstock that allow...

  11. Pest management practices aimed at curtailing citrus huanglongbing disease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB) is a devastating, insect-vectored disease of citrus sometimes referred to as citrus greening disease and putatively caused by phloem-limited bacteria within the genus Candidatus Liberibacter. Citrus trees infected by this disease decline in productivity; produce misshapen, inedib...

  12. Trailers transporting oranges to processing plants move Asian citrus psyllids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (citrus greening) is one of the most serious of citrus diseases. Movement of the disease occurs as a result of natural vector-borne infection and by movement of plant material. We demonstrate here that Diaphorina citri Kuwayama (vector of citrus greening pathogens) can be transported i...

  13. Current status of Citrus tristeza virus in Central California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Lindcove Research and Extension Center (LREC), Exeter, CA has 51 ha of citrus and is the field site and screenhouses for the University of California Citrus Clonal Protection Program (CCPP). LREC maintains a zero tolerance of Citrus tristeza virus (CTV) infected trees to protect the CCPP and re...

  14. Huanglongbing increases Diplodia Stem End Rot in Citrus sinensis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB), one of the most devastating diseases of citrus is caused by the a-Proteobacteria Candidatus Liberibacter. Diplodia natalensis Pole-Evans is a fungal pathogen which has been known to cause a postharvest stem-end rot of citrus, the pathogen infects citrus fruit under the calyx, an...

  15. Comparison of different detection methods for citrus greening disease based on airborne multispectral and hyperspectral imagery

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus greening or Huanglongbing (HLB) is a devastating disease spread in many citrus groves since first found in 2005 in Florida. Multispectral (MS) and hyperspectral (HS) airborne images of citrus groves in Florida were taken to detect citrus greening infected trees in 2007 and 2010. Ground truthi...

  16. The effect of ‘Candidatus Liberibacter asiaticus’ infection on the proteomic profiles and nutritional status of pre-symptomatic and symptomatic grapefruit (Citrus paradisi) plants

    PubMed Central

    2013-01-01

    Background Huanglongbing (HLB) is a highly destructive citrus disease which threatens citrus production worldwide and ‘Candidatus Liberibacter asiaticus’ (Las), a non-culturable phloem-limited bacterium, is an associated causal agent of the disease. To better understand the physiological and molecular processes involved in host responses to Las, 2-DE and mass spectrometry analyses, as well as ICP spectroscopy analysis were employed to elucidate the global protein expression profiles and nutrient concentrations in leaves of Las-infected grapefruit plants at pre-symptomatic or symptomatic stages for HLB. Results This study identified 123 protein spots out of 191 spots that showed significant changes in the leaves of grapefruit plants in response to Las infection and all identified spots matched to 69 unique proteins/peptides. A down-regulation of 56 proteins including those associated with photosynthesis, protein synthesis, and metabolism was correlated with significant reductions in the concentrations of Ca, Mg, Fe, Zn, Mn, and Cu in leaves of grapefruit plants in response to Las infection, particularly in symptomatic plants. Oxygen-evolving enhancer (OEE) proteins, a PSI 9 kDa protein, and a Btf3-like protein were among a small group of proteins that were down-regulated in both pre-symptomatic and symptomatic plants in response to Las infection. Furthermore, a Las-mediated up-regulation of 13 grapefruit proteins was detected, which included Cu/Zn superoxide dismutase, chitinases, lectin-related proteins, miraculin-like proteins, peroxiredoxins and a CAP 160 protein. Interestingly, a Las-mediated up-regulation of granule-bound starch synthase was correlated with an increase in the K concentrations of pre-symptomatic and symptomatic plants. Conclusions This study constitutes the first attempt to characterize the interrelationships between protein expression and nutritional status of Las-infected pre-symptomatic or symptomatic grapefruit plants and sheds light on

  17. Characterization of the microbial community structure in Candidatus Liberibacter asiaticus-infected citrus plants treated with antibiotics in the field

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB) is one of the most devastating diseases of citrus worldwide. There are no effective control measures for this newly emerging and century-old disease. Previously, we reported a combination of Penicillin G and Streptomycin were effective in eliminating or suppressing the HLB bacter...

  18. NADPH oxidases regulate septin-mediated cytoskeletal remodeling during plant infection by the rice blast fungus

    PubMed Central

    Ryder, Lauren S.; Dagdas, Yasin F.; Mentlak, Thomas A.; Kershaw, Michael J.; Thornton, Christopher R.; Schuster, Martin; Chen, Jisheng; Wang, Zonghua; Talbot, Nicholas J.

    2013-01-01

    The rice blast fungus Magnaporthe oryzae infects plants with a specialized cell called an appressorium, which uses turgor to drive a rigid penetration peg through the rice leaf cuticle. Here, we show that NADPH oxidases (Nox) are necessary for septin-mediated reorientation of the F-actin cytoskeleton to facilitate cuticle rupture and plant cell invasion. We report that the Nox2–NoxR complex spatially organizes a heteroligomeric septin ring at the appressorium pore, required for assembly of a toroidal F-actin network at the point of penetration peg emergence. Maintenance of the cortical F-actin network during plant infection independently requires Nox1, a second NADPH oxidase, which is necessary for penetration hypha elongation. Organization of F-actin in appressoria is disrupted by application of antioxidants, whereas latrunculin-mediated depolymerization of appressorial F-actin is competitively inhibited by reactive oxygen species, providing evidence that regulated synthesis of reactive oxygen species by fungal NADPH oxidases directly controls septin and F-actin dynamics. PMID:23382235

  19. Citrus Inventory

    NASA Technical Reports Server (NTRS)

    1994-01-01

    An aerial color infrared (CIR) mapping system developed by Kennedy Space Center enables Florida's Charlotte County to accurately appraise its citrus groves while reducing appraisal costs. The technology was further advanced by development of a dual video system making it possible to simultaneously view images of the same area and detect changes. An image analysis system automatically surveys and photo interprets grove images as well as automatically counts trees and reports totals. The system, which saves both time and money, has potential beyond citrus grove valuation.

  20. Citrus Inventory

    NASA Technical Reports Server (NTRS)

    1986-01-01

    Florida's Charlotte County Property Appraiser is using an aerial color infrared mapping system for inventorying citrus trees for valuation purposes. The ACIR system has significantly reduced the time and manpower required for appraisal. Aerial photographs are taken and interpreted by a video system which makes it possible to detect changes from previous years. Potential problems can be identified. KSC's TU Office has awarded a contract to the Citrus Research and Education Center to adapt a prototype system which would automatically count trees and report totals.

  1. Characterization of the microbial community structure in Candidatus Liberibacter asiaticus-infected citrus plants treated with antibiotics in the field

    PubMed Central

    2013-01-01

    Background Huanglongbing (HLB) is a worldwide devastating disease of citrus. There are no effective control measures for this newly emerging but century-old disease. Previously, we reported a combination of Penicillin G and Streptomycin was effective in eliminating or suppressing the associated bacterium, ‘Candidatus Liberibacter asiaticus’ (Las). Results Here we report the bacterial composition and community structure in HLB-affected citrus plants during a growing season and while being treated with antibiotic combinations PS (Penicillin G and Streptomycin) and KO (Kasugamycin and Oxytetracycline) using the Phylochip™ G3 array. Both antibiotic treatments resulted in significantly lower Las bacterial titers (Pr<0.05) and hybridization scores. Of the 50,000+ available operational taxonomic units (OTUs) on PhyloChip™ G3, 7,028 known OTUs were present in citrus leaf midribs. These OTUs were from 58 phyla, of which five contained 100 or more OTUs, Proteobacteria (44.1%), Firmicutes (23.5%), Actinobacteria (12.4%), Bacteroidetes (6.6%) and Cyanobacteria (3.2%). In the antibiotic treated samples, the number of OTUs decreased to a total of 5,599. The over-all bacterial diversity decreased with the antibiotic treatments, as did the abundance of 11 OTUs within Proteobacteria, Firmicutes, Bacteroidetes and Planctomycetes. Within the Proteobacteria, ten OTUs representing the class γ-proteobacteria increased in abundance after four months of treatment, when the Las bacterium was at its lowest level in the HLB-affected citrus field plants. Conclusions Our data revealed that Proteobacteria was constantly the dominant bacterial phylum recovered from citrus leaf midribs, with the α-proteobacterial and the γ-proteobacterial classes vying for prevalence. In addition, the level of bacterial diversity found in the leaf midribs of field citrus was greater than previously described. Bacterial cells in close proximity may be able to modify their microenvironment, making the

  2. Comparative ultrastructure of nonwounded Mexican lime and Yuzu leaves infected with the citrus canker bacterium Xanthomonas citri pv. citri.

    PubMed

    Lee, In Jung; Kim, Ki Woo; Hyun, Jae Wook; Lee, Yong Hoon; Park, Eun Woo

    2009-07-01

    Ultrastructural aspects of citrus canker development were investigated in nonwounded leaves of citrus species by transmission electron microscopy (TEM). A susceptible species Mexican lime and a resistant species Yuzu were spray-inoculated with a virulent strain of Xanthomonas citri pv. citri. Initial symptoms occurred on Mexican lime approximately 9 days after inoculation, whereas they appeared on Yuzu mostly 11 days after inoculation. In Mexican lime leaves, the bacterial invasion was usually accompanied by host cell wall dissolution and cellular disruption. Fibrillar materials from degenerated cell walls were usually found in intercellular spaces. Damaged host cells with necrotic cytoplasm showed the localized separation of plasma membrane from the cell wall. Bacterial multiplication and electron-transparent capsule-like structures around bacteria were commonly observed. Meanwhile, cell wall protuberances were prominent outside host cell walls in response to bacterial invasion in Yuzu leaves. Occlusion of intercellular spaces was also formed by the fusion of two or more individual cell wall protuberances originated from two adjacent host cells. Papillae-like materials accumulated locally within host cells in close proximity to bacteria. Some bacteria were found to be undergoing degeneration in xylem vessels. Also, the shrunken, inactive bacteria were surrounded by electron-translucent fibrillar materials in intercellular spaces, implying bacterial immobilization. These cellular responses are thought to be the consequences of defense responses of Yuzu leaves to invading bacteria. In both citrus species, X. citri pv. citri contained polyphosphate bodies showing electron-dense and elliptical structures in cytoplasm.

  3. RL-SAGE and microarray analysis of the rice transcriptome after Rhizoctonia solani infection.

    PubMed

    Venu, R C; Jia, Yulin; Gowda, Malali; Jia, Melissa H; Jantasuriyarat, Chatchawan; Stahlberg, Eric; Li, Huameng; Rhineheart, Andrew; Boddhireddy, Prashanth; Singh, Pratibha; Rutger, Neil; Kudrna, David; Wing, Rod; Nelson, James C; Wang, Guo-Liang

    2007-10-01

    Sheath blight caused by the fungal pathogen Rhizoctonia solani is an emerging problem in rice production worldwide. To elucidate the molecular basis of rice defense to the pathogen, RNA isolated from R. solani-infected leaves of Jasmine 85 was used for both RL-SAGE library construction and microarray hybridization. RL-SAGE sequence analysis identified 20,233 and 24,049 distinct tags from the control and inoculated libraries, respectively. Nearly half of the significant tags (> or =2 copies) from both libraries matched TIGR annotated genes and KOME full-length cDNAs. Among them, 42% represented sense and 7% antisense transcripts, respectively. Interestingly, 60% of the library-specific (> or =10 copies) and differentially expressed (>4.0-fold change) tags were novel transcripts matching genomic sequence but not annotated genes. About 70% of the genes identified in the SAGE libraries showed similar expression patterns (up or down-regulated) in the microarray data obtained from three biological replications. Some candidate RL-SAGE tags and microarray genes were located in known sheath blight QTL regions. The expression of ten differentially expressed RL-SAGE tags was confirmed with RT-PCR. The defense genes associated with resistance to R. solani identified in this study are useful genomic materials for further elucidation of the molecular basis of the defense response to R. solani and fine mapping of target sheath blight QTLs.

  4. Characterization of rice black-streaked dwarf virus- and rice stripe virus-derived siRNAs in singly and doubly infected insect vector Laodelphax striatellus.

    PubMed

    Li, Junmin; Andika, Ida Bagus; Shen, Jiangfeng; Lv, Yuanda; Ji, Yongqiang; Sun, Liying; Chen, Jianping

    2013-01-01

    Replication of RNA viruses in insect cells triggers an antiviral defense that is mediated by RNA interference (RNAi) which generates viral-derived small interfering RNAs (siRNAs). However, it is not known whether an antiviral RNAi response is also induced in insects by reoviruses, whose double-stranded RNA genome replication is thought to occur within core particles. Deep sequencing of small RNAs showed that when the small brown planthopper (Laodelphax striatellus) was infected by Rice black-streaked dwarf virus (RBSDV) (Reoviridae; Fijivirus), more viral-derived siRNAs accumulated than when the vector insect was infected by Rice stripe virus (RSV), a negative single-stranded RNA virus. RBSDV siRNAs were predominantly 21 and 22 nucleotides long and there were almost equal numbers of positive and negative sense. RBSDV siRNAs were frequently generated from hotspots in the 5'- and 3'-terminal regions of viral genome segments but these hotspots were not associated with any predicted RNA secondary structures. Under laboratory condition, L. striatellus can be infected simultaneously with RBSDV and RSV. Double infection enhanced the accumulation of particular genome segments but not viral coat protein of RBSDV and correlated with an increase in the abundance of siRNAs derived from RBSDV. The results of this study suggest that reovirus replication in its insect vector potentially induces an RNAi-mediated antiviral response.

  5. Citrus leprosis virus vectored by Brevipalpus phoenicis (Acari: Tenuipalpidae) on citrus in Brazil.

    PubMed

    Rodrigues, J C V; Kitajima, E W; Childers, C C; Chagas, C M

    2003-01-01

    Citrus leprosis is caused by Citrus leprosis virus (CiLV) that is transmitted by mites in the genus Brevipalpus (Acari: Tenuipalpidae). This disease directly reduces production and the life span of the citrus plant. The main symptoms of the disease include lesions on fruits, leaves, and twigs or small branches, causing premature fruit drop, defoliation, and death of the twigs or branches leading to serious tree decline. Leprosis is a highly destructive disease of citrus, wherever it occurs. The Brazilian citrus industry spends over 100 million US dollars annually on acaricides to control the vector, Brevipalpus phoenicis (Geijskes). This review contains information about the history of the etiology of citrus leprosis, its geographical distribution, host range, the role of the mite vectors, viral morphology and relationships with the infected cell, and transmissibility of the virus by the mite. In addition, data on the mite-virus-plant relationship, disease damage, and strategies for controlling disease spread are presented.

  6. Analysis of the Structure of the AVR1-CO39 Avirulence Locus in Virulent Rice-Infecting Isolates of Magnaporthe grisea

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The AVR1-CO39 gene that came from a Magnaporthe grisea isolate from weeping lovegrass controls avirulence on the rice cultivar CO39. AVR1-CO39 was not present in the genome of the rice-infecting M. grisea isolate Guy11 from French Guyana, suggesting that the gene had been deleted. Molecular analysis...

  7. Transcriptome Markers of Viral Persistence in Naturally-Infected Andes Virus (Bunyaviridae) Seropositive Long-Tailed Pygmy Rice Rats

    PubMed Central

    Campbell, Corey L.; Torres-Perez, Fernando; Acuna-Retamar, Mariana; Schountz, Tony

    2015-01-01

    Long-tailed pygmy rice rats (Oligoryzomys longicaudatus) are principal reservoir hosts of Andes virus (ANDV) (Bunyaviridae), which causes most hantavirus cardiopulmonary syndrome cases in the Americas. To develop tools for the study of the ANDV-host interactions, we used RNA-Seq to generate a de novo transcriptome assembly. Splenic RNA from five rice rats captured in Chile, three of which were ANDV-infected, was used to generate an assembly of 66,173 annotated transcripts, including noncoding RNAs. Phylogenetic analysis of selected predicted proteins showed similarities to those of the North American deer mouse (Peromyscus maniculatus), the principal reservoir of Sin Nombre virus (SNV). One of the infected rice rats had about 50-fold more viral burden than the others, suggesting acute infection, whereas the remaining two had levels consistent with persistence. Differential expression analysis revealed distinct signatures among the infected rodents. The differences could be due to 1) variations in viral load, 2) dimorphic or reproductive differences in splenic homing of immune cells, or 3) factors of unknown etiology. In the two persistently infected rice rats, suppression of the JAK-STAT pathway at Stat5b and Ccnot1, elevation of Casp1, RIG-I pathway factors Ppp1cc and Mff, and increased FC receptor-like transcripts occurred. Caspase-1 and Stat5b activation pathways have been shown to stimulate T helper follicular cell (TFH) development in other species. These data are also consistent with reports suggestive of TFH stimulation in deer mice experimentally infected with hantaviruses. In the remaining acutely infected rice rat, the apoptotic pathway marker Cox6a1 was elevated, and putative anti-viral factors Abcb1a, Fam46c, Spp1, Rxra, Rxrb, Trmp2 and Trim58 were modulated. Transcripts for preproenkephalin (Prenk) were reduced, which may be predictive of an increased T cell activation threshold. Taken together, this transcriptome dataset will permit rigorous

  8. Systemic Infection of Maize, Sorghum, Rice, and Beet Seedlings with Fumonisin-Producing and Nonproducing Fusarium verticillioides Strains

    PubMed Central

    Dastjerdi, Raana; Karlovsky, Petr

    2015-01-01

    Two fumonisin-nonproducing strains of Fusarium verticillioides and their fumonisin producing progenitors were tested for aggressiveness toward maize, sorghum, rice, and beetroot seedlings grown under greenhouse conditions. None of the plants showed obvious disease symptoms after root dip inoculation. Fungal biomass was determined by species-specific real-time PCR. No significant (P = 0.05) differences in systemic colonization were detected between the wild type strains and mutants not producing fumonisins. F. verticillioides was not detected in any of the non-inoculated control plants. The fungus grew from roots to the first two internodes/leaves of maize, rice and beet regardless of fumonisin production. The systemic growth of F. verticillioides in sorghum was limited. The results showed that fumonisin production was not required for the infection of roots of maize, rice and beet by F. verticillioides. PMID:26672472

  9. Citrus leprosis research update

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus leprosis is one of the oldest citrus diseases, but is also one of the most important emerging citrus diseases in South and Central America, and it is apparently spreading northward towards the U.S. Research in our labs and by others has shown that citrus leprosis disease is caused by a compl...

  10. Citrus blight research

    Technology Transfer Automated Retrieval System (TEKTRAN)

    With HLB now occurring throughout Florida citrus groves, citrus blight has been getting less attention even though the problem still exists. In fact, the combination of HLB and citrus blight has compounded the problem that the citrus industry is facing with decreased yields, small fruit size and tre...

  11. Two independent S-phase checkpoints regulate appressorium-mediated plant infection by the rice blast fungus Magnaporthe oryzae

    PubMed Central

    Osés-Ruiz, Míriam; Sakulkoo, Wasin; Littlejohn, George R.; Martin-Urdiroz, Magdalena

    2017-01-01

    To cause rice blast disease, the fungal pathogen Magnaporthe oryzae develops a specialized infection structure called an appressorium. This dome-shaped, melanin-pigmented cell generates enormous turgor and applies physical force to rupture the rice leaf cuticle using a rigid penetration peg. Appressorium-mediated infection requires septin-dependent reorientation of the F-actin cytoskeleton at the base of the infection cell, which organizes polarity determinants necessary for plant cell invasion. Here, we show that plant infection by M. oryzae requires two independent S-phase cell-cycle checkpoints. Initial formation of appressoria on the rice leaf surface requires an S-phase checkpoint that acts through the DNA damage response (DDR) pathway, involving the Cds1 kinase. By contrast, appressorium repolarization involves a novel, DDR-independent S-phase checkpoint, triggered by appressorium turgor generation and melanization. This second checkpoint specifically regulates septin-dependent, NADPH oxidase-regulated F-actin dynamics to organize the appressorium pore and facilitate entry of the fungus into host tissue. PMID:28028232

  12. Genetic Structure and Molecular Variability Analysis of Citrus sudden death-associated virus Isolates from Infected Plants Grown in Brazil

    PubMed Central

    Matsumura, Emilyn Emy; Coletta Filho, Helvécio Della; de Oliveira Dorta, Silvia; Nouri, Shahideh; Machado, Marcos Antonio

    2016-01-01

    Citrus sudden death-associated virus (CSDaV) is a monopartite positive-sense single-stranded RNA virus that was suggested to be associated with citrus sudden death (CSD) disease in Brazil. Here, we report the first study of the genetic structure and molecular variability among 31 CSDaV isolates collected from both symptomatic and asymptomatic trees in CSD-affected areas. Analyses of partial nucleotide sequences of five domains of the CSDaV genomic RNA, including those encoding for the methyltransferase, the multi-domain region (MDR), the helicase, the RNA-dependent RNA polymerase and the coat protein, showed that the MDR coding region was the most diverse region assessed here, and a possible association between this region and virus adaption to different host or plant tissues is considered. Overall, the nucleotide diversity (π) was low for CSDaV isolates, but the phylogenetic analyses revealed the predominance of two main groups, one of which showed a higher association with CSD-symptomatic plants. Isolates obtained from CSD-symptomatic plants, compared to those obtained from asymptomatic plants, showed higher nucleotide diversity, nonsynonymous and synonymous substitution rates and number of amino acid changes on the coding regions located closer to the 5’ end region of the genomic RNA. This work provides new insights into the genetic diversity of the CSDaV, giving support for further epidemiological studies. PMID:27999249

  13. Deciphering Cis-Regulatory Element Mediated Combinatorial Regulation in Rice under Blast Infected Condition

    PubMed Central

    Deb, Arindam; Kundu, Sudip

    2015-01-01

    Combinations of cis-regulatory elements (CREs) present at the promoters facilitate the binding of several transcription factors (TFs), thereby altering the consequent gene expressions. Due to the eminent complexity of the regulatory mechanism, the combinatorics of CRE-mediated transcriptional regulation has been elusive. In this work, we have developed a new methodology that quantifies the co-occurrence tendencies of CREs present in a set of promoter sequences; these co-occurrence scores are filtered in three consecutive steps to test their statistical significance; and the significantly co-occurring CRE pairs are presented as networks. These networks of co-occurring CREs are further transformed to derive higher order of regulatory combinatorics. We have further applied this methodology on the differentially up-regulated gene-sets of rice tissues under fungal (Magnaporthe) infected conditions to demonstrate how it helps to understand the CRE-mediated combinatorial gene regulation. Our analysis includes a wide spectrum of biologically important results. The CRE pairs having a strong tendency to co-occur often exhibit very similar joint distribution patterns at the promoters of rice. We couple the network approach with experimental results of plant gene regulation and defense mechanisms and find evidences of auto and cross regulation among TF families, cross-talk among multiple hormone signaling pathways, similarities and dissimilarities in regulatory combinatorics between different tissues, etc. Our analyses have pointed a highly distributed nature of the combinatorial gene regulation facilitating an efficient alteration in response to fungal attack. All together, our proposed methodology could be an important approach in understanding the combinatorial gene regulation. It can be further applied to unravel the tissue and/or condition specific combinatorial gene regulation in other eukaryotic systems with the availability of annotated genomic sequences and suitable

  14. A Rice Gene Homologous to Arabidopsis AGD2-LIKE DEFENSE1 Participates in Disease Resistance Response against Infection with Magnaporthe oryzae.

    PubMed

    Jung, Ga Young; Park, Ju Yeon; Choi, Hyo Ju; Yoo, Sung-Je; Park, Jung-Kwon; Jung, Ho Won

    2016-08-01

    ALD1 (ABERRANT GROWTH AND DEATH2 [AGD2]-LIKE DEFENSE1) is one of the key defense regulators in Arabidopsis thaliana and Nicotiana benthamiana. In these model plants, ALD1 is responsible for triggering basal defense response and systemic resistance against bacterial infection. As well ALD1 is involved in the production of pipecolic acid and an unidentified compound(s) for systemic resistance and priming syndrome, respectively. These previous studies proposed that ALD1 is a potential candidate for developing genetically modified (GM) plants that may be resistant to pathogen infection. Here we introduce a role of ALD1-LIKE gene of Oryza sativa, named as OsALD1, during plant immunity. OsALD1 mRNA was strongly transcribed in the infected leaves of rice plants by Magnaporthe oryzae, the rice blast fungus. OsALD1 proteins predominantly localized at the chloroplast in the plant cells. GM rice plants over-expressing OsALD1 were resistant to the fungal infection. The stable expression of OsALD1 also triggered strong mRNA expression of PATHOGENESIS-RELATED PROTEIN1 genes in the leaves of rice plants during infection. Taken together, we conclude that OsALD1 plays a role in disease resistance response of rice against the infection with rice blast fungus.

  15. Preparation of Chitosan nanoparticles and its effect on detached rice leaves infected with Pyricularia grisea.

    PubMed

    Manikandan, Appu; Sathiyabama, Muthukrishnan

    2016-03-01

    The aim of the present study was to prepare chitosan nanoparticles to evaluate their effect on protection of rice plants from blast fungus. Nanoparticles were prepared using the ionic gelation method by the interaction of Chitosan and sodium tripolyphosphate. The particle size, polydispersity index, zetapotential and structure was confirmed by DLS, FTIR, TEM and XRD. The Chitosan nanoparticle was evaluated for suppression of rice blast fungus (Pyricularia grisea) under the detached leaf condition. It is evident from our results that chitosan nanoparticle have potential in suppressing blast disease of rice which can be used further under field condition to protect rice plants from the devastating fungus.

  16. Sequential infection of Rice dwarf virus in the internal organs of its insect vector after ingestion of virus.

    PubMed

    Chen, Hongyan; Chen, Qian; Omura, Toshihiro; Uehara-Ichiki, Tamaki; Wei, Taiyun

    2011-09-01

    Confocal microscopy revealed that Rice dwarf virus (RDV) initially accumulated in epithelial cells of the filter chamber of leafhopper vector Nephotettix cincticeps 2 days after acquisition access feeding on diseased plants. Subsequently, RDV accumulation progressed to the anterior midgut, and then spread to the nervous system before infection of other organs. Furthermore, RDV accumulation progressed to the visceral muscles surrounding the anterior midgut. Later, RDV accumulation was detected in other parts of the alimentary canal, salivary glands and the follicular cells of the ovarioles in viruliferous insect vector. Our results suggest that RDV may use the muscle or neural tissues for viral dissemination from the infected vector's midgut into other tissues.

  17. Comparison of gene expression changes in susceptible, tolerant, and resistant hosts in response to infection with citrus tristeza virus and huanglongbing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The pathogens Candidatus Liberibacter asiaticus (Las) and Citrus tristeza virus (CTV) are both phloem limited and have significant economic impact on citrus production wherever they are found. Studies of host resistance have indicated that Poncirus trifoliata has tolerance or resistance to both path...

  18. Observations on the foliar nematode, Aphelenchoides besseyi, infecting tuberose and rice in India

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The foliar nematode Aphelenchoides besseyi causes white tip disease in rice (Oryza sativa L.) and floral malady in tuberose (Polianthes tuberosa L.). This nematode is widely distributed in the rice fields of many states of India, including West Bengal (WB), Andhra Pradesh (AP), Madhya Pradesh (MP) a...

  19. Morphological and morphometric variations in Aphelenchoides besseyi infecting tuberose and rice

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The foliar nematode Aphelenchoides besseyi causes the white tip disease of rice and a floral malady in tuberose in West Bengal (WB), India. The same nematode is widely distributed in many other states of India, including Andhra Pradesh (AP) and Madhya Pradesh (MP) rice fields. In order to generate i...

  20. Rice hull smoke extract inactivates Salmonella Typhimurium in laboratory media and protects infected mice against mortality

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A recently discovered and characterized rice hull liquid smoke extract was tested for bactericidal activity against Salmonella Typhimurium using the disc-agar method. The Minimum Inhibitory Concentration (MIC) value of rice hull smoke extract was found to be 0.822% (v/v). The in vivo antibacterial a...

  1. Dramatic Change in Citrus tristeza virus populations in the Dominican Republic

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) is the most destructive viral pathogen of citrus and has been an important concern for the citrus industry in the Dominican Republic. Earlier studies documented widespread distribution of mild isolates of the T30 genotype, which caused no disease in the infected trees, an...

  2. Bacterial brown leaf spot of citrus, a new disease caused by Burkholderia andropogonis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A new bacterial disease of citrus was recently identified in Florida and named as bacterial brown leaf spot (BBLS) of citrus. BBLS-infected citrus displayed flat, circular and brownish lesions with water-soaked margins surrounded by a chlorotic halo on leaves. Based on Biolog carbon source metabolic...

  3. Huanglongbing: An Overview of a Complex Pathosystem Ravaging the World's Citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus huanglongbing (HLB) has become a major disease and limiting factor of production in the citrus areas which have become infected. The disease was first reported in the Western Hemisphere in 2004 with devastating effects in major citrus producing areas such as Brazil and Florida. The destructi...

  4. Complete Genome sequence of citrus huanglongbing bacterium, ‘Candidatus Liberibacter asiaticus’ obtained through metagenomics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus huanglongbing (HLB) is the most destructive disease of citrus worldwide. It is spread by the citrus psyllids (Diaphorina citri and Trioza erytreae), and is associated with low-titer, phloem-limited infections by any of the three uncultured species of a-Proteobacteria: 'Candidatus Liberibact...

  5. Citrus leprosis virus C Infection Results in Hypersensitive-Like Response, Suppression of the JA/ET Plant Defense Pathway and Promotion of the Colonization of Its Mite Vector

    PubMed Central

    Arena, Gabriella D.; Ramos-González, Pedro L.; Nunes, Maria A.; Ribeiro-Alves, Marcelo; Camargo, Luis E. A.; Kitajima, Elliot W.; Machado, Marcos A.; Freitas-Astúa, Juliana

    2016-01-01

    Leprosis is a serious disease of citrus caused by Citrus leprosis virus C (CiLV-C, genus Cilevirus) whose transmission is mediated by false spider mites of the genus Brevipalpus. CiLV-C infection does not systemically spread in any of its known host plants, thus remaining restricted to local lesions around the feeding sites of viruliferous mites. To get insight into this unusual pathosystem, we evaluated the expression profiles of genes involved in defense mechanisms of Arabidopsis thaliana and Citrus sinensis upon infestation with non-viruliferous and viruliferous mites by using reverse-transcription qPCR. These results were analyzed together with the production of reactive oxygen species (ROS) and the appearance of dead cells as assessed by histochemical assays. After interaction with non-viruliferous mites, plants locally accumulated ROS and triggered the salicylic acid (SA) and jasmonate/ethylene (JA/ET) pathways. ERF branch of the JA/ET pathways was highly activated. In contrast, JA pathway genes were markedly suppressed upon the CiLV-C infection mediated by viruliferous mites. Viral infection also intensified the ROS burst and cell death, and enhanced the expression of genes involved in the RNA silencing mechanism and SA pathway. After 13 days of infestation of two sets of Arabidopsis plants with non-viruliferous and viruliferous mites, the number of mites in the CiLV-C infected Arabidopsis plants was significantly higher than in those infested with the non-viruliferous ones. Oviposition of the viruliferous mites occurred preferentially in the CiLV-C infected leaves. Based on these results, we postulated the first model of plant/Brevipalpus mite/cilevirus interaction in which cells surrounding the feeding sites of viruliferous mites typify the outcome of a hypersensitive-like response, whereas viral infection induces changes in the behavior of its vector. PMID:27933078

  6. Penicillium digitatum metabolites on synthetic media and citrus fruits.

    PubMed

    Ariza, Marta R; Larsen, Thomas O; Peterson, Bent O; Duus, Jens O; Barrero, Alejandro F

    2002-10-23

    Penicillium digitatum has been cultured on citrus fruits and yeast extract sucrose agar media (YES). Cultivation of fungal cultures on solid medium allowed the isolation of two novel tryptoquivaline-like metabolites, tryptoquialanine A (1) and tryptoquialanine B (2), also biosynthesized on citrus fruits. Their structural elucidation is described on the basis of their spectroscopic data, including those from 2D NMR experiments. The analysis of the biomass sterols led to the identification of 8-12. Fungal infection on the natural substrates induced the release of citrus monoterpenes together with fungal volatiles. The host-pathogen interaction in nature and the possible biological role of citrus volatiles are also discussed.

  7. Non-transgenic RNAi technology to control insects on citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This research demonstrated a non-transgenic delivery method for ribonucleic acid interference, RNAi, that reduced fitness as measured in increased mortality over time, of two insect pests of citrus, ie. psyllids and leafhoppers. The Asian citrus psyllid transmits a deadly plant-infecting bacterium o...

  8. Interactions of citrus variegated chlorosis (CVC) with endophytic bacteria

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus variegated chlorosis (CVC), is a disease of sweet orange [Citrus sinensis (L.)], is caused by Xylella fastidiosa subsp. pauca, a phytopathogenic bacterium that has been shown to infect all sweet orange cultivars. Xylella fastidiosa is a fastidious Gram negative, xylem-limited bacterium which ...

  9. Morphological and molecular characterization of Magnaporthe oryzae (fungus) from infected rice leaf samples

    NASA Astrophysics Data System (ADS)

    Muni, Nurulhidayah Mat; Nadarajah, Kalaivani

    2014-09-01

    Magnaporthe oryzae is a plant-pathogenic fungus that causes a serious disease affecting rice called rice blast. Outbreaks of rice blast have been a threat to the global production of rice. This fungal disease is estimated to cause production losses of US55 million each year in South and Southeast Asia. It has been used as a primary model for elucidating various aspects of the host-pathogen interaction with its host. We have isolated five isolates of Magnaporthe oryzae from diseased leaf samples obtained from the field at Kompleks Latihan MADA, Kedah, Malaysia. We have identified the isolates using morphological and microscopic studies on the fungal spores and the lesions on the diseased leaves. Amplification of the internal transcribed spacer (ITS) was carried out with universal primers ITS1 and ITS4. The sequence of each isolates showed at least 99% nucleotide identity with the corresponding sequence in GenBank for Magnaporthe oryzae.

  10. Clues into the metagenome of huanglongbing infected citrus by analysis of ancillary sequences from Ion Torrent whole genome Candidatus Liberibacter asiaticus sequencing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB) is a globally devastating disease of citrus. Presently, three etiological agents are associated with HLB and include; Candidatus Liberibacter asiaticus (CLas), Candidatus Liberibacter americanus; and Candidatus Liberibacter africanus. Attempts to determine alternate (non-Liberi...

  11. Rice Stripe Virus Infection Alters mRNA Levels of Sphingolipid-Metabolizing Enzymes and Sphingolipids Content in Laodelphax striatellus

    PubMed Central

    Li, Fei-Qiang; Bai, Yue-Liang; Shi, Xiao-Xiao; Zhu, Mu-Fei; Zhang, Min-Jing; Mao, Cun-Gui; Zhu, Zeng-Rong

    2017-01-01

    Sphingolipids and their metabolites have been implicated in viral infection and replication in mammal cells but how their metabolizing enzymes in the host are regulated by viruses remains largely unknown. Here we report the identification of 12 sphingolipid genes and their regulation by Rice stripe virus in the small brown planthopper (Laodelphax striatellus Fallén), a serious pest of rice throughout eastern Asia. According to protein sequence similarity, we identified 12 sphingolipid enzyme genes in L. striatellus. By comparing their mRNA levels in viruliferous versus nonviruliferous L. striatellus at different life stages by qPCR, we found that RSV infection upregulated six genes (LsCGT1, LsNAGA1, LsSGPP, LsSMPD4, LsSMS, and LsSPT) in most stages of L. striatellus. Especially, four genes (LsCGT1, LsSMPD2, LsNAGA1, and LsSMS) and another three genes (LsNAGA1, LsSGPP, and LsSMS) were significantly upregulated in viruliferous third-instar and fourth-instar nymphs, respectively. HPLC-MS/MS results showed that RSV infection increased the levels of various ceramides, such as Cer18:0, Cer20:0, and Cer22:0 species, in third and fourth instar L. striatellus nymphs. Together, these results demonstrate that RSV infection alters the transcript levels of various sphingolipid enzymes and the contents of sphingolipids in L. striatellus, indicating that sphingolipids may be important for RSV infection or replication in L. striatellus. PMID:28130458

  12. Bismerthiazol inhibits Xanthomonas citri subsp. citri growth and induces differential expression of citrus defense-related genes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker, caused by Xanthomonas citri ssp. citri (Xcc), is a serious disease and causes substantial economic losses to the citrus industry worldwide. The bactericide, bismerthiazol, has been widely used to control rice bacterial blight (Xanthomonas oryzae pv. oryzae). In this paper, we demonstr...

  13. Characteristic expression of twelve rice PR1 family genes in response to pathogen infection, wounding, and defense-related signal compounds (121/180)

    PubMed Central

    Iwai, Takayoshi; Seo, Shigemi; Yanagawa, Yuki; Kawahigasi, Hiroyuki; Hirose, Sakino; Ohkawa, Yasunobu; Ohashi, Yuko

    2008-01-01

    Pathogenesis-related (PR) proteins have been used as markers of plant defense responses, and are classified into 17 families. However, precise information on the majority members in specific PR families is still limited. We were interested in the individual characteristics of rice PR1 family genes, and selected 12 putatively active genes using rice genome databases for expressed genes. All were upregulated upon compatible and/or incompatible rice-blast fungus interactions; three were upregulated in the early infection period and four in the late infection period. Upon compatible rice–bacterial blight interaction, four genes were upregulated, six were not affected, and one was downregulated. These results are in striking contrast to those among 22 ArabidopsisPR1 genes where only one gene was pathogen-inducible. The responses of individual genes to salicylic acid, jasmonic acid, and ethylene induced defense signaling pathways in rice are likely to be different from those in dicot plants. Transcript levels in healthy leaves, roots, and flowers varied according to each gene. Analysis of the partially overlapping expression patterns of rice PR1 genes in healthy tissues and in response to pathogens and other stresses would be useful to understand their possible functions and for use as characteristic markers for defense-related studies in rice. PMID:18247056

  14. Phylogenomic analysis uncovers the evolutionary history of nutrition and infection mode in rice blast fungus and other Magnaporthales

    PubMed Central

    Luo, Jing; Qiu, Huan; Cai, Guohong; Wagner, Nicole E.; Bhattacharya, Debashish; Zhang, Ning

    2015-01-01

    The order Magnaporthales (Ascomycota, Fungi) includes devastating pathogens of cereals, such as the rice blast fungus Pyricularia (Magnaporthe) oryzae, which is a model in host-pathogen interaction studies. Magnaporthales also includes saprotrophic species associated with grass roots and submerged wood. Despite its scientific and economic importance, the phylogenetic position of Magnaporthales within Sordariomycetes and the interrelationships of its constituent taxa, remain controversial. In this study, we generated novel transcriptome data from 21 taxa that represent key Magnaporthales lineages of different infection and nutrition modes and phenotypes. Phylogenomic analysis of >200 conserved genes allowed the reconstruction of a robust Sordariomycetes tree of life that placed the monophyletic group of Magnaporthales sister to Ophiostomatales. Among Magnaporthales, three major clades were recognized: 1) an early diverging clade A comprised of saprotrophs associated with submerged woods; 2) clade B that includes the rice blast fungus and other pathogens that cause blast diseases of monocot plants. These species infect the above-ground tissues of host plants using the penetration structure, appressorium; and 3) clade C comprised primarily of root-associated species that penetrate the root tissue with hyphopodia. The well-supported phylogenies provide a robust framework for elucidating evolution of pathogenesis, nutrition modes, and phenotypic characters in Magnaporthales. PMID:25819715

  15. A host basal transcription factor is a key component for infection of rice by TALE-carrying bacteria

    PubMed Central

    Yuan, Meng; Ke, Yinggen; Huang, Renyan; Ma, Ling; Yang, Zeyu; Chu, Zhaohui; Xiao, Jinghua; Li, Xianghua; Wang, Shiping

    2016-01-01

    Transcription activator-like effectors (TALEs) are sequence-specific DNA binding proteins found in a range of plant pathogenic bacteria, where they play important roles in host-pathogen interactions. However, it has been unclear how TALEs, after they have been injected into the host cells, activate transcription of host genes required for infection success. Here, we show that the basal transcription factor IIA gamma subunit TFIIAγ5 from rice is a key component for infection by the TALE-carrying bacterium Xanthomonas oryzae pv. oryzae, the causal agent for bacterial blight. Direct interaction of several TALEs with TFIIAγ5 is required for activation of disease susceptibility genes. Conversely, reduced expression of the TFIIAγ5 host gene limits the induction of susceptibility genes and thus decreases bacterial blight symptoms. Suppression or mutation of TFIIAγ5 can also reduce bacterial streak, another devastating disease of rice caused by TALE-carrying X. oryzae pv. oryzicola. These results have important implications for formulating a widely applicable strategy with which to improve resistance of plants to TALE-carrying pathogens. DOI: http://dx.doi.org/10.7554/eLife.19605.001 PMID:27472897

  16. A host basal transcription factor is a key component for infection of rice by TALE-carrying bacteria.

    PubMed

    Yuan, Meng; Ke, Yinggen; Huang, Renyan; Ma, Ling; Yang, Zeyu; Chu, Zhaohui; Xiao, Jinghua; Li, Xianghua; Wang, Shiping

    2016-07-29

    Transcription activator-like effectors (TALEs) are sequence-specific DNA binding proteins found in a range of plant pathogenic bacteria, where they play important roles in host-pathogen interactions. However, it has been unclear how TALEs, after they have been injected into the host cells, activate transcription of host genes required for infection success. Here, we show that the basal transcription factor IIA gamma subunit TFIIAγ5 from rice is a key component for infection by the TALE-carrying bacterium Xanthomonas oryzae pv. oryzae, the causal agent for bacterial blight. Direct interaction of several TALEs with TFIIAγ5 is required for activation of disease susceptibility genes. Conversely, reduced expression of the TFIIAγ5 host gene limits the induction of susceptibility genes and thus decreases bacterial blight symptoms. Suppression or mutation of TFIIAγ5 can also reduce bacterial streak, another devastating disease of rice caused by TALE-carrying X. oryzae pv. oryzicola. These results have important implications for formulating a widely applicable strategy with which to improve resistance of plants to TALE-carrying pathogens.

  17. Phylogenomic analysis uncovers the evolutionary history of nutrition and infection mode in rice blast fungus and other Magnaporthales.

    PubMed

    Luo, Jing; Qiu, Huan; Cai, Guohong; Wagner, Nicole E; Bhattacharya, Debashish; Zhang, Ning

    2015-03-30

    The order Magnaporthales (Ascomycota, Fungi) includes devastating pathogens of cereals, such as the rice blast fungus Pyricularia (Magnaporthe) oryzae, which is a model in host-pathogen interaction studies. Magnaporthales also includes saprotrophic species associated with grass roots and submerged wood. Despite its scientific and economic importance, the phylogenetic position of Magnaporthales within Sordariomycetes and the interrelationships of its constituent taxa, remain controversial. In this study, we generated novel transcriptome data from 21 taxa that represent key Magnaporthales lineages of different infection and nutrition modes and phenotypes. Phylogenomic analysis of >200 conserved genes allowed the reconstruction of a robust Sordariomycetes tree of life that placed the monophyletic group of Magnaporthales sister to Ophiostomatales. Among Magnaporthales, three major clades were recognized: 1) an early diverging clade A comprised of saprotrophs associated with submerged woods; 2) clade B that includes the rice blast fungus and other pathogens that cause blast diseases of monocot plants. These species infect the above-ground tissues of host plants using the penetration structure, appressorium; and 3) clade C comprised primarily of root-associated species that penetrate the root tissue with hyphopodia. The well-supported phylogenies provide a robust framework for elucidating evolution of pathogenesis, nutrition modes, and phenotypic characters in Magnaporthales.

  18. Comparative Transcriptome and iTRAQ Proteome Analyses of Citrus Root Responses to Candidatus Liberibacter asiaticus Infection

    PubMed Central

    Jiang, Nong-hui; Jiang, Bo; Zhang, Yong-yan; Wu, Bo; Hu, Min-lun; Zeng, Ji-wu; Yan, Hua-xue; Yi, Gan-jun; Zhong, Guang-yan

    2015-01-01

    Root samples of ‘Sanhu’ red tangerine trees infected with and without Candidatus Liberibacter asiaticus (CLas) were collected at 50 days post inoculation and subjected to RNA-sequencing and isobaric tags for relative and absolute quantification (iTRAQ) to profile the differentially expressed genes (DEGs) and proteins (DEPs), respectively. Quantitative real-time PCR was subsequently used to confirm the expression of 16 selected DEGs. Results showed that a total of 3956 genes and 78 proteins were differentially regulated by HLB-infection. Among the most highly up-regulated DEPs were sperm specific protein 411, copper ion binding protein, germin-like proteins, subtilisin-like proteins and serine carboxypeptidase-like 40 proteins whose transcript levels were concomitantly up-regulated as shown by RNA-seq data. Comparison between our results and those of the previously reported showed that known HLB-modulated biological pathways including cell-wall modification, protease-involved protein degradation, carbohydrate metabolism, hormone synthesis and signaling, transcription activities, and stress responses were similarly regulated by HLB infection but different or root-specific changes did exist. The root unique changes included the down-regulation in genes of ubiquitin-dependent protein degradation pathway, secondary metabolism, cytochrome P450s, UDP-glucosyl transferases and pentatricopeptide repeat containing proteins. Notably, nutrient absorption was impaired by HLB-infection as the expression of the genes involved in Fe, Zn, N and P adsorption and transportation were significantly changed. HLB-infection induced some cellular defense responses but simultaneously reduced the biosynthesis of the three major classes of secondary metabolites, many of which are known to have anti-pathogen activities. Genes involved in callose deposition were up-regulated whereas those involved in callose degradation were also up-regulated, indicating that the sieve tube elements in

  19. Comparative Transcriptome and iTRAQ Proteome Analyses of Citrus Root Responses to Candidatus Liberibacter asiaticus Infection.

    PubMed

    Zhong, Yun; Cheng, Chun-Zhen; Jiang, Nong-Hui; Jiang, Bo; Zhang, Yong-Yan; Wu, Bo; Hu, Min-Lun; Zeng, Ji-Wu; Yan, Hua-Xue; Yi, Gan-Jun; Zhong, Guang-Yan

    2015-01-01

    Root samples of 'Sanhu' red tangerine trees infected with and without Candidatus Liberibacter asiaticus (CLas) were collected at 50 days post inoculation and subjected to RNA-sequencing and isobaric tags for relative and absolute quantification (iTRAQ) to profile the differentially expressed genes (DEGs) and proteins (DEPs), respectively. Quantitative real-time PCR was subsequently used to confirm the expression of 16 selected DEGs. Results showed that a total of 3956 genes and 78 proteins were differentially regulated by HLB-infection. Among the most highly up-regulated DEPs were sperm specific protein 411, copper ion binding protein, germin-like proteins, subtilisin-like proteins and serine carboxypeptidase-like 40 proteins whose transcript levels were concomitantly up-regulated as shown by RNA-seq data. Comparison between our results and those of the previously reported showed that known HLB-modulated biological pathways including cell-wall modification, protease-involved protein degradation, carbohydrate metabolism, hormone synthesis and signaling, transcription activities, and stress responses were similarly regulated by HLB infection but different or root-specific changes did exist. The root unique changes included the down-regulation in genes of ubiquitin-dependent protein degradation pathway, secondary metabolism, cytochrome P450s, UDP-glucosyl transferases and pentatricopeptide repeat containing proteins. Notably, nutrient absorption was impaired by HLB-infection as the expression of the genes involved in Fe, Zn, N and P adsorption and transportation were significantly changed. HLB-infection induced some cellular defense responses but simultaneously reduced the biosynthesis of the three major classes of secondary metabolites, many of which are known to have anti-pathogen activities. Genes involved in callose deposition were up-regulated whereas those involved in callose degradation were also up-regulated, indicating that the sieve tube elements in roots

  20. Use of digital PCR to improve early detection of CLas infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing is a devastating disease of citrus caused by the bacterium Candidatus Liberibacter asiaticus. Huanglongbing has devastated the Florida citrus industry and is threatening citrus in Texas and California. Detection of Candidatus Liberibacter asiaticus infections as early as possible is ...

  1. Bacterial Diversity Analysis of Huanglongbing Pathogen-Infected Citrus, Using PhyloChip Arrays and 16S rRNA Gene Clone Library Sequencing▿ †

    PubMed Central

    Sagaram, Uma Shankar; DeAngelis, Kristen M.; Trivedi, Pankaj; Andersen, Gary L.; Lu, Shi-En; Wang, Nian

    2009-01-01

    The bacterial diversity associated with citrus leaf midribs was characterized for citrus groves that contained the Huanglongbing (HLB) pathogen, which has yet to be cultivated in vitro. We employed a combination of high-density phylogenetic 16S rRNA gene microarrays and 16S rRNA gene clone library sequencing to determine the microbial community composition for symptomatic and asymptomatic citrus midribs. Our results revealed that citrus leaf midribs can support a diversity of microbes. PhyloChip analysis indicated that 47 orders of bacteria in 15 phyla were present in the citrus leaf midribs, while 20 orders in 8 phyla were observed with the cloning and sequencing method. PhyloChip arrays indicated that nine taxa were significantly more abundant in symptomatic midribs than in asymptomatic midribs. “Candidatus Liberibacter asiaticus” was detected at a very low level in asymptomatic plants but was over 200 times more abundant in symptomatic plants. The PhyloChip analysis results were further verified by sequencing 16S rRNA gene clone libraries, which indicated the dominance of “Candidatus Liberibacter asiaticus” in symptomatic leaves. These data implicate “Candidatus Liberibacter asiaticus” as the pathogen responsible for HLB disease. PMID:19151177

  2. Citrus Waste Biomass Program

    SciTech Connect

    Karel Grohman; Scott Stevenson

    2007-01-30

    Renewable Spirits is developing an innovative pilot plant bio-refinery to establish the commercial viability of ehtanol production utilizing a processing waste from citrus juice production. A novel process based on enzymatic hydrolysis of citrus processing waste and fermentation of resulting sugars to ethanol by yeasts was successfully developed in collaboration with a CRADA partner, USDA/ARS Citrus and Subtropical Products Laboratory. The process was also successfully scaled up from laboratory scale to 10,000 gal fermentor level.

  3. Gene expression in Citrus sinensis fruit tissues harvested from huanglongbing-infected trees: comparison with girdled fruit.

    PubMed

    Liao, Hui-Ling; Burns, Jacqueline K

    2012-05-01

    Distribution of viable Candidatus Liberibacter asiaticus (CaLas) in sweet orange fruit and leaves ('Hamlin' and 'Valencia') and transcriptomic changes associated with huanglongbing (HLB) infection in fruit tissues are reported. Viable CaLas was present in most fruit tissues tested in HLB trees, with the highest titre detected in vascular tissue near the calyx abscission zone. Transcriptomic changes associated with HLB infection were analysed in flavedo (FF), vascular tissue (VT), and juice vesicles (JV) from symptomatic (SY), asymptomatic (AS), and healthy (H) fruit. In SY 'Hamlin', HLB altered the expression of more genes in FF and VT than in JV, whereas in SY 'Valencia', the number of genes whose expression was changed by HLB was similar in these tissues. The expression of more genes was altered in SY 'Valencia' JV than in SY 'Hamlin' JV. More genes were also affected in AS 'Valencia' FF and VT than in AS 'Valencia' JV. Most genes whose expression was changed by HLB were classified as transporters or involved in carbohydrate metabolism. Physiological characteristics of HLB-infected and girdled fruit were compared to differentiate between HLB-specific and carbohydrate metabolism-related symptoms. SY and girdled fruit were smaller than H and ungirdled fruit, respectively, with poor juice quality. However, girdling did not cause misshapen fruit or differential peel coloration. Quantitative PCR analysis indicated that many selected genes changed their expression significantly in SY flavedo but not in girdled flavedo. Mechanisms regulating development of HLB symptoms may lie in the host disease response rather than being a direct consequence of carbohydrate starvation.

  4. Gene expression in Citrus sinensis fruit tissues harvested from huanglongbing-infected trees: comparison with girdled fruit

    PubMed Central

    Liao, Hui-Ling; Burns, Jacqueline K.

    2012-01-01

    Distribution of viable Candidatus Liberibacter asiaticus (CaLas) in sweet orange fruit and leaves (‘Hamlin’ and ‘Valencia’) and transcriptomic changes associated with huanglongbing (HLB) infection in fruit tissues are reported. Viable CaLas was present in most fruit tissues tested in HLB trees, with the highest titre detected in vascular tissue near the calyx abscission zone. Transcriptomic changes associated with HLB infection were analysed in flavedo (FF), vascular tissue (VT), and juice vesicles (JV) from symptomatic (SY), asymptomatic (AS), and healthy (H) fruit. In SY ‘Hamlin’, HLB altered the expression of more genes in FF and VT than in JV, whereas in SY ‘Valencia’, the number of genes whose expression was changed by HLB was similar in these tissues. The expression of more genes was altered in SY ‘Valencia’ JV than in SY ‘Hamlin’ JV. More genes were also affected in AS ‘Valencia’ FF and VT than in AS ‘Valencia’ JV. Most genes whose expression was changed by HLB were classified as transporters or involved in carbohydrate metabolism. Physiological characteristics of HLB-infected and girdled fruit were compared to differentiate between HLB-specific and carbohydrate metabolism-related symptoms. SY and girdled fruit were smaller than H and ungirdled fruit, respectively, with poor juice quality. However, girdling did not cause misshapen fruit or differential peel coloration. Quantitative PCR analysis indicated that many selected genes changed their expression significantly in SY flavedo but not in girdled flavedo. Mechanisms regulating development of HLB symptoms may lie in the host disease response rather than being a direct consequence of carbohydrate starvation. PMID:22407645

  5. Visual rating and the use of image analysis for assessing different symptoms of citrus canker on grapefruit leaves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker is caused by the bacterial pathogen Xanthomonas axonopodis pv citri (Xac) and infects several citrus species in wet tropical and subtropical citrus growing regions. Accurate, precise and reproducible disease assessment is needed for monitoring epidemics and disease response in breeding...

  6. Comparative proteomics reveals differential induction of both biotic and abiotic stress response associated proteins in rice during Xanthomonas oryzae pv. oryzae infection.

    PubMed

    Kumar, Anirudh; Bimolata, Waikhom; Kannan, Monica; Kirti, P B; Qureshi, Insaf Ahmed; Ghazi, Irfan Ahmad

    2015-07-01

    Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight disease in rice and brutally affects the yield up to 50 % of total production. Here, we report a comparative proteomics analysis of total foliar protein isolated from infected rice leaves of susceptible Pusa Basmati 1 (PB1) and resistant Oryza longistaminata genotypes. Two-dimensional gel electrophoresis (2-DE) coupled with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) approaches identified 29 protein spots encoding unique proteins from both the genotypes. Identified proteins belonged to a large number of biological and molecular functions related to biotic and abiotic stress proteins which are potentially involved during Xoo infection. Biotic and abiotic stress-related proteins were induced during Xoo infection, indicating the activation of common stress pathway during bacterial blight infection. Candidate genes conferring tolerance against bacterial blight, which include germin-like protein, putative r40c1, cyclin-dependent kinase C, Ent-isokaur-15-ene synthase and glutathione-dependent dehydroascorbate reductase 1 (GSH-DHAR1), were also induced, with germin-like proteins induced only in the resistant rice genotype O. longistaminata. Energy, metabolism and hypothetical proteins were common among both the genotypes. Further, host defence/stress-related proteins were mostly expressed in resistant genotype O. longistaminata, indicating possible co-evolution of the pathogen and the wild rice, O. longistaminata.

  7. Two Novel Transcriptional Regulators Are Essential for Infection-related Morphogenesis and Pathogenicity of the Rice Blast Fungus Magnaporthe oryzae

    PubMed Central

    Yan, Xia; Li, Ya; Yue, Xiaofeng; Wang, Congcong; Que, Yawei; Kong, Dandan; Ma, Zhonghua; Talbot, Nicholas J.; Wang, Zhengyi

    2011-01-01

    The cyclic AMP-dependent protein kinase A signaling pathway plays a major role in regulating plant infection by the rice blast fungus Magnaporthe oryzae. Here, we report the identification of two novel genes, MoSOM1 and MoCDTF1, which were discovered in an insertional mutagenesis screen for non-pathogenic mutants of M. oryzae. MoSOM1 or MoCDTF1 are both necessary for development of spores and appressoria by M. oryzae and play roles in cell wall differentiation, regulating melanin pigmentation and cell surface hydrophobicity during spore formation. MoSom1 strongly interacts with MoStu1 (Mstu1), an APSES transcription factor protein, and with MoCdtf1, while also interacting more weakly with the catalytic subunit of protein kinase A (CpkA) in yeast two hybrid assays. Furthermore, the expression levels of MoSOM1 and MoCDTF1 were significantly reduced in both Δmac1 and ΔcpkA mutants, consistent with regulation by the cAMP/PKA signaling pathway. MoSom1-GFP and MoCdtf1-GFP fusion proteins localized to the nucleus of fungal cells. Site-directed mutagenesis confirmed that nuclear localization signal sequences in MoSom1 and MoCdtf1 are essential for their sub-cellular localization and biological functions. Transcriptional profiling revealed major changes in gene expression associated with loss of MoSOM1 during infection-related development. We conclude that MoSom1 and MoCdtf1 functions downstream of the cAMP/PKA signaling pathway and are novel transcriptional regulators associated with cellular differentiation during plant infection by the rice blast fungus. PMID:22144889

  8. Digital PCR for detection of citrus pathogens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus trees are often infected with multiple pathogens of economic importance, especially those with insect or mite vectors. Real-time/quantitative PCR (qPCR) has been used for high-throughput detection and relative quantification of pathogens; however, target reference or standards are required. I...

  9. Differential proteomics profiling of the ova between healthy and Rice stripe virus-infected female insects of Laodelphax striatellus

    PubMed Central

    Liu, Beibei; Qin, Faliang; Liu, Wenwen; Wang, Xifeng

    2016-01-01

    Rice stripe virus-infected females of the small brown planthopper (SBPH, Laodelphax striatellus) usually lay fewer eggs with a longer hatch period, low hatchability, malformation and retarded or defective development compared with healthy females. To explore the molecular mechanism of those phenomena, we analyzed the differential proteomics profiling of the ova between viruliferous and healthy female insects using an isobaric tag for relative and absolute quantitation (iTRAQ) approach. We obtained 147 differentially accumulated proteins: 98 (66.7%) proteins increased, but 49 (33.3%) decreased in the ova of the viruliferous females. RT-qPCR was used to verify the 12 differential expressed proteins from iTRAQ, finding that trends in the transcriptional change for the 12 genes were consistent with those at the proteomic level. Differentially expressed proteins that were associated with meiosis (serine/threonine-protein phosphatase 2B and cyclin B3) and mitosis (cyclin B3 and dynein heavy chain) in viruliferous ova may contribute to low hatchability and defective or retarded development. Alterations in the abundance of proteins involved in the respiratory chain and nutrition metabolism may affect embryonic development. Our study begins to explain macroscopical developmental phenomena and explore the mechanisms by which Rice stripe virus impacts the development of SBPH. PMID:27277140

  10. The MET13 methylenetetrahydrofolate reductase gene is essential for infection-related morphogenesis in the rice blast fungus Magnaporthe oryzae.

    PubMed

    Yan, Xia; Que, Yawei; Wang, Hong; Wang, Congcong; Li, Ya; Yue, Xiaofeng; Ma, Zhonghua; Talbot, Nicholas J; Wang, Zhengyi

    2013-01-01

    Methylenetetrahydrofolate reductases (MTHFRs) play a key role in the biosynthesis of methionine in both prokaryotic and eukaryotic organisms. In this study, we report the identification of a novel T-DNA-tagged mutant WH672 in the rice blast fungus Magnaporthe oryzae, which was defective in vegetative growth, conidiation and pathogenicity. Analysis of the mutation confirmed a single T-DNA insertion upstream of MET13, which encodes a 626-amino-acid protein encoding a MTHFR. Targeted gene deletion of MET13 resulted in mutants that were non-pathogenic and significantly impaired in aerial growth and melanin pigmentation. All phenotypes associated with Δmet13 mutants could be overcome by addition of exogenous methionine. The M. oryzae genome contains a second predicted MTHFR-encoding gene, MET12. The deduced amino acid sequences of Met13 and Met12 share 32% identity. Interestingly, Δmet12 mutants produced significantly less conidia compared with the isogenic wild-type strain and grew very poorly in the absence of methionine, but were fully pathogenic. Deletion of both genes resulted in Δmet13Δmet12 mutants that showed similar phenotypes to single Δmet13 mutants. Taken together, we conclude that the MTHFR gene, MET13, is essential for infection-related morphogenesis by the rice blast fungus M. oryzae.

  11. The MET13 Methylenetetrahydrofolate Reductase Gene Is Essential for Infection-Related Morphogenesis in the Rice Blast Fungus Magnaporthe oryzae

    PubMed Central

    Wang, Hong; Wang, Congcong; Li, Ya; Yue, Xiaofeng; Ma, Zhonghua; Talbot, Nicholas J.; Wang, Zhengyi

    2013-01-01

    Methylenetetrahydrofolate reductases (MTHFRs) play a key role in the biosynthesis of methionine in both prokaryotic and eukaryotic organisms. In this study, we report the identification of a novel T-DNA-tagged mutant WH672 in the rice blast fungus Magnaporthe oryzae, which was defective in vegetative growth, conidiation and pathogenicity. Analysis of the mutation confirmed a single T-DNA insertion upstream of MET13, which encodes a 626-amino-acid protein encoding a MTHFR. Targeted gene deletion of MET13 resulted in mutants that were non-pathogenic and significantly impaired in aerial growth and melanin pigmentation. All phenotypes associated with Δmet13 mutants could be overcome by addition of exogenous methionine. The M. oryzae genome contains a second predicted MTHFR-encoding gene, MET12. The deduced amino acid sequences of Met13 and Met12 share 32% identity. Interestingly, Δmet12 mutants produced significantly less conidia compared with the isogenic wild-type strain and grew very poorly in the absence of methionine, but were fully pathogenic. Deletion of both genes resulted in Δmet13Δmet12 mutants that showed similar phenotypes to single Δmet13 mutants. Taken together, we conclude that the MTHFR gene, MET13, is essential for infection-related morphogenesis by the rice blast fungus M. oryzae. PMID:24116181

  12. Management of the Citrus Nematode, Tylenchulus semipenetrans

    PubMed Central

    Verdejo-Lucas, S.; McKenry, M. V.

    2004-01-01

    Of the many nematode species that parasitize citrus, Tylenchulus semipenetrans is the most important on a worldwide basis. Management of the citrus nematode remains problematic as no one tactic gives adequate control of the nematode. An overall management strategy must include such components as site selection, use of non-infected nursery stock, use of at lease one post-plant nematode control tactic, and careful management of other elements of the environment that may stress the trees. Nematicides continue to play a key role in management of this pest. Optimum results require careful attention to application techniques. PMID:19262822

  13. Wind speed effects on the quantity of Xanthomonas citri subsp. citri dispersed downwind from canopies of grapefruit trees infected with citrus canker

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The epidemic of citrus canker (Xanthomonas citri subsp. citri, Xcc) in Florida continues to expand since termination of the eradication program in 2006. Storms are known to be associated with disease spread, but little information exists on the interaction of fundamental physical and biological proc...

  14. Study on citrus response to huanglongbing highlights a down-regulation of defense-related proteins in lemon plants upon 'Ca Liberibacter asiaticus' infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB) is a highly destructive citrus disease; the bacterium, ‘Candidatus Liberibacter asiaticus’ (Las) is a presumed causal agent. In this study, comparative 2-DE and mass spectrometry, in addition to Inductively Coupled Plasma (ICP) spectroscopy analyses, were applied to elucidate dif...

  15. Certification Programs for Citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus certification programs designed to ensure that healthy plants of the highest genetic potential are being planted in the field are the basic building block of an integrated pest management program. Certification programs began for citrus began with the discovery that the diseases were graft t...

  16. Cercosporoid diseases of Citrus.

    PubMed

    Huang, Feng; Groenewald, J Z; Zhu, Li; Crous, P W; Li, Hongye

    2015-01-01

    Citrus leaves and fruits exhibiting disease symptoms ranging from greasy spot, yellow spot, small or large brown spot, black dot, and brown dot were sampled from Fujian, Guangdong, Guizhou, Hunan, Jiangxi, Yunnan, Zhejiang provinces and the Guangxi Zhuang Autonomous Region in China. In total 82 isolates representing various cercosporoid genera were isolated from these disease symptoms, which were supplemented with eight Citrus cercosporoid isolates collected from other countries. Based on a morphological and phylogenetic study using sequence data from the nuclear ribosomal DNA's ITS1-5.8S-ITS2 regions (ITS), and partial actin (act), β-tubulin (tub2), 28S nuclear ribosomal RNA (28S rDNA) and translation elongation factor 1-α (tef1) genes, these strains were placed in the following genera: Cercospora, Pallidocercospora, Passalora, Pseudocercospora, Verrucisporota and Zasmidium. All isolates tended to be sterile, except the Zasmidium isolates associated with citrus greasy spot-like symptoms, which subsequently were compared with phylogenetically similar isolates occurring on Citrus and other hosts elsewhere. From these results four Zasmidium species were recognized on Citrus, namely Z. indonesianum on Citrus in Indonesia, Z. fructicola and Z. fructigenum on Citrus in China and Z. citri-griseum, which appears to have a wide host range including Acacia, Citrus, Eucalyptus and Musa, as well as a global distribution.

  17. An Inexpensive and Safe Experiment to Demonstrate Koch's Postulates Using Citrus Fruit

    ERIC Educational Resources Information Center

    Jakobi, Steven

    2010-01-01

    Citrus fruit (oranges, tangerines, grapefruit or lemons) purchased in a grocery store can be experimentally infected with readily-available sources of "Penicillium digitatum" to demonstrate the four basic steps of Koch's postulates, also known as proof of pathogenicity. The mould is isolated from naturally-infected citrus fruit into pure culture…

  18. Viability of ‘Candidatus Liberibacter asiaticus’ prolonged by addition of citrus juice to culture medium

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing, or citrus greening disease, is associated with infection by the phloem-limited bacterium ‘Candidatus Liberibacter asiaticus’ (LAS). Infection with LAS, vectored by the Asian citrus psyllid (Diaphorina citri), is incurable; therefore, knowledge regarding LAS biology and pathogenesis is...

  19. Abundance of citrus leafminer larvae on citrus and citrus-related germplasm

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The citrus leafminer, Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae), is a key pest in most citrus growing regions worldwide. Adult citrus leafminers oviposit primarily on young elongating flush of citrus as well as other Rutaceae and some ornamental plants. Larvae feed on the epiderm...

  20. Quantitative distribution of 'Candidatus Liberibacter asiaticus' in citrus plants with citrus huanglongbing.

    PubMed

    Li, Wenbin; Levy, Laurene; Hartung, John S

    2009-02-01

    Citrus huanglongbing (HLB), or greening disease, is strongly associated with any of three nonculturable gram-negative bacteria belonging to 'Candidatus Liberibacter spp.' 'Ca. Liberibacter spp.' are transmitted by citrus psyllids to all commercial cultivars of citrus. The diseases can be lethal to citrus and have recently become widespread in both São Paulo, Brazil, and Florida, United States, the locations of the largest citrus industries in the world. Asiatic HLB, the form of the disease found in Florida, is associated with 'Ca. Liberibacter asiaticus' and is the subject of this report. The nonculturable nature of the pathogen has hampered research and little is known about the distribution of 'Ca. L. asiaticus' in infected trees. In this study, we have used a quantitative polymerase chain reaction assay to systematically quantify the distribution of 'Ca. L. asiaticus' genomes in tissues of six species of citrus either identified in the field during survey efforts in Florida or propagated in a greenhouse in Beltsville, MD. The populations of 'Ca. L. asiaticus' inferred from the distribution of 16S rDNA sequences specific for 'Ca. L. asiaticus' in leaf midribs, leaf blades, and bark samples varied by a factor of 1,000 among samples prepared from the six citrus species tested and by a factor of 100 between two sweet orange trees tested. In naturally infected trees, above-ground portions of the tree averaged 10(10) 'Ca. L. asiaticus' genomes per gram of tissue. Similar levels of 'Ca. L. asiaticus' genomes were observed in some but not all root samples from the same plants. In samples taken from greenhouse-inoculated trees, levels of 'Ca. L. asiaticus' genomes varied systematically from 10(4) genomes/g at the graft inoculation site to 10(10) genomes/g in some leaf petioles. Root samples from these trees also contained 'Ca. L. asiaticus' at 10(7) genomes/g. In symptomatic fruit tissues, 'Ca. L. asiaticus' genomes were also readily detected and quantified. The highest

  1. Correlation of electronic monitoring and stylet pathways further elucidate the role of sclerenchymatous ring as a barrier to phloem feeding on citrus leaves by Asian citrus psyllid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Asian citrus psyllid (ACP) feeding behaviors play a significant role in the transmission of the phloem-limited Candidatus Liberibacter asiaticus (CLas) bacteria that cause citrus greening disease. Sustained phloem ingestion by ACP on CLas infected plants is very important in pathogen acquisition and...

  2. Common Genetic Pathways Regulate Organ-Specific Infection-Related Development in the Rice Blast Fungus[W

    PubMed Central

    Tucker, Sara L.; Besi, Maria I.; Galhano, Rita; Franceschetti, Marina; Goetz, Stephan; Lenhert, Steven; Osbourn, Anne; Sesma, Ane

    2010-01-01

    Magnaporthe oryzae is the most important fungal pathogen of rice (Oryza sativa). Under laboratory conditions, it is able to colonize both aerial and underground plant organs using different mechanisms. Here, we characterize an infection-related development in M. oryzae produced on hydrophilic polystyrene (PHIL-PS) and on roots. We show that fungal spores develop preinvasive hyphae (pre-IH) from hyphopodia (root penetration structures) or germ tubes and that pre-IH also enter root cells. Changes in fungal cell wall structure accompanying pre-IH are seen on both artificial and root surfaces. Using characterized mutants, we show that the PMK1 (for pathogenicity mitogen-activated protein kinase 1) pathway is required for pre-IH development. Twenty mutants with altered pre-IH differentiation on PHIL-PS identified from an insertional library of 2885 M. oryzae T-DNA transformants were found to be defective in pathogenicity. The phenotypic analysis of these mutants revealed that appressorium, hyphopodium, and pre-IH formation are genetically linked fungal developmental processes. We further characterized one of these mutants, M1373, which lacked the M. oryzae ortholog of exportin-5/Msn5p (EXP5). Mutants lacking EXP5 were much less virulent on roots, suggesting an important involvement of proteins and/or RNAs transported by EXP5 during M. oryzae root infection. PMID:20348434

  3. Transmission Rates of ‘Ca. Liberibacter asiaticus’ by Asian Citrus Psyllid Are Enhanced by the Presence and Developmental Stage of Citrus Flush.

    PubMed

    Hall, David G; Albrecht, Ute; Bowman, Kim D

    2016-04-01

    Asian citrus psyllid (Diaphorina citri Kuwayama) transmits a bacterium ‘Candidatus Liberibacter asiaticus’ (CLas) putatively responsible for a devastating citrus disease known as Asiatic huanglongbing (HLB) (citrus greening disease). The psyllid and disease have invaded many citrus-growing regions including the United States, where the disease is seriously jeopardizing the Florida citrus industry. We recently concluded research that showed CLas transmission rates are increased when citrus flush is present. Flush is any new leaf growth ranging in development from first emergence up until the leaves are fully expanded yet still tender. In an experiment with seedlings of a rootstock cultivar ‘US-942’, a 1-wk infestation of 20 Asian citrus psyllids from an infected colony resulted in 53–60% of seedlings becoming infected when flush was present compared with only 7% when no flush was present. In a second experiment with ‘US-942’, 77–97% of seedlings became infected when flush was present compared with 40% when no flush was present. A similar experiment with ‘Valencia’ sweet orange resulted in 23, 80, and 3% seedlings becoming infected when young, older, or no flush was present, respectively. Young plants are therefore more likely to contract HLB if flush is present, with older flush promoting higher infection rates under the conditions of this study. Based on this finding, healthy citrus should be protected from Asian citrus psyllid infestations throughout a flush. To evaluate germplasm for CLas resistance, inoculations using infected Asian citrus psyllid would best be achieved if flush is present.

  4. Rearing migratory endoparasitic nematodes in citrus callus and roots produced from citrus leaves.

    PubMed

    Inserra, R N; O'Bannon, J H

    1975-07-01

    Radopholus similis and Pratylenchus coffeae were reared on callus and roots developed from citrus leaves. Callus formed best when leaf petioles were immersed in Astatula fine sand and the leaves were sprayed daily with 4 ppm 2,4-D solution and maintained at 25 or 30 C. The nematodes completed one generation in 20 days at 25 C. Highest populations of R. similis (1,127) occurred after 50 days, and the highest for P. coffeae (619) after 70 days. Leaf-callus cultures from R. similis-resistant citrus rootstocks showed the same degree of infection as susceptible rough lemon callus after 30 days.

  5. Citrus huanglongbing: a newly relevant disease presents unprecedented challenges.

    PubMed

    Wang, Nian; Trivedi, Pankaj

    2013-07-01

    Citrus huanglongbing (HLB) is one of the oldest citrus diseases and has been known for over a century. HLB is caused by 'Candidatus Liberibacter' spp. that are phloem-limited, fastidious α-proteobacteria and infect hosts in different Kingdoms (i.e., Animalia and Plantae). When compared with well-characterized, cultivatable plant-pathogenic Gram-negative bacteria, the interactions of uncultured insect-vectored plant-pathogenic bacteria, including 'Ca. Liberibacter' spp., with their hosts remain poorly understood. 'Ca. Liberibacter' spp. have been known to cause HLB, which has been rapidly spreading worldwide, resulting in dramatic economic losses. HLB presents an unprecedented challenge to citrus production. In this review, we focus on the most recent research on citrus, 'Candidatus Liberibacter asiaticus', and psyllid interactions, specifically considering the following topics: evolutionary relationships among 'Ca. Liberibacter' spp., genetic diversity, host range, genome analysis, transmission, virulence mechanisms, and the ecological importance of HLB. Currently, no efficient management strategy is available to control HLB, although some promising progress has been made. Further studies are needed to understand citrus, 'Ca. L. asiaticus', and psyllid interactions to design innovative management strategies. Although HLB has been problematic for over a century, we can only win the battle against HLB with a coordinated and deliberate effort by the citrus industry, citrus growers, researchers, legislatures, and governments.

  6. Understanding superinfection exclusion by complex populations of Citrus tristeza virus.

    PubMed

    Bergua, María; Kang, Sung-Hwan; Folimonova, Svetlana Y

    2016-12-01

    Superinfection exclusion (SIE) is a phenomenon in which a primary viral infection restricts a secondary infection with the same or closely related virus. Previously we showed that SIE by Citrus tristeza virus (CTV) occurs only between isolates of the same virus genotype. This work, however, was done using single genotype-containing isolates, while most field citrus trees harbor complex populations composed of different virus genotypes. Here we examined SIE in plants simultaneously infected with several CTV genotypes. The experiments showed that exclusion of a secondary infection by a CTV variant was triggered by the presence of another variant of the same genotype in the primary population, even under the conditions of its low-level accumulation, and was not affected by co-occurrence of additional heterologous genotypes. The same rule appeared to be in effect when SIE by mixed populations was tested in a series of different citrus varieties.

  7. Visualization of ‘Candidatus Liberibacter asiaticus’ cells in citrus seed coats with fluorescence in situ hybridization and transmission electron microscopy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    ‘Candidatus Liberibacter asiaticus’ is bacterium implicated as the causal agent of the economically damaging disease of citrus called huanglongbing (HLB). The bacterium is spread by movement of infected citrus propagation material and by the Asian citrus psyllid, Diaphorina citri Kuwayama. Seed tr...

  8. High incidence of preharvest colonization of huanglongbing-symptomatic Citrus sinensis fruit by Lasiodiplodia theobromae (Diplodia natalensis) and exacerbation of postharvest fruit decay by that fungus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB), presumably caused by bacterium Candidatus Liberibacter asiaticus (CLas), is a devastating citrus disease associated with excessive pre-harvest fruit drop. Lasiodiplodia theobromae (Diplodia) is the causal organism of citrus stem end rot (SER). The pathogen infects citrus fruit ...

  9. Citrus diseases with global ramifications including citrus canker and huanglongbing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Although there are a number of diseases that plague citrus production worldwide, two bacterial diseases are particularly problematic. Both are of Asian origin and currently cause severe economic damage: Asiatic citrus canker (ACC) and citrus huanglongbing (HLB). Although ACC has been found in the ...

  10. Developing Transgenic Citrus for Resistance to Huanglongbing and Citrus Canker

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB) and Citrus Bacterial Canker (CBC) are serious threats to citrus production, and resistant transgenic citrus is desirable. Genes for antimicrobial peptides (AMPs) with diverse promoters have been used to generate thousands of rootstock and scion transformants. D35S::D4E1 transfor...

  11. Effect of X-irradiation on Citrus Canker Pathogen Xanthomonas citri subsp. citri of Satsuma Mandarin Fruits

    PubMed Central

    Song, Min-A; Park, Jae Sin; Kim, Ki Deok; Jeun, Yong Chull

    2015-01-01

    Citrus canker caused by Xanthomonas citri subsp. citri (Xcc) is one of the most important bacterial diseases of citrus. Because citrus canker is not found in many countries including European Union and Australia, Xcc is strictly regulated in order to prevent its spread. In this study, the effects of X-irradiation on Xcc growth either in the suspension or on the surface of citrus fruits were investigated. The suspension containing 1×107 cfu/ml of Xcc was irradiated with different absorbed doses of X-irradiation ranging from 50 to 400 Gy. The results showed that Xcc was fully dead at 400 Gy of X-irradiation. To determine the effect of X-irradiation on quarantine, the Xcc-inoculated citrus fruits were irradiated with different X-ray doses at which Xcc was completely inhibited by an irradiation dose of 250 Gy. The D10 value for Xcc on citrus fruits was found to be 97 Gy, indicating the possibility of direct application on citrus quarantine without any side sterilizer. Beside, presence of Xcc on the surface of asymptomatic citrus fruits obtained from citrus canker-infected orchards was noted. It indicated that the exporting citrus fruits need any treatment so that Xcc on the citrus fruits should be completely eliminated. Based on these results, ionizing radiation can be considered as an alternative method of eradicating Xcc for export of citrus fruits. PMID:26672670

  12. Huanglongbing: Devastating disease of citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus greening or Huanglongbing (HLB) disease is devastating the citrus industry worldwide, from Asia to the Americas. The only major citrus regions free of this disease are the Mediterranean and Australia/New Zealand. To date there is no cure, no resistance and no therapy for HLB and production ra...

  13. The value of citrus genebanking

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Access to diverse citrus genetic resources is critical for breeding new citrus cultivars that have higher quality fruit and improved resistance to pathogens and changing environmental conditions. The USDA-ARS National Clonal Repository for Citrus and Dates (NCGRCD) maintains a very diverse collectio...

  14. Genome Assembly of Citrus Leprosis Virus Nuclear Type Reveals a Close Association with Orchid Fleck Virus

    PubMed Central

    Stone, Andrew; Otero-Colina, Gabriel; Wei, Gang; Choudhary, Nandlal; Achor, Diann; Shao, Jonathan; Levy, Laurene; Nakhla, Mark K.; Hollingsworth, Charla R.; Hartung, John S.; Schneider, William L.

    2013-01-01

    The complete genome of citrus leprosis virus nuclear type (CiLV-N) was identified by small RNA sequencing utilizing leprosis-affected citrus samples collected from the state of Querétaro, Mexico. The nucleotide identity and phylogenetic analysis indicate that CiLV-N is very closely related to orchid fleck virus, which typically infects Cymbidium species. PMID:23887919

  15. Genome assembly of citrus leprosis virus nuclear type reveals a close association with orchid fleck virus.

    PubMed

    Roy, Avijit; Stone, Andrew; Otero-Colina, Gabriel; Wei, Gang; Choudhary, Nandlal; Achor, Diann; Shao, Jonathan; Levy, Laurene; Nakhla, Mark K; Hollingsworth, Charla R; Hartung, John S; Schneider, William L; Brlansky, Ronald H

    2013-07-25

    The complete genome of citrus leprosis virus nuclear type (CiLV-N) was identified by small RNA sequencing utilizing leprosis-affected citrus samples collected from the state of Querétaro, Mexico. The nucleotide identity and phylogenetic analysis indicate that CiLV-N is very closely related to orchid fleck virus, which typically infects Cymbidium species.

  16. Molecular Marker Analysis of Citrus tristeza virus (CTV) isolates from the Dominican Republic

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Samples of citrus tissue infected with Citrus tristeza virus (CTV) were collected from Persian lime, mandarin, Washington navel, Valencia or grapefruit trees from various locations in the Dominican Republic. Desiccated tissue samples were re-hydrated and virions extracted by grinding samples in buff...

  17. Resistance of sweet orange Pera (Citrus sinensis) genotypes to Xanthomonas citri subsp. citri under field conditions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker control is based on protection measures and eradication of plants infected with Xanthomonas citri subsp. citri. Although these measures show satisfactory results, the use of resistant genotypes is an important alternative for citrus canker control. The aim of this study was to evaluate...

  18. Transmission and propagation of ‘Candidatus Liberibacter asiaticus’ by grafting with individual citrus leaves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB) is a chronic, progressive decline disease in citrus associated with systemic infection by the bacterium Candidatus Liberibacter asiaticus (CLas). Transmission in the field is by the Asian citrus psyllid (ACP), Diaphorina citri, Kuwayama. Experimental propagation of CLas is done c...

  19. Effectiveness of Citrus Fruits on Helicobacter pylori

    PubMed Central

    2017-01-01

    It is known that Helicobacter pylori infection is associated with chronic gastritis, peptic ulcer, and gastric carcinoma. Due to the increased side effects of the treatment regimens and the development of antimicrobial resistance, a number of natural compounds have been tested as potential alternatives. In this review, we will examine the current knowledge on the effect of Citrus fruits and their derivatives against H. pylori, highlighting the remaining outstanding questions on the development of novel therapeutic strategies.

  20. 75 FR 17289 - Citrus Seed Imports; Citrus Greening and Citrus Variegated Chlorosis

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-06

    ... Animal and Plant Health Inspection Service 7 CFR Part 319 RIN 0579-AD07 Citrus Seed Imports; Citrus... States of plants and plant parts, except fruit and seeds, of all genera, species, and varieties of the... of nursery stock to prohibit the importation of propagative seed of several Rutaceae (citrus...

  1. Phloem restriction of viroids in three citrus hosts is overcome by grafting with Etrog citron: potential involvement of a translocatable factor.

    PubMed

    Bani-Hashemian, Seyed Mehdi; Pensabene-Bellavia, Giovanni; Duran-Vila, Nuria; Serra, Pedro

    2015-08-01

    Viroid systemic spread involves cell-to-cell movement from initially infected cells via plasmodesmata, long-distance movement within the phloem and again cell-to-cell movement to invade distal tissues including the mesophyll. Citrus exocortis viroid (CEVd), hop stunt viroid, citrus bent leaf viroid, citrus dwarfing viroid, citrus bark cracking viroid and citrus viroid V remained phloem restricted when singly infecting Citrus karna, Citrus aurantium and Poncirus trifoliata, but not Etrog citron, where they were additionally detected in mesophyll protoplasts. However, when CEVd-infected C. karna was side-grafted with Etrog citron--with the resulting plants being composed of a C. karna stock and an Etrog citron branch--the viroid was detected in mesophyll protoplasts of the former, thus indicating that the ability of Etrog citron to support viroid invasion of non-vascular tissues was transferred to the stock. Further results suggest that a translocatable factor from Etrog citron mediates this viroid trafficking.

  2. 7 CFR 301.76-3 - Quarantined areas; citrus greening and Asian citrus psyllid.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 5 2011-01-01 2011-01-01 false Quarantined areas; citrus greening and Asian citrus...) ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE DOMESTIC QUARANTINE NOTICES Citrus Greening and Asian Citrus Psyllid § 301.76-3 Quarantined areas; citrus greening and Asian citrus...

  3. 7 CFR 301.76-3 - Quarantined areas; citrus greening and Asian citrus psyllid.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 5 2013-01-01 2013-01-01 false Quarantined areas; citrus greening and Asian citrus...) ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE DOMESTIC QUARANTINE NOTICES Citrus Greening and Asian Citrus Psyllid § 301.76-3 Quarantined areas; citrus greening and Asian citrus...

  4. 7 CFR 301.76-3 - Quarantined areas; citrus greening and Asian citrus psyllid.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 5 2014-01-01 2014-01-01 false Quarantined areas; citrus greening and Asian citrus...) ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE DOMESTIC QUARANTINE NOTICES Citrus Greening and Asian Citrus Psyllid § 301.76-3 Quarantined areas; citrus greening and Asian citrus...

  5. 7 CFR 301.76-3 - Quarantined areas; citrus greening and Asian citrus psyllid.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 5 2012-01-01 2012-01-01 false Quarantined areas; citrus greening and Asian citrus...) ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE DOMESTIC QUARANTINE NOTICES Citrus Greening and Asian Citrus Psyllid § 301.76-3 Quarantined areas; citrus greening and Asian citrus...

  6. Diversity of bacteriophages infecting Xanthomonas oryzae pv. oryzae in paddy fields and its potential to control bacterial leaf blight of rice.

    PubMed

    Chae, Jong-Chan; Hung, Nguyen Bao; Yu, Sang-Mi; Lee, Ha Kyung; Lee, Yong Hoon

    2014-06-28

    Bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae (Xoo) is a very serious disease in rice-growing regions of the world. In spite of their economic importance, there are no effective ways of protecting rice plants from this disease. Bacteriophages infecting Xoo affect the population dynamics of the pathogen and consequently the occurrence of the disease. In this study, we investigated the diversity, host range, and infectivity of Xoo phages, and their use as a bicontrol agent on BLB was tested. Among the 34 phages that were isolated from floodwater in paddy fields, 29 belonged to the Myoviridae family, which suggests that the dominant phage in the ecosystem was Myoviridae. The isolated phages were classified into two groups based on plaque size produced on the lawn of Xoo. In general, there was a negative relationship between plaque size and host range, and interestingly the phages having a narrow host range had low efficiency of infectivity. The deduced protein sequence analysis of htf genes indicated that the gene was not a determinant of host specificity. Although the difference in host range and infectivity depending on morphotype needs to be addressed, the results revealed deeper understanding of the interaction between the phages and Xoo strains in floodwater and damp soil environments. The phage mixtures reduced the occurrence of BLB when they were treated with skim milk. The results indicate that the Xoo phages could be used as an alternative control method to increase the control efficacy and reduce the use of agrochemicals.

  7. Antioxidant activity of Citrus fruits.

    PubMed

    Zou, Zhuo; Xi, Wanpeng; Hu, Yan; Nie, Chao; Zhou, Zhiqin

    2016-04-01

    Citrus is well-known for its nutrition and health-promotion values. This reputation is derived from the studies on the biological functions of phytochemicals in Citrus fruits and their derived products in the past decades. In recent years, the antioxidant activity of Citrus fruits and their roles in the prevention and treatment of various human chronic and degenerative diseases have attracted more and more attention. Citrus fruits are suggested to be a good source of dietary antioxidants. To have a better understanding of the mechanism underlying the antioxidant activity of Citrus fruits, we reviewed a study on the antioxidant activity of the phytochemicals in Citrus fruits, introduced methods for antioxidant activity evaluation, discussed the factors which influence the antioxidant activity of Citrus fruits, and summarized the underlying mechanism of action. Some suggestions for future study were also presented.

  8. Risk factors associated with white spot syndrome virus infection in a Vietnamese rice-shrimp farming system.

    PubMed

    Corsin, F; Turnbull, J F; Hao, N V; Mohan, C V; Phi, T T; Phuoc, L H; Tinh, N T; Morgan, K L

    2001-10-29

    White spot disease (WSD) is a pandemic disease caused by a virus commonly known as white spot syndrome virus (WSSV). Several risk factors for WSD outbreaks have been suggested. However, there have been very few studies to identify risk factors for WSD outbreaks in culture systems. This paper presents and discusses the risk factors for WSSV infection identified during a longitudinal observational study conducted in a Vietnamese rice-shrimp farming system. A total of 158 variables were measured comprising location, features of the pond, management practices, pond bottom quality, shrimp health and other animals in the pond. At the end of the study period WSSV was detected in 15 of the 24 ponds followed through the production cycle (62.5%). One hundred and thirty-nine variables were used in univariate analyses. All the variables with a p-value < or = 0.10 were used in unconditional logistic regression in a forward stepwise model. An effect of location was identified in both univariate and multivariate analyses showing that ponds located in the eastern portion of the study site, closer to the sea, were more likely to test positive for WSSV by 1-step PCR at harvest. Ponds with shrimp of a smaller average size 1 mo after stocking tended to be positive for WSSV at the end of the production cycle. Average weight at 1 mo was also highlighted in multivariate analyses when considered as either a risk factor or an outcome. Other risk factors identified in univariate analyses were earlier date of stocking and use of commercial feed. A number of variables also appeared to be associated with a reduced risk of WSSV at harvest including the presence of dead post larvae in the batch sampled at stocking, presence of Hemigrapsus spp. crabs during the first month of production, feeding vitamin premix or legumes, presence of high numbers of shrimp with bacterial infection and the presence of larger mud crabs or gobies at harvest. No associations were detected with WSSV at harvest and

  9. Influence of metalaxyl on three nematodes of citrus.

    PubMed

    Kaplan, D T

    1983-07-01

    Metalaxyl significantly reduced population of Pratylenchus coffeae, Radopholus similis, and Tylenchulus semipenetrans in roots of Citrus limon (rough lemon) under greenhouse conditions. Postinoculation treatment of rough lemon seedlings was not as effective i n reducing nematode populations as was treatment before inoculation. Fewer nematodes infected metalaxyl-treated roots than nontreated roots. However, incubation of nematodes in metalaxyl did not inhibit nematode motility or their ability to locate and infect roots. Cellular responses to nematode injection differed between treated and nontreated tissues. Metalaxyl appeared to confer nematode contraol by modifying citrus roots such that a normally susceptible rootstock became tolerant.

  10. The conundrum of a unique protein encoded by citrus tristeza virus that is dispensable for infection of most hosts yet shows characteristics of a viral movement protein.

    PubMed

    Bak, Aurélie; Folimonova, Svetlana Y

    2015-11-01

    Citrus tristeza virus (CTV), one of the most economically important viruses, produces a unique protein, p33, which is encoded only in the genomes of isolates of CTV. Recently, we demonstrated that membrane association of the p33 protein confers virus ability to extend its host range. In this work we show that p33 shares characteristics of viral movement proteins. Upon expression in a host cell, the protein localizes to plasmodesmata and displays the ability to form extracellular tubules. Furthermore, p33 appears to traffic via the cellular secretory pathway and the actin network to plasmodesmata locations and is likely being recycled through the endocytic pathway. Finally, our study reveals that p33 colocalizes with a putative movement protein of CTV, the p6 protein. These results suggest a potential role of p33 as a noncanonical viral movement protein, which mediates virus translocation in the specific hosts.

  11. LED Blue Light-induced changes in phenolics and ethylene in citrus fruit: Implication in elicited resistance against Penicillium digitatum infection.

    PubMed

    Ballester, Ana-Rosa; Lafuente, María T

    2017-03-01

    The objective was to investigate whether LED Blue Light (LBL) induces changes in phenolics and ethylene production of sweet oranges, and whether they participate in LBL-elicited resistance against the most important postharvest pathogen (Penicillium digitatum) of citrus fruit. The expression of relevant genes of the phenylpropanoid and ethylene biosynthetic pathways during elicitation of resistance was also determined. Different LBL (wavelength 450nm) quantum fluxes were used within the 60-630μmolm(-2)s(-1) range. The HPLC analysis showed that the most relevant increase in phenylpropanoids occurred in scoparone, which markedly increased 3days after exposing fruits to a very high quantum flux (630μmolm(-2)s(-1)) for 18h. However, phenylpropanoids, including scoparone, were not critical factors in LBL-induced resistance. The genes involved in ethylene biosynthesis were differentially regulated by LBL. Ethylene is not involved in elicited resistance, although high LBL levels increased ethylene production in only 1h.

  12. High Protective Efficacy of Probiotics and Rice Bran against Human Norovirus Infection and Diarrhea in Gnotobiotic Pigs

    PubMed Central

    Lei, Shaohua; Ramesh, Ashwin; Twitchell, Erica; Wen, Ke; Bui, Tammy; Weiss, Mariah; Yang, Xingdong; Kocher, Jacob; Li, Guohua; Giri-Rachman, Ernawati; Trang, Nguyen Van; Jiang, Xi; Ryan, Elizabeth P.; Yuan, Lijuan

    2016-01-01

    Probiotics have been recognized as vaccine adjuvants and therapeutic agents to treat acute gastroenteritis in children. We previously showed that rice bran (RB) reduced human rotavirus diarrhea in gnotobiotic pigs. Human noroviruses (HuNoVs) are the major pathogens causing non-bacterial acute gastroenteritis worldwide. In this study, Lactobacillus rhamnosus GG (LGG) and Escherichia coli Nissle 1917 (EcN) were first screened for their ability to bind HuNoV P particles and virions derived from clinical samples containing HuNoV genotype GII.3 and GII.4, then the effects of LGG+EcN and RB on HuNoV infection and diarrhea were investigated using the gnotobiotic pig model. While LGG+EcN colonization inhibited HuNoV shedding, probiotic cocktail regimens in which RB feeding started 7 days prior to or 1 day after viral inoculation in the LGG+EcN colonized gnotobiotic pigs exhibited high protection against HuNoV diarrhea and shedding, characterized by significantly reduced incidence (89 versus 20%) and shorter mean duration of diarrhea (2.2 versus 0.2 days), as well as shorter mean duration of virus shedding (3.2 versus 1.0 days). In both probiotic cocktail groups, the diarrhea reduction rates were 78% compared with the control group, and diarrhea severity was reduced as demonstrated by the significantly lower cumulative fecal scores. The high protective efficacy of the probiotic cocktail regimens was attributed to stimulation of IFN-γ+ T cell responses, increased production of intestinal IgA and IgG, and maintenance of healthy intestinal morphology (manifested as longer villi compared with the control group). Therefore, probiotic cocktail regimens containing LGG+EcN and RB may represent highly efficacious strategies to prevent and treat HuNoV gastroenteritis, and potentially other human enteric pathogens. PMID:27853451

  13. High Protective Efficacy of Probiotics and Rice Bran against Human Norovirus Infection and Diarrhea in Gnotobiotic Pigs.

    PubMed

    Lei, Shaohua; Ramesh, Ashwin; Twitchell, Erica; Wen, Ke; Bui, Tammy; Weiss, Mariah; Yang, Xingdong; Kocher, Jacob; Li, Guohua; Giri-Rachman, Ernawati; Trang, Nguyen Van; Jiang, Xi; Ryan, Elizabeth P; Yuan, Lijuan

    2016-01-01

    Probiotics have been recognized as vaccine adjuvants and therapeutic agents to treat acute gastroenteritis in children. We previously showed that rice bran (RB) reduced human rotavirus diarrhea in gnotobiotic pigs. Human noroviruses (HuNoVs) are the major pathogens causing non-bacterial acute gastroenteritis worldwide. In this study, Lactobacillus rhamnosus GG (LGG) and Escherichia coli Nissle 1917 (EcN) were first screened for their ability to bind HuNoV P particles and virions derived from clinical samples containing HuNoV genotype GII.3 and GII.4, then the effects of LGG+EcN and RB on HuNoV infection and diarrhea were investigated using the gnotobiotic pig model. While LGG+EcN colonization inhibited HuNoV shedding, probiotic cocktail regimens in which RB feeding started 7 days prior to or 1 day after viral inoculation in the LGG+EcN colonized gnotobiotic pigs exhibited high protection against HuNoV diarrhea and shedding, characterized by significantly reduced incidence (89 versus 20%) and shorter mean duration of diarrhea (2.2 versus 0.2 days), as well as shorter mean duration of virus shedding (3.2 versus 1.0 days). In both probiotic cocktail groups, the diarrhea reduction rates were 78% compared with the control group, and diarrhea severity was reduced as demonstrated by the significantly lower cumulative fecal scores. The high protective efficacy of the probiotic cocktail regimens was attributed to stimulation of IFN-γ(+) T cell responses, increased production of intestinal IgA and IgG, and maintenance of healthy intestinal morphology (manifested as longer villi compared with the control group). Therefore, probiotic cocktail regimens containing LGG+EcN and RB may represent highly efficacious strategies to prevent and treat HuNoV gastroenteritis, and potentially other human enteric pathogens.

  14. The β-1,3-glucanosyltransferases (Gels) affect the structure of the rice blast fungal cell wall during appressorium-mediated plant infection.

    PubMed

    Samalova, Marketa; Mélida, Hugo; Vilaplana, Francisco; Bulone, Vincent; Soanes, Darren M; Talbot, Nicholas J; Gurr, Sarah J

    2017-03-01

    The fungal wall is pivotal for cell shape and function, and in interfacial protection during host infection and environmental challenge. Here, we provide the first description of the carbohydrate composition and structure of the cell wall of the rice blast fungus Magnaporthe oryzae. We focus on the family of glucan elongation proteins (Gels) and characterize five putative β-1,3-glucan glucanosyltransferases that each carry the Glycoside Hydrolase 72 signature. We generated targeted deletion mutants of all Gel isoforms, that is, the GH72(+) , which carry a putative carbohydrate-binding module, and the GH72(-) Gels, without this motif. We reveal that M. oryzae GH72(+) GELs are expressed in spores and during both infective and vegetative growth, but each individual Gel enzymes are dispensable for pathogenicity. Further, we demonstrated that a Δgel1Δgel3Δgel4 null mutant has a modified cell wall in which 1,3-glucans have a higher degree of polymerization and are less branched than the wild-type strain. The mutant showed significant differences in global patterns of gene expression, a hyper-branching phenotype and no sporulation, and thus was unable to cause rice blast lesions (except via wounded tissues). We conclude that Gel proteins play significant roles in structural modification of the fungal cell wall during appressorium-mediated plant infection.

  15. Antimicrobial Nanoemulsion Formulation with Improved Penetration of Foliar Spray through Citrus Leaf Cuticles to Control Citrus Huanglongbing.

    PubMed

    Yang, Chuanyu; Powell, Charles A; Duan, Yongping; Shatters, Robert; Zhang, Muqing

    2015-01-01

    Huanglongbing (HLB) is the most serious disease affecting the citrus industry worldwide to date. The causal agent, Candidatus Liberibacter asiaticus (Las), resides in citrus phloem, which makes it difficult to effectively treat with chemical compounds. In this study, a transcuticular nanoemulsion formulation was developed to enhance the permeation of an effective antimicrobial compound (ampicillin; Amp) against HLB disease through the citrus cuticle into the phloem via a foliar spray. The results demonstrated that efficiency of cuticle isolation using an enzymatic method (pectinase and cellulase) was dependent on the citrus cultivar and Las-infection, and it was more difficult to isolate cuticles from valencia orange (Citrus sinensis) and HLB-symptomatic leaves. Of eight adjuvants tested, Brij 35 provided the greatest increase in permeability of the HLB-affected cuticle with a 3.33-fold enhancement of cuticular permeability over water control. An in vitro assay using Bacillus subtilis showed that nanoemulsion formulations containing Amp (droplets size = 5.26 ± 0.04 nm and 94 ± 1.48 nm) coupled with Brij 35 resulted in greater inhibitory zone diameters (5.75 mm and 6.66 mm) compared to those of Brij 35 (4.34 mm) and Amp solution (2.83 mm) alone. Furthermore, the nanoemulsion formulations eliminated Las bacteria in HLB-affected citrus in planta more efficiently than controls. Our study shows that a water in oil (W/O) nanoemulsion formulation may provide a useful model for the effective delivery of chemical compounds into citrus phloem via a foliar spray for controlling citrus HLB.

  16. Antimicrobial Nanoemulsion Formulation with Improved Penetration of Foliar Spray through Citrus Leaf Cuticles to Control Citrus Huanglongbing

    PubMed Central

    Yang, Chuanyu; Powell, Charles A.; Duan, Yongping; Shatters, Robert; Zhang, Muqing

    2015-01-01

    Huanglongbing (HLB) is the most serious disease affecting the citrus industry worldwide to date. The causal agent, Candidatus Liberibacter asiaticus (Las), resides in citrus phloem, which makes it difficult to effectively treat with chemical compounds. In this study, a transcuticular nanoemulsion formulation was developed to enhance the permeation of an effective antimicrobial compound (ampicillin; Amp) against HLB disease through the citrus cuticle into the phloem via a foliar spray. The results demonstrated that efficiency of cuticle isolation using an enzymatic method (pectinase and cellulase) was dependent on the citrus cultivar and Las-infection, and it was more difficult to isolate cuticles from valencia orange (Citrus sinensis) and HLB-symptomatic leaves. Of eight adjuvants tested, Brij 35 provided the greatest increase in permeability of the HLB-affected cuticle with a 3.33-fold enhancement of cuticular permeability over water control. An in vitro assay using Bacillus subtilis showed that nanoemulsion formulations containing Amp (droplets size = 5.26 ± 0.04 nm and 94 ± 1.48 nm) coupled with Brij 35 resulted in greater inhibitory zone diameters (5.75 mm and 6.66 mm) compared to those of Brij 35 (4.34 mm) and Amp solution (2.83 mm) alone. Furthermore, the nanoemulsion formulations eliminated Las bacteria in HLB-affected citrus in planta more efficiently than controls. Our study shows that a water in oil (W/O) nanoemulsion formulation may provide a useful model for the effective delivery of chemical compounds into citrus phloem via a foliar spray for controlling citrus HLB. PMID:26207823

  17. Insecticide sprays, natural enemy assemblages and predation on Asian citrus psyllid, Diaphorina citri (Hemiptera: Psyllidae).

    PubMed

    Monzo, C; Qureshi, J A; Stansly, P A

    2014-10-01

    The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama is considered a key citrus pest due to its role as vector of 'huanglongbing' (HLB) or citrus greening, probably the most economically damaging disease of citrus. Insecticidal control of the vector is still considered a cornerstone of HLB management to prevent infection and to reduce reinoculation of infected trees. The severity of HLB has driven implementation of intensive insecticide programs against ACP with unknown side effects on beneficial arthropod fauna in citrus agroecosystems. We evaluated effects of calendar sprays directed against this pest on natural enemy assemblages and used exclusion to estimate mortality they imposed on ACP populations in citrus groves. Predator exclusion techniques were used on nascent colonies of D. citri in replicated large untreated and sprayed plots of citrus during the four major flushing periods over 2 years. Population of spiders, arboreal ants and ladybeetles were independently assessed. Monthly sprays of recommended insecticides for control of ACP, adversely affected natural enemy populations resulting in reduced predation on ACP immature stages, especially during the critical late winter/early spring flush. Consequently, projected growth rates of the ACP population were greatest where natural enemies had been adversely affected by insecticides. Whereas, this result does not obviate the need for insecticidal control of ACP, it does indicate that even a selective regimen of sprays can impose as yet undetermined costs in terms of reduced biological control of this and probably other citrus pests.

  18. Transmission efficiency of Xylella fastidiosa by sharpshooters (Hemiptera: Cicadellidae) in coffee and citrus.

    PubMed

    Marucci, Rosangela C; Lopes, João R S; Cavichioli, Rodney R

    2008-08-01

    Xylella fastidiosa (Wells, Raju, Hung, Weisburg, Mandelco-Paul, and Brenner) is a bacterial pathogen transmitted by several sharpshooters in two tribes of Cicadellinae (Proconiini and Cicadellini). Here, we compared the transmission efficiency of X. fastidiosa in coffee (Coffea arabica L.) and citrus [Citrus sinensis (L.) Osbeck] by Cicadellini [Bucephalogonia xanthophis (Berg) and Dilobopterus costalimai Young] and Proconiini [Homalodisca ignorata Melichar and Oncometopia facialis (Signoret)] sharpshooters that occur in both crops. At different seasons, healthy adults of each species were submitted to a 48-h acquisition access period on citrus or coffee source plants infected with X. fastidiosa isolates that cause Citrus variegated chlorosis (CVC) and Coffee leaf scorch (CLS), respectively, and then confined on healthy seedlings of the corresponding host plant for a 48-h inoculation access period. No significant effect of inoculation season was observed when comparing infection rates of citrus or coffee plants inoculated by vectors at different times of the year. In citrus, the transmission rate by single insects was significantly higher for H. ignorata (30%) in relation to B. xanthophis (5%) and O. facialis (1.1%), but there was no difference among vector species in coffee, whose transmission rates ranged from 1.2 to 7.2%. Comparing host plants, H. ignorata was more effective in transmitting X. fastidiosa to citrus (30%) in relation to coffee (2.2%), whereas the other vectors transmitted the bacterium to both hosts with similar efficiencies. Despite these variations, vector efficiency in coffee and citrus is lower than that reported in other hosts.

  19. Direct detection of the plant pathogens Burkholderia glumae, Burkholderia gladioli pv. gladioli, and Erwinia chrysanthemi pv. zeae in infected rice seedlings using matrix assisted laser desorption/ionization time-of-flight mass spectrometry.

    PubMed

    Kajiwara, Hideyuki

    2016-01-01

    The plant pathogens Burkholderia glumae, Burkholderia gladioli pv. gladioli, and Erwinia chrysanthemi pv. zeae were directly detected in extracts from infected rice seedlings by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). This method did not require culturing of the pathogens on artificial medium. In the MALDI-TOF MS analysis, peaks originating from bacteria were found in extracts from infected rice seedlings. The spectral peaks showed significantly high scores, in spite of minor differences in spectra. The spectral peaks originating from host plant tissues did not affect this direct MALDI-TOF MS analysis for the rapid identification of plant pathogens.

  20. Evaluation of four phloem-specific promoters in vegetative tissues of transgenic citrus plants.

    PubMed

    Dutt, M; Ananthakrishnan, G; Jaromin, M K; Brlansky, R H; Grosser, J W

    2012-01-01

    'Mexican' lime (Citrus aurantifolia Swingle) was transformed with constructs that contained chimeric promoter-gus gene fusions of phloem-specific rolC promoter of Agrobacterium rhizogenes, Arabidopsis thaliana sucrose-H(+) symporter (AtSUC2) gene promoter of Arabidopsis thaliana, rice tungro bacilliform virus (RTBV) promoter and sucrose synthase l (RSs1) gene promoter of Oryza sativa (rice). Histochemical β-glucuronidase (GUS) analysis revealed vascular-specific expression of the GUS protein in citrus. The RTBV promoter was the most efficient promoter in this study while the RSs1 promoter could drive low levels of gus gene expression in citrus. These results were further validated by reverse transcription real-time polymerase chain reaction and northern blotting. Southern blot analysis confirmed stable transgene integration, which ranged from a single insertion to four copies per genome. The use of phloem-specific promoters in citrus will allow targeted transgene expression of antibacterial constructs designed to battle huanglongbing disease (HLB or citrus greening disease), associated with a phloem-limited Gram-negative bacterium.

  1. Ectopic expression of ubiquitin-conjugating enzyme gene from wild rice, OgUBC1, confers resistance against UV-B radiation and Botrytis infection in Arabidopsis thaliana

    SciTech Connect

    Jeon, En Hee; Pak, Jung Hun; Kim, Mi Jin; Kim, Hye Jeong; Shin, Sang Hyun; Lee, Jai Heon; Kim, Doh Hoon; Oh, Ju Sung; Oh, Boung-Jun; Jung, Ho Won; Chung, Young Soo

    2012-10-19

    Highlights: Black-Right-Pointing-Pointer We isolated a novel E2 ubiquitin-conjugating enzyme from leaves of wild rice plants. Black-Right-Pointing-Pointer The OgUBC1 was highly expressed in leaves treated with SA and UV-B radiation. Black-Right-Pointing-Pointer The recombinant OgUBC1 has an enzymatic activity of E2 in vitro. Black-Right-Pointing-Pointer The OgUBC1 could protect disruption of plant cells by UV-B radiation. Black-Right-Pointing-Pointer OgUBC1 confers disease resistance and UV-B tolerance in transgenic Arabidopsis plants. -- Abstract: A previously unidentified gene encoding ubiquitin-conjugating enzyme was isolated from leaves of wild rice plant treated with wounding and microbe-associated molecular patterns. The OgUBC1 gene was composed of 148 amino acids and contained a typical active site and 21 ubiquitin thioester intermediate interaction residues and 4 E3 interaction residues. Both exogenous application of salicylic acid and UV-B irradiation triggered expression of OgUBC1 in leaves of wild rice. Recombinant OgUBC1 proteins bound to ubiquitins in vitro, proposing that the protein might act as E2 enzyme in planta. Heterologous expression of the OgUBC1 in Arabidopsis thaliana protected plants from cellular damage caused by an excess of UV-B radiation. A stable expression of chalcone synthase gene was detected in leaves of OgUBC1-expressing Arabidopsis, resulting in producing higher amounts of anthocyanin than those in wild-type Col-0 plants. Additionally, both pathogenesis-related gene1 and 5 were transcribed in the transgenic Arabidopsis in the absence of pathogen infection. The OgUBC1-expressing plants were resistant to the infection of Botrytis cinerea. Taken together, we suggested that the OgUBC1 is involved in ubiquitination process important for cellular response against biotic and abiotic stresses in plants.

  2. 7 CFR 301.76-2 - Regulated articles for Asian citrus psyllid and citrus greening.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 5 2014-01-01 2014-01-01 false Regulated articles for Asian citrus psyllid and citrus...) ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE DOMESTIC QUARANTINE NOTICES Citrus Greening and Asian Citrus Psyllid § 301.76-2 Regulated articles for Asian citrus psyllid and...

  3. 7 CFR 301.76-2 - Regulated articles for Asian citrus psyllid and citrus greening.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 5 2013-01-01 2013-01-01 false Regulated articles for Asian citrus psyllid and citrus...) ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE DOMESTIC QUARANTINE NOTICES Citrus Greening and Asian Citrus Psyllid § 301.76-2 Regulated articles for Asian citrus psyllid and...

  4. 7 CFR 301.76-2 - Regulated articles for Asian citrus psyllid and citrus greening.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 5 2012-01-01 2012-01-01 false Regulated articles for Asian citrus psyllid and citrus...) ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE DOMESTIC QUARANTINE NOTICES Citrus Greening and Asian Citrus Psyllid § 301.76-2 Regulated articles for Asian citrus psyllid and...

  5. Chemical compounds effective against the citrus Huanglongbing bacterium 'Candidatus Liberibacter asiaticus' in planta.

    PubMed

    Zhang, Muqing; Powell, Charles A; Zhou, Lijuan; He, Zhenli; Stover, Ed; Duan, Yongping

    2011-09-01

    Citrus Huanglongbing (HLB) is one of the most destructive diseases of citrus worldwide and is threatening the survival of the Floridian citrus industry. Currently, there is no established cure for this century-old and emerging disease. As a possible control strategy for citrus HLB, therapeutic compounds were screened using a propagation test system with 'Candidatus Liberibacter asiaticus'-infected periwinkle and citrus plants. The results demonstrated that the combination of penicillin and streptomycin (PS) was effective in eliminating or suppressing the 'Ca. L. asiaticus' bacterium and provided a therapeutically effective level of control for a much longer period of time than when administering either antibiotic separately. When treated with the PS, 'Ca. L. asiaticus'-infected periwinkle cuttings achieved 70% of regeneration rates versus <50% by other treatments. The 'Ca. L. asiaticus' bacterial titers in the infected periwinkle plants, as measured by quantitative real-time polymerase chain reaction, decreased significantly following root soaking or foliar spraying with PS. Application of the PS via trunk injection or root soaking also eliminated or suppressed the 'Ca. L. asiaticus' bacterium in the HLB-affected citrus plants. This may provide a useful tool for the management of citrus HLB and other Liberibacter-associated diseases.

  6. Role Bending: Complex Relationships Between Viruses, Hosts, and Vectors Related to Citrus Leprosis, an Emerging Disease.

    PubMed

    Roy, Avijit; Hartung, John S; Schneider, William L; Shao, Jonathan; Leon, Guillermo; Melzer, Michael J; Beard, Jennifer J; Otero-Colina, Gabriel; Bauchan, Gary R; Ochoa, Ronald; Brlansky, Ronald H

    2015-07-01

    Citrus leprosis complex is an emerging disease in the Americas, associated with two unrelated taxa of viruses distributed in South, Central, and North America. The cytoplasmic viruses are Citrus leprosis virus C (CiLV-C), Citrus leprosis virus C2 (CiLV-C2), and Hibiscus green spot virus 2, and the nuclear viruses are Citrus leprosis virus N (CiLV-N) and Citrus necrotic spot virus. These viruses cause local lesion infections in all known hosts, with no natural systemic host identified to date. All leprosis viruses were believed to be transmitted by one species of mite, Brevipalpus phoenicis. However, mites collected from CiLV-C and CiLV-N infected citrus groves in Mexico were identified as B. yothersi and B. californicus sensu lato, respectively, and only B. yothersi was detected from CiLV-C2 and CiLV-N mixed infections in the Orinoco regions of Colombia. Phylogenetic analysis of the helicase, RNA-dependent RNA polymerase 2 domains and p24 gene amino acid sequences of cytoplasmic leprosis viruses showed a close relationship with recently deposited mosquito-borne negevirus sequences. Here, we present evidence that both cytoplasmic and nuclear viruses seem to replicate in viruliferous Brevipalpus species. The possible replication in the mite vector and the close relationship with mosquito borne negeviruses are consistent with the concept that members of the genus Cilevirus and Higrevirus originated in mites and citrus may play the role of mite virus vector.

  7. Chemical control of the Asian citrus psyllid and of huanglongbing disease in citrus.

    PubMed

    Boina, Dhana Raj; Bloomquist, Jeffrey R

    2015-06-01

    By 2014, huanglongbing (HLB), the most destructive disease of citrus, and its insect vector, the Asian citrus psyllid (ACP), Diaphorina citri (Kuwayama), became established in all major citrus-growing regions of the world, including the United States, with the exception of California. At present, application of insecticides is the most widely followed option for reducing ACP populations, while application of antibiotics for suppressing HLB disease/symptoms is being practiced in some citrus-growing regions. Application of insecticides during the dormant winter season, along with cultivation of HLB-free seedlings and early detection and removal of symptomatic and asymptomatic trees, has been very effective in managing ACP. Area-wide management of ACP by application of insecticides at low volume in large areas of citrus cultivation has been shown to be effective in managing HLB and reducing management costs. As insecticide resistance is a major problem in sustainable management of ACP, rotation/alternation of insecticides with different chemistries and modes of action needs to be followed. Besides control of the insect vector, use of antibiotics has temporarily suppressed the symptoms of HLB in diseased trees. Recent efforts to discover and screen existing as well as new compounds for their antibiotic and antimicrobial activities have identified some promising molecules for HLB control. There is an urgent need to find a sustainable solution to the HLB menace through chemical control of ACP populations and within HLB-infected trees through the judicious use of labeled insecticides (existing and novel chemistries) and antibiotics in area-wide management programs with due consideration to the insecticide resistance problem.

  8. Monoclonal antibody-based serological methods for detecting Citrus tristeza virus in citrus groves.

    PubMed

    Liu, Zhen; Chen, Zhe; Hong, Jian; Wang, Xuefeng; Zhou, Changyong; Zhou, Xueping; Wu, Jianxiang

    2016-08-01

    Citrus tristeza virus (CTV) is one of the most economically important citrus viruses and harms the citrus industry worldwide. To develop reliable and effective serological detection assays of CTV, the major capsid protein (CP) gene of CTV was expressed in Escherichia coli BL21 (DE3) using the expression vector pET-28a and purified through Ni+-NTA affinity chromatography. The recombinant protein was used to immunize BALB/c mice. Four hybridoma cell lines (14B10, 14H11, 20D5, and 20G12) secreting monoclonal antibodies (MAbs) against CTV were obtained through conventional hybridoma technology. The titers of MAb-containing ascitic fluids secreted by the four hybridoma lines ranged from 10(-6) to 10(-7) in indirect enzyme-linked immunosorbent assay (ELISA). Western blots showed that all four MAbs could specifically react with CTV CP. Using the prepared MAbs, dot-ELISA, Tissue print-ELISA, and triple antibody sandwich (TAS)-ELISA were developed to detect CTV in tree nurseries and epidemiological studies. The developed dot-ELISA and TAS-ELISA methods could detect CTV in crude extracts of infected citrus leaves with dilutions of 1:2560 and 1:10, 240 (w/v, g/mL), respectively. Tissue print-ELISA was particularly useful for large-scale field sample detection, mainly owing to its simplicity and lack of sample preparation requirements. The field survey revealed that CTV is prevalent on citrus trees in the Chongqing Municipality, Jiangxi Province, and Zhejiang Province of China. The coincidence rate of serological and RT-PCR test results reached more than 99.5%. The prepared MAbs against CTV and established sensitive and specific serological assays have a significant role in the detection and prevention and control of CTV in our country.

  9. Huanglongbing: An overview of a complex pathosystem ravaging the world's citrus.

    PubMed

    da Graça, John V; Douhan, Greg W; Halbert, Susan E; Keremane, Manjunath L; Lee, Richard F; Vidalakis, Georgios; Zhao, Hongwei

    2016-04-01

    Citrus huanglongbing (HLB) has become a major disease and limiting factor of production in citrus areas that have become infected. The destruction to the affected citrus industries has resulted in a tremendous increase to support research that in return has resulted in significant information on both applied and basic knowledge concerning this important disease to the global citrus industry. Recent research indicates the relationship between citrus and the causal agent of HLB is shaped by multiple elements, in which host defense responses may also play an important role. This review is intended to provide an overview of the importance of HLB to a wider audience of plant biologists. Recent advances on host-pathogen interactions, population genetics and vectoring of the causal agent are discussed.

  10. Agreement: Citrus College Faculty Association and Citrus Community College District.

    ERIC Educational Resources Information Center

    Citrus Community Coll. District, Glendora, CA.

    The collective bargaining agreement between the Citrus Community College District Board of Trustees and the Citrus College Faculty/California Teachers Association/National Education Association is presented. This contract, covering the period from July 1988 through June 1990, deals with the following topics: bargaining agent recognition and…

  11. Citrus Huanglongbing tolerance in Australian Citrus Relatives, Microcitrus and Eremocirus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tolerance, or resistance to citrus huanglongbing will be important as a long term solution for this disease. In a field trial conducted with over 1000 plants belonging to different genera in the sub-family Aurantioideae, we observed field tolerance in many Australian citrus relatives. To confirm the...

  12. Lateral organ boundaries 1 is a disease susceptibility gene for citrus bacterial canker disease.

    PubMed

    Hu, Yang; Zhang, Junli; Jia, Hongge; Sosso, Davide; Li, Ting; Frommer, Wolf B; Yang, Bing; White, Frank F; Wang, Nian; Jones, Jeffrey B

    2014-01-28

    Citrus bacterial canker (CBC) disease occurs worldwide and incurs considerable costs both from control measures and yield losses. Bacteria that cause CBC require one of six known type III transcription activator-like (TAL) effector genes for the characteristic pustule formation at the site of infection. Here, we show that Xanthomonas citri subspecies citri strain Xcc306, with the type III TAL effector gene pthA4 or with the distinct yet biologically equivalent gene pthAw from strain XccA(w), induces two host genes, CsLOB1 and CsSWEET1, in a TAL effector-dependent manner. CsLOB1 is a member of the Lateral Organ Boundaries (LOB) gene family of transcription factors, and CsSWEET1 is a homolog of the SWEET sugar transporter and rice disease susceptibility gene. Both TAL effectors drive expression of CsLOB1 and CsSWEET1 promoter reporter gene fusions when coexpressed in citrus or Nicotiana benthamiana. Artificially designed TAL effectors directed to sequences in the CsLOB1 promoter region, but not the CsSWEET1 promoter, promoted pustule formation and higher bacterial leaf populations. Three additional distinct TAL effector genes, pthA*, pthB, and pthC, also direct pustule formation and expression of CsLOB1. Unlike pthA4 and pthAw, pthB and pthC do not promote the expression of CsSWEET1. CsLOB1 expression was associated with the expression of genes associated with cell expansion. The results indicate that CBC-inciting species of Xanthomonas exploit a single host disease susceptibility gene by altering the expression of an otherwise developmentally regulated gene using any one of a diverse set of TAL effector genes in the pathogen populations.

  13. Lateral organ boundaries 1 is a disease susceptibility gene for citrus bacterial canker disease

    PubMed Central

    Hu, Yang; Zhang, Junli; Jia, Hongge; Sosso, Davide; Li, Ting; Frommer, Wolf B.; Yang, Bing; White, Frank F.; Wang, Nian; Jones, Jeffrey B.

    2014-01-01

    Citrus bacterial canker (CBC) disease occurs worldwide and incurs considerable costs both from control measures and yield losses. Bacteria that cause CBC require one of six known type III transcription activator-like (TAL) effector genes for the characteristic pustule formation at the site of infection. Here, we show that Xanthomonas citri subspecies citri strain Xcc306, with the type III TAL effector gene pthA4 or with the distinct yet biologically equivalent gene pthAw from strain XccAw, induces two host genes, CsLOB1 and CsSWEET1, in a TAL effector-dependent manner. CsLOB1 is a member of the Lateral Organ Boundaries (LOB) gene family of transcription factors, and CsSWEET1 is a homolog of the SWEET sugar transporter and rice disease susceptibility gene. Both TAL effectors drive expression of CsLOB1 and CsSWEET1 promoter reporter gene fusions when coexpressed in citrus or Nicotiana benthamiana. Artificially designed TAL effectors directed to sequences in the CsLOB1 promoter region, but not the CsSWEET1 promoter, promoted pustule formation and higher bacterial leaf populations. Three additional distinct TAL effector genes, pthA*, pthB, and pthC, also direct pustule formation and expression of CsLOB1. Unlike pthA4 and pthAw, pthB and pthC do not promote the expression of CsSWEET1. CsLOB1 expression was associated with the expression of genes associated with cell expansion. The results indicate that CBC-inciting species of Xanthomonas exploit a single host disease susceptibility gene by altering the expression of an otherwise developmentally regulated gene using any one of a diverse set of TAL effector genes in the pathogen populations. PMID:24474801

  14. Validation of high-throughput real time polymerase chain reaction assays for simultaneous detection of invasive citrus pathogens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A number of economically important citrus pathogens are spread by nursery propagation, arthropod vector transmission and in advertent importation and dissemination of infected plants. For these reasons, citrus disease management and clean stock programs need to employ an economical and sensitive p...

  15. Growth measurements and disease severity metrics of citrus identifies resistance and tolerance to Huanglongbing (HLB) in a Florida field planting

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In a Ft. Pierce, Florida field planting, growth and Huanglongbing (HLB) severity were assessed as indicators of HLB-tolerance on progenies of 83 seed-source genotypes (henceforth “parent genotypes”) of citrus and citrus-relatives from the Riverside, California genebank. Trees were exposed to infecte...

  16. Overexpression of a citrus NDR1 ortholog increases disease resistance in Arabidopsis

    PubMed Central

    Lu, Hua; Zhang, Chong; Albrecht, Ute; Shimizu, Rena; Wang, Guanfeng; Bowman, Kim D.

    2013-01-01

    Emerging devastating diseases, such as Huanglongbing (HLB) and citrus canker, have caused tremendous losses to the citrus industry worldwide. Genetic engineering is a powerful approach that could allow us to increase citrus resistance against these diseases. The key to the success of this approach relies on a thorough understanding of defense mechanisms of citrus. Studies of Arabidopsis and other plants have provided a framework for us to better understand defense mechanisms of citrus. Salicylic acid (SA) is a key signaling molecule involved in basal defense and resistance (R) gene-mediated defense against broad-spectrum pathogens. The Arabidopsis gene NDR1 (NON-RACE-SPECIFIC DISEASE RESISTANCE 1) is a positive regulator of SA accumulation and is specifically required for signaling mediated by a subset of R genes upon recognition of their cognate pathogen effectors. Our bioinformatic analysis identified an ortholog of NDR1 from citrus, CsNDR1. Overexpression of CsNDR1 complemented susceptibility conferred by the Arabidopsis ndr1-1 mutant to Pseudomonas syringae strains and also led to enhanced resistance to an oomycete pathogen Hyaloperonospora arabidopsidis. Such heightened resistance is associated with increased SA production and expression of the defense marker gene PATHOGENESIS RELATED 1 (PR1). In addition, we found that expression of PR1 and accumulation of SA were induced to modest levels in citrus infected with Candidatus Liberibacter asiaticus, the bacterial pathogen associated with HLB disease. Thus, our data suggest that CsNDR1 is a functional ortholog of Arabidopsis NDR1. Since Ca. L. asiaticus infection only activates modest levels of defense responses in citrus, we propose that genetically increasing SA/NDR1-mediated pathways could potentially lead to enhanced resistance against HLB, citrus canker, and other destructive diseases challenging global citrus production. PMID:23761797

  17. Molecular detection of Xanthomonas oryzae pv. oryzae, Xanthomonas oryzae pv. oryzicola, and Burkholderia glumae in infected rice seeds and leaves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Polymerase chain reaction (PCR) is particularly useful for plant pathogen detection. In the present study, multiplex PCR and SYBR green real-time PCR were developed to facilitate simultaneous detection of three important rice pathogens, Xanthomonas oryzae pv. oryzae, X. oryzae pv. oryzicola, and Bur...

  18. Ustilaginoidea virens infection of rice in Arkansas: Toxicity of false smut galls, their extracts and the Ustiloxin fraction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cool, wet conditions in the southern U.S. during the maturing stages of rice in 1998 contributed to outbreaks of false smut caused by Ustilaginoidea virens. Water extracts of false smut galls in Asia have been reported to contain ustiloxin toxins, cyclic peptide antibiotics that interfered with mi...

  19. The potential for citrus cryotherapy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus collections of pathogen-free plants are needed for breeding, research, and distribution to the user community. The Citrus Research Board funded research project “Development of cryotherapy as an improved method of eliminating graft transmissible pathogens in Citrus” sought to use cryotherapy,...

  20. Widespread applications of citrus cryopreservation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus genetic resources can now be successfully cryopreserved, which means that they can be placed into long-term storage at liquid nitrogen temperatures. This cryopreservation technology was specifically developed to address the immediate need to have secure long-term back-up storage for citrus co...

  1. Integrated Management of Citrus Canker

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fruit losses due to citrus canker, caused by Xanthomonas citri subsp. citri (Xcc), vary each crop season depending on citrus variety, tree age, flushing condition, leafminer control, and coincidence of weather events with occurrence of susceptible fruit and foliage. In 2012, crop losses in Hamlin f...

  2. The flavor of citrus fruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus is the largest cultivated fruit tree crop in the world, with total production of more than 100 million tons per year. The genus Citrus consists of different species, including several producing economically important crops, such as oranges, mandarins, grapefruit, pummelo, lemons and limes, c...

  3. Disease stress detection on citrus using a leaf optical model and field spectroscopy

    NASA Astrophysics Data System (ADS)

    Badnakhe, Mrunalini R.; Durbha, Surya; Adinarayana, J.

    2015-10-01

    As citrus is progressively contributing to horticultural production, wealth and economy of a country, it is necessary to understand the factors impacting citrus production. Gummosis is one of the most serious diseases causing considerable loss of overall citrus production and yield quality. A qualitative and quantitative analysis of citrus leaf biochemical properties are necessary to monitor the crop health, disease /pest stress and production. Total leaf chlorophyll content (Cab) represents one of the key biochemical factors which contributes in water, carbon, and energy exchange processes. Photosynthesis process in citrus will be disturbed as gummosis disease life cycle progresses. It is important to study Cab to evaluate the photosynthesis rate and disease stress. In this study the potential of Radiative Transfer (RT) PROSPECT model to retrieve Cab in citrus orchards was undertaken at different sites. The main goal is to evaluate the relationship between Cab and gummosis disease stress for citrus at various phenological stages. Inversion of PROSPECT model on measured hyperspectral data is carried out to extract the leaf level parameters influencing the disease. This model was inverted with the ground truth hyperspectral reading. The testing was separately initiated for healthy and infected plant leaves. This can lead to understand the disease stress on citrus leaves. For accuracy, raw spectra are filtered and processed which is an input parameter for Inversion PROSPECT model. Here, retrieved Cab content was correlated with gummosis disease stress in terms of oozing with R2 = 0.6021 and RMSE= 0.481272.

  4. Antagonists of Plant-parasitic Nematodes in Florida Citrus

    PubMed Central

    Walter, David Evans; Kaplan, David T.

    1990-01-01

    In a survey of antagonists of nematodes in 27 citrus groves, each with a history of Tylenchulus semipenetrans infestation, and 17 noncitrus habitats in Florida, approximately 24 species of microbial antagonists capable of attacking vermiform stages of Radopholus citrophilus were recovered. Eleven of these microbes and a species of Pasteuria also were observed attacking vermiform stages of T. semipenetrans. Verticillium chlamydosporium, Paecilomyces lilacinus, P. marquandii, Streptomyces sp., Arthrobotrys oligospora, and Dactylella ellipsospora were found infecting T. semipenetrans egg masses. Two species of nematophagous amoebae, five species of predatory nematodes, and 29 species of nematophagous arthropods also were detected. Nematode-trapping fungi and nematophagous arthropods were common inhabitants of citrus groves with a history of citrus nematode infestation; however, obligate parasites of nematodes were rare. PMID:19287759

  5. Antagonists of Plant-parasitic Nematodes in Florida Citrus.

    PubMed

    Walter, D E; Kaplan, D T

    1990-10-01

    In a survey of antagonists of nematodes in 27 citrus groves, each with a history of Tylenchulus semipenetrans infestation, and 17 noncitrus habitats in Florida, approximately 24 species of microbial antagonists capable of attacking vermiform stages of Radopholus citrophilus were recovered. Eleven of these microbes and a species of Pasteuria also were observed attacking vermiform stages of T. semipenetrans. Verticillium chlamydosporium, Paecilomyces lilacinus, P. marquandii, Streptomyces sp., Arthrobotrys oligospora, and Dactylella ellipsospora were found infecting T. semipenetrans egg masses. Two species of nematophagous amoebae, five species of predatory nematodes, and 29 species of nematophagous arthropods also were detected. Nematode-trapping fungi and nematophagous arthropods were common inhabitants of citrus groves with a history of citrus nematode infestation; however, obligate parasites of nematodes were rare.

  6. Development of loop-mediated isothermal amplification and SYBR green real-time PCR methods for the detection of Citrus yellow mosaic badnavirus in citrus species.

    PubMed

    Anthony Johnson, A M; Dasgupta, I; Sai Gopal, D V R

    2014-07-01

    Citrus yellow mosaic badnavirus (CMBV) is an important pathogen in southern India spread by infected citrus propagules. One of the measures to arrest the spread of CMBV is to develop methods to screen and certify citrus propagules as CMBV-free. The methods loop-mediated isothermal amplification (LAMP) and SYBR green real-time PCR (SGRTPCR) have been developed for the efficient detection of CMBV in citrus propagules. This paper compares the sensitivities of LAMP and SGRTPCR with polymerase chain reaction (PCR) for the detection of CMBV. Whereas PCR and LAMP were able to detect CMBV from a minimum of 10 ng of total DNA of infected leaf samples, SGRTPCR could detect the same from 1 ng of total DNA. Using SGRTPCR, the viral titres were estimated to be the highest in rough lemon and lowest in Nagpur Mandarin of the five naturally infected citrus species tested. The results will help in designing suitable strategies for the sensitive detection of CMBV from citrus propagules.

  7. A stable RNA virus-based vector for citrus trees

    SciTech Connect

    Folimonov, Alexey S.; Folimonova, Svetlana Y.; Bar-Joseph, Moshe; Dawson, William O.

    2007-11-10

    Virus-based vectors are important tools in plant molecular biology and plant genomics. A number of vectors based on viruses that infect herbaceous plants are in use for expression or silencing of genes in plants as well as screening unknown sequences for function. Yet there is a need for useful virus-based vectors for woody plants, which demand much greater stability because of the longer time required for systemic infection and analysis. We examined several strategies to develop a Citrus tristeza virus (CTV)-based vector for transient expression of foreign genes in citrus trees using a green fluorescent protein (GFP) as a reporter. These strategies included substitution of the p13 open reading frame (ORF) by the ORF of GFP, construction of a self-processing fusion of GFP in-frame with the major coat protein (CP), or expression of the GFP ORF as an extra gene from a subgenomic (sg) mRNA controlled either by a duplicated CTV CP sgRNA controller element (CE) or an introduced heterologous CE of Beet yellows virus. Engineered vector constructs were examined for replication, encapsidation, GFP expression during multiple passages in protoplasts, and for their ability to infect, move, express GFP, and be maintained in citrus plants. The most successful vectors based on the 'add-a-gene' strategy have been unusually stable, continuing to produce GFP fluorescence after more than 4 years in citrus trees.

  8. Detection of mechanical and disease stresses in citrus plants by fluorescence spectroscopy

    NASA Astrophysics Data System (ADS)

    Belasque, J., Jr.; Gasparoto, M. C. G.; Marcassa, L. G.

    2008-04-01

    We have investigated the detection of mechanical and disease stresses in citrus plants (Citrus limonia [L.] Osbeck) using laser-induced fluorescence spectroscopy. Due to its economic importance we have chosen to investigate the citrus canker disease, which is caused by the Xanthomonas axonopodis pv. citri bacteria. Mechanical stress was also studied because it plays an important role in the plant's infection by such bacteria. A laser-induced fluorescence spectroscopy system, composed of a spectrometer and a 532 nm10 mW excitation laser was used to perform fluorescence spectroscopy. The ratio of two chlorophyll fluorescence bands allows us to detect and discriminate between mechanical and disease stresses. This ability to discriminate may have an important application in the field to detect citrus canker infected trees.

  9. Candidate gene markers for Candidatus Liberibacter asiaticus for detecting citrus greening disease.

    PubMed

    Nageswara-Rao, Madhugiri; Irey, Mike; Garnsey, Stephen M; Gowda, Siddarame

    2013-06-01

    Citrus Huanglongbing (HLB) also known as citrus greening is one of the most devastating diseases of citrus worldwide. The disease is caused by Candidatus Liberibacter bacterium, vectored by the psyllid Diaphorina citri Kuwayama and Trioza erytreae Del Guercio. Citrus plants infected by the HLB bacterium may not show visible symptoms sometimes for years following infection. The aim of this study was to develop effective gene-specific primer pairs for polymerase chain reaction based method for quick screening of HLB disease. Thirty-two different gene-specific primer pairs, across the Ca. Liberibacter asiaticus genome, were successfully developed. The possibility of these primer pairs for cross-genome amplification across 'Ca. Liberibacter africanus' and 'Ca. Liberibacter americanus' were tested. The applicability of these primer pairs for detection and differentiation of Ca Liberibacter spp. is discussed.

  10. Enhancement of anti-complementary and radical scavenging activities in the submerged culture of Cordyceps sinensis by addition of citrus peel.

    PubMed

    Choi, Jang Won; Ra, Kyung Soo; Kim, Seong Yeong; Yoon, Taek Joon; Yu, Kwang-Won; Shin, Kwang-Soon; Lee, Sam Pin; Suh, Hyung Joo

    2010-08-01

    To investigate the optimal conditions for the production of Cordyceps sinensis by the submerged culture method, glucosamine and exopolysaccharide (EPS) productivities were determined in culture broth containing different carbon sources, principally rice bran and citrus peel. An optimal medium composition (1.5% rice bran, 0.5% molasses, 3% CSL, 0.1% KH(2)PO(4), and 0.05% MgSO(4)) and the optimal condition (25 degrees C and 5-6 d culture time) for high EPS productivity with potent immune-stimulating activities were obtained. The addition of citrus peel to the culture of C. sinensis under the optimized conditions improved EPS productivity and glucosamine content. Furthermore, anti-complementary activity was higher (58.0-80.8%) using citrus peel as compared to no addition of citrus peel (48.2-68.7%). Antioxidant activity (AEAC value) of the citrus peel culture was high (284.3-384.6 mg/100g) compared to that of the culture without citrus peel (142.8-219.5mg/100g), indicating that the citrus peel helped enhance the anti-complementary and antioxidant activities of C. sinensis.

  11. Citrus tristeza virus: a pathogen that changed the course of the citrus industry.

    PubMed

    Moreno, Pedro; Ambrós, Silvia; Albiach-Martí, Maria R; Guerri, José; Peña, Leandro

    2008-03-01

    shaping CTV populations. Measures to control CTV damage include quarantine and budwood certification programmes, elimination of infected trees, use of tristeza-tolerant rootstocks, or cross protection with mild isolates, depending on CTV incidence and on the virus strains and host varieties predominant in each region. Incorporating resistance genes into commercial varieties by conventional breeding is presently unfeasible, whereas incorporation of pathogen-derived resistance by plant transformation has yielded variable results, indicating that the CTV-citrus interaction may be more specific and complex than initially thought. A deep understanding of the interactions between viral proteins and host and vector factors will be necessary to develop reliable and sound control measures.

  12. Identification and profiling of conserved and novel microRNAs in Laodelphax striatellus in response to rice black-streaked dwarf virus (RBSDV) infection

    PubMed Central

    Li, Jun-Min; Zhou, Yan-Ru; Sun, Zong-Tao; Wang, Xu; Xie, Li; Chen, Jian-Ping

    2014-01-01

    MicroRNAs (miRNAs) are small non-coding endogenous RNA molecules that play important roles in various biological processes. This study examined microRNA profiles of Laodelphax striatellus using the small RNA libraries derived from virus free (VF) and rice black-streaked dwarf virus (RBSDV) infected (RB) insects. A total of 59 mature miRNAs (46 miRNA families) were identified as conserved insect miRNAs in both VF and RB libraries. Among these conserved miRNAs, 24 were derived from the two arms of 12 miRNA precursors. Nine conserved L. striatellus miRNAs were up-regulated and 12 were down-regulated in response to RBSDV infection. In addition, a total of 20 potential novel miRNA candidates were predicted in the VF and RB libraries. The miRNA transcriptome profiles and the identification of L. striatellus miRNAs differentially expressed in response to RBSDV infection will contribute to future studies to elucidate the complex miRNA-mediated regulatory network activated by pathogen challenge in insect vectors. PMID:26484150

  13. Agrobacterium-mediated transformation of Mexican lime (Citrus aurantifolia Swingle) using optimized systems for epicotyls and cotelydons

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Epicotyl and internodal stem segments provide the predominantly used explants for regeneration of transgenic citrus plants following co-cultivation with Agrobacterium. Previous reports using epicotyls segments from Mexican lime have shown low affinity for Agrobacterium tumefaciens infection which re...

  14. Comparison of α-glucosyl hesperidin of citrus fruits and epigallocatechin gallate of green tea on the Loss of Rotavirus Infectivity in Cell Culture

    PubMed Central

    Lipson, Steven M.; Ozen, Fatma S.; Louis, Samantha; Karthikeyan, Laina

    2015-01-01

    A number of secondary plant metabolites (e.g., flavonoids) possess antiviral/antimicrobial activity. Most flavonoids, however, are difficult to study, as they are immiscible in water-based systems. The relatively new semisynthetic α-glucosyl hesperitin (GH), and the natural plant product epigallocatechin gallate (EGCG) are unique among most flavonoids, as these flavonoids are highly soluble. The antiviral activity of these plant metabolites were investigated using the rotavirus as a model enteric virus system. Direct loss of virus structural integrity in cell-free suspension and titration of amplified RTV in host cell cultures was measured by a quantitative enzyme-linked immunosorbent assay (qEIA). After 30 min. 100 × 103 μg/ml GH reduced RTV antigen levels by ca. 90%. The same compound reduced infectivity (replication in cell culture) by a similar order of magnitude 3 to 4 days post inoculation. After 3 days in culture, EGCG concentrations of 80, 160, and 320 μg/ml reduced RTV infectivity titer levels to ca. 50, 20, and 15% of the control, respectively. Loss of RTV infectivity titers occurred following viral treatment by parallel testing of both GH and EGCG, with the latter, markedly more effective. Cytotoxicity testing showed no adverse effects by the phenolic concentrations used in this study. The unique chemical structure of each flavonoid rather than each phenolic’s inherent solubility may be ascribed to those marked differences between each molecule’s antiviral (anti-RTV) effects. The solubility of EGCG and GH obviated our need to use potentially confounding or obfuscating carrier molecules (e.g., methanol, ethanol, DMSO) denoting our use of a pure system environ. Our work further denotes the need to address the unique chemical nature of secondary plant metabolites before any broad generalizations in flavonoid (antiviral) activity may be proposed. PMID:25972850

  15. Past and future of a century old Citrus Tristeza virus collection: A California citrus germplasm tale

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The California Citrus Clonal Protection Program (CCPP) provides a mechanism for introduction and distribution of pathogen-free citrus varieties to California for use in research, variety improvement, or commercial production. Citrus tristeza virus (CTV) is a serious citrus pathogen worldwide. The pr...

  16. Past and future of a century old Citrus Tristeza Virus collection: A California citrus germplasm tale

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The California Citrus Clonal Protection Program (CCPP), Riverside, CA provides a mechanism for introduction and distribution of citrus germplasm from any citrus-growing area of the world to California for use in research, variety improvement, or by industry. Citrus tristeza virus (CTV) is a serious ...

  17. 78 FR 63369 - Citrus Canker, Citrus Greening, and Asian Citrus Psyllid; Interstate Movement of Regulated...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-24

    ... and through this area. Regulated articles are plants and plant parts of all species, clones, cultivars, strains, varieties, or hybrids of the genera Citrus and Fortunella, and all clones, cultivars,...

  18. Citrus Grove Mapping

    NASA Technical Reports Server (NTRS)

    1980-01-01

    Citrus growers have long used aerial photography to inventory the number of groves in production. A new development at Kennedy Space Center, aerial mapping of groves with color infrared (CIR) film, affords an important advance in grove management by detecting and locating unhealthy trees long before they could be detected by ground survey methods. Aerial CIR photography picks up light reflected from foliage-- light not visible to the human eye--and enables differentiation between healthy and "stressed" (diseased) trees of a Florida orange/grapefruit grove. Computer aided photo interpretation techniques permit grading diseased trees lightly, moderately or severely stressed or dead. Method of grove mapping has offered advantage to growers in early disease warning, possible savings through water regulation and provision of a permanent record of grove growth patterns.

  19. Control of virus diseases of citrus.

    PubMed

    Lee, Richard F

    2015-01-01

    Citrus is thought to have originated in Southeast Asia and horticulturally desirable clonal selections have been clonally cultivated for hundreds of years. While some citrus species have nucellar embryony, most cultivation of citrus has been by clonal propagation to ensure that propagated plants have the same traits as the parent selection. Clonal propagation also avoids juvenility, and the propagated plants produce fruit sooner. Because of the clonal propagation of citrus, citrus has accumulated a large number of viruses; many of these viruses are asymptomatic until a susceptible rootstock and/or scion is encountered. The viruses reported to occur in citrus will be summarized in this review. Methods of therapy to clean selected clones from viruses will be reviewed; the use of quarantine, clean stock, and certification programs for control of citrus viruses and other strategies to control insect spread citrus viruses, such as mild strain cross-protection and the use of pest management areas will be discussed.

  20. Functional characterization of electron-transferring flavoprotein and its dehydrogenase required for fungal development and plant infection by the rice blast fungus

    PubMed Central

    Li, Ya; Zhu, Jindong; Hu, Jiexiong; Meng, Xiuli; Zhang, Qi; Zhu, Kunpeng; Chen, Xiaomin; Chen, Xuehang; Li, Guangpu; Wang, Zonghua; Lu, Guodong

    2016-01-01

    Electron-transferring flavoprotein (ETF) and its dehydrogenase (ETFDH) are highly conserved electron carriers which mainly function in mitochondrial fatty acid β oxidation. Here, we report the identification and characterization of ETF α and β subunit encoding genes (ETFA and ETFB) and ETFDH encoding gene (ETFDH) in the rice blast fungus Magnaporthe oryzae. It was demonstrated that, by impacting fatty acid metabolism, ETF and ETFDH mutations led to severe growth and conidiation defects, which could be largely rescued by exogenous acetate or carbonate. Furthermore, although conidium germination and appressorium formation appeared to be normal in ETF and ETFDH mutants, most appressoria failed to penetrate the host epidermis due to low turgor pressure. The few appressoria that succeeded in penetration were severely restricted in invasive growth and consequently failed to cause disease. Moreover, ETF mutant etfb− induced ROS accumulation in infected host cells and exogenous antioxidant GSH accelerated mutant invading growth without increasing the penetration rate. In addition, mutant etfb− displayed elevated lipid body accumulation and reduced ATP synthesis. Taken together, ETF and ETFDH play an important role in fungal development and plant infection in M. oryzae by regulation of fatty acid metabolism, turgor establishment and induction of host ROS accumulation. PMID:27113712

  1. Precocious flowering of juvenile citrus induced by a viral vector based on Citrus leaf blotch virus: a new tool for genetics and breeding.

    PubMed

    Velázquez, Karelia; Agüero, Jesús; Vives, María C; Aleza, Pablo; Pina, José A; Moreno, Pedro; Navarro, Luis; Guerri, José

    2016-10-01

    The long juvenile period of citrus trees (often more than 6 years) has hindered genetic improvement by traditional breeding methods and genetic studies. In this work, we have developed a biotechnology tool to promote transition from the vegetative to the reproductive phase in juvenile citrus plants by expression of the Arabidopsis thaliana or citrus FLOWERING LOCUS T (FT) genes using a Citrus leaf blotch virus-based vector (clbvINpr-AtFT and clbvINpr-CiFT, respectively). Citrus plants of different genotypes graft inoculated with either of these vectors started flowering within 4-6 months, with no alteration of the plant architecture, leaf, flower or fruit morphology in comparison with noninoculated adult plants. The vector did not integrate in or recombine with the plant genome nor was it pollen or vector transmissible, albeit seed transmission at low rate was detected. The clbvINpr-AtFT is very stable, and flowering was observed over a period of at least 5 years. Precocious flowering of juvenile citrus plants after vector infection provides a helpful and safe tool to dramatically speed up genetic studies and breeding programmes.

  2. History and Diversity of Citrus leprosis virus Recorded in Herbarium Specimens.

    PubMed

    Hartung, John S; Roy, Avijit; Fu, Shimin; Shao, Jonathan; Schneider, William L; Brlansky, Ronald H

    2015-09-01

    Leprosis refers to two diseases of citrus that present similar necrotic local lesions, often surrounded by chlorotic haloes on citrus. Two distinct viruses are associated with this disease, one that produces particles primarily in the nucleus of infected plant cells (Citrus leprosis virus nuclear type [CiLV-N]; Dichorhavirus) and another type that produces particles in the cytoplasm of infected plant cells (Citrus leprosis virus cytoplasmic type [CiLV-C]; Cilevirus). Both forms are transmitted by Brevipalpid mites and have bipartite, single-stranded, RNA genomes. CiLV-C and CiLV-N are present in South and Central America and as far north as parts of Mexico. Although leprosis disease was originally described from Florida, it disappeared from there in the 1960s. The United States Department of Agriculture-Agricultural Research Service maintains preserved citrus specimens identified at inspection stations 50 or more years ago with symptoms of citrus leprosis. We isolated RNA from these samples and performed degradome sequencing. We obtained nearly full-length genome sequences of both a typical CiLV-C isolate intercepted from Argentina in 1967 and a distinct CiLV-N isolate obtained in Florida in 1948. The latter is a novel form of CiLV-N, not known to exist anywhere in the world today. We have also documented the previously unreported presence of CiLV-N in Mexico in the mid-20th century.

  3. Rapid screening for citrus canker resistance employing pathogen-associated molecular pattern-triggered immunity responses

    PubMed Central

    Pitino, Marco; Armstrong, Cheryl M; Duan, Yongping

    2015-01-01

    Citrus canker, caused by the bacterial pathogen Xanthomonas citri ssp. citri (Xcc), has been attributed to millions of dollars in loss or damage to commercial citrus crops in subtropical production areas of the world. Since identification of resistant plants is one of the most effective methods of disease management, the ability to screen for resistant seedlings plays a key role in the production of a long-term solution to canker. Here, an inverse correlation between reactive oxygen species (ROS) production by the plant and the ability of Xcc to grow and form lesions on infected plants is reported. Based on this information, a novel screening method that can rapidly identify citrus seedlings that are less susceptible to early infection by Xcc was devised by measuring ROS accumulation triggered by a 22-amino acid sequence of the conserved N-terminal part of flagellin (flg22) from X. citri ssp. citri (Xcc-flg22). In addition to limiting disease symptoms, ROS production was also correlated with the expression of basal defense-related genes such as the pattern recognition receptors LRR8 and FLS2, the leucine-rich repeat receptor-like protein RLP12, and the defense-related gene PR1, indicating an important role for pathogen-associated molecular pattern-triggered immunity (PTI) in determining resistance to citrus canker. Moreover, the differential expression patterns observed amongst the citrus seedlings demonstrated the existence of genetic variations in the PTI response among citrus species/varieties. PMID:26504581

  4. Weeping dragon, a unique ornamenal citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    ‘Weeping Dragon’ is a new ornamental citrus cultivar developed by intercrossing of two unusual and unique citrus types, Poncirus trifoliata cultivated variety (cv.) Flying Dragon, and Citrus sinensis cv. ‘Cipo’. This new hybrid cultivar combines strongly contorted and weeping growth traits in a smal...

  5. Citrus tristeza virus-aphid interactions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A review chapter on aphid transmission of Citrus tristeza virus is provided for a book on “Vector-Mediated Transmission of Plant Pathogens”. Earliest uses of citrus goes back over two millennia as items of trade, gifts and medicinal compounds. Citrus propagation during this period was by seed and si...

  6. Citrus tristeza virus-host interactions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) is a phloem-limited virus whose natural host range is restricted to citrus and related species. Although the virus has killed millions of trees, almost destroying whole industries, and continually limits production in many citrus growing areas, most isolates are mild or s...

  7. Citrus Limonoids: Analysis, Bioactivity, and Biomedical Prospects

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This publication is a review of the chemistry, biochemistry and bioactivity of limonoids occurring in citrus. The review chronologically relates the evolution of research in citrus limonoids beginning with their association with bitterness development in citrus juices. The chemical and biochemical...

  8. Deep sequencing of small RNAs from citrus affected by graft-transmissible diseases of unknown etiology leads to discovery of two novel viruses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus is susceptible to graft-transmissible pathogens; some of which are arthropod-transmitted. Citrus cultivars vary in susceptibility or tolerance to these pathogens and may remain symptomless when infected. Established tests for known disease agents are used in certification and disease manage...

  9. Evaluation of the effects of light source and plant materials in psyllid trapping levels in the traps for citrus shipping containers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Asian citrus psyllid (ACP), the principle vector for Huanglongbing (HLB), has been reported to be transported in truckloads of oranges in Florida. Citrus, especially Key limes and lemons, are shipped to the U.S. from Mexican states that are heavily infested with HLB and live, infected psyllids c...

  10. Leaf-disc grafting for the transmission of Candidatus Liberibacter asiaticus in citrus (Citrus sinensis; Rutaceae) seedlings1

    PubMed Central

    Tabay Zambon, Flavia; Plant, Karen; Etxeberria, Ed

    2017-01-01

    Premise of the study: The search for resistance/tolerance to the devastating citrus huanglongbing disease (syn. HLB or citrus greening) is generating an increasing number of new plants of diverse genetic makeup. As the increasing number of new plants require more space, resources, and time, the need for faster and more efficient HLB screening tests becomes crucial. Methods and Results: The leaf-disc grafting system described here consists in replacing a disc of leaf tissue with a similar disc from an infected plant. This can be performed in young seedlings not yet big enough to endure other types of grafting. Graft success and infection rates average approximately 80%. Conclusions: We describe the successful adaptation of leaf-disc grafting as a powerful screening tool for HLB. The system requires minimal plant material and can be performed in seedlings at a very young age with increased efficiency in terms of time, space, and resources. PMID:28090406

  11. Characterization of Citrus sinensis transcription factors closely associated with the non-host response to Xanthomonas campestris pv. vesicatoria.

    PubMed

    Daurelio, Lucas D; Romero, María S; Petrocelli, Silvana; Merelo, Paz; Cortadi, Adriana A; Talón, Manuel; Tadeo, Francisco R; Orellano, Elena G

    2013-07-01

    Plants, when exposed to certain pathogens, may display a form of genotype-independent resistance, known as non-host response. In this study, the response of Citrus sinensis (sweet orange) leaves to Xanthomonas campestris pv. vesicatoria (Xcv), a pepper and tomato pathogenic bacterium, was analyzed through biochemical assays and cDNA microarray hybridization and compared with Asiatic citrus canker infection caused by Xanthomonas citri subsp. citri. Citrus leaves exposed to the non-host bacterium Xcv showed hypersensitive response (HR) symptoms (cell death), a defense mechanism common in plants but poorly understood in citrus. The HR response was accompanied by differentially expressed genes that are associated with biotic stress and cell death. Moreover, 58 transcription factors (TFs) were differentially regulated by Xcv in citrus leaves, including 26 TFs from the stress-associated families AP2-EREBP, bZip, Myb and WRKY. Remarkably, in silico analysis of the distribution of expressed sequence tags revealed that 10 of the 58 TFs, belonging to C2C2-GATA, C2H2, CCAAT, HSF, NAC and WRKY gene families, were specifically over-represented in citrus stress cDNA libraries. This study identified candidate TF genes for the regulation of key steps during the citrus non-host HR. Furthermore, these TFs might be useful in future strategies of molecular breeding for citrus disease resistance.

  12. Effective disinfection of rough rice using infrared radiation heating

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to investigate the effect of infrared (IR) heating and tempering treatments on disinfection of Aspergillus flavus in freshly harvested rough rice and storage rice. Rice samples with initial moisture contents (IMCs) of 14.1 to 27.0% (wet basis) were infected with A. fl...

  13. Genome-wide analysis of citrus R2R3MYB genes and their spatiotemporal expression under stresses and hormone treatments.

    PubMed

    Xie, Rangjin; Li, Yongjie; He, Shaolan; Zheng, Yongqiang; Yi, Shilai; Lv, Qiang; Deng, Lie

    2014-01-01

    The R2R3MYB proteins represent one of the largest families of transcription factors, which play important roles in plant growth and development. Although genome-wide analysis of this family has been conducted in many species, little is known about R2R3MYB genes in citrus, In this study, 101 R2R3MYB genes has been identified in the citrus (Citrus sinesis and Citrus clementina) genomes, which are almost equal to the number of rice. Phylogenetic analysis revealed that they could be subdivided into 21 subgroups. The evolutionary relationships and the intro-exon organizations were also analyzed, revealing strong gene conservation but also the expansions of particular functional genes during the plant evolution. Tissue-specific expression profiles showed that 95 citrus R2R3MYB genes were expressed in at least one tissue and the other 6 genes showed very low expression in all tissues tested, suggesting that citrus R2R3MYB genes play important roles in the development of all citrus organs. The transcript abundance level analysis during abiotic conditions (NaCl, abscisic acid, jasmonic acid, drought and low temperature) identified a group of R2R3MYB genes that responded to one or multiple treatments, which showed a promising for improving citrus adaptation to stresses. Our results provided an essential foundation for the future selection of the citrus R2R3MYB genes for cloning and functional dissection with an aim of uncovering their roles in citrus growth and development.

  14. Distinctively variable sequence-based nuclear DNA markers for multilocus phylogeography of the soybean- and rice-infecting fungal pathogen Rhizoctonia solani AG-1 IA

    PubMed Central

    2009-01-01

    A series of multilocus sequence-based nuclear DNA markers was developed to infer the phylogeographical history of the Basidiomycetous fungal pathogen Rhizoctonia solani AG-1 IA infecting rice and soybean worldwide. The strategy was based on sequencing of cloned genomic DNA fragments (previously used as RFLP probes) and subsequent screening of fungal isolates to detect single nucleotide polymorphisms (SNPs). Ten primer pairs were designed based on these sequences, which resulted in PCR amplification of 200-320 bp size products and polymorphic sequences in all markers analyzed. By direct sequencing we identified both homokaryon and heterokaryon (i.e. dikaryon) isolates at each marker. Cloning the PCR products effectively estimated the allelic phase from heterokaryotic isolates. Information content varied among markers from 0.5 to 5.9 mutations per 100 bp. Thus, the former RFLP codominant probes were successfully converted into six distinctively variable sequence-based nuclear DNA markers. Rather than discarding low polymorphism loci, the combination of these distinctively variable anonymous nuclear markers would constitute an asset for the unbiased estimate of the phylogeographical parameters such as population sizes and divergent times, providing a more reliable species history that shaped the current population structure of R. solani AG-1 IA. PMID:21637462

  15. Magnesium Uptake by CorA Transporters Is Essential for Growth, Development and Infection in the Rice Blast Fungus Magnaporthe oryzae

    PubMed Central

    Reza, Md. Hashim; Shah, Hiral; Manjrekar, Johannes; Chattoo, Bharat B.

    2016-01-01

    Magnaporthe oryzae, the causative organism of rice blast, infects cereal crops and grasses at various stages of plant development. A comprehensive understanding of its metabolism and the implications on pathogenesis is necessary for countering this devastating crop disease. We present the role of the CorA magnesium transporters, MoAlr2 and MoMnr2, in development and pathogenicity of M. oryzae. The MoALR2 and MoMNR2 genes individually complement the Mg2+ uptake defects of a S. cerevisiae CorA transporter double mutant. MoALR2 and MoMNR2 respond to extracellular Mg2+ and Ca2+ levels and their expression is elevated under Mg2+ scarce conditions. RNA silencing mediated knockdown of MoALR2 (WT+siALR2, Δmnr2+siALR2 and ALR2+MNR2 simultaneous silencing) drastically alters intracellular cation concentrations and sensitivity to metal ions. MoALR2 silencing is detrimental to vegetative growth and surface hydrophobicity of mycelia, and the transformants display loss of cell wall integrity. MoALR2 is required for conidiogenesis and appressorium development, and is essential for infection. Investigation of knockdown transformants reveal low cAMP levels and altered expression of genes encoding proteins involved in MoMps1 cell wall integrity and cAMP MoPmk1 driven MAP Kinase signaling pathways. In contrast to MoALR2 knockdowns, the MoMNR2 deletion (Δmnr2) shows increased sensitivity to CorA inhibitors as well as altered cation sensitivity, but has limited effect on surface hydrophobicity and severity of plant infection. Interestingly, MoALR2 expression is elevated in Δmnr2. Impairment of development and infectivity of knockdown transformants and altered intracellular cation composition suggest that CorA transporters are essential for Mg2+ homeostasis within the cell, and are crucial to maintaining normal gene expression associated with cell structure, signal transduction and surface hydrophobicity in M. oryzae. We suggest that CorA transporters, and especially MoALR2

  16. Refinement of the Citrus tristeza virus resistance gene (Ctv) positional map in Poncirus trifoliata and generation of transgenic grapefruit (Citrus paradisi) plant lines with candidate resistance genes in this region.

    PubMed

    Rai, Mamta

    2006-06-01

    Citrus tristeza virus (CTV) is a major pathogen of Citrus. A single dominant gene Ctv present in the trifoliate relative of Citrus, Poncirus trifoliata confers broad spectrum resistance against CTV. Refinement of genetic maps has delimited this gene to a 121 kb region, comprising of ten candidate Ctv resistance genes. The ten candidate genes were individually cloned in Agrobacterium based binary vector and transformed into three CTV susceptible grapefruit varieties. Two of the candidate R-genes, R-2 and R-3 are exclusively expressed in transgenic plants and in Poncirus trifoliata, while five other genes are also expressed in non-transformed Citrus controls. Northern blotting with a CTV derived probe for assessment of infection in virus inoculated plants over a span of three growth periods, each comprising of six to eight weeks, indicates either an absence of initiation of infection or it's slow spread in R-2 plant lines or an initial appearance of infection and it's subsequent obliteration in some R-1 and R-4 plant lines. Limited genome walk up- and downstream form R-1 gene, based on it's 100% sequence identity between Poncirus and Citrus, indicates promoter identity of 92% between the two varieties. Further upstream and downstream sequencing indicates the presence of an O-methyl transferase and a Copia like gene respectively in Citrus instead of the amino acid transporter like gene upstream and a sugar transporter like gene downstream in Poncirus. The possibility of recombinations in the resistance locus of Citrus and the need for consistent monitoring for virus infection and gene expression in the transgenic Citrus trees is discussed.

  17. Site Plan & Site Section of Citrus Landscape (Showing Relationship ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    Site Plan & Site Section of Citrus Landscape (Showing Relationship of Victoria Avenue to Citrus Groves) - Arlington Heights Citrus Landscape, Southwestern portion of city of Riverside, Riverside, Riverside County, CA

  18. Chipping citrus wood for gasification

    SciTech Connect

    Churchill, D.B.; Hedden, S.L.; Whitney, J.D.; Shaw, L.N.

    1985-01-01

    Non-productive citrus trees were chipped with a portable fly-wheel-type chipper powered by a 45 kW engine. Chips were air dried under an open shed to 14% (w.b.) moisture content. By weight, approximately 50% of the total tree could be made into usable chips. The root system averaged 36% of the total tree weight.

  19. Exogenous treatment with salicylic acid attenuates occurrence of citrus canker in susceptible navel orange (Citrus sinensis Osbeck).

    PubMed

    Wang, Yin; Liu, Ji-Hong

    2012-08-15

    Citrus canker caused by Xanthomonas axonopodis pv. citri (Xac) is a devastating bacterial disease threatening the citrus industry. Salicylic acid (SA) plays a key role in plant defense response to biotic stress, but information is scarce concerning the application of SA to enhancing Xac resistance. In the present research attempts were made to investigate how exogenous application of SA influenced canker disease outbreak in navel orange (Citrus sinensis). Exogenously applied SA at 0.25 mM significantly enhanced the endogenous free and bound SA, particularly the latter. Upon exposure to Xac, lower disease incidence rate and smaller lesion sites were observed in the samples pre-treated with SA, accompanied by repression of bacterial growth at the lesion sites. Concurrent with the augmented disease resistance, SA-treated leaves had higher H₂O₂ level and smaller stomata apertures before or after Xac infection when compared with their counterparts pre-treated with water (control). SA treatment elevated the activities of phenylalanine ammonia-lyase and β-1,3-glucanase, but only the latter was higher in the SA-treated samples after Xac infection. In addition, mRNA levels of two pathogenesis-related genes, CsCHI and CsPR4A, were higher in the SA-treated samples relative to the control. Taken together, our results strongly suggest that the exogenously applied SA has evoked a cascade of physiological and molecular events that function singly or in concert to confer resistance to Xac invasion.

  20. Genome of Cnaphalocrocis medinalis Granulovirus, the First Crambidae-Infecting Betabaculovirus Isolated from Rice Leaffolder to Sequenced

    PubMed Central

    Han, Guangjie; Xu, Jian; Liu, Qin; Li, Chuanming; Xu, Hongxing; Lu, Zhongxian

    2016-01-01

    Cnaphalocrocis medinalis is a major pest of rice in South and South-East Asia. Insecticides are the major means farmers use for management. A naturally occurring baculovirus, C. medinalis granulovirus (CnmeGV), has been isolated from the larvae and this has the potential for use as microbial agent. Here, we described the complete genome sequence of CnmeGV and compared it to other baculovirus genomes. The genome of CnmeGV is 112,060 base pairs in length, has a G+C content of 35.2%. It contains 133 putative open reading frames (ORFs) of at least 150 nucleotides. A hundred and one (101) of these ORFs are homologous to other baculovirus genes including 37 baculovirus core genes. Thirty-two (32) ORFs are unique to CnmeGV with no homologues detected in the GeneBank and 53 tandem repeats (TRs) with sequence length from 25 to 551 nt intersperse throughout the genome of CnmeGV. Six (6) homologous regions (hrs) were identified interspersed throughout the genome. Hr2 contains 11 imperfect palindromes and a high content of AT sequence (about 73%). The unique ORF28 contains a coiled-coil region and a zinc finger-like domain of 4–50 residues specialized by two C2C2 zinc finger motifs that putatively bound two atoms of zinc. ORF21 encoding a chit-1 protein suggesting a horizontal gene transfer from alphabaculovirus. The putative protein presents two carbohydrate-binding module family 14 (CBM_14) domains rather than other homologues detected from betabaculovirus that only contains one chit-binding region. Gene synteny maps showed the colinearity of sequenced betabaculovirus. Phylogenetic analysis indicated that CnmeGV grouped in the betabaculovirus, with a close relation to AdorGV. The cladogram obtained in this work grouped the 17 complete GV genomes in one monophyletic clade. CnmeGV represents a new crambidae host-isolated virus species from the genus Betabaculovirus and is most closely relative of AdorGV. The analyses and information derived from this study will provide a

  1. Structural and biochemical characteristics of citrus flowers associated with defence against a fungal pathogen

    PubMed Central

    Marques, João Paulo Rodrigues; Amorim, Lilian; Silva-Junior, Geraldo José; Spósito, Marcel Bellato; Appezzato-da Gloria, Beatriz

    2015-01-01

    The constitutive characters of plants can be structural or biochemical and play an important role in their defence against pathogens. Citrus postbloom fruit drop (PFD) caused by Colletotrichum spp. is one of the most important fungal diseases of citrus. The pathogen infects the flowers, leading to premature fruit drop and reducing citrus production. However, flower buds smaller than 8 mm long are usually not infected by Colletotrichum spp. Thus, this study investigated whether there are constitutive mechanisms in flower buds related to Colletotrichum spp. infection. We studied flower buds that were 2, 3, 4, 8, 12 and 15 mm long and petals, after anthesis, of sweet orange ‘Valência’ using light and scanning electron microscopy and histochemistry. We evaluated the effect of volatile organic compounds (VOCs) in flowers (R-limonene and linalool) on the in vitro growth of Colletotrichum acutatum. We found that the arrangement of the epidermal papillae in the petal primordia, the occurrence of prismatic crystals and the distribution of oil glands are the main differences between buds smaller than 8 mm and buds 8–15 mm long. Osmophores at the tips of petals produced and accumulated phenols, terpenes and lipophilic compounds. Flower buds smaller than 8 mm long have constitutive structural and biochemical barriers to Colletotrichum spp. infection. In addition, this is the first time that osmophores have been reported in citrus. Our study shows that natural terpenes of Citrus flowers inhibit the fungal growth in vitro, highlighting the potential use of terpenes for the chemical control of PFD in citrus. PMID:25535209

  2. 76 FR 23449 - Citrus Canker, Citrus Greening, and Asian Citrus Psyllid; Interstate Movement of Regulated...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-27

    ... conditions, the articles are treated with methyl bromide and shipped in a container that has been sealed with... of any regulated article, including citrus nursery stock, provided that: The article is treated with methyl bromide in accordance with 7 CFR part 305. That part contains our phytosanitary...

  3. 78 FR 8435 - Importation of Fresh Citrus Fruit From Uruguay, Including Citrus Hybrids and Fortunella

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-02-06

    ... of citrus canker). In a previous revision of the PRA, citrus black spot (Guignardia citricarpa Kiely... of citrus black spot because the combination of conditions required for disease transmission from... compounds. However, if the rind is thin or damaged, or existing oviposition puncture holes are...

  4. Citrus Research Board-sponsored review of the University of California Riverside citrus breeding

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In October 2015 the Citrus Research Board (CRB) assembled a panel of experts to review the Citrus Research Board-sponsored Citrus Research and Genetics Programs at University of California Riverside (UCR). The panel consisted of: Gennaro Fazio, USDA/ARS, Geneva, NY; Maria Angeles Forner-Giner, Insti...

  5. Application of natural colorants on citrus fruit as alternatives to Citrus Red II

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Warm field temperatures can often result in poor peel color of some citrus varieties, especially early in the harvest season. Under these conditions, Florida oranges, temples, tangelos, and K-Early citrus fruit are allowed to be treated with Citrus Red No.2 (CR2) to help produce a more acceptable pe...

  6. Evaluation of resistance to asiatic citrus canker among selections of pera sweet orange (Citrus sinensis)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Asiatic citrus canker (ACC, caused by the bacterium Xanthomonas citri subsp. citri) is a destructive disease of citrus in Brazil and in several other citrus-producing countries. ACC management is problematic, and bactericides such as copper can be reasonably efficacious but do not completely control...

  7. Field ID guide to citrus relative hosts of Asian citrus psyllid & Huanglongbing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Rutaceae family of plants includes not only species within the genus Citrus, but also several other genera and species that may not be easily recognized as having any relationship to citrus at all. However, many of these citrus relatives are used for ornamental, culinary, or religious purposes. ...

  8. Production of transgenic citrus resistant to citrus canker and Huanglongbing diseases

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB or citrus greening disease) caused by Candidatus Liberibacter asiaticus (Las) is a great threat to the U.S. citrus industry. There are no proven strategies to eliminate HLB disease and no cultivars identified with strong HLB resistance. Citrus canker is also an economically import...

  9. Huanglongbing, a Systemic Disease, Restructures the Bacterial Community Associated with Citrus Roots▿

    PubMed Central

    Trivedi, Pankaj; Duan, Yongping; Wang, Nian

    2010-01-01

    To examine the effect of pathogens on the diversity and structure of plant-associated bacterial communities, we carried out a molecular analysis using citrus and huanglongbing as a host-disease model. 16S rRNA gene clone library analysis of citrus roots revealed shifts in microbial diversity in response to pathogen infection. The clone library of the uninfected root samples has a majority of phylotypes showing similarity to well-known plant growth-promoting bacteria, including Caulobacter, Burkholderia, Lysobacter, Pantoea, Pseudomonas, Stenotrophomonas, Bacillus, and Paenibacillus. Infection by “Candidatus Liberibacter asiaticus” restructured the native microbial community associated with citrus roots and led to the loss of detection of most phylotypes while promoting the growth of bacteria such as Methylobacterium and Sphingobacterium. In pairwise comparisons, the clone library from uninfected roots contained significantly higher 16S rRNA gene diversity, as reflected in the higher Chao 1 richness estimation (P ≤ 0.01) of 237.13 versus 42.14 for the uninfected and infected clone libraries, respectively. Similarly, the Shannon index of the uninfected clone library (4.46) was significantly higher than that of the infected clone library (2.61). Comparison of the uninfected clone library with the infected clone library using LIBSHUFF statistics showed a significant difference (P ≤ 0.05). Quantitative PCR analysis revealed that the bacterial community changes not only qualitatively but also quantitatively. The relative proportions of different groups of bacteria changed significantly after infection with the pathogen. These data indicate that infection of citrus by “Ca. Liberibacter asiaticus” has a profound effect on the structure and composition of the bacterial community associated with citrus roots. PMID:20382817

  10. Development and validation of a multiplex reverse transcription quantitative PCR (RT-qPCR) assay for the rapid detection of Citrus tristeza virus, Citrus psorosis virus, and Citrus leaf blotch virus.

    PubMed

    Osman, Fatima; Hodzic, Emir; Kwon, Sun-Jung; Wang, Jinbo; Vidalakis, Georgios

    2015-08-01

    A single real-time multiplex reverse transcription quantitative polymerase chain reaction (RT-qPCR) assay for the simultaneous detection of Citrus tristeza virus (CTV), Citrus psorosis virus (CPsV), and Citrus leaf blotch virus (CLBV) was developed and validated using three different fluorescently labeled minor groove binding qPCR probes. To increase the detection reliability, coat protein (CP) genes from large number of different isolates of CTV, CPsV and CLBV were sequenced and a multiple sequence alignment was generated with corresponding CP sequences from the GenBank and a robust multiplex RT-qPCR assay was designed. The capacity of the multiplex RT-qPCR assay in detecting the viruses was compared to singleplex RT-qPCR designed specifically for each virus and was assessed using multiple virus isolates from diverse geographical regions and citrus species as well as graft-inoculated citrus plants infected with various combination of the three viruses. No significant difference in detection limits was found and specificity was not affected by the inclusion of the three assays in a multiplex RT-qPCR reaction. Comparison of the viral load for each virus using singleplex and multiplex RT-qPCR assays, revealed no significant differences between the two assays in virus detection. No significant difference in Cq values was detected when using one-step and two-step multiplex RT-qPCR detection formats. Optimizing the RNA extraction technique for citrus tissues and testing the quality of the extracted RNA using RT-qPCR targeting the cytochrome oxidase citrus gene as an RNA specific internal control proved to generate better diagnostic assays. Results showed that the developed multiplex RT-qPCR can streamline viruses testing of citrus nursery stock by replacing three separate singleplex assays, thus reducing time and labor while retaining the same sensitivity and specificity. The three targeted RNA viruses are regulated pathogens for California's mandatory "Section 3701

  11. Photographic Remote Sensing of Sick Citrus Trees

    NASA Technical Reports Server (NTRS)

    Gausman, H. W.

    1971-01-01

    Remote sensing with infrared color aerial photography (Kodak Ektachrome Infrared Aero 8443 film) for detecting citrus tree anomalies is described. Illustrations and discussions are given for detecting nutrient toxicity symptoms, for detecting foot rot and sooty mold fungal diseases, and for distinguishing among citrus species. Also, the influence of internal leaf structure on light reflectance, transmittance, and absorptance are considered; and physiological and environmental factors that affect citrus leaf light reflectance are reviewed briefly and illustrated.

  12. Principles of Carbon Catabolite Repression in the Rice Blast Fungus: Tps1, Nmr1-3, and a MATE–Family Pump Regulate Glucose Metabolism during Infection

    PubMed Central

    Hartline, David; Quispe, Cristian F.; Madayiputhiya, Nandakumar; Wilson, Richard A.

    2012-01-01

    Understanding the genetic pathways that regulate how pathogenic fungi respond to their environment is paramount to developing effective mitigation strategies against disease. Carbon catabolite repression (CCR) is a global regulatory mechanism found in a wide range of microbial organisms that ensures the preferential utilization of glucose over less favourable carbon sources, but little is known about the components of CCR in filamentous fungi. Here we report three new mediators of CCR in the devastating rice blast fungus Magnaporthe oryzae: the sugar sensor Tps1, the Nmr1-3 inhibitor proteins, and the multidrug and toxin extrusion (MATE)–family pump, Mdt1. Using simple plate tests coupled with transcriptional analysis, we show that Tps1, in response to glucose-6-phosphate sensing, triggers CCR via the inactivation of Nmr1-3. In addition, by dissecting the CCR pathway using Agrobacterium tumefaciens-mediated mutagenesis, we also show that Mdt1 is an additional and previously unknown regulator of glucose metabolism. Mdt1 regulates glucose assimilation downstream of Tps1 and is necessary for nutrient utilization, sporulation, and pathogenicity. This is the first functional characterization of a MATE–family protein in filamentous fungi and the first description of a MATE protein in genetic regulation or plant pathogenicity. Perturbing CCR in Δtps1 and MDT1 disruption strains thus results in physiological defects that impact pathogenesis, possibly through the early expression of cell wall–degrading enzymes. Taken together, the importance of discovering three new regulators of carbon metabolism lies in understanding how M. oryzae and other pathogenic fungi respond to nutrient availability and control development during infection. PMID:22570632

  13. Citrus Allergy from Pollen to Clinical Symptoms

    PubMed Central

    Iorio, Rosa Anna; Del Duca, Stefano; Calamelli, Elisabetta; Pula, Chiara; Lodolini, Magda; Scamardella, Fortuna; Pession, Andrea; Ricci, Giampaolo

    2013-01-01

    Allergy to citrus fruits is often associated with pollinosis and sensitization to other plants due to a phenomenon of cross-reactivity. The aims of the present study were to highlight the cross-reactivity among citrus and the major allergenic pollens/fruits, throughout clinical and molecular investigations, and to evaluate the sensitization frequency to citrus fruits in a population of children and adults with pollinosis. We found a relevant percentage of sensitisation (39%) to citrus fruits in the patients recruited and in all of them the IgE-mediated mechanism has been confirmed by the positive response to the prick-to-prick test. RT-PCR experiments showed the expression of Cit s 1, Cit s 3 and a profilin isoform, already described in apple, also in Citrus clementine pollen. Data of multiple sequence alignments demonstrated that Citrus allergens shared high percentage identity values with other clinically relevant species (i.e. Triticum aestivum, Malus domestica), confirming the possible cross-allergenicity citrus/grasses and citrus/apple. Finally, a novelty of the present work has been the expression of two phospholipaseA2 isoforms (PLA2 α and β) in Citrus as well as in Triticum pollens; being PLA2 able to generate pro-inflammatory factors, this enzyme could participate in the activation of the allergenic inflammatory cascade. PMID:23308273

  14. Symptoms induced by transgenic expression of p23 from Citrus tristeza virus in phloem-associated cells of Mexican lime mimic virus infection without the aberrations accompanying constitutive expression.

    PubMed

    Soler, Nuria; Fagoaga, Carmen; López, Carmelo; Moreno, Pedro; Navarro, Luis; Flores, Ricardo; Peña, Leandro

    2015-05-01

    Citrus tristeza virus (CTV) is phloem restricted in natural citrus hosts. The 23-kDa protein (p23) encoded by the virus is an RNA silencing suppressor and a pathogenicity determinant. The expression of p23, or its N-terminal 157-amino-acid fragment comprising the zinc finger and flanking basic motifs, driven by the constitutive 35S promoter of cauliflower mosaic virus, induces CTV-like symptoms and other aberrations in transgenic citrus. To better define the role of p23 in CTV pathogenesis, we compared the phenotypes of Mexican lime transformed with p23-derived transgenes from the severe T36 and mild T317 CTV isolates under the control of the phloem-specific promoter from Commelina yellow mottle virus (CoYMV) or the 35S promoter. Expression of the constructs restricted to the phloem induced a phenotype resembling CTV-specific symptoms (vein clearing and necrosis, and stem pitting), but not the non-specific aberrations (such as mature leaf epinasty and yellow pinpoints, growth cessation and apical necrosis) observed when p23 was ectopically expressed. Furthermore, vein necrosis and stem pitting in Mexican lime appeared to be specifically associated with p23 from T36. Phloem-specific accumulation of the p23Δ158-209(T36) fragment was sufficient to induce the same anomalies, indicating that the region comprising the N-terminal 157 amino acids of p23 is responsible (at least in part) for the vein clearing, stem pitting and, possibly, vein corking in this host.

  15. Huanglongbing alters the structure and functional diversity of microbial communities associated with citrus rhizosphere

    PubMed Central

    Trivedi, Pankaj; He, Zhili; Van Nostrand, Joy D; Albrigo, Gene; Zhou, Jizhong; Wang, Nian

    2012-01-01

    The diversity and stability of bacterial communities present in the rhizosphere heavily influence soil and plant quality and ecosystem sustainability. The goal of this study is to understand how ‘Candidatus Liberibacter asiaticus' (known to cause Huanglongbing, HLB) influences the structure and functional potential of microbial communities associated with the citrus rhizosphere. Clone library sequencing and taxon/group-specific quantitative real-time PCR results showed that ‘Ca. L. asiaticus' infection restructured the native microbial community associated with citrus rhizosphere. Within the bacterial community, phylum Proteobacteria with various genera typically known as successful rhizosphere colonizers were significantly greater in clone libraries from healthy samples, whereas phylum Acidobacteria, Actinobacteria and Firmicutes, typically more dominant in the bulk soil were higher in ‘Ca. L. asiaticus'-infected samples. A comprehensive functional microarray GeoChip 3.0 was used to determine the effects of ‘Ca. L. asiaticus' infection on the functional diversity of rhizosphere microbial communities. GeoChip analysis showed that HLB disease has significant effects on various functional guilds of bacteria. Many genes involved in key ecological processes such as nitrogen cycling, carbon fixation, phosphorus utilization, metal homeostasis and resistance were significantly greater in healthy than in the ‘Ca. L. asiaticus'-infected citrus rhizosphere. Our results showed that the microbial community of the ‘Ca. L. asiaticus'-infected citrus rhizosphere has shifted away from using more easily degraded sources of carbon to the more recalcitrant forms. Overall, our study provides evidence that the change in plant physiology mediated by ‘Ca. L. asiaticus' infection could elicit shifts in the composition and functional potential of rhizosphere microbial communities. In the long term, these fluctuations might have important implications for the productivity and

  16. In vitro and in vivo effect of Citrus limon essential oil against sarcoptic mange in rabbits

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effect of lemon oil (Citrus limon) on Sarcoptes scabiei var. cuniculi was evaluated in vitro and in vivo. The mite samples were collected from naturally infected rabbits. The lemon oil was prepared in six concentrations by dilution with distilled water (2.5, 5, 10, 20, 50, and 100 %). In vitro a...

  17. History and diversity of Citrus leprosis virus recorded in herbarium specimens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus leprosis disease is at the same time an emerging, a current and a historical problem. Two distinct viruses are associated with this disease, one that produces particles primarily in the nucleus of infected plant cells (CiLV-N; Dichorhavirus) and another, much more widespread type that produc...

  18. Recovery of pectic hydrocolloids and phenolics from Huanglongbing related dropped citrus fruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus fruit drop has dramatically increased concomitantly with continued declining tree health and crop harvest size caused by HLB infection. This loss of harvestable fruit has been detrimental to both growers and processors. If this fruit could be recovered and converted to alternative value added...

  19. Rice SAPs are responsive to multiple biotic stresses and overexpression of OsSAP1, an A20/AN1 zinc-finger protein, enhances the basal resistance against pathogen infection in tobacco.

    PubMed

    Tyagi, Himani; Jha, Shweta; Sharma, Meenakshi; Giri, Jitender; Tyagi, Akhilesh K

    2014-08-01

    Eukaryotic A20/AN1 zinc-finger proteins (ZFPs) play an important role in the regulation of immune and stress response. After elucidation of the role of first such protein, OsSAP1, in abiotic stress tolerance, 18 rice stress associated protein (SAP) genes have been shown to be regulated by multiple abiotic stresses. In the present study, expression pattern of all the 18 OsSAP genes have been analysed in response to different biotic stress simulators, in order to get insights into their possible involvement in biotic stress tolerance. Our results showed the upregulation of OsSAP1 and OsSAP11 by all biotic stress simulator treatments. Furthermore, the functional role of OsSAP1 in plant defence responses has been explored through overexpression in transgenic plants. Constitutive expression of OsSAP1 in transgenic tobacco resulted into enhanced disease resistance against virulent bacterial pathogen, together with the upregulation of known defence-related genes. Present investigation suggests that rice SAPs are responsive to multiple biotic stresses and OsSAP1 plays a key role in basal resistance against pathogen infection. This strongly supports the involvement of rice SAPs in cross-talk between biotic and abiotic stress signalling pathways, which makes them ideal candidate to design strategies for protecting crop plants against multiple stresses.

  20. Green synthesis of gold nanoparticles using Citrus fruits (Citrus limon, Citrus reticulata and Citrus sinensis) aqueous extract and its characterization.

    PubMed

    Sujitha, Mohanan V; Kannan, Soundarapandian

    2013-02-01

    This study reports the biological synthesis of gold nanoparticles by the reduction of HAuCl(4) by using citrus fruits (Citrus limon, Citrus reticulata and Citrus sinensis) juice extract as the reducing and stabilizing agent. A various shape and size of gold nanoparticles were formed when the ratio of the reactants were altered with respect to 1.0mM chloroauric acid solution. The gold nanoparticles obtained were characterized by UV-visible spectra, transmission electron microscopy (TEM) and X-ray diffraction (XRD). TEM studies showed the particles to be of various shapes and sizes and particle size ranges from 15 to 80 nm. Selected-area electron diffraction (SAED) pattern confirmed fcc phase and crystallinity of the particles. The X-ray diffraction analysis revealed the distinctive facets (111, 200, 220 and 222 planes) of gold nanoparticles. Dynamic light scattering (DLS) studies revealed that the average size for colloid gp(3) of C. limon, C. reticulata and C. sinensis are 32.2 nm, 43.4 nm and 56.7 nm respectively. The DLS graph showed that the particles size was larger and more polydispersed compared to the one observed by TEM due to the fact that the measured size also includes the bio-organic compounds enveloping the core of the Au NPs. Zeta potential value for gold nanoparticles obtained from colloid gp(3) of C. limon, C. reticulata and C. sinensis are -45.9, -37.9 and -31.4 respectively indicating the stability of the synthesized nanoparticles. Herein we propose a novel, previously unexploited method for the biological syntheses of polymorphic gold nanoparticles with potent biological applications.

  1. Green synthesis of gold nanoparticles using Citrus fruits (Citrus limon, Citrus reticulata and Citrus sinensis) aqueous extract and its characterization

    NASA Astrophysics Data System (ADS)

    Sujitha, Mohanan V.; Kannan, Soundarapandian

    2013-02-01

    This study reports the biological synthesis of gold nanoparticles by the reduction of HAuCl4 by using citrus fruits (Citrus limon, Citrus reticulata and Citrus sinensis) juice extract as the reducing and stabilizing agent. A various shape and size of gold nanoparticles were formed when the ratio of the reactants were altered with respect to 1.0 mM chloroauric acid solution. The gold nanoparticles obtained were characterized by UV-visible spectra, transmission electron microscopy (TEM) and X-ray diffraction (XRD). TEM studies showed the particles to be of various shapes and sizes and particle size ranges from 15 to 80 nm. Selected-area electron diffraction (SAED) pattern confirmed fcc phase and crystallinity of the particles. The X-ray diffraction analysis revealed the distinctive facets (1 1 1, 2 0 0, 2 2 0 and 2 2 2 planes) of gold nanoparticles. Dynamic light scattering (DLS) studies revealed that the average size for colloid gp3 of C. limon, C. reticulata and C. sinensis are 32.2 nm, 43.4 nm and 56.7 nm respectively. The DLS graph showed that the particles size was larger and more polydispersed compared to the one observed by TEM due to the fact that the measured size also includes the bio-organic compounds enveloping the core of the Au NPs. Zeta potential value for gold nanoparticles obtained from colloid gp3 of C. limon, C. reticulata and C. sinensis are -45.9, -37.9 and -31.4 respectively indicating the stability of the synthesized nanoparticles. Herein we propose a novel, previously unexploited method for the biological syntheses of polymorphic gold nanoparticles with potent biological applications.

  2. 7 CFR 319.56-38 - Citrus from Chile.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 5 2012-01-01 2012-01-01 false Citrus from Chile. 319.56-38 Section 319.56-38... SERVICE, DEPARTMENT OF AGRICULTURE FOREIGN QUARANTINE NOTICES Fruits and Vegetables § 319.56-38 Citrus from Chile. Clementines (Citrus reticulata Blanco var. Clementine), mandarins (Citrus reticulata...

  3. 7 CFR 319.56-38 - Citrus from Chile.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 5 2014-01-01 2014-01-01 false Citrus from Chile. 319.56-38 Section 319.56-38... SERVICE, DEPARTMENT OF AGRICULTURE FOREIGN QUARANTINE NOTICES Fruits and Vegetables § 319.56-38 Citrus from Chile. Clementines (Citrus reticulata Blanco var. Clementine), mandarins (Citrus reticulata...

  4. 7 CFR 319.56-38 - Citrus from Chile.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 5 2013-01-01 2013-01-01 false Citrus from Chile. 319.56-38 Section 319.56-38... SERVICE, DEPARTMENT OF AGRICULTURE FOREIGN QUARANTINE NOTICES Fruits and Vegetables § 319.56-38 Citrus from Chile. Clementines (Citrus reticulata Blanco var. Clementine), mandarins (Citrus reticulata...

  5. Cooperation in the Conservation of Citrus Genetic Resources: Riverside, California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A consortium of cooperating programs for the conservation and utilization of citrus genetic resources is centered at the University of California, Riverside (UCR). University units include the Citrus Clonal Protection Program (CCPP), Citrus Variety Collection (CVC), and Citrus Breeding Program (CBP...

  6. A comparison of plant species for rearing Asian citrus psyllid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Five plant genotypes were compared with respect to Asian citrus psyllid (ACP) reproduction potential: Bergera koenigii, Citrus aurantiifolia, C. macrophylla, C. taiwanica and Murraya paniculata. Asian citrus psyllid reproduction is dependent on young flush and thus Asian citrus psyllid production po...

  7. Investigating alternatives to traditional insecticides: effectiveness of entomopathogenic fungi and Bacillus thuringiensis against citrus thrips and avocado thrips (Thysanoptera: Thripidae).

    PubMed

    Zahn, Deane K; Morse, Joseph G

    2013-02-01

    Citrus thrips, Scirtothrips citri (Moulton) (Thysanoptera: Thripidae), is a plant-feeding pest most widely recognized for causing damage to citrus (Citrus spp. L. [Rutaceae]) and mango (Mangifera indica L. [Anacardiaceae]) fruits. This insect has recently broadened its known host range to become a significant pest of California grown blueberries. Avocado thrips, Scirtothrips. perseae Nakahara (Thysanoptera: Thripidae), is a recent, invasive pest of California avocados, Persea americana Mill. (Laurales: Lauraceae). Effective alternatives to traditional pesticides are desirable for both pests to reduce impacts on natural enemies and broaden control options in an effort to minimize pesticide resistance via rotation of control materials. We evaluated Bacillus thuringiensis (Bt) subsp. israelensis proteins (Cyt 1A and Cry 11A, activated and inactivated) and multiple strains (GHA, 1741ss, SFBb1, S44ss, NI1ss, and 3769ss) of Beauveria bassiana (Balsamo) Vuillemin against both species. Avocado thrips and citrus thrips were not susceptible to either Bt protein tested, regardless of activation status. All strains of B. bassiana were able to infect both avocado thrips and citrus thrips. However, the commercially available GHA strain was the most effective strain against both species and had a faster rate of infection then the other strains tested. Citrus thrips were more susceptible than avocado thrips to all B. bassiana strains (LC50 and LC95 of 8.6 x 10(4) and 4.8 x 10(6) conidia per ml for citrus thrips, respectively). Investigation of citrus thrips field control using the GHA strain of B. bassiana is therefore justified.

  8. Rapid cyling plant breeding in citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Resistance or tolerance to huanglongbing (HLB) and other important traits have been identified in several citrus types and relatives and associated markers should be identified soon. What is urgently needed in addition is an accelerated strategy for citrus variety breeding. Identification and use of...

  9. Functional analysis of block deesterified citrus pectins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    After removal of soluble sugars and other compounds by washing, citrus peel is largely composed of pectin, cellulose and hemicellulose. In order to utilize the greatest amount of citrus peel product, it would appear reasonable that one or all three of these polysaccharides be converted to a useful ...

  10. Citrus Community College District Mentor Program.

    ERIC Educational Resources Information Center

    Hinds, Diane; Sprague, Caroline

    The Citrus College faculty and staff addressed the problem of lower career aspirations among women by establishing a support group, Citrus Women in Higher Education (CWHE). In addition to group meetings and special programs of interest, the CWHE has developed a mentor program to address the problem of blocked careers faced by women. The goals of…

  11. Resistance and tolerance to Huanglongbing in citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Stover-led research team focuses on developing improved citrus scion varieties, through both conventional breeding and transgenics, with special attention to confronting the threat of citrus huanglongbing (HLB). Collaborations are underway with more than 20 research groups around the world to ha...

  12. Cryopreservation and Cryotherapy of Citrus Cultivars

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Long-term conservation of Citrus clones can be accomplished by cryopreservation. Shoot tips will survive liquid nitrogen exposure and storage when appropriately desiccated and treated with cryoprotectant solutions. In our research, vegetative Citrus budwood is shipped from Riverside to Fort Collin...

  13. Conservation of citrus germplasm - an international survey

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus is an economically important tree fruit crop in many subtropical and tropical areas. Most cultivated species likely originated in Southern China, Northeast India and Southeast Asia. Many species are inter-fertile and some cultivated citrus types including sweet orange, lemon and grapefruit, ...

  14. Characterization of Pseudomonas syringae pv. syringae, Causal Agent of Citrus Blast of Mandarin in Montenegro.

    PubMed

    Ivanović, Žarko; Perović, Tatjana; Popović, Tatjana; Blagojević, Jovana; Trkulja, Nenad; Hrnčić, Snježana

    2017-02-01

    Citrus blast caused by bacterium Pseudomonas syringae is a very important disease of citrus occuring in many areas of the world, but with few data about genetic structure of the pathogen involved. Considering the above fact, this study reports genetic characterization of 43 P. syringae isolates obtained from plant tissue displaying citrus blast symptoms on mandarin (Citrus reticulata) in Montenegro, using multilocus sequence analysis of gyrB, rpoD, and gap1 gene sequences. Gene sequences from a collection of 54 reference pathotype strains of P. syringae from the Plant Associated and Environmental Microbes Database (PAMDB) was used to establish a genetic relationship with our isolates obtained from mandarin. Phylogenetic analyses of gyrB, rpoD, and gap1 gene sequences showed that P. syringae pv. syringae causes citrus blast in mandarin in Montenegro, and belongs to genomospecies 1. Genetic homogeneity of isolates suggested that the Montenegrian population might be clonal which indicates a possible common source of infection. These findings may assist in further epidemiological studies of this pathogen and for determining mandarin breeding strategies for P. syringae control.

  15. Combination of Kluyveromyces marxianus and sodium bicarbonate for controlling green mold of citrus fruit.

    PubMed

    Geng, Peng; Chen, Shaohua; Hu, Meiying; Rizwan-Ul-Haq, Muhammad; Lai, Kaiping; Qu, Fei; Zhang, Yanbo

    2011-12-02

    Biocontrol efficacy of an antagonistic yeast Kluyveromyces marxianus was evaluated individually or in combination with sodium bicarbonate (SBC) against green mold of citrus fruit caused by Penicillium digitatum. Their effects on postharvest quality of citrus fruit were also investigated. The results indicated that the antagonistic activity of K. marxianus at 1×10⁸ CFU/mL on green mold of citrus fruit was enhanced by 2% SBC treatment. In artificial inoculation trials, disease control after 3 and 6 days, respectively, with the mixture of K. marxianus and 2% SBC (18.33%, 58.33%) was significantly improved over that obtained with K. marxianus (41.67%, 70.00%) or SBC (43.33%, 81.67%) alone. The combination of K. marxianus with SBC was as effective as the imazalil treatment in natural infection trials, which gave about 90% control of green mold. Addition of 2% SBC significantly stimulated the growth of K. marxianus in citrus fruit wounds after 72 h. Moreover, K. marxianus, SBC and their combination did not impair quality parameters including weight loss, fruit firmness, total soluble solids, titratable acidity and ascorbic acid at 4 °C for 30 days followed by 20 °C for 15 days. These results suggested that the use of SBC is a useful approach to improve the efficacy of K. marxianus for the postharvest green mold of citrus fruit.

  16. Characterization of Pseudomonas syringae pv. syringae, Causal Agent of Citrus Blast of Mandarin in Montenegro

    PubMed Central

    Ivanović, Žarko; Perović, Tatjana; Popović, Tatjana; Blagojević, Jovana; Trkulja, Nenad; Hrnčić, Snježana

    2017-01-01

    Citrus blast caused by bacterium Pseudomonas syringae is a very important disease of citrus occuring in many areas of the world, but with few data about genetic structure of the pathogen involved. Considering the above fact, this study reports genetic characterization of 43 P. syringae isolates obtained from plant tissue displaying citrus blast symptoms on mandarin (Citrus reticulata) in Montenegro, using multilocus sequence analysis of gyrB, rpoD, and gap1 gene sequences. Gene sequences from a collection of 54 reference pathotype strains of P. syringae from the Plant Associated and Environmental Microbes Database (PAMDB) was used to establish a genetic relationship with our isolates obtained from mandarin. Phylogenetic analyses of gyrB, rpoD, and gap1 gene sequences showed that P. syringae pv. syringae causes citrus blast in mandarin in Montenegro, and belongs to genomospecies 1. Genetic homogeneity of isolates suggested that the Montenegrian population might be clonal which indicates a possible common source of infection. These findings may assist in further epidemiological studies of this pathogen and for determining mandarin breeding strategies for P. syringae control. PMID:28167885

  17. Citrus quarantine, sanitary and certification programs in the USA. Prevention of introduction and distribution of citrus diseases

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus germplasm originated in Australasia, the Far East, and Africa, thus all citrus grown in the New World was imported. This importation of citrus also resulted in importation of graft transmissible pathogens of citrus, many of which are latent in their original host but can cause epidemics of t...

  18. Antennal and behavioral response of the Asian citrus psyllid (Diaphorina citri Kuwayama) to degradation products of citrus volatiles

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae) vectors the bacterial causal pathogen of the deadly citrus disease, Huanglongbing (citrus greening) which is a major threat to citrus industry worldwide. We studied antennal and behavioral responses to principal components of head...

  19. Tolerance of the trifoliate citrus hybrid US-897 (Citrus reticulata Blanco x Poncirus trifoliata L. Raf.) to Huanglongbing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB) is a destructive disease of citrus in most citrus-producing countries worldwide. The disease, presumably caused by phloem-limited bacteria of the genus Candidatus Liberibacter, affects all known citrus species and citrus relatives with little known resistance. Typical disease s...

  20. The WRKY Transcription Factor Family in Citrus: Valuable and Useful Candidate Genes for Citrus Breeding.

    PubMed

    Ayadi, M; Hanana, M; Kharrat, N; Merchaoui, H; Marzoug, R Ben; Lauvergeat, V; Rebaï, A; Mzid, R

    2016-10-01

    WRKY transcription factors belong to a large family of plant transcriptional regulators whose members have been reported to be involved in a wide range of biological roles including plant development, adaptation to environmental constraints and response to several diseases. However, little or poor information is available about WRKY's in Citrus. The recent release of completely assembled genomes sequences of Citrus sinensis and Citrus clementina and the availability of ESTs sequences from other citrus species allowed us to perform a genome survey for Citrus WRKY proteins. In the present study, we identified 100 WRKY members from C. sinensis (51), C. clementina (48) and Citrus unshiu (1), and analyzed their chromosomal distribution, gene structure, gene duplication, syntenic relation and phylogenetic analysis. A phylogenetic tree of 100 Citrus WRKY sequences with their orthologs from Arabidopsis has distinguished seven groups. The CsWRKY genes were distributed across all ten sweet orange chromosomes. A comprehensive approach and an integrative analysis of Citrus WRKY gene expression revealed variable profiles of expression within tissues and stress conditions indicating functional diversification. Thus, candidate Citrus WRKY genes have been proposed as potentially involved in fruit acidification, essential oil biosynthesis and abiotic/biotic stress tolerance. Our results provided essential prerequisites for further WRKY genes cloning and functional analysis with an aim of citrus crop improvement.

  1. Infection

    DTIC Science & Technology

    2010-09-01

    standing, diagnosis, and treatment of musculoskeletal infections. Key Words: musculoskeletal infection, biofilm , bacteria, biomaterial (J Orthop Trauma...form a biofilm , or slime layer.1 The recurrence of infections is often the result of microbial biofilm formation on the implant, enabling the persistence...Klebsiella pneumoniae). Staphylococcus species is by far the most studied pathogen in musculoskeletal infections and can produce a multilayered biofilm

  2. Annotation of the Asian citrus psyllid genome reveals a reduced innate immune system

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus production worldwide is currently facing significant losses due to citrus greening disease, also known as huanglongbing. The citrus greening bacteria, Candidatus Liberibacter asiaticus (CLas), is a persistent propagative pathogen transmitted by the Asian citrus psyllid, Diaphorina citri Kuway...

  3. Small RNA profiling reveals phosphorus deficiency as a contributing factor in symptom expression for citrus huanglongbing disease.

    PubMed

    Zhao, Hongwei; Sun, Ruobai; Albrecht, Ute; Padmanabhan, Chellappan; Wang, Airong; Coffey, Michael D; Girke, Thomas; Wang, Zonghua; Close, Timothy J; Roose, Mikeal; Yokomi, Raymond K; Folimonova, Svetlana; Vidalakis, Georgios; Rouse, Robert; Bowman, Kim D; Jin, Hailing

    2013-03-01

    Huanglongbing (HLB) is a devastating citrus disease that is associated with bacteria of the genus 'Candidatus Liberibacter' (Ca. L.). Powerful diagnostic tools and management strategies are desired to control HLB. Host small RNAs (sRNA) play a vital role in regulating host responses to pathogen infection and are used as early diagnostic markers for many human diseases, including cancers. To determine whether citrus sRNAs regulate host responses to HLB, sRNAs were profiled from Citrus sinensis 10 and 14 weeks post grafting with Ca. L. asiaticus (Las)-positive or healthy tissue. Ten new microRNAs (miRNAs), 76 conserved miRNAs, and many small interfering RNAs (siRNAs) were discovered. Several miRNAs and siRNAs were highly induced by Las infection, and can be potentially developed into early diagnosis markers of HLB. miR399, which is induced by phosphorus starvation in other plant species, was induced specifically by infection of Las but not Spiroplasma citri that causes citrus stubborn-a disease with symptoms similar to HLB. We found a 35% reduction of phosphorus in Las-positive citrus trees compared to healthy trees. Applying phosphorus oxyanion solutions to HLB-positive sweet orange trees reduced HLB symptom severity and significantly improved fruit production during a 3-year field trial in south-west Florida. Our molecular, physiological, and field data suggest that phosphorus deficiency is linked to HLB disease symptomology.

  4. Postharvest decay control of citrus fruit by preharvest pyrimethanil spray.

    PubMed

    D'Aquino, S; Angioni, A; Suming, D; Palma, A; Schirra, M

    2013-01-01

    Preharvest infections or conidia load on fruit surface by Penicillium digitatum, P. italicum, Alternaria citri and other filamentous fungi can cause important postharvest losses of citrus fruit. Reduction in pruning frequency occurred in the last decade together with un-picked yield that eventually rots on the trees have increased the risk of postharvest decay especially when environmental conditions at picking time are favourable to pathogens' development. Sanitation procedures in the packinghouses, alternate use of postharvest fungicides with different modes of action, along with fungicide application before harvest could be an effective approach to minimize postharvest decay in citrus fruit. The present study investigated the effectiveness of a preharvest treatment with pyrimethanil (PYR), a broad spectrum fungicide, recently registered in different citrus-producing countries for postharvest treatments of citrus fruit and widely used worldwide as a preharvest treatment to control various diseases in different crops. PYR (750 mg/L) was sprayed by a hand-back sprayer at run-off on 'Fremont' mandarins. The day after the treatment, half of the trees were sprayed with a 10(4) conidial suspension of P. digitatum at run-off. Fruit were harvested following 2 or 4 weeks from treatments. Sound or either wounded 2-mm-deep and 2-mm-wide or superficial wound-scratched fruit were stored at 20 degrees C and 90% RH and inspected for decay after 1, 2 or 3 weeks of storage. In fruit harvested after 2 weeks from field treatment, PYR remarkably reduced decay development during two weeks of storage in sound fruit and in wound-scratched fruit and was fairly effective even after 4 weeks from treatment, but was ineffective in fruit wounded 2 mm deep and 2 mm wide. PYR was also effective in reducing preharvest decay incited by P. digitatum, P. italicum and Botrytis cinerea, but not by other pathogens. Results show that preharvest treatment with PYR could be a feasible approach to reduce

  5. Transcriptome Profiling of Citrus Fruit Response to Huanglongbing Disease

    PubMed Central

    Martinelli, Federico; Uratsu, Sandra L.; Albrecht, Ute; Reagan, Russell L.; Phu, My L.; Britton, Monica; Buffalo, Vincent; Fass, Joseph; Leicht, Elizabeth; Zhao, Weixiang; Lin, Dawei; D'Souza, Raissa; Davis, Cristina E.; Bowman, Kim D.; Dandekar, Abhaya M.

    2012-01-01

    Huanglongbing (HLB) or “citrus greening” is the most destructive citrus disease worldwide. In this work, we studied host responses of citrus to infection with Candidatus Liberibacter asiaticus (CaLas) using next-generation sequencing technologies. A deep mRNA profile was obtained from peel of healthy and HLB-affected fruit. It was followed by pathway and protein-protein network analysis and quantitative real time PCR analysis of highly regulated genes. We identified differentially regulated pathways and constructed networks that provide a deep insight into the metabolism of affected fruit. Data mining revealed that HLB enhanced transcription of genes involved in the light reactions of photosynthesis and in ATP synthesis. Activation of protein degradation and misfolding processes were observed at the transcriptomic level. Transcripts for heat shock proteins were down-regulated at all disease stages, resulting in further protein misfolding. HLB strongly affected pathways involved in source-sink communication, including sucrose and starch metabolism and hormone synthesis and signaling. Transcription of several genes involved in the synthesis and signal transduction of cytokinins and gibberellins was repressed while that of genes involved in ethylene pathways was induced. CaLas infection triggered a response via both the salicylic acid and jasmonic acid pathways and increased the transcript abundance of several members of the WRKY family of transcription factors. Findings focused on the fruit provide valuable insight to understanding the mechanisms of the HLB-induced fruit disorder and eventually developing methods based on small molecule applications to mitigate its devastating effects on fruit production. PMID:22675433

  6. Chipping citrus wood for gasifiction

    SciTech Connect

    Churchill, D.B.; Hedden, S.L.; Whitney, J.D.; Shaw, L.N.

    1984-01-01

    Both green and dead citrus trees were used for chipping. Chip moisture content, fuel analysis, drying time, and data on fuel/tonne of chips were obtained. The average moisture contents of green and dead trees when chipped were 25% and 16% (wet basis) respectively. Chips were sized to a minimum of 0.32 squared cm x 0.32 cm thick to a maximum of 5.0 cm squared x 0.32 cm thick and normally required 4 weeks to air dry to 14% (wet basis) moisture content before use. Approximately 50% of the total tree by weight could be made into usable chips. 9 references.

  7. Citrus breeding, genetics and genomics in Japan

    PubMed Central

    Omura, Mitsuo; Shimada, Takehiko

    2016-01-01

    Citrus is one of the most cultivated fruits in the world, and satsuma mandarin (Citrus unshiu Marc.) is a major cultivated citrus in Japan. Many excellent cultivars derived from satsuma mandarin have been released through the improvement of mandarins using a conventional breeding method. The citrus breeding program is a lengthy process owing to the long juvenility, and it is predicted that marker-assisted selection (MAS) will overcome the obstacle and improve the efficiency of conventional breeding methods. To promote citrus molecular breeding in Japan, a genetic mapping was initiated in 1987, and the experimental tools and resources necessary for citrus functional genomics have been developed in relation to the physiological analysis of satsuma mandarin. In this paper, we review the progress of citrus breeding and genome researches in Japan and report the studies on genetic mapping, expression sequence tag cataloguing, and molecular characterization of breeding characteristics, mainly in terms of the metabolism of bio-functional substances as well as factors relating to, for example, fruit quality, disease resistance, polyembryony, and flowering. PMID:27069387

  8. CIBMAN: Database exploring Citrus biodiversity of Manipur

    PubMed Central

    Sanabam, Rakesh; Somkuwar, Bharat Gopalrao; Thingnam, Gourshyam; Moirangthem, Sila; Handique, Pratap Jyoti; Huidrom, Sunitibala

    2012-01-01

    The rich wealth of Citrus genetic resources makes India to enjoy a remarkable position in the “Citrus belt of the world”. We have developed CIBMAN, a unique database on Citrus biodiversity of Manipur which comprises 33 accessions collected through extensive survey for more than three years. CIBMAN provides integrated access to Citrus species through sophisticated web interface which has following capabilities a) morphological details, b) socio-economic details, c) taxonomic details and d) geographical distribution. Morphological variability among Citrus accessions is due to variance in their genome which contributes to diverse agronomical traits and diverse bioactive compounds of high value. This diverse gene pool can be potential source for genetic improvement of existing cultivars and rootstocks. Systematic collection, characterization and conservation of the underutilized or lesser exploited varieties is required for incorporating in breeding program and conserve the germplasm from ever going on genetic erosion. This database will be useful for scientific validations and updating of traditional wisdom in bioprospecting aspects especially industrialization of Citrus found in the state. Further, the features will be suited for detailed investigation on potential medicinal and edible Citrus that make CIBMAN a powerful tool for sustainable management. Availability http://ibsd.gov.in/cibman PMID:23139595

  9. GC-MS metabolomic differentiation of selected citrus varieties with different sensitivity to citrus huanglongbing.

    PubMed

    Cevallos-Cevallos, Juan M; Futch, David B; Shilts, Turksen; Folimonova, Svetlana Y; Reyes-De-Corcuera, José I

    2012-04-01

    Huanglongbing (HLB) is the most destructive disease of citrus worldwide. The rapid identification of tolerant varieties is considered a critical step towards controlling HLB. GC-MS metabolite profiles were used to differentiate HLB-tolerant citrus varieties 'Poncirus trifoliata' (TR) and 'Carrizo citrange' (CAR) from HLB-sensitive varieties 'Madam Vinous sweet orange' (MV) and 'Duncan' grapefruit (DG). PCR analyses revealed that MV was the most sensitive variety followed by DG and the tolerant varieties CAR and TR. Metabolomic multivariate analysis allowed classification of the cultivars in apparent agreement with PCR results. Higher levels of the amino acids l-proline, l-serine, and l-aspartic acid, as well as the organic acids butanedioic and tetradecanoic acid, and accumulation of galactose in healthy plants were characteristic of the most sensitive variety MV when compared to all other varieties. Only galactose was significantly higher in DG when compared to the tolerant varieties TR and CAR. The tolerant varieties showed higher levels of l-glycine and mannose when compared to sensitive varieties MV and DG. Profiling of the sensitive varieties MV and DG over a 20-week period after inoculation of those with the HLB-containing material revealed strong responses of metabolites to HLB infection that differed from the response of the tolerant varieties. Significant changes of l-threonine level in the leaves from old mature flushes and l-serine, l-threonine, scyllo-inositol, hexadecanoic acid, and mannose in the leaves from young developing flushes were observed in MV. Significant changes in myo-inositol in old flushes and l-proline, indole, and xylose in new flushes were observed in DG.

  10. Genome assembly of the fungus Cochliobolus miyabeanus, and transcriptome analysis during early stages of infection on American wild rice (Zizania palustris L.)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cochliobolus miyabeanus causes a severe, yield-reducing leaf spot disease on rice (Oryza sativa) and two North American specialty crops, American wildrice (Zizania palustris) and switchgrass (Panicum virgatum). Despite the importance of the pathogen in wildrice, little is known about mechanisms of p...

  11. Comprehensive analysis of CCCH-type zinc finger gene family in citrus (Clementine mandarin) by genome-wide characterization.

    PubMed

    Liu, Shengrui; Khan, Muhammad Rehman Gul; Li, Yongping; Zhang, Jinzhi; Hu, Chungen

    2014-10-01

    The CCCH-type zinc finger proteins comprise a large gene family of regulatory proteins and are widely distributed in eukaryotic organisms. The CCCH proteins have been implicated in multiple biological processes and environmental responses in plants. Little information is available, however, about CCCH genes in plants, especially in woody plants such as citrus. The release of the whole-genome sequence of citrus allowed us to perform a genome-wide analysis of CCCH genes and to compare the identified proteins with their orthologs in model plants. In this study, 62 CCCH genes and a total of 132 CCCH motifs were identified, and a comprehensive analysis including the chromosomal locations, phylogenetic relationships, functional annotations, gene structures and conserved motifs was performed. Distribution mapping revealed that 54 of the 62 CCCH genes are unevenly dispersed on the nine citrus chromosomes. Based on phylogenetic analysis and gene structural features, we constructed 5 subfamilies of 62 CCCH members and integrative subfamilies from citrus, Arabidopsis, and rice, respectively. Importantly, large numbers of SNPs and InDels in 26 CCCH genes were identified from Poncirus trifoliata and Fortunella japonica using whole-genome deep re-sequencing. Furthermore, citrus CCCH genes showed distinct temporal and spatial expression patterns in different developmental processes and in response to various stress conditions. Our comprehensive analysis of CleC3Hs is a valuable resource that further elucidates the roles of CCCH family members in plant growth and development. In addition, variants and comparative genomics analyses deepen our understanding of the evolution of the CCCH gene family and will contribute to further genetics and genomics studies of citrus and other plant species.

  12. Transcriptome Profiling of Huanglongbing (HLB) Tolerant and Susceptible Citrus Plants Reveals the Role of Basal Resistance in HLB Tolerance

    PubMed Central

    Wang, Yunsheng; Zhou, Lijuan; Yu, Xiaoyue; Stover, Ed; Luo, Feng; Duan, Yongping

    2016-01-01

    Huanglongbing (HLB) is currently the most destructive disease of citrus worldwide. Although there is no immune cultivar, field tolerance to HLB within citrus and citrus relatives has been observed at the USDA Picos farm at Ft. Pierce, Florida, where plants have been exposed to a very high level of HLB pressure since 2006. In this study, we used RNA-Seq to evaluate expression differences between two closely related cultivars after HLB infection: HLB-tolerant “Jackson” grapefruit-like-hybrid trees and HLB susceptible “Marsh” grapefruit trees. A total of 686 genes were differentially expressed (DE) between the two cultivars. Among them, 247 genes were up-expressed and 439 were down-expressed in tolerant citrus trees. We also identified a total of 619 genes with significant differential expression of alternative splicing isoforms between HLB tolerant and HLB susceptible citrus trees. We analyzed the functional categories of DE genes using two methods, and revealed that multiple pathways have been suppressed or activated in the HLB tolerant citrus trees, which lead to the activation of the basal resistance or immunity of citrus plants. We have experimentally verified the expressions of 14 up-expressed genes and 19 down-expressed genes on HLB-tolerant “Jackson” trees and HLB-susceptible “Marsh” trees using real time PCR. The results showed that the expression of most genes were in agreement with the RNA-Seq results. This study provided new insights into HLB-tolerance and useful guidance for breeding HLB-tolerant citrus in the future. PMID:27446161

  13. Creation of transgenic rice plants producing small interfering RNA of Rice tungro spherical virus.

    PubMed

    Le, Dung Tien; Chu, Ha Duc; Sasaya, Takahide

    2015-01-01

    Rice tungro spherical virus (RTSV), also known as Rice waika virus, does not cause visible symptoms in infected rice plants. However, the virus plays a critical role in spreading Rice tungro bacilliform virus (RTBV), which is the major cause of severe symptoms of rice tungro disease. Recent studies showed that RNA interference (RNAi) can be used to develop virus-resistance transgenic rice plants. In this report, we presented simple procedures and protocols needed for the creation of transgenic rice plants capable of producing small interfering RNA specific against RTSV sequences. Notably, our study showed that 60 out of 64 individual hygromycin-resistant lines (putative transgenic lines) obtained through transformation carried transgenes designed for producing hairpin double-stranded RNA. Northern blot analyses revealed the presence of small interfering RNA of 21- to 24-mer in 46 out of 56 confirmed transgenic lines. Taken together, our study indicated that transgenic rice plants carrying an inverted repeat of 500-bp fragments encoding various proteins of RTSV can produce small interfering RNA from the hairpin RNA transcribed from that transgene. In light of recent studies with other viruses, it is possible that some of these transgenic rice lines might be resistant to RTSV.

  14. Characterization of the Asian Citrus Psyllid Transcriptome

    PubMed Central

    Reese, Justin; Christenson, Matthew K.; Leng, Nan; Saha, Surya; Cantarel, Brandi; Lindeberg, Magdalen; Tamborindeguy, Cecilia; MacCarthy, Justin; Weaver, Daniel; Trease, Andrew J.; Ready, Steven V.; Davis, Vincent M.; McCormick, Courtney; Haudenschild, Christian; Han, Shunsheng; Johnson, Shannon L.; Shelby, Kent S.; Huang, Hong; Bextine, Blake R.; Shatters, Robert G.; Hall, David G.; Davis, Paul H.; Hunter, Wayne B.

    2014-01-01

    The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is a vector for the causative agents of Huanglongbing, which threatens citrus production worldwide. This study reports and discusses the first D. citri transcriptomes, encompassing the three main life stages of D. citri, egg, nymph and adult. The transcriptomes were annotated using Gene Ontology (GO) and insecticide-related genes within each life stage were identified to aid the development of future D. citri insecticides. Transcriptome assemblies and other sequence data are available for download at the International Asian Citrus Psyllid Genome Consortium website [http://psyllid.org/download] and at NCBI [http://www.ncbi.nlm.nih.gov/bioproject/29447]. PMID:24511328

  15. Isolation and identification of citrus psorosis virus Egyptian isolate (CPsV-EG).

    PubMed

    Ghazal, S A; El-Dougdoug, Kh A; Mousa, A A; Fahmy, H; Sofy, A R

    2008-01-01

    Citrus psorosis ophiovirus (CPsV), is considered to be of the most serious and deter mental virus pathogen's citrus species trees in Egypt. CPsV-EG was isolated from infected citrus grapefruit (C. paradisi Macf.) at Agric. Res. Centre (ARC). The grapefruit which used for CPsV-EG isolate was found to be free from CTV, CEVd and Spiroplasma citri where as gave -ve results with DTBIA, tissue print hybridization and Diene's stain respectively. CPsV-EG was detected on the basis of biological indexing by graft inoculation which gave oak leaf pattern (OLP) on Dweet tangor and serological assay by DAS-ELISA using Mab specific CPsV. CPsV-EG was reacted with variable responses on 16 host plants belonging to 6 families. Only 8 host plants are susceptible and showed visible external symptoms which appeared as local, systemic and local followed by systemic infections. CPsV-EG isolate was transmitted from infected citrus to citrus by syringe and grafting and herbaceous plants by forefinger inoculation and syringe. The woody indicators and rootstocks were differed in response to CPsV-EG isolate which appeared as no-response, response, sensitivity and hypersensitivity. The serological characters represented as the antigenic determinants of CPsV-EG isolate related to monoclonal antibodies specific CPsV strain where as appeared precipitation reaction by DAS-ELISA and DTBIA. The partial fragment of RNA3 (coat protein gene) of CPsV-EG (-1140bp and -571bp) was amplified by reverse transcription-polymerase chain reaction (RT-PCR) from grapefruit tissues using two sets primers specific CPsV (CPV3 and CPV4) and (PS66 and PS65) respectively. The virus under study was identified as CPsV-EG isolate according to biological, serological and molecular characters.

  16. Estimation of transgene copy number in transformed citrus plants by quantitative multiplex real-time PCR.

    PubMed

    Omar, Ahmad A; Dekkers, Marty G H; Graham, James H; Grosser, Jude W

    2008-01-01

    Quantitative real-time PCR (qRT-PCR) was adapted to estimate transgene copy number of the rice Xa21 gene in transgenic citrus plants. This system used TaqMan qRT-PCR and the endogenous citrus gene encoding for lipid transfer protein (LTP). Transgenic "Hamlin" sweet orange plants were generated using two different protoplast-GFP transformation systems: cotransformation and single plasmid transformation. A dilution series of genomic DNA from one of the transgenic lines was used to generate a standard curve for the endogenous LTP and the transgene Xa21. This standard curve was used for relative quantification of the endogenous gene and the transgene. Copy numbers of the transgene Xa21 detected from qRT-PCR analysis correlated with that from Southern blot analysis (r = 0.834). Thus, qRT-PCR is an efficient means of estimating copy number in transgenic citrus plants. This analysis can be performed at much earlier stages of transgenic plant development than southern blot analysis, which expedites investigation of transgenes in slow-growing woody plants.

  17. Diversity of endophytic bacterial populations and their interaction with Xylella fastidiosa in citrus plants.

    PubMed

    Araújo, Welington L; Marcon, Joelma; Maccheroni, Walter; Van Elsas, Jan Dirk; Van Vuurde, Jim W L; Azevedo, João Lúcio

    2002-10-01

    Citrus variegated chlorosis (CVC) is caused by Xylella fastidiosa, a phytopathogenic bacterium that can infect all Citrus sinensis cultivars. The endophytic bacterial communities of healthy, resistant, and CVC-affected citrus plants were studied by using cultivation as well as cultivation-independent techniques. The endophytic communities were assessed in surface-disinfected citrus branches by plating and denaturing gradient gel electrophoresis (DGGE). Dominant isolates were characterized by fatty-acid methyl ester analysis as Bacillus pumilus, Curtobacterium flaccumfaciens, Enterobacter cloacae, Methylobacterium spp. (including Methylobacterium extorquens, M. fujisawaense, M. mesophilicum, M. radiotolerans, and M. zatmanii), Nocardia sp., Pantoea agglomerans, and Xanthomonas campestris. We observed a relationship between CVC symptoms and the frequency of isolation of species of Methylobacterium, the genus that we most frequently isolated from symptomatic plants. In contrast, we isolated C. flaccumfaciens significantly more frequently from asymptomatic plants than from those with symptoms of CVC while P. agglomerans was frequently isolated from tangerine (Citrus reticulata) and sweet-orange (C. sinensis) plants, irrespective of whether the plants were symptomatic or asymptomatic or showed symptoms of CVC. DGGE analysis of 16S rRNA gene fragments amplified from total plant DNA resulted in several bands that matched those from the bacterial isolates, indicating that DGGE profiles can be used to detect some endophytic bacteria of citrus plants. However, some bands had no match with any isolate, suggesting the occurrence of other, nonculturable or as yet uncultured, endophytic bacteria. A specific band with a high G+C ratio was observed only in asymptomatic plants. The higher frequency of C. flaccumfaciens in asymptomatic plants suggests a role for this organism in the resistance of plants to CVC.

  18. Rice hull smoke extract protects mice against a salmonella lipopolysaccharide-induced endotoxemia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rice hulls accounting for 20% of the rice crop are a byproduct of post-harvest rice processing. Endotoxemia (sepsis, septic shock) is an inflammatory, virulent often fatal disease that results mainly from infection with Salmonella and other Gram-negative bacteria. The present study investigated the...

  19. Antihyperlipidemic effects of Citrus sinensis, Citrus paradisi, and their combinations

    PubMed Central

    Mallick, Neelam; Khan, Rafeeq Alam

    2016-01-01

    Objective: Hyperlipidemia, extensively contributes in the progression of coronary heart diseases and atherosclerosis, but may be managed through alterations in the nutritional pattern. Several studies show that diet rich in polyphenols and antioxidants have antiatherogenic effects. Citrus sinensis and Citrus paradisi are widely known for health benefits and have found to produce antioxidant, anti-inflammatory, and hypolipidemic effects, hence current research was planned to determine the hypolipidemic effects of C. sinensis and C. paradisi in rats receiving diet rich in cholesterol. Materials and Methods: All rats were divided into 11 groups each comprising 10 animals: Normal control group and hyperlipidemic control. C. sinensis treated three groups, C. paradisi treated three groups, C. sinensis and C. paradisi combination treated two groups, and group treated atorvastatin. All rats in the respective groups were treated orally with sterile water, juices, and standard drug for 8 weeks and lipid profile was estimated at the end of dosing. Results: Cholesterol, triglycerides (TGs), and low-density lipoprotein (LDL) were decreased at all the three doses of C. sinensis and C. paradisi but rise in high-density lipoprotein (HDL) was only significant at 8 ml/kg, and 0.3 ml/kg, respectively. Animals received the combination doses of C. sinensis and C. paradisi also showed a highly significant fall in cholesterol, LDL, and TGs, however HDL level was significantly elevated by SPJ-2 combination. Conclusion: Results suggest that C. sinensis and C. paradisi possess antihyperlipidemic activity due to phytochemicals and other essential nutrients, hence may serve as cardioprotective by preventing thrombosis. PMID:27134462

  20. Rice ( Oryza) hemoglobins

    PubMed Central

    Arredondo-Peter, Raúl; Moran, Jose F.; Sarath, Gautam

    2014-01-01

    Hemoglobins (Hbs) corresponding to non-symbiotic (nsHb) and truncated (tHb) Hbs have been identified in rice ( Oryza). This review discusses the major findings from the current studies on rice Hbs. At the molecular level, a family of the nshb genes, consisting of hb1, hb2, hb3, hb4 and hb5, and a single copy of the thb gene exist in Oryza sativa var. indica and O. sativa var. japonica, Hb transcripts coexist in rice organs and Hb polypeptides exist in rice embryonic and vegetative organs and in the cytoplasm of differentiating cells. At the structural level, the crystal structure of rice Hb1 has been elucidated, and the structures of the other rice Hbs have been modeled. Kinetic analysis indicated that rice Hb1 and 2, and possibly rice Hb3 and 4, exhibit a very high affinity for O 2, whereas rice Hb5 and tHb possibly exhibit a low to moderate affinity for O 2. Based on the accumulated information on the properties of rice Hbs and data from the analysis of other plant and non-plant Hbs, it is likely that Hbs play a variety of roles in rice organs, including O 2-transport, O 2-sensing, NO-scavenging and redox-signaling. From an evolutionary perspective, an outline for the evolution of rice Hbs is available. Rice nshb and thb genes vertically evolved through different lineages, rice nsHbs evolved into clade I and clade II lineages and rice nshbs and thbs evolved under the effect of neutral selection. This review also reveals lacunae in our ability to completely understand rice Hbs. Primary lacunae are the absence of experimental information about the precise functions of rice Hbs, the properties of modeled rice Hbs and the cis-elements and trans-acting factors that regulate the expression of rice hb genes, and the partial understanding of the evolution of rice Hbs. PMID:25653837

  1. Rice ( Oryza) hemoglobins.

    PubMed

    Arredondo-Peter, Raúl; Moran, Jose F; Sarath, Gautam

    2014-01-01

    Hemoglobins (Hbs) corresponding to non-symbiotic (nsHb) and truncated (tHb) Hbs have been identified in rice ( Oryza). This review discusses the major findings from the current studies on rice Hbs. At the molecular level, a family of the nshb genes, consisting of hb1, hb2, hb3, hb4 and hb5, and a single copy of the thb gene exist in Oryza sativa var. indica and O. sativa var. japonica, Hb transcripts coexist in rice organs and Hb polypeptides exist in rice embryonic and vegetative organs and in the cytoplasm of differentiating cells. At the structural level, the crystal structure of rice Hb1 has been elucidated, and the structures of the other rice Hbs have been modeled. Kinetic analysis indicated that rice Hb1 and 2, and possibly rice Hb3 and 4, exhibit a very high affinity for O 2, whereas rice Hb5 and tHb possibly exhibit a low to moderate affinity for O 2. Based on the accumulated information on the properties of rice Hbs and data from the analysis of other plant and non-plant Hbs, it is likely that Hbs play a variety of roles in rice organs, including O 2-transport, O 2-sensing, NO-scavenging and redox-signaling. From an evolutionary perspective, an outline for the evolution of rice Hbs is available. Rice nshb and thb genes vertically evolved through different lineages, rice nsHbs evolved into clade I and clade II lineages and rice nshbs and thbs evolved under the effect of neutral selection. This review also reveals lacunae in our ability to completely understand rice Hbs. Primary lacunae are the absence of experimental information about the precise functions of rice Hbs, the properties of modeled rice Hbs and the cis-elements and trans-acting factors that regulate the expression of rice hb genes, and the partial understanding of the evolution of rice Hbs.

  2. A mathematical model of transmission of rice tungro disease by Nephotettix Virescens

    NASA Astrophysics Data System (ADS)

    Blas, Nikki T.; Addawe, Joel M.; David, Guido

    2016-11-01

    One of the major threats in rice agriculture is the Tungro virus, which is transmitted semi-persistently to rice plants via green rice leafhoppers called Nephotettix Virescens. Tungro is polycyclic and complex disease of rice associated by dual infection with Rice Tungro Bacilliform Virus (RTBV) and Rice Tungro Spherical Virus (RTSV). Interaction of the two viruses results in the degeneration of the host. In this paper, we used a plant-vector system of ordinary differential equations to model the spread of the disease in a model rice field. Parameter values were obtained from studies on the entomology of Nephotettix Virescens and infection rates of RTSV and RTBV. The system was analyzed for equilibrium solutions, and solved numerically for susceptible rice varieties (Taichung Native 1).

  3. Brassica cover cropping for management of sheath blight of rice

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sheath blight, caused by Rhizoctonia solani, is the most important disease limiting rice production in Texas and other rice-producing states. The fungal pathogen survives between crops as soilborne sclerotia and mycelium in infected plant debris. These sclerotia and colonized plant debris float on t...

  4. Effect of Cyantraniliprole, a Novel Insecticide, on the Inoculation of Candidatus Liberibacter Asiaticus Associated with Citrus Huanglongbing by the Asian Citrus Psyllid (Hemiptera: Liviidae).

    PubMed

    Ammar, El-Desouky; Hall, David G; Alvarez, Juan M

    2015-04-01

    The Asian citrus psyllid (Diaphorina citri Kuwayama) is the principal vector of 'Candidatus Liberibacter asiaticus' (CLas) associated with huanglongbing (HLB), the most serious citrus disease worldwide. New control measures including pesticides are urgently needed to combat HLB, especially to protect young or newly planted citrus trees from CLas-inoculation by vector psyllids. Here, we tested CLas-inoculation by D. citri adults (CLas-exposed, reared on infected plants) by feeding them for 7 d on excised healthy citrus leaves with dry residues of cyantraniliprole (Exirel), a novel insecticide, in comparison with fenpropathrin (Danitol 2.4EC), an insecticide commonly used against D. citri. Fewer adults settled (putatively feeding or probing) on leaves treated with cyantraniliprole than those treated with fenpropathrin or water controls. Also, psyllid adults died at a slower rate on leaves treated with cyantraniliprole than those treated with fenpropathrin, although the final cumulative mortality did not differ between the two treatments. In quantitative real-time polymerase chain reaction tests, 59.0-65.3% of the CLas-exposed psyllid adults were proven to be CLas-positive. Inoculation rates of CLas (using 10 adults per leaf) into untreated healthy citrus leaves (47.5-85%) were significantly higher than rates into leaves treated with cyantraniliprole or fenpropathrin (2.5-12.5%). Reduced inoculation rates to leaves treated with cyantraniliprole probably occurred as a result of reduced feeding or probing by D. citri. The excised leaf assay method, which took only a few weeks compared with up to a year or longer using whole plants, can be an effective tool for testing the effect of new pesticides or other treatments in reducing CLas inoculation or transmission by psyllid vectors.

  5. Evaluation of natural colorants and their application on citrus fruit as alternatives to citrus red II

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The poor peel color of some varieties of oranges and the hybrids, especially for early season fruits, is caused by the subtropical climate of Florida, and has resulted in the use of a red dye on the peel to improve fruit appearance and marketability. Citrus Red II (CR2), the commercial citrus color ...

  6. Citrus County Schools Copyright Guidelines Recommended by the Citrus County Association of School Media Specialists.

    ERIC Educational Resources Information Center

    Citrus County School District, Inverness, FL.

    This document contains copyright guidelines determined appropriate for the Citrus County School System (Florida) by the Citrus County Association of School Media Specialists in May, 1992. These guidelines are based on interpretation and understanding of current copyright law as applied to education and implemented in school districts in the United…

  7. Apparent tolerance to huanglongbing in citrus and citrus-related germplasm

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In a Ft. Pierce, Florida field planting, growth and huanglongbing (HLB) severity were assessed as indicators of HLB-tolerance on progenies of 83 seed-source accessions of citrus and citrus-relatives from the Riverside, California genebank. The HLB-pathogen (Candidatus Liberibacter asiaticus) and vec...

  8. Rice Responses and Resistance to Planthopper-Borne Viruses at Transcriptomic and Proteomic Levels.

    PubMed

    Cui, Feng; Zhao, Wan; Luo, Lan; Kang, Le

    2016-01-01

    Rice (Oryza sativa) is one of the most important cereal crops in the world, especially in Asian areas. Rice virus diseases are considered as the most serious threat to rice yields. Most rice viruses are transmitted by hemipteran insects such as planthoppers and leafhoppers. In Asia five rice viruses are transmitted mainly by three planthopper species in a persistent manner: Rice stripe virus, Rice black-streaked dwarf virus, Rice ragged stunt virus, Rice grassy stunt virus, and Southern rice black-streaked dwarf virus. In rice antivirus studies, several individual genes have been shown to function in rice resistance to viruses. Since plant responses to viral infection are complex, system-level omic studies are required to fully understand the responses. Recently more and more omic studies have appeared in the literatures on relationships between planthoppers and viruses, employing microarray, RNA-Seq, small RNA deep sequencing, degradome sequencing, and proteomic analysis. In this paper, we review the current knowledge and progress of omic studies in rice plant responses and resistance to four planthopper-borned viruses. We also discuss progress in the omic study of the interactions of planthoppers and rice viruses. Future research directions and translational applications of fundamental knowledge of virus-vector-rice interactions are proposed.

  9. Infection

    MedlinePlus

    ... 23(4):251-69. Association for Professionals in Infection Control and Epidemiology (APIC) guideline. Back to Top Administration ... : Hospital Scope | Glossary | References | Site Map | Credits Freedom of ...

  10. Detection of citrus Huanglongbing by Fourier transform infrared-attenuated total reflection spectroscopy.

    PubMed

    Hawkins, Samantha A; Park, Bosoon; Poole, Gavin H; Gottwald, Timothy; Windham, William R; Lawrence, Kurt C

    2010-01-01

    Citrus Huanglongbing (HLB, also known as citrus greening disease) was discovered in Florida in 2005 and is spreading rapidly amongst the citrus growing regions of the state. Detection via visual symptoms of the disease is not a long-term viable option. New techniques are being developed to test for the disease in its earlier presymptomatic stages. Fourier transform infrared-attenuated total reflection (FT-IR-ATR) spectroscopy is a candidate for rapid, inexpensive, early detection of the disease. The mid-infrared region of the spectrum reveals dramatic changes that take place in the infected leaves when compared to healthy non-infected leaves. The carbohydrates that give rise to peaks in the 900-1180 cm(-1) range are reliable in distinguishing leaves from infected plants versus non-infected plants. A model based on chemometrics was developed using the spectra from 179 plants of known disease status. This model then correctly predicted the status of >95% of the plants tested.

  11. A novel virus of the genus Cilevirus causing symptoms similar to citrus leprosis.

    PubMed

    Roy, Avijit; Choudhary, Nandlal; Guillermo, Leon M; Shao, Jonathan; Govindarajulu, Ananthakrishnan; Achor, Diann; Wei, G; Picton, D D; Levy, L; Nakhla, M K; Hartung, John S; Brlansky, R H

    2013-05-01

    Citrus leprosis in Colombia was previously shown to be caused by cytoplasmic Citrus leprosis virus (CiLV-C). In 2011, enzyme-linked immunosorbent assay and reverse-transcription polymerase chain reaction (RT-PCR)-based diagnostic methods failed to identify CiLV-C from citrus samples with symptoms similar to citrus leprosis; however, virions similar to CiLV-C were observed in the cytoplasm of the symptomatic leaves by transmission electron microscopy. Furthermore, the causal organism was transmitted by the false spider mite, Brevipalpus phoenicis, to healthy citrus seedlings. A library of small RNAs was constructed from symptomatic leaves and used as the template for Illumina high-throughput parallel sequencing. The complete genome sequence and structure of a new bipartite RNA virus was determined. RNA1 (8,717 nucleotides [nt]) contained two open reading frames (ORFs). ORF1 encoded the replication module, consisting of five domains: namely, methyltransferase (MTR), cysteine protease-like, FtsJ-MTR, helicase (Hel), and RNA-dependent RNA polymerase (RdRp); whereas ORF2 encoded the putative coat protein. RNA2 (4,989 nt) contained five ORFs that encode the movement protein (MP) and four hypothetical proteins (p7, p15, p24, and p61). The structure of this virus genome resembled that of CiLV-C except that it contained a long 3' untranslated terminal region and an extra ORF (p7) in RNA2. Both the RNA1 and RNA2 of the new virus had only 58 and 50% nucleotide identities, respectively, with known CiLV-C sequences and, thus, it appears to be a novel virus infecting citrus. Phylogenetic analyses of the MTR, Hel, RdRp, and MP domains also indicated that the new virus was closely related to CiLV-C. We suggest that the virus be called Citrus leprosis virus cytoplasmic type 2 (CiLV-C2) and it should be unambiguously classified as a definitive member of the genus Cilevirus. A pair of CiLV-C2 genome-specific RT-PCR primers was designed and validated to detect its presence in citrus

  12. Postphloem, Nonvascular Transfer in Citrus

    PubMed Central

    Koch, Karen E.; Avigne, Wayne T.

    1990-01-01

    Postphloem, nonvascular assimilate transport occurs over an unusually long area in citrus fruit and thus facilitates investigation of this process relative to sugar entry into many sink structures. Labeled photosynthates moving into juice tissues of grapefruit (Citrus paradisi Macf.) slowed dramatically after entering the postphloem transport path (parenchyma cells, narrow portions of segment epidermis, and hair-like, parenchymatous stalks of juice sacs). Kinetic, metabolic, and compositional data indicated that transfer through the nonvascular area was delayed many hours by temporary storage and/or equilibration with sugars in compartments along the postphloem path. Labeled assimilates were generally recovered as sucrose throughout the path, and extent of hexose formation enroute bore no apparent relationship to the assimilate transfer process. Even after 24 hours, radiolabel was restricted to discrete, highly localized areas directly between vascular bundles and juice sacs. Postphloem transfer occurred against an ascending sucrose concentration gradient in young fruit, whereas a descending gradient (favoring diffusion/cytoplasmic streaming) developed only later in maturation. Involvement of a postphloem bulk flow is complicated in the present instance by the extremely limited water loss from juice sacs either via transpiration or fluid backflow. Nonetheless, tissue expansion can account for a collective water inflow of at least 1.0 milliliter per day throughout the majority of juice sac development, thus providing a modest, but potentially important means of nonvascular solution flow. Overall, data indicate postphloem transfer (a) can follow highly localized paths through sizable nonvascular areas (up to 3.0 centimeters total), (b) appears to involve temporary storage and/or equilibration with compartmentalized sugars enroute, (c) can occur either against an overall up-hill sugar gradient (young tissues) or along a descending gradient (near full expansion), and

  13. Site Plan & Site Section of Citrus Landscape (Showing Relationship ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    Site Plan & Site Section of Citrus Landscape (Showing Relationship of Groves & Irrigation System to Grove Canal) - Arlington Heights Citrus Landscape, Southwestern portion of city of Riverside, Riverside, Riverside County, CA

  14. Observations on the entomopathogenic fungus Hirsutella citriformis attacking adult Diaphorina citri (Hemiptera: Psyllid) in a managed citrus grove

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A two-year field study was conducted in an orange grove (0.7 ha) in Florida to characterize the phenology of the entomopathogen Hirsutella citriformis Speare infecting adults of the Asian citrus psyllid, Diaphorina citri Kuwayama. On the average over the two-year study, 23 percent of adults observed...

  15. Seasonal prevalence of the insect pathogenic fungus Colletotrichum nymphaeae in Brazilian citrus groves under different chemical pesticide regimes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In a field study, we identified an endemic fungal entomopathogen, known as the 'salmão' fungus infecting populations of citrus scale, Praelongorthezia praelonga Douglas. The identification of this fungus is close to Colletotrichum nymphaeae (Sordariomycetes: Glomerellales) based on morphological sim...

  16. STUDIES ON THE BACTERIAL LEAF BLIGHT OF RICE PLANT. ON THE DISTRIBUTION OF BACTERIUM ORYZAE (UEDA ET ISHIYAMA) NAKATA UPON THE RICE PLANTS (PRELIMINARY REPORT),

    DTIC Science & Technology

    The report contains a study of an outbreak of bacterial leaf blight of rice plants. An investigation of the primary source was made. Knowledge of the...distribution of infective agent on rice plants, when the primary infection occurs and before the appearance of the blight is recognized, was thought

  17. Preparation of an in-house reference material containing fumonisins in Thai rice and matrix extension of the analytical method for Japanese rice.

    PubMed

    Awaludin, Norhafniza; Nagata, Reiko; Kawasaki, Tomomi; Kushiro, Masayo

    2009-12-01

    Mycotoxin contamination in rice is less reported, compared to that in wheat or maize, however, some Fusarium fungi occasionally infect rice in the paddy field. Fumonisins are mycotoxins mainly produced by Fusarium verticillioides, which often ruins maize. Rice adherent fungus Gibberella fujikuroi is taxonomically near to F. verticillioides, and there are sporadic reports of fumonisin contamination in rice from Asia, Europe and the United States. Therefore, there exists the potential risk of fumonisin contamination in rice as well as the need for the validated analytical method for fumonisins in rice. Although both natural and spiked reference materials are available for some Fusarium mycotoxins in matrices of wheat and maize, there are no reference materials for Fusarium mycotoxins in rice. In this study, we have developed a method for the preparation of a reference material containing fumonisins in Thai rice. A ShakeMaster grinding machine was used for the preparation of a mixed material of blank Thai rice and F. verticillioides-infected Thai rice. The homogeneity of the mixed material was confirmed by one-way analysis of variance, which led this material to serve as an in-house reference material. Using this reference material, several procedures to extract fumonisins from Thai rice were compared. Accordingly, we proved the applicability of an effective extraction procedure for the determination of fumonisins in Japanese rice.

  18. Distinct Carotenoid and Flavonoid Accumulation in a Spontaneous Mutant of Ponkan (Citrus reticulata Blanco) Results in Yellowish Fruit and Enhanced Postharvest Resistance.

    PubMed

    Luo, Tao; Xu, Kunyang; Luo, Yi; Chen, Jiajing; Sheng, Ling; Wang, Jinqiu; Han, Jingwen; Zeng, Yunliu; Xu, Juan; Chen, Jianmin; Wu, Qun; Cheng, Yunjiang; Deng, Xiuxin

    2015-09-30

    As the most important fresh fruit worldwide, citrus is often subjected to huge postharvest losses caused by abiotic and biotic stresses. As a promising strategy to reduce postharvest losses, enhancing natural defense by potential metabolism reprogramming in citrus mutants has rarely been reported. The yellowish spontaneous mutant of Ponkan (Citrus reticulata Blanco) (YP) was used to investigate the influence of metabolism reprogramming on postharvest performance. Our results show that reduced xanthophyll accumulation is the cause of yellowish coloring of YP and might be attributed to the reduced carotenoid sequestration capacity and upregulated expression of carotenoid cleavage dioxygenase genes. Constantly higher levels of polymethoxylated flavones (PMFs) during the infection and the storage stage might make significant contribution to the more strongly induced resistance against Penicillium digitatum and lower rotting rate. The present study demonstrates the feasibility of applying bud mutants to improve the postharvest performance of citrus fruits.

  19. Impacts of Horticultural Mineral Oils and Two Insecticide Practices on Population Fluctuation of Diaphorina citri and Spread of Huanglongbing in a Citrus Orchard in Sarawak

    PubMed Central

    Leong, Stephen Chan Teck; Abang, Fatimah; Beattie, Andrew; Kueh, Roland Jui Heng; Wong, Sing King

    2012-01-01

    Aspects of the incidence and spread of the citrus disease huanglongbing (HLB) in relation to the vector Diaphorina citri population fluctuation were studied from January 1999 to December 2001 seasons in a 0.8 ha citrus orchard at Jemukan (1° 33′N, 110° 41′E), Southwest Sarawak in Malaysia. In relation to insecticide and horticultural mineral oils (HMOs) use, levels of HLB infection rose quite rapidly over the next 3 years in the unsprayed control and less rapidly in the other treatments such as imidacloprid, nC24HMO, and triazophos/cypermethrin/chlorpyrifos. Levels of HLB as determined by Polymerase Chain Reaction (PCR) were 42.2%, 9.4%, 11.4%, and 22.7%, respectively. The effects of nC24HMO and conventional pesticides on the citrus psyllid population and parasitoids in citrus orchard were also determined. PMID:22629178

  20. Annotation of the Asian Citrus Psyllid Genome Reveals a Reduced Innate Immune System

    PubMed Central

    Arp, Alex P.; Hunter, Wayne B.; Pelz-Stelinski, Kirsten S.

    2016-01-01

    Citrus production worldwide is currently facing significant losses due to citrus greening disease, also known as Huanglongbing. The citrus greening bacteria, Candidatus Liberibacter asiaticus (CLas), is a persistent propagative pathogen transmitted by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae). Hemipterans characterized to date lack a number of insect immune genes, including those associated with the Imd pathway targeting Gram-negative bacteria. The D. citri draft genome was used to characterize the immune defense genes present in D. citri. Predicted mRNAs identified by screening the published D. citri annotated draft genome were manually searched using a custom database of immune genes from previously annotated insect genomes. Toll and JAK/STAT pathways, general defense genes Dual oxidase, Nitric oxide synthase, prophenoloxidase, and cellular immune defense genes were present in D. citri. In contrast, D. citri lacked genes for the Imd pathway, most antimicrobial peptides, 1,3-β-glucan recognition proteins (GNBPs), and complete peptidoglycan recognition proteins. These data suggest that D. citri has a reduced immune capability similar to that observed in A. pisum, P. humanus, and R. prolixus. The absence of immune system genes from the D. citri genome may facilitate CLas infections, and is possibly compensated for by their relationship with their microbial endosymbionts. PMID:27965582

  1. Annotation of the Asian Citrus Psyllid Genome Reveals a Reduced Innate Immune System.

    PubMed

    Arp, Alex P; Hunter, Wayne B; Pelz-Stelinski, Kirsten S

    2016-01-01

    Citrus production worldwide is currently facing significant losses due to citrus greening disease, also known as Huanglongbing. The citrus greening bacteria, Candidatus Liberibacter asiaticus (CLas), is a persistent propagative pathogen transmitted by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae). Hemipterans characterized to date lack a number of insect immune genes, including those associated with the Imd pathway targeting Gram-negative bacteria. The D. citri draft genome was used to characterize the immune defense genes present in D. citri. Predicted mRNAs identified by screening the published D. citri annotated draft genome were manually searched using a custom database of immune genes from previously annotated insect genomes. Toll and JAK/STAT pathways, general defense genes Dual oxidase, Nitric oxide synthase, prophenoloxidase, and cellular immune defense genes were present in D. citri. In contrast, D. citri lacked genes for the Imd pathway, most antimicrobial peptides, 1,3-β-glucan recognition proteins (GNBPs), and complete peptidoglycan recognition proteins. These data suggest that D. citri has a reduced immune capability similar to that observed in A. pisum, P. humanus, and R. prolixus. The absence of immune system genes from the D. citri genome may facilitate CLas infections, and is possibly compensated for by their relationship with their microbial endosymbionts.

  2. Genome wide selection in Citrus breeding.

    PubMed

    Gois, I B; Borém, A; Cristofani-Yaly, M; de Resende, M D V; Azevedo, C F; Bastianel, M; Novelli, V M; Machado, M A

    2016-10-17

    Genome wide selection (GWS) is essential for the genetic improvement of perennial species such as Citrus because of its ability to increase gain per unit time and to enable the efficient selection of characteristics with low heritability. This study assessed GWS efficiency in a population of Citrus and compared it with selection based on phenotypic data. A total of 180 individual trees from a cross between Pera sweet orange (Citrus sinensis Osbeck) and Murcott tangor (Citrus sinensis Osbeck x Citrus reticulata Blanco) were evaluated for 10 characteristics related to fruit quality. The hybrids were genotyped using 5287 DArT_seq(TM) (diversity arrays technology) molecular markers and their effects on phenotypes were predicted using the random regression - best linear unbiased predictor (rr-BLUP) method. The predictive ability, prediction bias, and accuracy of GWS were estimated to verify its effectiveness for phenotype prediction. The proportion of genetic variance explained by the markers was also computed. The heritability of the traits, as determined by markers, was 16-28%. The predictive ability of these markers ranged from 0.53 to 0.64, and the regression coefficients between predicted and observed phenotypes were close to unity. Over 35% of the genetic variance was accounted for by the markers. Accuracy estimates with GWS were lower than those obtained by phenotypic analysis; however, GWS was superior in terms of genetic gain per unit time. Thus, GWS may be useful for Citrus breeding as it can predict phenotypes early and accurately, and reduce the length of the selection cycle. This study demonstrates the feasibility of genomic selection in Citrus.

  3. First report of citrus leprosis virus nuclear type in Colombia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus leprosis is a difficult viral disease causing significant damage to citrus fruit in South America and Central America. The disease is marked by dramatic lesions on fruit, leaves and stems resulting in unmarketable product. Citrus leprosis virus cytoplasmic types (CiLV-C and CiLV-C2) wer edete...

  4. Cryopreservation of Citrus shoot tips using micrografting for recovery

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The USDA-ARS National Plant Germplasm System (NPGS) and the University of California Citrus Variety Collection maintain more than 888 unique accessions representing 132 taxa of Citrus, Fortunella, and Citrus wild species relatives within field, screenhouse, and greenhouse collections. We have ident...

  5. 21 CFR 74.302 - Citrus Red No. 2.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 1 2010-04-01 2010-04-01 false Citrus Red No. 2. 74.302 Section 74.302 Food and... ADDITIVES SUBJECT TO CERTIFICATION Foods § 74.302 Citrus Red No. 2. (a) Identity. (1) The color additive Citrus Red No. 2 is principally 1-(2,5-dimethoxyphenylazo)-2-naphthol. (2) The following diluents may...

  6. 7 CFR 319.56-41 - Citrus from Peru.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 5 2011-01-01 2011-01-01 false Citrus from Peru. 319.56-41 Section 319.56-41... SERVICE, DEPARTMENT OF AGRICULTURE FOREIGN QUARANTINE NOTICES Fruits and Vegetables § 319.56-41 Citrus from Peru. Grapefruit (Citrus paradisi), limes (C. aurantiifolia), mandarins or tangerines...

  7. 7 CFR 319.56-41 - Citrus from Peru.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 5 2014-01-01 2014-01-01 false Citrus from Peru. 319.56-41 Section 319.56-41... SERVICE, DEPARTMENT OF AGRICULTURE FOREIGN QUARANTINE NOTICES Fruits and Vegetables § 319.56-41 Citrus from Peru. Grapefruit (Citrus paradisi), limes (C. aurantiifolia), mandarins or tangerines...

  8. 7 CFR 319.56-41 - Citrus from Peru.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 5 2012-01-01 2012-01-01 false Citrus from Peru. 319.56-41 Section 319.56-41... SERVICE, DEPARTMENT OF AGRICULTURE FOREIGN QUARANTINE NOTICES Fruits and Vegetables § 319.56-41 Citrus from Peru. Grapefruit (Citrus paradisi), limes (C. aurantiifolia), mandarins or tangerines...

  9. 7 CFR 319.56-41 - Citrus from Peru.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 5 2013-01-01 2013-01-01 false Citrus from Peru. 319.56-41 Section 319.56-41... SERVICE, DEPARTMENT OF AGRICULTURE FOREIGN QUARANTINE NOTICES Fruits and Vegetables § 319.56-41 Citrus from Peru. Grapefruit (Citrus paradisi), limes (C. aurantiifolia), mandarins or tangerines...

  10. 21 CFR 74.302 - Citrus Red No. 2.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 1 2011-04-01 2011-04-01 false Citrus Red No. 2. 74.302 Section 74.302 Food and... ADDITIVES SUBJECT TO CERTIFICATION Foods § 74.302 Citrus Red No. 2. (a) Identity. (1) The color additive Citrus Red No. 2 is principally 1-(2,5-dimethoxyphenylazo)-2-naphthol. (2) The following diluents may...

  11. Susceptibility of sixteen citrus genotypes to Candidatus Liberibacter asiaticus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB) disease is the most serious threat to citrus production worldwide and in the last decade has devastated the Florida citrus industry. HLB is associated with a phloem-limited a-proteobacter Candidatus Liberibacter asiaticus (CLas) and its insect vector the Asian citrus psyllid (Dia...

  12. Long term field evaluation reveals HLB resistance in Citrus relatives

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus huanglongbing (HLB) is a destructive disease with no known cure. To identify sources of HLB resistance in the subfamily Aurantioideae to which citrus belongs, we conducted a six-year field trial under natural disease challenge conditions in an HLB endemic region. The study included 65 Citrus ...

  13. 21 CFR 74.302 - Citrus Red No. 2.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 1 2012-04-01 2012-04-01 false Citrus Red No. 2. 74.302 Section 74.302 Food and... ADDITIVES SUBJECT TO CERTIFICATION Foods § 74.302 Citrus Red No. 2. (a) Identity. (1) The color additive Citrus Red No. 2 is principally 1-(2,5-dimethoxyphenylazo)-2-naphthol. (2) The following diluents may...

  14. 21 CFR 74.302 - Citrus Red No. 2.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 1 2014-04-01 2014-04-01 false Citrus Red No. 2. 74.302 Section 74.302 Food and... ADDITIVES SUBJECT TO CERTIFICATION Foods § 74.302 Citrus Red No. 2. (a) Identity. (1) The color additive Citrus Red No. 2 is principally 1-(2,5-dimethoxyphenylazo)-2-naphthol. (2) The following diluents may...

  15. 21 CFR 74.302 - Citrus Red No. 2.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 1 2013-04-01 2013-04-01 false Citrus Red No. 2. 74.302 Section 74.302 Food and... ADDITIVES SUBJECT TO CERTIFICATION Foods § 74.302 Citrus Red No. 2. (a) Identity. (1) The color additive Citrus Red No. 2 is principally 1-(2,5-dimethoxyphenylazo)-2-naphthol. (2) The following diluents may...

  16. Current California Drought: Impact on Citrus Trees and Potential Mitigation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    California is in another cycle of extended drought. The article reviews and discusses likely impact of the current drought on citrus growers and potential mitigation techniques. Citrus physiological responses to water stress is briefly reviewed. The direct impact of drought on citrus is reduced frui...

  17. Evaluation of natural colorants and their application on citrus fruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Warm temperatures can often result in poor peel color of some citrus varieties, especially early in the harvest season. Under these conditions, Florida oranges, temples, tangelos, and K-Early citrus fruit are allowed to be treated with Citrus Red No.2 (CR2) to help produce a more acceptable peel col...

  18. Preharvest measures for postharvest improvement in marketable fresh citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker, caused by the bacterium Xanthomonas citri supsp. citri, (Xcc),was once the scourge of the Florida citrus industry from the early 1900’s on and off until the early 1990’s. It has been replaced, for the most part, by citrus greening but still continues to be a problem for Florida fres...

  19. Preharvest measures for postharvest improvement in marketable fresh citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker caused by the bacterium Xanthomonas citri supsp. citri, (Xcc), was once the scourge of the Florida citrus industry from the early 1900’s on and off until the early 1990’s. It has been replaced, for the most part, by citrus greening but still continues to be a problem for Florida fresh ...

  20. Molecular analysis of citrus rust (Phyllocoptruta oleivora) populations in Texas

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phyllocoptruta olevoria (Acari: Eriophyidae) or the citrus rust mite (CRM) is one of the most economically important mite pests of citrus worldwide. CRM originated from Asia, however it is found in most citrus producing areas, such as, Brazil, Morocco and the United States. Though CRM is a major pes...

  1. Intercropping of citrus and guava trees for management of Huanglongbing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recent studies conducted in Viet Nam by Vietnamese, Japanese and Australian scientists indicate that interplanting citrus with guava negated infestations of Asian citrus psyllid and consequently huanglongbing, a serious disease caused by a bacterium vectored by the psyllid. Young citrus interplanted...

  2. Incidence of invasive Asian citrus psyllid, Diaphorina citri (Hemiptera: Psyllidae) and its introduced parasitoid Tamarixia radiata (Hymenoptera: Eulophidae) in Florida citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Psyllidae) vectors the bacterium Candidatus Liberibacter asiaticus, one of the causal organisms of Huanglongbing or citrus greening, a devastating disease of citrus. A eulophid parasitoid, Tamarixia radiata Waterson, was imported ...

  3. Citrus tristeza virus (CTV) Causing Proteomic and Enzymatic Changes in Sweet Orange Variety “Westin”

    PubMed Central

    Dória, Milena Santos; de Sousa, Aurizângela Oliveira; Barbosa, Cristiane de Jesus; Costa, Márcio Gilberto Cardoso; Gesteira, Abelmon da Silva; Souza, Regina Martins; Freitas, Ana Camila Oliveira; Pirovani, Carlos Priminho

    2015-01-01

    Citrus Tristeza disease, caused by CTV (Citrus tristeza virus), committs citrus plantations around the world and specifically attacks phloem tissues of the plant. The virus exists as a mixture of more or less severe variants, which may or may not cause symptoms of Tristeza. The objective of this study was to analyze the changes caused by CTV in the proteome of stems of sweet orange, as well as in the activity and gene expression of antioxidant enzymes. The CTV-infected sweet orange displayed mild symptoms, which were characterized by the presence of sparse stem pitting throughout their stems. The presence of virus was confirmed by RT-PCR. Proteomic analysis by 2DE-PAGE-MS / MS revealed the identity of 40 proteins differentially expressed between CTV- infected and -non-infected samples. Of these, 33 were up-regulated and 7 were down-regulated in CTV-infected samples. Among the proteins identified stands out a specific from the virus, the coat protein. Other proteins identified are involved with oxidative stress and for this their enzymatic activity was measured. The activity of superoxide dismutase (SOD) was higher in CTV-infected samples, as catalase (CAT) showed higher activity in uninfected samples. The activity of guaiacol peroxidase (GPX) did not vary significantly between samples. However, ascorbate peroxidase (APX) was more active in the infected samples. The relative expression of the genes encoding CAT, SOD, APX and GPX was analyzed by quantitative real time PCR (RT-qPCR). The CTV-infected samples showed greater accumulation of transcripts, except for the CAT gene. This gene showed higher expression in the uninfected samples. Taken together, it can be concluded that the CTV affects the protein profile and activity and gene expression of antioxidant enzymes in plants infected by this virus. PMID:26207751

  4. Roles of plant hormones and their interplay in rice immunity.

    PubMed

    Yang, Dong-Lei; Yang, Yinong; He, Zuhua

    2013-05-01

    Plant hormones have been extensively studied for their importance in innate immunity particularly in the dicotyledonous model plant Arabidopsis thaliana. However, only in the last decade, plant hormones were demonstrated to play conserved and divergent roles in fine-tuning immune in rice (Oryza sativa L.), a monocotyledonous model crop plant. Emerging evidence showed that salicylic acid (SA) plays a role in rice basal defense but is differentially required by rice pattern recognition receptor (PRR) and resistance (R) protein-mediated immunity, and its function is likely dependent on the signaling pathway rather than the change of endogenous levels. Jasmonate (JA) plays an important role in rice basal defense against bacterial and fungal infection and may be involved in the SA-mediated resistance. Ethylene (ET) can act as a positive or negative modulator of disease resistance, depending on the pathogen type and environmental conditions. Brassinosteroid (BR) signaling and abscisic acid (ABA) either promote or defend against infection of pathogens with distinct infection/colonization strategies. Auxin and gibberellin (GA) are generally thought of as negative regulators of innate immunity in rice. Moreover, GA interacts antagonistically with JA signaling in rice development and immunity through the DELLA protein as a master regulator of the two hormone pathways. In this review, we summarize the roles of plant hormones in rice immunity and discuss their interplay/crosstalk mechanisms and the complex regulatory network of plant hormone pathways in fine-tuning rice immunity and growth.

  5. Production of mono- and polyclonal antibodies to Citrus leprosis virus C2 and their application in triple antibody sandwich ELISA and immunocapture RT-PCR diagnostic assays.

    PubMed

    Choudhary, Nandlal; Roy, Avijit; Leon, M G; Wei, G; Nakhla, M K; Levy, L; Brlansky, R H

    2017-05-01

    The newly discovered Citrus leprosis virus cytoplasmic type 2 (CiLV-C2) is one of the causal virus of citrus leprosis disease complex; which leads to substantial loss of citrus production in the states of Meta and Casanare of Colombia. Specific and sensitive detection methods are needed to monitor the dissemination of CiLV-C2 in Colombia, and to prevent introduction of CiLV-C2 to other citrus growing countries. Toward this end, putative coat protein gene (CPG) of CiLV-C2 was amplified from CiLV-C2 infected citrus tissues. The CPG was cloned, expressed and purified a recombinant coat protein of ∼31kDa which used to generate monoclonal antibodies and polyclonal antisera. Four monoclonal antibodies and two polyclonal antisera were selected as being specific following Western blotting. The monoclonal antibody MAb E5 and polyclonal antiserum PAb UF715 were selected testing with an extract of CiLV-C2 infected leaves using triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA). In addition, an immunocapture RT-PCR was standardized using MAb E5 for specific and sensitive detection of CiLV-C2. The standardized TAS-ELISA and IC-RT-PCR were able to detect CiLV-C2 in the extracts of symptomatic citrus leprosis tissues up to the dilutions of 1:160 and 1:2580, respectively. Result demonstrated that CiLV-C2 is present in citrus orchards in Meta and Casanare citrus growing areas of Colombia. TAS-ELISA could be used for routine detection of CiLV-C2, epidemiological studies, and for border inspections for quarantine purposes. IC-RT-PCR could be valuable for CiLV-C2 validation and viral genome analysis.

  6. Infection,

    DTIC Science & Technology

    1980-10-16

    inapparent infection. A refeeding program may thus become complicated by the sudden appearance of a life-threatening infectious illness (3). (3) The...Beisel, W. R. 23 Unusually low serum concentrations of inorganic phosphate have been reported in patients with gram-negative sepsis and in Reye’s syndrome ...infection should be corrected by a well-managed program of convalescent-period refeeding . This aspect of nutritional support is too often ignored. On the

  7. Evaluation of natural colorants and their application on citrus fruit as alternatives to Citrus Red No. 2

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Warm field temperatures can often result in poor peel color of some citrus varieties, especially early in the harvest season. Under these conditions, Florida oranges, temples, tangelos, and K-Early citrus fruit are allowed to be treated with Citrus Red No.2 (CR2) to help produce a more acceptable pe...

  8. Antimicrobial nanoemulsion formulation with improved penetration of foliar spray through citrus leaf cuticles to control citrus Huanglongbing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB) is one of the most serious citrus diseases that threaten citrus industry worldwide. Because Candidatus Liberibacter asiaticus (Las) resides in citrus phloem, it is difficult to deliver an effective chemical compound into the phloem for control of HLB. In this study, a transcuticu...

  9. Overexpression of a modified plant thionin enhances disease resistance to citrus canker and huanglongbing (HLB, citrus greening)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB or citrus greening disease) caused by Candidatus Liberibacter asiaticus (Las) is a great threat to the United States citrus industry. Citrus canker is also an economically important disease associated with a bacterial pathogen (Xanthomonas citri). In this study, we characterized e...

  10. Degradation products of citrus volatile organic compounds (VOCs) acting as phagostimulants that increase probing behavior of Asian citrus psyllid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Volatile phytochemicals play a role in orientation by phytophagous insects. We studied antennal and behavioral responses of the Asian citrus psyllid, Diaphorina citri Kuwayama, vector of the citrus greening disease pathogen. Little or no response to citrus leaf volatiles was detected by electroanten...

  11. Does citrus leaf miner impair hydraulics and fitness of citrus host plants?

    PubMed

    Raimondo, Fabio; Trifilò, Patrizia; Gullo, Maria A Lo

    2013-12-01

    Gas exchange and hydraulic features were measured in leaves of three different Citrus species (Citrus aurantium L., Citrus limon L., Citrus  ×  paradisii Macfad) infested by Phyllocnistis citrella Staiton, with the aim to quantify the impact of this pest on leaf hydraulics and, ultimately, on plant fitness. Infested leaves were characterized by the presence on the leaf blade of typical snake-shaped mines and, in some cases, of a crumpled leaf blade. Light microscopy showed that leaf crumpling was induced by damage to the cuticular layer. In all three Citrus species examined: (a) the degree of infestation did not exceed 10% of the total surface area of infested plants; (b) control and infested leaves showed similar values of minimum diurnal leaf water potential, leaf hydraulic conductance and functional vein density; and (c) maximum diurnal values of stomatal conductance to water vapour, transpiration rate and photosynthetic rate (An) were similar in both control leaves and the green areas of infested leaves. A strong reduction of An was recorded only in mined leaf areas. Our data suggest that infestation with P. citrella does not cause conspicuous plant productivity reductions in young Citrus plants, at least not in the three Citrus species studied here.

  12. Detection of citrus canker and Huanglongbing using fluorescence imaging spectroscopy and support vector machine technique.

    PubMed

    Wetterich, Caio Bruno; Felipe de Oliveira Neves, Ruan; Belasque, José; Marcassa, Luis Gustavo

    2016-01-10

    Citrus canker and Huanglongbing (HLB) are citrus diseases that represent a serious threat to the citrus production worldwide and may cause large economic losses. In this work, we combined fluorescence imaging spectroscopy (FIS) and a machine learning technique to discriminate between these diseases and other ordinary citrus conditions that may be present at citrus orchards, such as citrus scab and zinc deficiency. Our classification results are highly accurate when discriminating citrus canker from citrus scab (97.8%), and HLB from zinc deficiency (95%). These results show that it is possible to accurately identify citrus diseases that present similar symptoms.

  13. Rapid and sensitive detection of Citrus Bacterial Canker by loop-mediated isothermal amplification combined with simple visual evaluation methods

    PubMed Central

    2010-01-01

    Background Citrus Bacterial Canker (CBC) is a major, highly contagious disease of citrus plants present in many countries in Asia, Africa and America, but not in the Mediterranean area. There are three types of Citrus Bacterial Canker, named A, B, and C that have different genotypes and posses variation in host range within citrus species. The causative agent for type A CBC is Xanthomonas citri subsp. citri, while Xanthomonas fuscans subsp. aurantifolii, strain B causes type B CBC and Xanthomonas fuscans subsp. aurantifolii strain C causes CBC type C. The early and accurate identification of those bacteria is essential for the protection of the citrus industry. Detection methods based on bacterial isolation, antibodies or polymerase chain reaction (PCR) have been developed previously; however, these approaches may be time consuming, laborious and, in the case of PCR, it requires expensive laboratory equipment. Loop-mediated isothermal amplification (LAMP), which is a novel isothermal DNA amplification technique, is sensitive, specific, fast and requires no specialized laboratory equipment. Results A loop-mediated isothermal amplification assay for the diagnosis of Citrus Bacterial Canker (CBC-LAMP) was developed and evaluated. DNA samples were obtained from infected plants or cultured bacteria. A typical ladder-like pattern on gel electrophoresis was observed in all positive samples in contrast to the negative controls. In addition, amplification products were detected by visual inspection using SYBRGreen and using a lateral flow dipstick, eliminating the need for gel electrophoresis. The sensitivity and specificity of the assay were evaluated in different conditions and using several sample sources which included purified DNA, bacterium culture and infected plant tissue. The sensitivity of the CBC-LAMP was 10 fg of pure Xcc DNA, 5 CFU in culture samples and 18 CFU in samples of infected plant tissue. No cross reaction was observed with DNA of other phytopathogenic

  14. Noninvasive maturity detection of citrus with machine vision

    NASA Astrophysics Data System (ADS)

    Ying, Yibin; Xu, Zhenggang; Fu, Xiaping; Liu, Yande

    2004-03-01

    A computer vision system was established to explore a method for citrus maturity detection. The surface color information and the ratio of total soluble solid to titratable acid (TSS/TA) were used as maturity indexes of citrus. The spectral reflectance properties with different color were measured by UV-240 ultraviolet and visible spectrophotometer. The biggest discrepancy of gray levels between citrus pixels and background pixels was in blue component image by image background segmentation. Dynamic threshold method for background segmentation had best result in blue component image. Methods for citrus image color description were studied. The citrus spectral reflectance experiments showed that green surface and saffron surface of citrus were of highest spectral reflectance at the wavelength of 700nm, the difference between them reached to maximum, about 53%, and the image acquired at this wavelength was of more color information for maturity detection. A triple-layer feed forward network was established to map citrus maturity from the hue frequency sequence by the mean of artificial neural network. After training, the network mapper was used to detect the maturity of the test sample set, which was composed of 252 Weizhang citrus with different maturity. The identification accuracy of mature citrus reached 79.1%, that of immature citrus was 63.6%, and the mean identification accuracy was 77.8%. This study suggested that it is feasible to detect citrus maturity non-invasively by using the computer vision system and hue frequency sequence method.

  15. Horsfall-Barratt recalibration and replicated severity estimates of citrus canker

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker is a serious disease of citrus in tropical and subtropical citrus growing regions. Accurate and precise assessment of citrus canker and other plant pathogens is needed to obtain good quality data. Citrus canker assessment data were used to ascertain some of the mechanics of the Horsfal...

  16. Population structure and diversity of citrus tristeza virus (CTV) isolates in Hunan province, China.

    PubMed

    Xiao, Cui; Yao, Run-Xian; Li, Fang; Dai, Su-Ming; Licciardello, Grazia; Catara, Antonino; Gentile, Alessandra; Deng, Zi-Niu

    2017-02-01

    Stem-pitting (SP) is the main type of citrus tristeza virus (CTV) that causes severe damage to citrus trees, especially those of sweet orange, in Hunan province, China. Understanding the local CTV population structure should provide clues for effective mild strain cross-protection (MSCP) of the SP strain of CTV. In this study, markers for the p23 gene, multiple molecular markers (MMMs), and sequence analysis of the three silencing suppressor genes (p20, p23 and p25) were employed to analyze the genetic diversity and genotype composition of the CTV population based on 51 CTV-positive samples collected from 14 citrus orchards scattered around six major citrus-growing areas of Hunan. The results indicated that the CTV population structure was extremely complex and that infection was highly mixed. In total, p23 gene markers resulted in six profiles, and MMMs demonstrated 25 profiles. The severe VT and T3 types appeared to be predominantly associated with SP, while the mild T30 and RB types were related to asymptomatic samples. Based on phylogenetic analysis of the amino acid sequences of p20, p23 and p25, 19 representative CTV samples were classified into seven recently established CTV groups and a potentially novel one. A high level of genetic diversity, as well as potential recombination, was revealed among different CTV isolates. Five pure SP severe and two pure mild strains were identified by genotype composition analysis. Taken together, the results update the genetic diversity of CTV in Hunan with the detection of one possible novel strain, and this information might be applicable for the selection of appropriate mild CTV strains for controlling citrus SP disease through cross-protection.

  17. Global efforts in managing rice blast disease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rice blast disease caused by the fungus Magnaporthe oryzae is a major destructive disease threatening global food security. Resistance (R) genes to M. oryzae are effective in preventing infections by strains of M. oryzae carry the corresponding avirulence (AVR) genes. Effectiveness of genetic resist...

  18. Effective Antibiotics against ‘Candidatus Liberibacter asiaticus’ in HLB-Affected Citrus Plants Identified via the Graft-Based Evaluation

    PubMed Central

    Powell, Charles A.; Doud, Melissa S.; Yang, Chuanyu; Duan, Yongping

    2014-01-01

    Citrus huanglongbing (HLB), caused by three species of fastidious, phloem-limited ‘Candidatus Liberibacter’, is one of the most destructive diseases of citrus worldwide. To date, there is no established cure for this century-old and yet, newly emerging disease. As a potential control strategy for citrus HLB, 31 antibiotics were screened for effectiveness and phytotoxicity using the optimized graft-based screening system with ‘Candidatus Liberibacter asiaticus’ (Las)-infected citrus scions. Actidione and Oxytetracycline were the most phytotoxic to citrus with less than 10% of scions surviving and growing; therefore, this data was not used in additional analyses. Results of principal component (PCA) and hierarchical clustering analyses (HCA) demonstrated that 29 antibiotics were clustered into 3 groups: highly effective, partly effective, and not effective. In spite of different modes of actions, a number of antibiotics such as, Ampicillin, Carbenicillin, Penicillin, Cefalexin, Rifampicin and Sulfadimethoxine were all highly effective in eliminating or suppressing Candidatus Liberibacter asiaticus indicated by both the lowest Las infection rate and titers of the treated scions and inoculated rootstock. The non-effective group, including 11 antibiotics alone with three controls, such as Amikacin, Cinoxacin, Gentamicin, Kasugamycin, Lincomycin, Neomycin, Polymixin B and Tobramycin, did not eliminate or suppress Las in the tested concentrations, resulting in plants with increased titers of Las. The other 12 antibiotics partly eliminated or suppressed Las in the treated and graft-inoculated plants. The effective and non-phytotoxic antibiotics could be potential candidates for control of citrus HLB, either for the rescue of infected citrus germplasm or for restricted field application. PMID:25372135

  19. Biogenic emissions from Citrus species in California

    NASA Astrophysics Data System (ADS)

    Fares, Silvano; Gentner, Drew R.; Park, Jeong-Hoo; Ormeno, Elena; Karlik, John; Goldstein, Allen H.

    2011-09-01

    Biogenic Volatile Organic Compounds (BVOC) emitted from plants are the dominant source of reduced carbon chemicals to the atmosphere and are important precursors to the photochemical production of ozone and secondary organic aerosols. Considering the extensive land used for agriculture, cultivated Citrus plantations may play an important role in the chemistry of the atmosphere especially in regions such as the Central Valley of California. Moreover, the BVOC emissions from Citrus species have not been characterized in detail and more species-specific inputs for regional models of BVOC emissions are needed. In this study, we measured the physiological parameters and emissions of the most relevant BVOC (oxygenated compounds, monoterpenes, and sesquiterpenes) for four predominant Citrus species planted in California ( Citrus sinensis var. 'Parent Navel', Citrus limon var. 'Meyer', Citrus reticulata var. 'W. Murcott' and 'Clementine'). We used two analytical techniques to measure a full range of BVOC emitted: Proton Transfer Reaction Mass Spectrometry (PTR-MS) and gas chromatography with mass spectrometry. Methanol, followed by acetone and acetaldehyde, were the dominant BVOC emitted from lemon and mandarin trees (basal emission rates up to 300 ng(C) g(DW) -1 h -1), while oxygenated monoterpenes, monoterpenes, and sesquiterpenes were the main BVOC emitted from orange trees (basal emission rates up to = 2500 ng(C) g(DW) -1 h -1). Light and temperature-dependent algorithms were better predictors of methanol, acetaldehyde, acetone, isoprene and monoterpenes for all the Citrus species. Whereas, temperature-dependent algorithms were better predictors of oxygenated monoterpenes, and sesquiterpenes. We observed that flowering increased emissions from orange trees by an order of magnitude with the bulk of BVOC emissions being comprised of monoterpenes, sesquiterpenes, and oxygenated monoterpenes. Chemical speciation of BVOC emissions show that the various classes of terpene

  20. 7 CFR 301.76-2 - Regulated articles for Asian citrus psyllid and citrus greening.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... and plant parts (including leaves), except fruit, of: Aegle marmelos, Aeglopsis chevalieri, Afraegle... asiatica, Triphasia trifolia, Vepris (=Toddalia) lanceolata, and Zanthoxylum fagara. (b) Propagative seed..., the movement of propagative seed of these species from an area quarantined for citrus greening...

  1. Southern rice black-streaked dwarf virus: a white-backed planthopper-transmitted fijivirus threatening rice production in Asia.

    PubMed

    Zhou, Guohui; Xu, Donglin; Xu, Dagao; Zhang, Maoxin

    2013-09-09

    Southern rice black-streaked dwarf virus (SRBSDV), a non-enveloped icosahedral virus with a genome of 10 double-stranded RNA segments, is a novel species in the genus Fijivirus (family Reoviridae) first recognized in 2008. Rice plants infected with this virus exhibit symptoms similar to those caused by Rice black-streaked dwarf virus. Since 2009, the virus has rapidly spread and caused serious rice losses in East and Southeast Asia. Significant progress has been made in recent years in understanding this disease, especially about the functions of the viral genes, rice-virus-insect interactions, and epidemiology and control measures. The virus can be efficiently transmitted by the white-backed planthopper (WBPH, Sogatella furcifera) in a persistent circulative propagative manner but cannot be transmitted by the brown planthopper (Nilaparvata lugens) and small brown planthopper (Laodelphax striatellus). Rice, maize, Chinese sorghum (Coix lacryma-jobi) and other grass weeds can be infected via WBPH. However, only rice plays a major role in the virus infection cycle because of the vector's preference. In Southeast Asia, WBPH is a long-distance migratory rice pest. The disease cycle can be described as follows: SRBSDV and its WBPH vector overwinter in warm tropical or sub-tropical areas; viruliferous WBPH adults carry the virus from south to north via long-distance migration in early spring, transmit the virus to rice seedlings in the newly colonized areas, and lay eggs on the infected seedlings; the next generation of WBPHs propagate on infected seedlings, become viruliferous, disperse, and cause new disease outbreaks. Several molecular and serological methods have been developed to detect SRBSDV in plant tissues and individual insects. Control measures based on protection from WBPH, including seedbed coverage, chemical seed treatments, and chemical spraying of seedlings, have proven effective in China.

  2. The resistance of sour orange to Citrus tristeza virus is mediated by both the salicylic acid and RNA silencing defence pathways.

    PubMed

    Gómez-Muñoz, Neus; Velázquez, Karelia; Vives, María Carmen; Ruiz-Ruiz, Susana; Pina, José Antonio; Flores, Ricardo; Moreno, Pedro; Guerri, José

    2016-09-02

    Citrus tristeza virus (CTV) induces in the field the decline and death of citrus varieties grafted on sour orange (SO) rootstock, which has forced the use of alternative decline-tolerant rootstocks in affected countries, despite the highly desirable agronomic features of the SO rootstock. Declining citrus plants display phloem necrosis below the bud union. In addition, SO is minimally susceptible to CTV compared with other citrus varieties, suggesting partial resistance of SO to CTV. Here, by silencing different citrus genes with a Citrus leaf blotch virus-based vector, we have examined the implication of the RNA silencing and salicylic acid (SA) defence pathways in the resistance of SO to CTV. Silencing of the genes RDR1, NPR1 and DCL2/DCL4, associated with these defence pathways, enhanced virus spread and accumulation in SO plants in comparison with non-silenced controls, whereas silencing of the genes NPR3/NPR4, associated with the hypersensitive response, produced a slight decrease in CTV accumulation and reduced stunting of SO grafted on CTV-infected rough lemon plants. We also found that the CTV RNA silencing suppressors p20 and p23 also suppress the SA signalling defence, with the suppressor activity being higher in the most virulent isolates.

  3. Volatile constituents of wild citrus Mangshanyegan (Citrus nobilis Lauriro) peel oil.

    PubMed

    Liu, Cuihua; Cheng, Yunjiang; Zhang, Hongyan; Deng, Xiuxin; Chen, Feng; Xu, Juan

    2012-03-14

    Volatiles of a wild mandarin, Mangshanyegan (Citrus nobilis Lauriro), were characterized by GC-MS, and their aroma active compounds were identified by aroma extract dilution analysis (AEDA) and gas chromatography-olfactometry (GC-O). The volatile profile of Mangshanyegan was compared with those of other four citrus species, Kaopan pummelo (Citrus grandis), Eureka lemon (Citrus limon), Huangyanbendizao tangerine (Citrus reticulata), and Seike navel orange (Citrus sinensis). Monoterpene hydrocarbons predominated in Mangshanyegan, in particular d-limonene and β-myrcene, which accounted for 85.75 and 10.89% of total volatiles, respectively. Among the 12 compounds with flavor dilution factors (FD) = 27, 8 oxygenated compounds, including (Z)- and (E)-linalool oxides, were present only in Mangshanyegan. The combined results of GC-O, quantitative analysis, odor activity values (OAVs), and omission tests revealed that β-myrcene and (Z)- and (E)-linalool oxides were the characteristic aroma compounds of Mangshanyegan, contributing to the balsamic and floral notes of its aroma.

  4. Research in rice fields

    USGS Publications Warehouse

    ,

    2000-01-01

    Between 1987 and 1999, 2.4-3 million acres of rice were planted annually nationwide. Rice fields are a major component of the contemporary landscapes in the Gulf Coastal Plain, the Mississippi Alluvial Valley, and Central Valley of California. In 1998, approximately 600,000 acres of rice were planted in Louisiana. In the Louisiana plant commodities report for 1998, total value for rice was over $350 million; sugarcane was the only plant commodity that exceeded this value. Louisiana has over 2,000 rice farmers supporting over 12,000 jobs in the state. Rice fields in the United States receive high use by wildlife, especially shorebirds, wading birds, and waterfowl. Waterbirds use rice fields for food, shelter, and breeding habitat.

  5. Phenolic Phytoalexins in Rice: Biological Functions and Biosynthesis

    PubMed Central

    Cho, Man-Ho; Lee, Sang-Won

    2015-01-01

    Phytoalexins are inducible secondary metabolites possessing antimicrobial activity against phytopathogens. Rice produces a wide array of phytoalexins in response to pathogen attacks and environmental stresses. With few exceptions, most phytoalexins identified in rice are diterpenoid compounds. Until very recently, flavonoid sakuranetin was the only known phenolic phytoalexin in rice. However, recent studies have shown that phenylamides are involved in defense against pathogen attacks in rice. Phenylamides are amine-conjugated phenolic acids that are induced by pathogen infections and abiotic stresses including ultra violet (UV) radiation in rice. Stress-induced phenylamides, such as N-trans-cinnamoyltryptamine, N-p-coumaroylserotonin and N-cinnamoyltyramine, have been reported to possess antimicrobial activities against rice bacterial and fungal pathogens, an indication of their direct inhibitory roles against invading pathogens. This finding suggests that phenylamides act as phytoalexins in rice and belong to phenolic phytoalexins along with sakuranetin. Phenylamides also have been implicated in cell wall reinforcement for disease resistance and allelopathy of rice. Synthesis of phenolic phytoalexins is stimulated by phytopathogen attacks and abiotic challenges including UV radiation. Accumulating evidence has demonstrated that biosynthetic pathways including the shikimate, phenylpropanoid and arylmonoamine pathways are coordinately activated for phenolic phytoalexin synthesis, and related genes are induced by biotic and abiotic stresses in rice. PMID:26690131

  6. Complete genome sequence of citrus huanglongbing bacterium, 'Candidatus Liberibacter asiaticus' obtained through metagenomics.

    PubMed

    Duan, Yongping; Zhou, Lijuan; Hall, David G; Li, Wenbin; Doddapaneni, Harshavardhan; Lin, Hong; Liu, Li; Vahling, Cheryl M; Gabriel, Dean W; Williams, Kelly P; Dickerman, Allan; Sun, Yijun; Gottwald, Tim

    2009-08-01

    Citrus huanglongbing is the most destructive disease of citrus worldwide. It is spread by citrus psyllids and is associated with a low-titer, phloem-limited infection by any of three uncultured species of alpha-Proteobacteria, 'Candidatus Liberibacter asiaticus', 'Ca. L. americanus', and 'Ca. L. africanus'. A complete circular 'Ca. L. asiaticus' genome has been obtained by metagenomics, using the DNA extracted from a single 'Ca. L. asiaticus'-infected psyllid. The 1.23-Mb genome has an average 36.5% GC content. Annotation revealed a high percentage of genes involved in both cell motility (4.5%) and active transport in general (8.0%), which may contribute to its virulence. 'Ca. L. asiaticus' appears to have a limited ability for aerobic respiration and is likely auxotrophic for at least five amino acids. Consistent with its intracellular nature, 'Ca. L. asiaticus' lacks type III and type IV secretion systems as well as typical free-living or plant-colonizing extracellular degradative enzymes. 'Ca. L. asiaticus' appears to have all type I secretion system genes needed for both multidrug efflux and toxin effector secretion. Multi-protein phylogenetic analysis confirmed 'Ca. L. asiaticus' as an early-branching and highly divergent member of the family Rhizobiaceae. This is the first genome sequence of an uncultured alpha-proteobacteria that is both an intracellular plant pathogen and insect symbiont.

  7. Identification and characterization of chitin synthase genes in the postharvest citrus fruit pathogen Penicillium digitatum.

    PubMed

    Gandía, Mónica; Harries, Eleonora; Marcos, Jose F

    2012-06-01

    In this study, we carried out the isolation and characterization of chitin synthase genes (CHS) of the main citrus fruit postharvest pathogen Penicillium digitatum. Using distinct sets of degenerate primers designed from conserved regions of CHS genes of yeast and filamentous fungi, PCR methods, and a DNA genomic library, five putative CHS genes (PdigCHSI, PdigCHSII, PdigCHSIII, PdigCHSV, and PdigCHSVII) were identified, isolated, sequenced, and characterized. Phylogenetic analyses, sequence identity, and domain conservation support the annotation as CHS. A very high sequence identity and strong synteny were found with corresponding regions from the genome of Penicillium chrysogenum. Gene expression of P. digitatum CHS genes during mycelium axenic growth, under oxidative and osmotic stress conditions, and during infection of citrus fruits was confirmed and quantified using quantitative RT-PCR (qRT-PCR). PdigCHSIII had the highest expression among the five genes by one order of magnitude, while PdigCHSII had the lowest. However, PdigCHSII was strongly induced coincident with conidial production, suggesting a role in conidiogenesis. The expression of PdigCHSI, PdigCHSIII, PdigCHSV, and PdigCHSVII was upregulated during infection of citrus fruit. PdigCHSV and PdigCHSVII coexpressed in most of the experiments carried out, and they are separated by a 1.77 kb intergenic region and arranged in opposite directions.

  8. ‘Candidatus Liberibacter asiaticus’, causal agent of citrus Huanglongbing, is reduced by treatment with Brassinosteroids

    SciTech Connect

    Canales, Eduardo; Coll, Yamilet; Hernández, Ingrid; Portieles, Roxana; Rodríguez García, Mayra; López, Yunior; Aranguren, Miguel; Alonso, Eugenio; Delgado, Roger; Luis, Maritza; Batista, Lochy; Paredes, Camilo; Rodríguez, Meilyn; Pujol, Merardo; Ochagavia, María Elena; Falcón, Viviana; Terauchi, Ryohei; Matsumura, Hideo; Ayra-Pardo, Camilo; Llauger, Raixa; Pérez, María del Carmen; Núñez, Mirian; Borrusch, Melissa S.; Walton, Jonathan D.; Silva, Yussuan; Pimentel, Eulogio; Borroto, Carlos; Borrás-Hidalgo, Orlando

    2016-01-05

    Huanglongbing (HLB) constitutes the most destructive disease of citrus worldwide, yet no established efficient management measures exist for it. Brassinosteroids, a family of plant steroidal compounds, are essential for plant growth, development and stress tolerance. As a possible control strategy for HLB, epibrassinolide was applied to as a foliar spray to citrus plants infected with the causal agent of HLB, ‘Candidatus Liberibacter asiaticus’. The bacterial titers were reduced after treatment with epibrassinolide under both greenhouse and field conditions but were stronger in the greenhouse. Known defense genes were induced in leaves by epibrassinolide. With the SuperSAGE technology combined with next generation sequencing, induction of genes known to be associated with defense response to bacteria and hormone transduction pathways were identified. Lastly, the results demonstrate that epibrassinolide may provide a useful tool for the management of HLB.

  9. ‘Candidatus Liberibacter asiaticus’, causal agent of citrus Huanglongbing, is reduced by treatment with Brassinosteroids

    DOE PAGES

    Canales, Eduardo; Coll, Yamilet; Hernández, Ingrid; ...

    2016-01-05

    Huanglongbing (HLB) constitutes the most destructive disease of citrus worldwide, yet no established efficient management measures exist for it. Brassinosteroids, a family of plant steroidal compounds, are essential for plant growth, development and stress tolerance. As a possible control strategy for HLB, epibrassinolide was applied to as a foliar spray to citrus plants infected with the causal agent of HLB, ‘Candidatus Liberibacter asiaticus’. The bacterial titers were reduced after treatment with epibrassinolide under both greenhouse and field conditions but were stronger in the greenhouse. Known defense genes were induced in leaves by epibrassinolide. With the SuperSAGE technology combined with nextmore » generation sequencing, induction of genes known to be associated with defense response to bacteria and hormone transduction pathways were identified. Lastly, the results demonstrate that epibrassinolide may provide a useful tool for the management of HLB.« less

  10. ‘Candidatus Liberibacter asiaticus’, Causal Agent of Citrus Huanglongbing, Is Reduced by Treatment with Brassinosteroids

    PubMed Central

    Hernández, Ingrid; Portieles, Roxana; Rodríguez García, Mayra; López, Yunior; Aranguren, Miguel; Alonso, Eugenio; Delgado, Roger; Luis, Maritza; Batista, Lochy; Paredes, Camilo; Rodríguez, Meilyn; Pujol, Merardo; Ochagavia, María Elena; Falcón, Viviana; Terauchi, Ryohei; Matsumura, Hideo; Ayra-Pardo, Camilo; Llauger, Raixa; Pérez, María del Carmen; Núñez, Mirian; Borrusch, Melissa S.; Walton, Jonathan D.; Silva, Yussuan; Pimentel, Eulogio; Borroto, Carlos; Borrás-Hidalgo, Orlando

    2016-01-01

    Huanglongbing (HLB) constitutes the most destructive disease of citrus worldwide, yet no established efficient management measures exist for it. Brassinosteroids, a family of plant steroidal compounds, are essential for plant growth, development and stress tolerance. As a possible control strategy for HLB, epibrassinolide was applied to as a foliar spray to citrus plants infected with the causal agent of HLB, ‘Candidatus Liberibacter asiaticus’. The bacterial titers were reduced after treatment with epibrassinolide under both greenhouse and field conditions but were stronger in the greenhouse. Known defense genes were induced in leaves by epibrassinolide. With the SuperSAGE technology combined with next generation sequencing, induction of genes known to be associated with defense response to bacteria and hormone transduction pathways were identified. The results demonstrate that epibrassinolide may provide a useful tool for the management of HLB. PMID:26731660

  11. Changes in Anthocyanin Production during Domestication of Citrus1[OPEN

    PubMed Central

    Garcia-Lor, Andrés; Licciardello, Concetta; Las Casas, Giuseppina; Ramadugu, Chandrika; Krueger, Robert; Fanciullino, Anne-Laure; Froelicher, Yann

    2017-01-01

    Mandarin (Citrus reticulata), citron (Citrus medica), and pummelo (Citrus maxima) are important species of the genus Citrus and parents of the interspecific hybrids that constitute the most familiar commercial varieties of Citrus: sweet orange, sour orange, clementine, lemon, lime, and grapefruit. Citron produces anthocyanins in its young leaves and flowers, as do species in genera closely related to Citrus, but mandarins do not, and pummelo varieties that produce anthocyanins have not been reported. We investigated the activity of the Ruby gene, which encodes a MYB transcription factor controlling anthocyanin biosynthesis, in different accessions of a range of Citrus species and in domesticated cultivars. A white mutant of lemon lacks functional alleles of Ruby, demonstrating that Ruby plays an essential role in anthocyanin production in Citrus. Almost all the natural variation in pigmentation by anthocyanins in Citrus species can be explained by differences in activity of the Ruby gene, caused by point mutations and deletions and insertions of transposable elements. Comparison of the allelic constitution of Ruby in different species and cultivars also helps to clarify many of the taxonomic relationships in different species of Citrus, confirms the derivation of commercial varieties during domestication, elucidates the relationships within the subgenus Papeda, and allows a new genetic classification of mandarins. PMID:28196843

  12. New Pseudomonas spp. Are Pathogenic to Citrus

    PubMed Central

    Beiki, Farid; Busquets, Antonio; Gomila, Margarita; Rahimian, Heshmat; Lalucat, Jorge; García-Valdés, Elena

    2016-01-01

    Five putative novel Pseudomonas species shown to be pathogenic to citrus have been characterized in a screening of 126 Pseudomonas strains isolated from diseased citrus leaves and stems in northern Iran. The 126 strains were studied using a polyphasic approach that included phenotypic characterizations and phylogenetic multilocus sequence analysis. The pathogenicity of these strains against 3 cultivars of citrus is demonstrated in greenhouse and field studies. The strains were initially grouped phenotypically and by their partial rpoD gene sequences into 11 coherent groups in the Pseudomonas fluorescens phylogenetic lineage. Fifty-three strains that are representatives of the 11 groups were selected and analyzed by partial sequencing of their 16S rRNA and gyrB genes. The individual and concatenated partial sequences of the three genes were used to construct the corresponding phylogenetic trees. The majority of the strains were identified at the species level: P. lurida (5 strains), P. monteilii (2 strains), P. moraviensis (1 strain), P. orientalis (16 strains), P. simiae (7 strains), P. syringae (46 strains, distributed phylogenetically in at least 5 pathovars), and P. viridiflava (2 strains). This is the first report of pathogenicity on citrus of P. orientalis, P. simiae, P. lurida, P. moraviensis and P. monteilii strains. The remaining 47 strains that could not be identified at the species level are considered representatives of at least 5 putative novel Pseudomonas species that are not yet described. PMID:26919540

  13. Asian citrus psyllid genome (Diaphorina citri, Hemiptera)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Psyllid genome is a scientific breakthrough in that it opens the psyllid genetic blueprint to investigations of all questions ranging from taxonomic origins to the understanding of developmental biology, to the acquisition and transmission of pathogens. The Asian citrus psyllid, Diaphorina citri...

  14. Reasons for inconsistent citrus canker control

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Crop losses from citrus canker in 2014 for Hamlin due to premature fruit drop, or for grapefruit from unacceptable severity of fruit lesions, were highly variable due to periodic rains that in certain locations were coincident with grapefruit flushes in February-March or with early Hamlin fruit deve...

  15. Ecology of the Asian citrus pysllid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Host selection by psyllids in general appears to involve taste rather than olfaction. Adults are often less discriminating than nymphs. A priori, there is good reason to doubt that Asian citrus psyllid (ACP) uses a long-distance sex pheromone or that ACP orients to host plant volatiles over large (m...

  16. New Pseudomonas spp. Are Pathogenic to Citrus.

    PubMed

    Beiki, Farid; Busquets, Antonio; Gomila, Margarita; Rahimian, Heshmat; Lalucat, Jorge; García-Valdés, Elena

    2016-01-01

    Five putative novel Pseudomonas species shown to be pathogenic to citrus have been characterized in a screening of 126 Pseudomonas strains isolated from diseased citrus leaves and stems in northern Iran. The 126 strains were studied using a polyphasic approach that included phenotypic characterizations and phylogenetic multilocus sequence analysis. The pathogenicity of these strains against 3 cultivars of citrus is demonstrated in greenhouse and field studies. The strains were initially grouped phenotypically and by their partial rpoD gene sequences into 11 coherent groups in the Pseudomonas fluorescens phylogenetic lineage. Fifty-three strains that are representatives of the 11 groups were selected and analyzed by partial sequencing of their 16S rRNA and gyrB genes. The individual and concatenated partial sequences of the three genes were used to construct the corresponding phylogenetic trees. The majority of the strains were identified at the species level: P. lurida (5 strains), P. monteilii (2 strains), P. moraviensis (1 strain), P. orientalis (16 strains), P. simiae (7 strains), P. syringae (46 strains, distributed phylogenetically in at least 5 pathovars), and P. viridiflava (2 strains). This is the first report of pathogenicity on citrus of P. orientalis, P. simiae, P. lurida, P. moraviensis and P. monteilii strains. The remaining 47 strains that could not be identified at the species level are considered representatives of at least 5 putative novel Pseudomonas species that are not yet described.

  17. Founder lines for improved citrus biotechnology

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This article discusses the research needed to develop the RMCE strategy and molecular assays for site-specific recombinases as tools for genome manipulation. Explanation of genetic engineering used to generate transgenic citrus plants to exhibit a novel phenotype, but not to contain the recombinase...

  18. Rheology and composition of processed citrus fiber

    Technology Transfer Automated Retrieval System (TEKTRAN)

    While fibrous byproducts are abundant in supply, using them in food products in such a way to not degrade taste or texture can be challenging. Citrus fibers have been shown to have high water holding and viscous properties. However, to better incorporate dried orange pulp into foods, their propert...

  19. Engineering canker-resistant plants through CRISPR/Cas9-targeted editing of the susceptibility gene CsLOB1 promoter in citrus.

    PubMed

    Peng, Aihong; Chen, Shanchun; Lei, Tiangang; Xu, Lanzhen; He, Yongrui; Wu, Liu; Yao, Lixiao; Zou, Xiuping

    2017-03-29

    Citrus canker, caused by Xanthomonas citri subsp. citri (Xcc), is severely damaging to the global citrus industry. Targeted editing of host disease-susceptibility genes represents an interesting and potentially durable alternative in plant breeding for resistance. Here, we report improvement of citrus canker resistance through CRISPR/Cas9-targeted modification of the susceptibility gene CsLOB1 promoter in citrus. Wanjincheng orange (Citrus sinensis Osbeck) harbors at least three copies of the CsLOB1(G) allele and one copy of the CsLOB1(-) allele. The promoter of both alleles contains the effector binding element (EBEPthA4 ), which is recognized by the main effector PthA4 of Xcc to activate CsLOB1 expression to promote citrus canker development. Five pCas9/CsLOB1sgRNA constructs were designed to modify the EBEPthA4 of the CsLOB1 promoter in Wanjincheng orange. Among these constructs, mutation rates were 11.5% to 64.7%. Homozygous mutants were generated directly from citrus explants. Sixteen lines that harbored EBEPthA4 modifications were identified from 38 mutant plants. Four mutation lines (S2-5, S2-6, S2-12 and S5-13), in which promoter editing disrupted CsLOB1 induction in response to Xcc infection, showed enhanced resistance to citrus canker compared with the wild type. No canker symptoms were observed in the S2-6 and S5-13 lines. Promoter editing of CsLOB1(G) alone was sufficient to enhance citrus canker resistance in Wanjincheng orange. Deletion of the entire EBEPthA4 sequence from both CsLOB1 alleles conferred a high degree of resistance to citrus canker. The results demonstrate that CRISPR/Cas9-mediated promoter editing of CsLOB1 is an efficient strategy for generation of canker-resistant citrus cultivars. This article is protected by copyright. All rights reserved.

  20. Simultaneous visualization of two Citrus tristeza virus genotypes provides new insights into the structure of multi-component virus populations in a host.

    PubMed

    Bergua, María; Phelan, Dane M; Bak, Aurélie; Bloom, David C; Folimonova, Svetlana Y

    2016-04-01

    Complex Citrus tristeza virus (CTV) populations composed of mixtures of different strains of the virus are commonly found in citrus trees in the field. At present, little is known about how these populations are formed, maintained, and how they are structured within a host. Here we used a novel in situ hybridization approach allowing simultaneous visualization of two different RNA targets with high sensitivity and specificity to examine the distribution of two isolates, T36 and T68-1, representing phylogenetically distinct strains of CTV, in a citrus host in single and mixed infections. Remarkably, in doubly inoculated plants the two virus variants appeared to be well mixed within the infected tissue and showed no spatial segregation. In addition, both CTV variants were often found occupying the same cells. Possible mechanisms involved in shaping CTV populations and the biological significance of the observed lack of structural separation of the individual components are discussed.

  1. Transmission rates of ‘Ca. Liberibacter asiaticus’ by Asian citrus psyllid are enhanced by the presence and developmental stage of citrus flush

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Asian citrus psyllid (ACP) transmits a bacterium ‘Candidatus Liberibacter asiaticus’ (CLas) putatively responsible for a devastating citrus disease known as Asiatic huanglongbing (HLB) (= citrus greening disease). The psyllid and disease have invaded many citrus-growing regions around the world incl...

  2. Cooked rice texture and rice flour pasting properties; impacted by rice temperature during milling.

    PubMed

    Saleh, Mohammed; Meullenet, Jean-Francois

    2015-03-01

    Rice milling plays a key factor in determining rice quality and value. Therefore accurate quality assessments are critical to the rice industry. This study was undertaken to assess the effect of exposing rice to elevated temperatures during milling, on cooked rice texture and rice flour pasting properties. Two long (Cybonnett and Francis) and one medium (Jupiter) rice (oryzae sativa L.) cultivars were milled using McGill laboratory mill for 30 and 40 s after warmed up the mill before milling. Four different milling temperatures per milling duration were achieved. Cooked rice texture properties were assessed using a uniaxial compression test and rice flour pasting properties measured using a TA-2000 rheometer. Results of this study showed that exposure of rice to high temperatures during milling significantly decreased cooked rice firmness. An increase in milled rice temperature after milling from 10.0 to 13.3 °C resulted in a 5.4 and 8.1 N decrease in cooked rice firmness. Although not always significant, the increase in milled rice temperature during milling resulted in an increase in cooked rice stickiness. The increase in milling temperature also showed significant increase in rice flour pasting properties. Changes in rice functional characteristics were attributed to the changes occurring to rice chemical constituents due to temperature exposure as indicated by the increase in rice protein hydrophobicity. Proteins are known to affect rice starch water holding capacity and other starch gelatinization properties.

  3. Metabolic variations in different citrus rootstock cultivars associated with different responses to Huanglongbing.

    PubMed

    Albrecht, Ute; Fiehn, Oliver; Bowman, Kim D

    2016-10-01

    Huanglongbing (HLB) is one of the most destructive bacterial diseases of citrus. No resistant cultivars have been identified, although tolerance has been observed in the genus Poncirus and some of its hybrids with Citrus that are commonly used as rootstocks. In this study we exploited this tolerance by comparing five different tolerant hybrids with a cultivar that shows pronounced HLB sensitivity to discern potential contributing metabolic factors. Whole leaves of infected and non-infected greenhouse-grown seedlings were extracted and subjected to untargeted GC-TOF MS based metabolomics. After BinBase data filtering, 342 (experiment 1) and 650 (experiment 2) unique metabolites were quantified, of which 122 and 195, respectively, were assigned by chemical structures. The number of metabolites found to be differently regulated in the infected state compared with the non-infected state varied between the cultivars and was largest (166) in the susceptible cultivar Cleopatra mandarin (Citrus reticulata) and lowest (3) in the tolerant cultivars US-897 (C. reticulata 'Cleopatra' × Poncirus trifoliata) and US-942 (C. reticulata 'Sunki' × P. trifoliata) from experiment 2. Tolerance to HLB did not appear to be associated with accumulation of higher amounts of protective metabolites in response to infection. Many metabolites were found in higher concentrations in the tolerant cultivars compared with susceptible Cleopatra mandarin and may play important roles in conferring tolerance to HLB. Lower availability of specific sugars necessary for survival of the pathogen may also be a contributing factor in the decreased disease severity observed for these cultivars.

  4. Slight Fermentation with Lactobacillus fermentium Improves the Taste (Sugar:Acid Ratio) of Citrus (Citrus reticulata cv. chachiensis) Juice.

    PubMed

    Yu, Yuanshan; Xiao, Gengsheng; Xu, Yujuan; Wu, Jijun; Fu, Manqin; Wen, Jing

    2015-11-01

    The aim of this study was to evaluate the hypothesis that fermentation with Lactobacillus fermentium, which can metabolize citric acid, could be applied in improving the taste (sugar:acid ratio) of citrus juice. During fermentation, the strain of L. fermentium can preferentially utilize citric acid of citrus (Citrus reticulata cv. Chachiensis) juice to support the growth without the consumption of sugar. After 6 h of fermentation with L. fermentium at 30 °C, the sugar:acid ratio of citrus juice increased to 22:1 from 12:1, which resulted in that the hedonic scores of sweetness, acidity and overall acceptability of fermented-pasteurized citrus juice were higher than the unfermented-pasteurized citrus juice. Compared with unfermented-pasteurized citrus juice, the ORAC value and total amino acid showed a reduction, and no significant change (P > 0.05) in the L*, a*, b*, total soluble phenolics and ascorbic acid (Vc) content in the fermented-pasteurized citrus juice was observed as compared with unfermented-pasteurized citrus juice. Hence, slight fermentation with L. fermentium can be used for improving the taste (sugar:acid ratio) of citrus juice with the well retaining of quality.

  5. Lactobacillus paraplantarum 11-1 Isolated from Rice Bran Pickles Activated Innate Immunity and Improved Survival in a Silkworm Bacterial Infection Model.

    PubMed

    Nishida, Satoshi; Ishii, Masaki; Nishiyama, Yayoi; Abe, Shigeru; Ono, Yasuo; Sekimizu, Kazuhisa

    2017-01-01

    Lactic acid bacteria (LAB) have high immune system-stimulating activity and are considered beneficial for human health as probiotics in the gut. The innate immune system is highly conserved between mammals and insects. Microbe-associated molecular patterns (e.g., peptidoglycan and β-glucan) induce cytokine maturation, which, in silkworm larvae, leads to muscle contraction. The purpose of this study is to find a novel probiotic by using silkworm muscle contraction assay. In the present study, we isolated LAB derived from rice bran pickles. We selected highly active LAB to activate the innate immune system of the silkworm, which was assayed based on silkworm muscle contraction. Of various LAB, L. paraplantarum 11-1 strongly stimulated innate immunity in the silkworm, leading to stronger silkworm contraction than a dairy-based LAB. Silkworms fed a diet containing L. paraplantarum 11-1 exhibited tolerance against the pathogenicity of Pseudomonas aeruginosa. These findings suggest that L. paraplantarum 11-1 could be a useful probiotic for activating innate immunity.

  6. Lactobacillus paraplantarum 11-1 Isolated from Rice Bran Pickles Activated Innate Immunity and Improved Survival in a Silkworm Bacterial Infection Model

    PubMed Central

    Nishida, Satoshi; Ishii, Masaki; Nishiyama, Yayoi; Abe, Shigeru; Ono, Yasuo; Sekimizu, Kazuhisa

    2017-01-01

    Lactic acid bacteria (LAB) have high immune system-stimulating activity and are considered beneficial for human health as probiotics in the gut. The innate immune system is highly conserved between mammals and insects. Microbe-associated molecular patterns (e.g., peptidoglycan and β-glucan) induce cytokine maturation, which, in silkworm larvae, leads to muscle contraction. The purpose of this study is to find a novel probiotic by using silkworm muscle contraction assay. In the present study, we isolated LAB derived from rice bran pickles. We selected highly active LAB to activate the innate immune system of the silkworm, which was assayed based on silkworm muscle contraction. Of various LAB, L. paraplantarum 11-1 strongly stimulated innate immunity in the silkworm, leading to stronger silkworm contraction than a dairy-based LAB. Silkworms fed a diet containing L. paraplantarum 11-1 exhibited tolerance against the pathogenicity of Pseudomonas aeruginosa. These findings suggest that L. paraplantarum 11-1 could be a useful probiotic for activating innate immunity. PMID:28373863

  7. Direct suppression of a rice bacterial blight (Xanthomonas oryzae pv. oryzae) by monoterpene (S)-limonene.

    PubMed

    Lee, Gun Woong; Chung, Moon-Soo; Kang, Mihyung; Chung, Byung Yeoup; Lee, Sungbeom

    2016-05-01

    Rice bacterial blight, caused by Xanthomonas oryzae pv. oryzae (Xoo), is a severe disease of rice plants. Upon pathogen infection, rice biosynthesizes phytoalexins, including diterpenoids such as momilactones, phytocassanes, and oryzalexins. However, information on headspace volatiles in response to Xoo infection is limited. We have examined headspace volatile terpenes, induced by the infection of Xoo, and investigated their biological roles in the rice plant. Monoterpenes α-thujene, α-pinene, sabinene, myrcene, α-terpene, and (S)-limonene and sesquiterpenes cyclosativene, α-copaene, and β-elemene were detected from 1-week-old Xoo-infected rice seedlings, by solid-phase microextraction-gas chromatography-mass spectrometry. All monoterpenes were constitutively released from rice seedlings before Xoo infection. However, (S)-limonene emission was further elicited after exposure of the seedlings to Xoo in coincidence with upregulation of limonene synthase gene (OsTPS20) transcripts. Only the stereospecific (S)-limonene [and not (R)-limonene or other monoterpenes] severely inhibited Xoo growth, as confirmed by disc diffusion and liquid culture assays. Rice seedlings showed suppressed pathogenic symptoms suggestive of resistance to Xoo infection after foliar treatment with (S)-limonene. Collectively, our findings suggest that (S)-limonene is a volatile phytoanticipin, which plays a significant role in suppressing Xoo growth in rice seedlings.

  8. A genetic system for Citrus Tristeza Virus using the non-natural host Nicotiana benthamiana: an update

    PubMed Central

    Ambrós, Silvia; Ruiz-Ruiz, Susana; Peña, Leandro; Moreno, Pedro

    2013-01-01

    In nature Citrus tristeza virus (CTV), genus Closterovirus, infects only the phloem cells of species of Citrus and related genera. Finding that the CTV T36 strain replicated in Nicotiana benthamiana (NB) protoplasts and produced normal virions allowed development of the first genetic system based on protoplast transfection with RNA transcribed from a full-genome cDNA clone, a laborious and uncertain system requiring several months for each experiment. We developed a more efficient system based on agroinfiltration of NB leaves with CTV-T36-based binary plasmids, which caused systemic infection in this non-natural host within a few weeks yielding in the upper leaves enough CTV virions to readily infect citrus by slash inoculation. Stem agroinoculation of citrus and NB plants with oncogenic strains of Agrobacterium tumefaciens carrying a CTV-T36 binary vector with a GUS marker, induced GUS positive galls in both species. However, while most NB tumors were CTV positive and many plants became systemically infected, no coat protein or viral RNA was detected in citrus tumors, even though CTV cDNA was readily detected by PCR in the same galls. This finding suggests (1) strong silencing or CTV RNA processing in transformed cells impairing infection progress, and (2) the need for using NB as an intermediate host in the genetic system. To maintain CTV-T36 in NB or assay other CTV genotypes in this host, we also tried to graft-transmit the virus from infected to healthy NB, or to mechanically inoculate NB leaves with virion extracts. While these trials were mostly unsuccessful on non-treated NB plants, agroinfiltration with silencing suppressors enabled for the first time infecting NB plants by side-grafting and by mechanical inoculation with virions, indicating that previous failure to infect NB was likely due to virus silencing in early infection steps. Using NB as a CTV host provides new possibilities to study virus-host interactions with a simple and reliable system. PMID

  9. Citrus alongside the sinking wreckage of MV Pacific Star in ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    Citrus alongside the sinking wreckage of MV Pacific Star in the Pacific Ocean. Pacific Star's captain used his vessel to ram the cutter after he was ordered to stop and submit to inspection by a boarding team. Citrus was not seriously damaged in the collision. U.S. Coast Guard personnel recovered a large amount of marijuana from the wreckage - U.S. Coast Guard Cutter CITRUS, Coos Bay, Coos County, OR

  10. Comparative influence of Radopholus similis and Pratylenchus coffeae on citrus.

    PubMed

    O'Bannon, J H; Radewald, J D; Tomerlin, A T; Inserra, R N

    1976-01-01

    Pratylenchus coffeae was as pathogenic as Radopholus sirnilis to commercial citrus rootstocks. No rootstock resistant to R. similis was resistant to P. coffeae. Both nematodes stunted citrus in three soil types. Seedling damage by P. coffeae and R. similis was greatest in fine- and coarse-textured soils, respectively. Reproduction and survival on citrus were greater for P. coffeae than for R. similis. Mixed inoculations with R. similis and P. coffeae resulted in lower populations of each species than did separate inoculations.

  11. Synergism between southern rice black-streaked dwarf virus and rice ragged stunt virus enhances their insect vector acquisition.

    PubMed

    Li, Shu; Wang, Han; Zhou, Guohui

    2014-07-01

    Southern rice black-streaked dwarf virus (SRBSDV), a tentative species in the genus Fijivirus, family Reoviridae, is a novel rice virus transmitted by the white-backed planthopper (Sogatella furcifera). Since its discovery in 2001, SRBSDV has spread rapidly throughout eastern and southeastern Asia and caused large rice losses in China and Vietnam. Rice ragged stunt virus (RRSV) (genus Oryzavirus, family Reoviridae) is a common rice virus vectored by the brown planthopper (Nilaparvata lugens). RRSV is also widely distributed in eastern and southeastern Asia but has not previously caused serious problems in China owing to its low incidence. With SRBSDV's spread, however, RRSV has become increasingly common in China, and is frequently found in co-infection with SRBSDV. In this study, we show that SRBSDV and RRSV interact synergistically, the first example of synergism between plant viruses in the family Reoviridae. Rice plants co-infected with both viruses displayed enhanced stunting, earlier symptoms, and higher virus titers compared with singly infected plants. Furthermore, white-backed and brown planthoppers acquired SRBSDV and RRSV, respectively, from co-infected plants at higher rates. We propose that increased RRSV incidence in Chinese fields is partly due to synergism between SRBSDV and RRSV.

  12. Discrimination of Citrus reticulata Blanco and Citrus reticulata 'Chachi' by gas chromatograph-mass spectrometry based metabolomics approach.

    PubMed

    Duan, Li; Guo, Long; Dou, Li-Li; Zhou, Chang-Lin; Xu, Feng-Guo; Zheng, Guo-Dong; Li, Ping; Liu, E-Hu

    2016-12-01

    Citri Reticulatae Pericarpium, mainly including the pericarp of Citrus reticulata Blanco and the pericarp of Citrus reticulata 'Chachi', has been consumed daily as food and dietary supplement for centuries. In this study, GC-MS based metabolomics was employed to compare comprehensively the volatile constituents in Citrus reticulata Blanco and Citrus reticulata 'Chachi'. Principal component analysis and orthogonal partial least squares discrimination analysis indicated that samples could be distinguished effectively from one another. Fifteen metabolites were finally identified for use as chemical markers in discrimination of Citri Reticulatae Pericarpium samples. The antimicrobial activity against Gram-negative and Gram-positive bacteria of the volatile oil from Citrus reticulata Blanco and Citrus reticulata 'Chachi' was investigated preliminarily.

  13. Contextual view showing northeastern eucalyptus windbreak and portion of citrus ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    Contextual view showing northeastern eucalyptus windbreak and portion of citrus orchard. Camera facing 118" east-southeast. - Goerlitz House, 9893 Highland Avenue, Rancho Cucamonga, San Bernardino County, CA

  14. Identification of putative TAL effector targets of the citrus canker pathogens shows functional convergence underlying disease development and defense response

    PubMed Central

    2014-01-01

    Background Transcriptional activator-like (TAL) effectors, formerly known as the AvrBs3/PthA protein family, are DNA-binding effectors broadly found in Xanthomonas spp. that transactivate host genes upon injection via the bacterial type three-secretion system. Biologically relevant targets of TAL effectors, i.e. host genes whose induction is vital to establish a compatible interaction, have been reported for xanthomonads that colonize rice and pepper; however, citrus genes modulated by the TAL effectors PthA“s” and PthC“s” of the citrus canker bacteria Xanthomonas citri (Xc) and Xanthomonas aurantifolii pathotype C (XaC), respectively, are poorly characterized. Of particular interest, XaC causes canker disease in its host lemon (Citrus aurantifolia), but triggers a defense response in sweet orange. Results Based on, 1) the TAL effector-DNA binding code, 2) gene expression data of Xc and XaC-infiltrated sweet orange leaves, and 3) citrus hypocotyls transformed with PthA2, PthA4 or PthC1, we have identified a collection of Citrus sinensis genes potentially targeted by Xc and XaC TAL effectors. Our results suggest that similar with other strains of Xanthomonas TAL effectors, PthA2 and PthA4, and PthC1 to some extent, functionally converge. In particular, towards induction of genes involved in the auxin and gibberellin synthesis and response, cell division, and defense response. We also present evidence indicating that the TAL effectors act as transcriptional repressors and that the best scoring predicted DNA targets of PthA“s” and PthC“s” in citrus promoters predominantly overlap with or localize near to TATA boxes of core promoters, supporting the idea that TAL effectors interact with the host basal transcriptional machinery to recruit the RNA pol II and start transcription. Conclusions The identification of PthA“s” and PthC“s” targets, such as the LOB (LATERAL ORGAN BOUNDARY) and CCNBS genes that we report here, is key for the understanding

  15. The role as inoculum sources of Xanthomonas citri pv. citri surviving on the infected Satsuma mandarin fruits.

    PubMed

    Kang, So Young; Kim, Ki Deok; Hong, Jeum Kyu; Hyun, He Nam; Jeun, Yong Chull

    2014-05-01

    Importing citrus fruits infected by Asiatic citrus canker caused by Xanthomonas citri pv. citri (Xcc) can act as an inoculum source for the disease epidemic in citrus canker-free countries. In this study, the pathogenicity of the causal agent of Asiatic citrus canker surviving on infected Satsuma mandarin fruits was evaluated. The washing solution of infected Satsuma mandarin fruits did not cause lesion formation on the citrus leaves. However, a typical citrus canker lesion was formed on the leaves after inoculation with higher concentrations of the inoculum from the washing solution (washing solution II). It indicated that the pathogenicity of the citrus canker surviving on the symptomatic Satsuma mandarin fruits was not changed. Scanning electron microscopic observation showed that the numbers of bacterial cells on the leaves of Satsuma mandarin which inoculated with the washing solution directly (washing solution I) was less compared to those of leaves inoculated with the washing solution II. This result spports that the pathogenicity of Xcc surviving on Satsuma mandarin fruits may not be changed but that the sucessful infection of citrus caker may depend on the concentration of the inoculum.

  16. Correlation of Diplodia (Lasiodiplodia theobromae) infection, huanglongbing, ethylene production, fruit removal force and pre-harvest fruit drop

    Technology Transfer Automated Retrieval System (TEKTRAN)

    One symptom of citrus huanglongbing (HLB) disease is excessive pre-harvest fruit drop. As the severity of HLB has progressed through commercial citrus groves in Florida, pre-harvest fruit drop has increased, resulting in significant loss of yield. A greater incidence of Diplodia infection was recent...

  17. MicroRNAs in Rice Innate Immunity.

    PubMed

    Baldrich, Patricia; San Segundo, Blanca

    2016-12-01

    MicroRNAs (miRNAs) are short regulatory non-coding RNAs that guide gene silencing in most eukaryotes. They regulate gene expression by triggering sequence-specific cleavage or translational repression of target transcripts. Plant miRNAs are known to play important roles in a wide range of developmental processes. Increasing evidence also supports that the modulation of miRNA levels plays an important role in reprogramming plant responses to abiotic stress (drought, cold, salinity and nutrient deficiency) and biotic stress (antibacterial resistance). Most of these studies were carried out in the model plant Arabidopsis thaliana. During the last years, the adoption of high-throughput sequencing technologies has significantly contributed to uncover multiple miRNAs while allowing miRNA profiling in plants. However, although a plethora of rice miRNAs have been shown to be regulated by pathogen infection, the biological function remains largely unknown for most of them. In this review, we summarize our current understanding on the contribution of miRNAs to rice immunity and discuss their potential applications in rice biotechnology. A better understanding of the miRNA species controlling rice immunity may lead to practical biotechnological applications leading to the development of appropriate strategies for rice protection.

  18. Determinants for grading Malaysian rice

    NASA Astrophysics Data System (ADS)

    ChePa, Noraziah; Yusoff, Nooraini; Ahmad, Norhayati

    2016-08-01

    Due to un-uniformity of rice grading practices in Malaysia, zones which actively producing rice in Malaysia are using their own way of grading rice. Rice grading is important in determining rice quality and its subsequent price in the market. It is an important process applied in the rice production industry with the purpose of ensuring that the rice produced for the market meets the quality requirements of consumer. Two important aspects that need to be considered in determining rice grades are grading technique and determinants to be used for grading (usually referred as rice attributes). This article proposes the list of determinants to be used in grading Malaysian rice. Determinants were explored through combination of extensive literature review and series of interview with the domain experts and practitioners. The proposed determinants are believed to be beneficial to BERNAS in improving the current Malaysian rice grading process.

  19. Rapid Assessment of the Citrus Tristeza Virus Isolates Detected in Spring 2007 at the Lindcove Research and Extension Center, Exeter, Calif.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) was detected in at least 50 trees at the 71 ha Lindcove Research and Extension Center (LREC) near Exeter, Calif. in spring 2007. In the previous 3 years, 3, 1, and 5 trees were infected. The purpose of this research was to assess the aphid transmissibility and molecular ...

  20. RNAi suppression of rice endogenous storage proteins enhances the production of rice-based Botulinum neutrotoxin type A vaccine.

    PubMed

    Yuki, Yoshikazu; Mejima, Mio; Kurokawa, Shiho; Hiroiwa, Tomoko; Kong, Il Gyu; Kuroda, Masaharu; Takahashi, Yoko; Nochi, Tomonori; Tokuhara, Daisuke; Kohda, Tomoko; Kozaki, Shunji; Kiyono, Hiroshi

    2012-06-13

    Mucosal vaccines based on rice (MucoRice) offer a highly practical and cost-effective strategy for vaccinating large populations against mucosal infections. However, the limitation of low expression and yield of vaccine antigens with high molecular weight remains to be overcome. Here, we introduced RNAi technology to advance the MucoRice system by co-introducing antisense sequences specific for genes encoding endogenous rice storage proteins to minimize storage protein production and allow more space for the accumulation of vaccine antigen in rice seed. When we used RNAi suppression of a combination of major rice endogenous storage proteins, 13 kDa prolamin and glutelin A in a T-DNA vector, we could highly express a vaccine comprising the 45 kDa C-terminal half of the heavy chain of botulinum type A neurotoxin (BoHc), at an average of 100 μg per seed (MucoRice-BoHc). The MucoRice-Hc was water soluble, and was expressed in the cytoplasm but not in protein body I or II of rice seeds. Thus, our adaptation of the RNAi system improved the yield of a vaccine antigen with a high molecular weight. When the mucosal immunogenicity of the purified MucoRice-BoHc was examined, the vaccine induced protective immunity against a challenge with botulinum type A neurotoxin in mice. These findings demonstrate the efficiency and utility of the advanced MucoRice system as an innovative vaccine production system for generating highly immunogenic mucosal vaccines of high-molecular-weight antigens.

  1. Combination of multispectral remote sensing, variable rate technology and environmental modeling for citrus pest management.

    PubMed

    Du, Qian; Chang, Ni-Bin; Yang, Chenghai; Srilakshmi, Kanth R

    2008-01-01

    The Lower Rio Grande Valley (LRGV) of south Texas is an agriculturally rich area supporting intensive production of vegetables, fruits, grain sorghum, and cotton. Modern agricultural practices involve the combined use of irrigation with the application of large amounts of agrochemicals to maximize crop yields. Intensive agricultural activities in past decades might have caused potential contamination of soil, surface water, and groundwater due to leaching of pesticides in the vadose zone. In an effort to promote precision farming in citrus production, this paper aims at developing an airborne multispectral technique for identifying tree health problems in a citrus grove that can be combined with variable rate technology (VRT) for required pesticide application and environmental modeling for assessment of pollution prevention. An unsupervised linear unmixing method was applied to classify the image for the grove and quantify the symptom severity for appropriate infection control. The PRZM-3 model was used to estimate environmental impacts that contribute to nonpoint source pollution with and without the use of multispectral remote sensing and VRT. Research findings using site-specific environmental assessment clearly indicate that combination of remote sensing and VRT may result in benefit to the environment by reducing the nonpoint source pollution by 92.15%. Overall, this study demonstrates the potential of precision farming for citrus production in the nexus of industrial ecology and agricultural sustainability.

  2. Phylogenetic and Molecular Variability Studies Reveal a New Genetic Clade of Citrus leprosis virus C

    PubMed Central

    Ramos-González, Pedro Luis; Chabi-Jesus, Camila; Guerra-Peraza, Orlene; Breton, Michèle Claire; Arena, Gabriella Dias; Nunes, Maria Andreia; Kitajima, Elliot Watanabe; Machado, Marcos Antonio; Freitas-Astúa, Juliana

    2016-01-01

    Citrus leprosis virus C (CiLV-C) causes a severe disease affecting citrus orchards in the Western hemisphere. This study reveals the molecular variability of the virus by analyzing four genomic regions (p29, p15, MP and RNA2-intergenic region) distributed over its two RNAs. Nucleotide diversity (π) values were relatively low but statistically different over the analyzed genes and subpopulations, indicating their distinct evolutionary history. Values of πp29 and πMP were higher than those of πp15 and πRNA2–IR, whereas πMP was increased due to novel discovered isolates phylogenetically clustered in a divergent clade that we called SJP. Isolate BR_SP_SJP_01 RNA1 and RNA2 sequences, clade SJP, showed an identity of 85.6% and 88.4%, respectively, with those corresponding to CiLV-C, the type member of the genus Cilevirus, and its RNA2 5′-proximal region was revealed as a minor donor in a putative inter-clade recombination event. In addition to citrus, BR_SP_SJP_01 naturally infects the weed Commelina benghalensis and is efficiently transmitted by Brevipalpus yothersi mites. Our data demonstrated that negative selection was the major force operating in the evaluated viral coding regions and defined amino acids putatively relevant for the biological function of cilevirus proteins. This work provides molecular tools and sets up a framework for further epidemiological studies. PMID:27275832

  3. Immunodiagnosis of Citrus leprosis virus C using a polyclonal antibody to an expressed putative coat protein.

    PubMed

    Choudhary, Nandlal; Roy, Avijit; Guillermo, Leon M; Picton, D D; Wei, G; Nakhla, M K; Levy, L; Brlansky, R H

    2013-11-01

    Citrus leprosis virus C (CiLV-C), a causal agent for citrus leprosis disease, is present in South and Central America and is a threat for introduction into the U.S. citrus industry. A specific, inexpensive and reliable antibody based detection system is needed for the rapid identification of CiLV-C. The CiLV-C is very labile and has not been purified in sufficient amount for antibody production. The p29 gene of CiLV-C genome that codes for the putative coat protein (PCP) was codon optimized for expression in Escherichia coli and synthesized in vitro. The optimized gene was sub-cloned into the bacterial expression vector pDEST17 and transferred into E. coli BL21AI competent cells. The expression of PCP containing N-terminal His-tag was optimized by induction with l-arabinose. Induced cells were disrupted by sonication and expressed PCP was purified by affinity chromatography using Ni-NTA agarose. The purified expressed PCP was then used as an immunogen for injections into rabbits to produce polyclonal antibody (PAb). The PAb specific to the expressed PCP was identified using Western blotting. The antibody was successfully used to detect CiLV-C in the symptomatic CiLV-C infected tissues using double antibody sandwich-enzyme-linked-immunosorbent (DAS-ELISA), indirect ELISA and dot-blot immunoassay (DBIA) formats.

  4. Selection of reference genes for expression analyses of red-fleshed sweet orange (Citrus sinensis).

    PubMed

    Pinheiro, T T; Nishimura, D S; De Nadai, F B; Figueira, A; Latado, R R

    2015-12-28

    Red-fleshed oranges (Citrus sinensis) contain high levels of carotenoids and lycopene. The growing consumer demand for products with health benefits has increased interest in these types of Citrus cultivars as a potential source of nutraceuticals. However, little is known about the physiology of these cultivars under Brazilian conditions. Transcriptome and gene expression analyses are important tools in the breeding and management of red-fleshed sweet orange cultivars. Reverse transcription quantitative polymerase chain reaction is a method of quantifying gene expression, but various standardizations are required to obtain precise, accurate, and specific results. Among the standardizations required, the choice of suitable stable reference genes is fundamental. The objective of this study was to evaluate the stability of 11 candidate genes using various tissue and organ samples from healthy plants or leaves from citrus greening disease (Huanglongbing)-symptomatic plants of a Brazilian red-fleshed cultivar ('Sanguínea de Mombuca'), in order to select the most suitable reference gene for investigating gene expression under these conditions. geNorm and NormFinder identified genes that encoded translation initiation factor 3, ribosomal protein L35, and translation initiation factor 5A as the most stable genes under the biological conditions tested, and genes coding actin (ACT) and the subunit of the PSI reaction center subunit III were the least stable. Phosphatase, malate dehydrogenase, and ACT were the most stable genes in the leaf samples of infected plants.

  5. Viability of 'Candidatus Liberibacter asiaticus' prolonged by addition of citrus juice to culture medium.

    PubMed

    Parker, Jennifer K; Wisotsky, Sarah R; Johnson, Evan G; Hijaz, Faraj M; Killiny, Nabil; Hilf, Mark E; De La Fuente, Leonardo

    2014-01-01

    Huanglongbing, or citrus greening disease, is associated with infection by the phloem-limited bacterium 'Candidatus Liberibacter asiaticus'. Infection with 'Ca. L. asiaticus' is incurable; therefore, knowledge regarding 'Ca. L. asiaticus' biology and pathogenesis is essential to develop a treatment. However, 'Ca. L. asiaticus' cannot currently be successfully cultured, limiting its study. To gain insight into the conditions conducive for growth of 'Ca. L. asiaticus' in vitro, 'Ca. L. asiaticus' inoculum obtained from seed of fruit from infected pomelo trees (Citrus maxima 'Mato Buntan') was added to different media, and cell viability was monitored for up to 2 months using quantitative polymerase chain reaction in conjunction with ethidium monoazide. Media tested included one-third King's B (K), K with 50% juice from the infected fruit, K with 50% commercially available grapefruit juice, and 100% commercially available grapefruit juice. Results show that juice-containing media dramatically prolong viability compared with K in experiments reproduced during 2 years using different juice sources. Furthermore, biofilm formed at the air-liquid interface of juice cultures contained 'Ca. L. asiaticus' cells, though next-generation sequencing indicated that other bacterial genera were predominant. Chemical characterization of the media was conducted to discuss possible factors sustaining 'Ca. L. asiaticus' viability in vitro, which will contribute to future development of a culture medium for 'Ca. L. asiaticus'.

  6. Proteome profile of maize (Zea Mays L.) leaf tissue at the flowering stage after long-term adjustment to rice black-streaked dwarf virus infection.

    PubMed

    Li, Kunpeng; Xu, Changzheng; Zhang, Juren

    2011-10-10

    Maize rough dwarf disease (MRDD) is a viral disease and causes great yield loss. To better understand the effects of MRDD on plant growth and metabolism, comparative proteomic analysis of leaves from virus-infected and normal plants was performed. In order to eliminate the interference of Ribulose-1, 5-bisphosphate carboxylase with low-abundance proteins, total proteins were pre-fractionated by 15% PEG and the proteins from supernatant and precipitated fractions were analyzed by 2-DE, subsequently. Out of approximately 1200 protein spots detected, less than 2% of the spots on the gels were overlapping between the fractions of precipitation and supernatant. We identified 91 differentially accumulated proteins that belong to multiple metabolic/biochemical pathways in plants. Further analysis of these identified proteins indicated that MRDD resulted in dramatic changes in the fundamental metabolism, including glycolysis and starch metabolism, and eventually the significant differences in morphology and development between virus-infected and normal plants. Moreover, MRDD occurrence increased the demands for G-proteins, antioxidant enzymes, lipoxygenases and UDP-glucosyltransferase BX9, which may play important roles in response of plant against virus infection. The results also suggested that MRDD is a complicated disease controlled by multigene participating in different pathways.

  7. Subacute toxicity assessment of carotenoids extracted from citrus peel (Nanfengmiju, Citrus reticulata Blanco) in rats.

    PubMed

    Xue, Feng; Li, Chen; Pan, Siyi

    2012-02-01

    The mixture of carotenoids extracted from citrus peel (Nanfengmiju, Citrus reticulata Blanco) was tested for subacute oral toxicity. In this study, dose levels of 0, 200, 500 and 2000 mg/kg body weight/day were administered by gavage to 10 Wistar rats/sex/group for 28 days. No statistically significant, dose-related effect on food consumption, food efficiency, body weight gain, clinical signs or ophthalmoscopic parameters was observed in any treatment group. Urinalysis, hematological, blood coagulation and serum biochemical examination as well as necropsy or histopathology showed that no observed adverse effect was found. These findings suggested that the No-Observed-Adverse-Effect Level for the mixture of carotenoids extracted from citrus peel was at least 2000 mg/kg body weight/day.

  8. Chemical and biological comparison of the fruit extracts of Citrus wilsonii Tanaka and Citrus medica L.

    PubMed

    Zhao, Pan; Duan, Li; Guo, Long; Dou, Li-Li; Dong, Xin; Zhou, Ping; Li, Ping; Liu, E-Hu

    2015-04-15

    Citri Fructus (CF), the mature fruit of Citrus wilsonii Tanaka (CWT) or Citrus medica L. (CML), is an important citrus by-product with health promoting and nutritive properties. The present study compares the chemical and biological differences of CWT and CML. Thin layer chromatography and high performance liquid chromatography, coupled with quadrupole time-of-flight tandem mass spectrometry techniques, were employed to compare the chemical profiles of CWT and CML. A total of 25 compounds were identified and the results indicated that there were significant differences in chemical composition between the two CF species. The quantitative results obtained by HPLC coupled with diode array detector method demonstrated that naringin was present in the highest amounts in CWT, whilst nomilin was the most dominant constituent in CML. It was also found that CWT had significantly higher free radical-scavenging activity than CML.

  9. Citrus transformation using juvenile tissue explants.

    PubMed

    Orbović, Vladimir; Grosser, Jude W

    2015-01-01

    The most frequently used method for production of citrus transgenic plants is via Agrobacterium-mediated transformation of tissues found on explants obtained from juvenile seedlings. Within the last decade and especially within the last 5-6 years, this robust method was employed to produce thousands of transgenic plants. With the newly applied screening methods that allow easier and faster detection of transgenic shoots, estimates of transformation rate for some cultivars have gone up making this approach even more attractive. Although adjustments have to be made regarding the (varietal) source of the starting material and Agrobacterium strain used in each experiment preformed, the major steps of this procedure have not changed significantly if at all. Transgenic citrus plants produced this way belong to cultivars of rootstocks, sweet oranges, grapefruits, mandarins, limes, and lemons.

  10. Cyclic Lipopeptides from Bacillus subtilis ABS–S14 Elicit Defense-Related Gene Expression in Citrus Fruit

    PubMed Central

    Waewthongrak, Waewruedee; Leelasuphakul, Wichitra; McCollum, Greg

    2014-01-01

    Effects of cyclic lipopeptides (CLPs) obtained from Bacillus subtilis ABS-S14 on eliciting defense-related gene transcription and activity of defense-related enzymes; glucanase (GLU), chitinase (CHI), peroxidase (POX) and lipoxygenase (LOX) in Citrus sinensis cv. Valencia fruit were determined. The maximum level of GLU transcripts induced in fruit treated with fengycin was significantly greatest among treatments at 48 h. Surfactin enhanced the LOX and POX transcripts. In parallel, corresponding enzyme activities were correlated with changes in gene expression observed in fruit inoculated with Penicillium digitatum following treatment with individual CLPs. Synergistic effects of fengycin and iturin A, fengycin and surfactin were shown in gene transcript of GLU and CHI, respectively, and surfactin induced POX and LOX gene expression of citrus flavedo without pathogen infection. These results suggest that fengycin and surfactin act as elicitors of defense-related gene expression in “Valencia” fruit following infection. PMID:25329301

  11. Cyclic LIPopeptides from Bacillus subtilis ABS-S14 elicit defense-related gene expression in citrus fruit.

    PubMed

    Waewthongrak, Waewruedee; Leelasuphakul, Wichitra; McCollum, Greg

    2014-01-01

    Effects of cyclic lipopeptides (CLPs) obtained from Bacillus subtilis ABS-S14 on eliciting defense-related gene transcription and activity of defense-related enzymes; glucanase (GLU), chitinase (CHI), peroxidase (POX) and lipoxygenase (LOX) in Citrus sinensis cv. Valencia fruit were determined. The maximum level of GLU transcripts induced in fruit treated with fengycin was significantly greatest among treatments at 48 h. Surfactin enhanced the LOX and POX transcripts. In parallel, corresponding enzyme activities were correlated with changes in gene expression observed in fruit inoculated with Penicillium digitatum following treatment with individual CLPs. Synergistic effects of fengycin and iturin A, fengycin and surfactin were shown in gene transcript of GLU and CHI, respectively, and surfactin induced POX and LOX gene expression of citrus flavedo without pathogen infection. These results suggest that fengycin and surfactin act as elicitors of defense-related gene expression in "Valencia" fruit following infection.

  12. Relating raw rice color and composition to cooked rice color.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Traditionally, the color of milled rice is economically important. The whiter the rice the more it is preferred by consumers and the more value it has in the market place. Little attention has been given to relating raw rice color to cooked milled rice color and, specifically, to determining the i...

  13. New depside from Citrus reticulata Blanco.

    PubMed

    Phetkul, Uraiwan; Phongpaichit, Souwalak; Watanapokasin, Ramida; Mahabusarakam, Wilawan

    2014-01-01

    A new depside, named depcitrus A (1), and 31 known compounds were isolated from the peels, leaves and branch barks of Citrus reticulata Blanco. Methylation of the high polarity fractions from the branch barks and peels gave one new methylated compound named depcitrus B (14) and five known compounds. Their structures were established based on spectroscopic evidence. The antioxidant, antimicrobial and cytotoxic activities of some pure compounds were evaluated.

  14. Comparison of evapotranspiration rates for flatwoods and ridge citrus

    USGS Publications Warehouse

    Jia, X.; Swancar, A.; Jacobs, J.M.; Dukes, M.D.; Morgan, K.

    2007-01-01

    Florida citrus groves are typically grown in two regions of the state: flatwoods and ridge. The southern flatwoods citrus area has poorly drained fine textured sands with low organic matter in the shallow root zone. Ridge citrus is located in the northern ridge citrus zone and has fine to coarse textured sands with low water-holding capacity. Two commercial citrus groves, selected from each region, were studied from 15 July 2004 to 14 July 2005. The flatwoods citrus (FC) grove had a grass cover and used drainage ditches to remove excess water from the root zone. The ridge citrus (RC) grove had a bare soil surface with weeds periodically eliminated by tillage. Citrus crop evapotranspiration (ETc) rates at the two citrus groves were measured by the eddy correlation method, and components in the energy balance were also examined and compared. The study period had higher than average rainfall, and as a result, the two locations had similar annual ETc rates (1069 and 1044 mm for RC and FC, respectively). The ETc rates were 59% (RC) and 47% (FC) of the rainfall amounts during the study period. The annual reference crop evapotranspiration (ETo) rates were 1180 mm for RC and 1419 mm for FC, estimated using the standardized reference evapotranspiration equation. The citrus crop coefficients (Kc, ratio of ETc to ET o) were different between the two locations because of differences in latitude, ground cover, and rainfall amounts. The Kc values ranged from 0.70 between December and March to 1.05 between July and November for RC, and from 0.65 between November and May to 0.85 between June and October for FC. The results are consistent with other Kc values reported from field studies on citrus in both Florida and elsewhere using these and alternate methods.

  15. The citrus postharvest pathogen Penicillium digitatum depends on the PdMpkB kinase for developmental and virulence functions.

    PubMed

    Ma, Haijie; Sun, Xuepeng; Wang, Mingshuang; Gai, Yunpeng; Chung, Kuang-Ren; Li, Hongye

    2016-11-07

    The postharvest pathogen Penicillium digitatum causes green mold decay on citrus fruit, resulting in severe economic losses. To explore possible factors involved in fungal pathogenesis, phenotypic characterization of the budding yeast Fus3/Kiss1 mitogen-activated protein (MAP) kinase homolog was carried out. The P. digitatum MAP kinase B coding gene, designated PdMpkB, was functionally inactivated via homologous recombination. The fungal strain (∆PdMpkB) carrying a PdMpkBdeletion demonstrated altered gene expression profiles, reduced growth and conidiogenesis, elevated resistance to osmotic stress, and failed to induce green mold decay on citrus fruit. ∆PdMpkB was more resistant to CaCl2, NaCl and sorbitol than its progenitor strain, indicating a negative regulatory function of PdMpkB in osmotic stress adaptation. Fungal infection assays on citrus fruit revealed that ∆PdMpkB proliferated poorly within host tissues, induced water-soaking lesions, failed to break through host cuticle layers and thus, failed to produce aerial hyphae and conidia. Introduction of a functional copy of PdMpkB into a null mutant restored all defective phenotypes. Transcriptome analysis revealed that inactivation of PdMpkB impacted expression of the genes associated with cell wall-degrading enzyme activities, carbohydrate and amino acid metabolisms, conidial formation, and numerous metabolic processes. Our results define pivotal roles of the PdMpkB-mediated signaling pathway in developmental and pathological functions in the citrus postharvest pathogen P. digitatum.

  16. Rice (Oryza) hemoglobins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Hemoglobins (Hbs) corresponding to non-symbiotic (nsHb) and truncated (tHb) Hbs have been identified in rice (Oryza). This review discusses the major findings from the current studies on rice Hbs. At the molecular level, a family of the nshb genes, consisting of hb1, hb2, hb3, hb4 and hb5, and a sin...

  17. Making rice even healthier!

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rice is a naturally healthy food, but what if it could be made even healthier? Would Americans eat more rice if it could be advertised to be a 'New and Improved' source of calcium to promote bone growth, or iron to prevent anemia? Grocery stores are full of foods that are vitamin enhanced to attract...

  18. Candidatus Liberibacter americanus induces significant reprogramming of the transcriptome of the susceptible citrus genotype

    PubMed Central

    2013-01-01

    Background Citrus huanglongbing (HLB) disease is caused by endogenous, phloem-restricted, Gram negative, uncultured bacteria named Candidatus Liberibacter africanus (CaLaf), Ca. L. asiaticus (CaLas), and Ca. L. americanus (CaLam), depending on the continent where the bacteria were first detected. The Asian citrus psyllid vector, Diaphorina citri, transmits CaLas and CaLam and both Liberibacter species are present in Brazil. Several studies of the transcriptional response of citrus plants manifesting HLB symptoms have been reported, but only for CaLas infection. This study evaluated the transcriptional reprogramming of a susceptible genotype of sweet orange challenged with CaLam, using a customized 385K microarray containing approximately 32,000 unigene transcripts. We analyzed global changes in gene expression of CaLam-infected leaves of sweet orange during the symptomatic stage of infection and compared the results with previously published microarray studies that used CaLas-infected plants. Twenty candidate genes were selected to validate the expression profiles in symptomatic and asymptomatic PCR-positive leaves infected with CaLas or CaLam. Results The microarray analysis identified 633 differentially expressed genes during the symptomatic stage of CaLam infection. Among them, 418 (66%) were upregulated and 215 (34%) were down regulated. Five hundred and fourteen genes (81%) were orthologs of genes from Arabidopsis thaliana. Gene set enrichment analysis (GSEA) revealed that several of the transcripts encoded transporters associated with the endomembrane system, especially zinc transport. Among the most biologically relevant gene transcripts in GSEA were those related to signaling, metabolism and/or stimulus to hormones, genes responding to stress and pathogenesis, biosynthesis of secondary metabolites, oxidative stress and transcription factors belonging to different families. Real time PCR of 20 candidate genes validated the expression pattern of some genes in

  19. 76 FR 8603 - Citrus Seed Imports; Citrus Greening and Citrus Variegated Chlorosis

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-15

    ... of seedlings from infected seed tested positive for HLB via polymerase chain reaction (PCR) analysis... tested for HLB via PCR analysis at time periods ranging from 7 weeks to 9 months after sowing. Five of... months after the initial test yielded negative PCR results. \\4\\ Albrect, Ute and Kim D. Bowman....

  20. Cytological and molecular analysis of nonhost resistance in rice to wheat powdery mildew and leaf rust pathogens.

    PubMed

    Cheng, Yulin; Yao, Juanni; Zhang, Hongchang; Huang, Lili; Kang, Zhensheng

    2015-07-01

    Cereal powdery mildews caused by Blumeria graminis and cereal rusts caused by Puccinia spp. are constant disease threats that limit the production of almost all important cereal crops. Rice is an intensively grown agricultural cereal that is atypical because of its immunity to all powdery mildew and rust fungi. We analyzed the nonhost interactions between rice and the wheat powdery mildew fungus B. graminis f. sp. tritici (Bgt) and the wheat leaf rust fungus Puccinia triticina (Ptr) to identify the basis of nonhost resistance (NHR) in rice against cereal powdery mildew and rust fungi at cytological and molecular levels. No visible symptoms were observed on rice leaves inoculated with Bgt or Ptr. Microscopic observations showed that both pathogens exhibited aberrant differentiation and significantly reduced penetration frequencies on rice compared to wheat. The development of Bgt and Ptr was also completely arrested at early infection stages in cases of successful penetration into rice leaves. Attempted infection of rice by Bgt and Ptr induced similar defense responses, including callose deposition, accumulation of reactive oxygen species, and hypersensitive response in rice epidermal and mesophyll cells, respectively. Furthermore, a set of defense-related genes were upregulated in rice against Bgt and Ptr infection. Rice is an excellent monocot model for genetic and molecular studies. Therefore, our results demonstrate that rice is a useful model to study the mechanisms of NHR to cereal powdery mildew and rust fungi, which provides useful information for the development of novel and durable strategies to control these important pathogens.

  1. 7 CFR 93.5 - Fees for citrus product analyses set by cooperative agreement.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 3 2013-01-01 2013-01-01 false Fees for citrus product analyses set by cooperative... (CONTINUED) COMMODITY LABORATORY TESTING PROGRAMS PROCESSED FRUITS AND VEGETABLES Citrus Juices and Certain Citrus Products § 93.5 Fees for citrus product analyses set by cooperative agreement. The fees for...

  2. 7 CFR 93.5 - Fees for citrus product analyses set by cooperative agreement.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 3 2014-01-01 2014-01-01 false Fees for citrus product analyses set by cooperative... (CONTINUED) COMMODITY LABORATORY TESTING PROGRAMS PROCESSED FRUITS AND VEGETABLES Citrus Juices and Certain Citrus Products § 93.5 Fees for citrus product analyses set by cooperative agreement. The fees for...

  3. 7 CFR 93.5 - Fees for citrus product analyses set by cooperative agreement.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 3 2012-01-01 2012-01-01 false Fees for citrus product analyses set by cooperative... (CONTINUED) COMMODITY LABORATORY TESTING PROGRAMS PROCESSED FRUITS AND VEGETABLES Citrus Juices and Certain Citrus Products § 93.5 Fees for citrus product analyses set by cooperative agreement. The fees for...

  4. Screening Citrus germplasm for tolerance to HLB and Diaphorina citri--progress

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This article summarizes a project that is being funded in part by the California Citrus Research Board. In a previous field trial, 87 genotypes of citrus were planted in the field in Florida and the different genotypes were evaluated for tolerance to Asian citrus psyllid, citrus leaf miner, and hua...

  5. Continuous process for enhanced release and recovery of pectic hydrocolloids and phenolics from citrus biomass

    Technology Transfer Automated Retrieval System (TEKTRAN)

    During the 2012/2013 harvesting season the Florida citrus processing industry produced 0.55 × 106 metric tons (MT) of dried citrus pellets from citrus processing waste (CPW). The citrus pellets were marketed as a low value animal feed which may sell for $0.22 per kg or less. This biomass also contai...

  6. Evaluating citrus germplasm for huanglongbing (HLB) resistance: USDA-ARS Inoculation Program

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Asian citrus psyllid (ACP), Diaphorina citri, is an important pest because it vectors bacteria responsible for a serious disease of citrus known as huanglongbing (citrus greening disease). USDA-ARS researchers recently established a program for screening citrus germplasm for resistance to the di...

  7. Quantitative PCR Method for Diagnosis of Citrus Bacterial Canker†

    PubMed Central

    Cubero, J.; Graham, J. H.; Gottwald, T. R.

    2001-01-01

    For diagnosis of citrus bacterial canker by PCR, an internal standard is employed to ensure the quality of the DNA extraction and that proper requisites exist for the amplification reaction. The ratio of PCR products from the internal standard and bacterial target is used to estimate the initial bacterial concentration in citrus tissues with lesions. PMID:11375206

  8. Synephrine Content of Juice from Satsuma Mandarins (Citrus unshiu Marcovitch)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Synephrine, the main protoalkaloid in Citrus species, is commonly analyzed as the active component in citrus peel-containing herbal supplements, but the edible parts of mandarins have been largely ignored. We determined the synephrine concentration in the juices of C. unshiu mandarins harvested fro...

  9. Sensory and quality evaluation of selected citrus hybrids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Increasingly consumers are becoming more sophisticated in their demand for diversity of products, greater health potential and good eating quality. The evaluation of a population of mandarin citrus (Citrus reticulata) and mandarin hybrids, was initiated in 2006-2007 with the goal of establishing bas...

  10. Consideration for alternative outlet for new citrus hybrids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus sinensis (sweet orange, ex. Hamlin, Midsweet, Valencia) is the source of “orange juice” and juice must contain no less than 90% C. sinensis to be marketed as such. Juice produced from Citrus reticulata (mandarins) and C. reticulata hybrids (Orlando, Murcott, Fallglo, Sunburst, Minneola) can b...

  11. Characterization of constituents in the peel of Citrus kawachiensis (Kawachibankan).

    PubMed

    Amakura, Yoshiaki; Yoshimura, Morio; Ouchi, Kazusa; Okuyama, Satoshi; Furukawa, Yoshiko; Yoshida, Takashi

    2013-01-01

    Fifteen constituents, including a new compound, were isolated from an ethanolic extract of the peel of Citrus kawachiensis Hort. ex. Y. Tanaka (Japanese brand name, kawachibankan) which is one of the citrus products specific to Ehime, Japan. The new compound was characterized as 4'-dihydrophaseic acid β-glucopyranose ester (15) on the basis of spectral and chemical evidence.

  12. Microwave extraction of citrus peel to release pectin

    Technology Transfer Automated Retrieval System (TEKTRAN)

    After removal of soluble sugars and other compounds by washing, citrus peel is largely composed of pectin, cellulose and hemicellulose. In order to utilize the greatest amount of citrus peel product, it would appear reasonable that one or all three of these polysaccharides be converted to a useful m...

  13. Asian citrus psyllid, huanglongbing and the orange jasmine conundrum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Asian citrus psyllid (ACP) is an important invasive pest in Florida because it vectors a bacterium ‘Ca. Liberibacter asiaticus’ (LAS) responsible for a devastating citrus disease known as huanglongbing (HLB). Soon after finding HLB in Florida, concerns arose that orange jasmine in urban areas wo...

  14. Behavioral assay on Asian citrus psyllid attraction to orange jasmine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Asian citrus psyllid (ACP) is an important pest because it transmits a bacterium putatively responsible for huanglongbing, a devastating citrus disease. Research on ACP chemical ecology is of interest with respect to identifying attractants and repellents for managing the psyllid. We report on a...

  15. An evaluation of candidate repellents against Asian citrus psyllid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Asian citrus psyllid (ACP) is an important pest because it transmits a bacterium putatively responsible for huanglongbing, a devastating citrus disease. Research on ACP chemical ecology is of interest with respect to identifying attractants and repellents for managing the psyllid. With respect t...

  16. Characterizing the citrus cultivar Carrizo genome through 454 shotgun sequencing.

    PubMed

    Belknap, William R; Wang, Yi; Huo, Naxin; Wu, Jiajie; Rockhold, David R; Gu, Yong Q; Stover, Ed

    2011-12-01

    The citrus cultivar Carrizo is the single most important rootstock to the US citrus industry and has resistance or tolerance to a number of major citrus diseases, including citrus tristeza virus, foot rot, and Huanglongbing (HLB, citrus greening). A Carrizo genomic sequence database providing approximately 3.5×genome coverage (haploid genome size approximately 367 Mb) was populated through 454 GS FLX shotgun sequencing. Analysis of the repetitive DNA fraction indicated a total interspersed repeat fraction of 36.5%. Assembly and characterization of abundant citrus Ty3/gypsy elements revealed a novel type of element containing open reading frames encoding a viral RNA-silencing suppressor protein (RNA binding protein, rbp) and a plant cytokinin riboside 5′-monophosphate phosphoribohydrolase-related protein (LONELY GUY, log). Similar gypsy elements were identified in the Populus trichocarpa genome. Gene-coding region analysis indicated that 24.4% of the nonrepetitive reads contained genic regions. The depth of genome coverage was sufficient to allow accurate assembly of constituent genes, including a putative phloem-expressed gene. The development of the Carrizo database (http://citrus.pw.usda.gov/) will contribute to characterization of agronomically significant loci and provide a publicly available genomic resource to the citrus research community.

  17. Behavioral plasticity in Diaphorina citri feeding on different citrus tissues

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Asian Citrus Psyllid (Diaphorina citri) is a vector of Candidatus Liberibacter asiaticus (CLas), a phloem-limited bacterial pathogen of citrus. While psyllid nymphs are obligate feeders on young flush leaves, adults can be found on leaves of any age. To understand risk of transmission of CLas, i...

  18. What's the latest on USDA citrus scion breeding?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In its 120 year history, the USDA citrus scion breeding program has provided the mainstays of the Florida specialty citrus industry. In the last few years, the USDA has released US Early Pride mandarin, US Seedless Pineapple sweet orange and most recently, US Furr/US Furr-ST mandarin. Clearly huangl...

  19. Citrus fruit quality assessment; producer and consumer perspectives

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Consumption of citrus fruit and juices is popular with consumers worldwide and makes an important contribution to a healthy diet. Nevertheless, consumer preferences for citrus have undergone significant changes over the last twenty years and it is important to understand what consumers are looking ...

  20. Development of a global conservation strategy for citrus genetic resources

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus is an economically important world tree fruit crop with production in more than 146 countries. The center of origin for citrus is considered to be Southeastern Asia including southern China, northeastern India, and Malaysia, with secondary centers in surrounding areas. Novel and commercially ...

  1. Gene expression analysis to understand cold tolerance in citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus cultivars show a wide range of tolerance to cold temperatures. Lemons and limes are known to be sensitive to cold while certain mandarins and trifoliate oranges can endure severe winters. To understand the mechanism of cold tolerance in citrus, we selected three known cold-sensitive and three...

  2. A phagostimulant blend for the Asian citrus psyllid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Chemical cues that condition orientation by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), are of great interest because it is the primary vector of the causal pathogen of citrus greening disease. Previous work in our lab identified a blend of formic and acetic acids as s...

  3. Utilization of founder lines for improved Citrus biotechnology via RMCE

    Technology Transfer Automated Retrieval System (TEKTRAN)

    On October 1st 2011 the CRB chose to fund a unique research project, the development of citrus cultivars specifically for genetic engineering (GE). The objective of this research was to develop GE citrus ‘Founder Lines’ containing DNA sequences that will allow the precise insertion of genes for de...

  4. Changes in anthocyanin production during domestication of citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mandarin (C. reticulata), citron (C. medica) and pummelo (C. maxima) are imortant fruit species of the genus Citrus and parents of the interspecific hybrids that constitute the most familiar commercial varities of citrus: sweet orange, sour orange, clementine, lemon lme and grapefruit. Citron and it...

  5. Tolerance of trifoliate citrus rootstock hybrids to Candidatus Liberibacter asiaticus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Candidatus Liberibacter asiaticus (Las) is the suspected causal agent of Huanglongbing (HLB) in Florida and other citrus producing countries around the world. Although little resistance to HLB is found within commercial citrus varieties, tolerance has been reported for some varieties that are common...

  6. Molecular diversity of Citrus tristeza virus in California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) is a serious citrus pathogen worldwide. Recent genetic studies have identified five standard CTV genotypic groups: T30, VT, T36, T3, and B165/T68. Field surveys performed in California in 2008-2010 identified primarily MCA13-negative CTV isolates with T30-like genotype. C...

  7. Insights regarding sensory evaluation of bitterness development in citrus juice

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Delayed bitterness is a well-known phenomenon in citrus juice and has a negative impact on juice quality. Bitterness results when the tasteless limonoic acid A-ring lactone (LARL) in juice is converted to the bitter compound limonin after juicing. Citrus varieties that produce juice that becomes bit...

  8. Developing cryotherapy to eliminate graft-transmissible pathogens in citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This article summarizes research being conducted as part of a project funded by the California Citrus Research Board to develop cryotherapy (freezing buds in liquid nitrogen, and then recovering them) as a viable method for elimination of graft transmissible pathogens from Citrus. There are current...

  9. Citrus production systems to survive greening – horticultural practices

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fruit yield is a critical component in the long-term profitability of citrus growers in Florida. Increasingly, two factors outside the control of the growers are forcing Florida citrus growers to re-evaluate the sustainability of their current operations. These factors are: 1) impact of canker and ...

  10. A novel method for citrus propagation: Seed Grafting

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We have developed a technique for grafting citrus seeds to a citrus stock plant that allows the apparent development of a successful vascular connection between seed and stock plant and germination and development of a seedling which appears indistinguishable from seedlings germinated from seeds pla...

  11. Differential analyses of major allergen proteins in wild-type rice and rice producing a fragment of anti-rotavirus antibody.

    PubMed

    Yuki, Yoshikazu; Kurokawa, Shiho; Kozuka-Hata, Hiroko; Tokuhara, Daisuke; Mejima, Mio; Kuroda, Masaharu; Oyama, Masaaki; Nishimaki-Mogami, Tomoko; Teshima, Reiko; Kiyono, Hiroshi

    2016-04-01

    To develop oral antibody therapy against rotavirus infection, we previously produced a recombinant fragment of llama heavy-chain antibody to rotavirus (ARP1) in rice seeds (MucoRice-ARP1). We intend to use a purification-free rice powder for clinical application but needed to check whether MucoRice-ARP1 had increased levels of known allergen proteins. For this purpose, we used two-dimensional fluorescence difference gel electrophoresis to compare the allergen protein levels in MucoRice-ARP1 and wild-type rice. We detected no notable differences, except in the levels of α-amylase/trypsin inhibitor-like family proteins. Because by this approach we could not completely separate ARP1 from the proteins of this family, we confirmed the absence of changes in the levels of these allergens by using shotgun mass spectrometry as well as immunoblot. By using immunoelectron microscopy, we also showed that RAG2, a member of the α-amylase/trypsin inhibitor-like protein family, was relocated from protein bodies II to the plasma membrane or cell wall in MucoRice-ARP1 seed. The relocation did not affect the level of RAG2. We demonstrated that most of the known rice allergens were not considerably upregulated by the genetic modification in MucoRice-ARP1. Our data suggest that MucoRice-ARP1 is a potentially safe oral antibody for clinical application.

  12. Rice oxalate oxidase gene driven by green tissue-specific promoter increases tolerance to sheath blight pathogen (Rhizoctonia solani) in transgenic rice.

    PubMed

    Molla, Kutubuddin A; Karmakar, Subhasis; Chanda, Palas K; Ghosh, Satabdi; Sarkar, Sailendra N; Datta, Swapan K; Datta, Karabi

    2013-12-01

    Rice sheath blight, caused by the necrotrophic fungus Rhizoctonia solani, is one of the most devastating and intractable diseases of rice, leading to a significant reduction in rice productivity worldwide. In this article, in order to examine sheath blight resistance, we report the generation of transgenic rice lines overexpressing the rice oxalate oxidase 4 (Osoxo4) gene in a green tissue-specific manner which breaks down oxalic acid (OA), the pathogenesis factor secreted by R. solani. Transgenic plants showed higher enzyme activity of oxalate oxidase (OxO) than nontransgenic control plants, which was visualized by histochemical assays and sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Transgenic rice leaves were more tolerant than control rice leaves to exogenous OA. Transgenic plants showed a higher level of expression of other defence-related genes in response to pathogen infection. More importantly, transgenic plants exhibited significantly enhanced durable resistance to R. solani. The overexpression of Osoxo4 in rice did not show any detrimental phenotypic or agronomic effect. Our findings indicate that rice OxO can be utilized effectively in plant genetic manipulation for sheath blight resistance, and possibly for resistance to other diseases caused by necrotrophic fungi, especially those that secrete OA. This is the first report of the expression of defence genes in rice in a green tissue-specific manner for sheath blight resistance.

  13. Somatic Embryogenesis: Still a Relevant Technique in Citrus Improvement.

    PubMed

    Omar, Ahmad A; Dutt, Manjul; Gmitter, Frederick G; Grosser, Jude W

    2016-01-01

    The genus Citrus contains numerous fresh and processed fruit cultivars that are economically important worldwide. New cultivars are needed to battle industry threatening diseases and to create new marketing opportunities. Citrus improvement by conventional methods alone has many limitations that can be overcome by applications of emerging biotechnologies, generally requiring cell to plant regeneration. Many citrus genotypes are amenable to somatic embryogenesis, which became a key regeneration pathway in many experimental approaches to cultivar improvement. This chapter provides a brief history of plant somatic embryogenesis with focus on citrus, followed by a discussion of proven applications in biotechnology-facilitated citrus improvement techniques, such as somatic hybridization, somatic cybridization, genetic transformation, and the exploitation of somaclonal variation. Finally, two important new protocols that feature plant regeneration via somatic embryogenesis are provided: protoplast transformation and Agrobacterium-mediated transformation of embryogenic cell suspension cultures.

  14. Nutritional and physicochemical characteristic of commercial Spanish citrus juices.

    PubMed

    Alvarez, J; Pastoriza, S; Alonso-Olalla, R; Delgado-Andrade, C; Rufián-Henares, J A

    2014-12-01

    Citrus juices are perceived as healthy foods by consumers due to their richness in antioxidant compounds. Despite the large number of papers about the antioxidant activity of citrus juices, less is known about the relationship with physicochemical properties. This paper shows that the overall antioxidant activity of citrus juices is underestimated with the standard methodologies, being up to 10-times higher with the GAR method (including an in vitro gastrointestinal digestion). 70% of the antioxidant activity was found in the soluble fraction and citrus juices contributed up to 12% of the overall antioxidant intake within the Spanish diet. Physicochemical parameters, such as colour, fluorescence, 5-hydroxymethylfurfural and furfural contents, were correlated with nutritional parameters in some samples. The intake of HMF was negligible from commercial citrus juices and was absent in freshly squeezed ones. Finally, a mathematical model is developed to classify juices depending on their nature or storage conditions.

  15. Comparative transcriptome analysis of two rice varieties in response to rice stripe virus and small brown planthoppers during early interaction.

    PubMed

    Zheng, Wenjing; Ma, Li; Zhao, Jiaming; Li, Zhiqiang; Sun, Fuyu; Lu, Xiaochun

    2013-01-01

    Rice stripe, a virus disease, transmitted by a small brown planthopper (SBPH), has greatly reduced production of japonica rice in East Asia, especially in China. Although we have made great progress in mapping resistance genes, little is known about the mechanism of resistance. By de novo transcriptome assembling, we gained sufficient transcript data to analyze changes in gene expression of early interaction in response to SBPH and RSV infection in rice. Respectively 648 and 937 DEGs were detected from the disease-resistant (Liaonong 979) and the susceptible (Fengjin) varieties, most of which were up-regulated. We found 37 genes related to insect resistance, which mainly included genes for jasmonate-induced protein, TIFY protein, lipoxygenase, as well as trypsin inhibitor genes and transcription factor genes. In the interaction process between RSV and rice, 87 genes were thought to be related to RSV resistance; these primarily included 12 peroxidase biosynthesis genes, 12 LRR receptor-like protein kinase genes, 6 genes coding pathogenesis-related proteins, 4 glycine-rich cell wall structural protein genes, 2 xyloglucan hydrolase genes and a cellulose synthase. The results indicate that the rice-pathogen interaction happened both in disease-resistant and susceptible varieties, and some genes related to JA biosynthesis played key roles in the interaction between SBPHs and rice. When rice was infected by RSV a hypersensitive reaction (HR) in the disease-resistant variety was suppressed, which resulted from an increase in peroxidase expression and down-regulation of LRR receptor-like protein kinase and pathogenesis-related proteins, while, the changes of peroxidase biosynthesis, glycine-rich cell wall structural protein, cellulose synthase and xyloglucan endotransglucosylase/hydrolase could lead to the strengthening of physical barriers of rice, which may be an important resistance mechanism to RSV in rice.

  16. Fourier transform infra-red spectroscopy using an attenuated total reflection probe to distinguish between Japanese larch, pine and citrus plants in healthy and diseased states

    NASA Astrophysics Data System (ADS)

    Gandolfo, D. S.; Mortimer, H.; Woodhall, J. W.; Boonham, N.

    2016-06-01

    FTIR spectroscopy coupled with an Attenuated Total Reflection (ATR) sampling probe has been demonstrated as a technique for detecting disease in plants. Spectral differences were detected in Japanese Larch (Larix kaempferi) infected with Phytophthora ramorum at 3403 cm-1 and 1730 cm-1, from pine (Pinus spp.) infected with Bursaphelenchus xylophilus at 1070 cm-1, 1425 cm-1, 1621 cm-1 and 3403 cm-1 and from citrus (Citrus spp.) infected with 'Candidatus liberibacter' at 960 cm-1, 1087 cm-1, 1109 cm-1, 1154 cm-1, 1225 cm-1, 1385 cm-1, 1462 cm-1, 1707 cm-1, 2882 cm-1, 2982 cm-1 and 3650 cm-1. A spectral marker in healthy citrus has been identified as Pentanone but is absent from the diseased sample spectra. This agrees with recent work by Aksenov, 2014. Additionally, the spectral signature of Cutin was identified in the spectra of Pinus spp. and Citrus spp. and is consistent with work by Dubis, 1999 and Heredia-Guerrero, 2014.

  17. MoVrp1, a putative verprolin protein, is required for asexual development and infection in the rice blast fungus Magnaporthe oryzae

    PubMed Central

    Huang, Lin; Zhang, Shengpei; Yin, Ziyi; Liu, Muxing; Li, Bing; Zhang, Haifeng; Zheng, Xiaobo; Wang, Ping; Zhang, Zhengguang

    2017-01-01

    Endocytosis is a crucial cellular process in eukaryotic cells which involves clathrin and/or adaptor proteins, lipid kinases, phosphatases and the actin cytoskeleton. Verprolin proteins, such as Vrp1 in Saccharomyces cerevisiae, are conserved family proteins that regulate actin binding and endocytosis. Here, we identified and characterized MoVrp1 as the yeast Vrp1 homolog in Magnaporthe oryzae. Deletion of the MoVRP1 gene resulted in defects in vegetative growth, asexual development, and infection of the host plant. The ∆Movrp1 mutants also exhibited decreased extracellular peroxidase and laccase activities and showed defects in colony pigmentation, hyphal surface hydrophobicity, cell wall integrity, autophagy, endocytosis, and secretion of avirulent effector. Our studies provided new evidences that MoVrp1 involved in actin cytoskeleton is important for growth, morphogenesis, cellular trafficking, and fungal pathogenesis. PMID:28117435

  18. Metabolic interplay between the Asian citrus psyllid and its Profftella symbiont: An Achilles’ heel of the citrus greening insect vector

    Technology Transfer Automated Retrieval System (TEKTRAN)

    ‘Candidatus Liberibacter asiaticus’ (CLas), the bacterial pathogen associated with citrus greening disease, is transmitted by Diaphorina citri, the Asian citrus psyllid. Interactions among D. citri and its microbial endosymbionts, including ‘Candidatus Profftella armatura’, are likely to impact tra...

  19. Transmission of Citrus leprosis virus C by Brevipalpus phoenicis (Geijskes) to Alternative Host Plants Found in Citrus Orchards

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The equivalent of US$ 75 million is spent each year in Brazil to control Brevipalpus phoenicis, a mite vector of Citrus leprosis virus C (CiLV-C). In this study we investigated the possibility that hedgerows, windbreaks, and weeds normally found in citrus orchards could host CiLV-C. Mites reared on ...

  20. Pathogenicity of Heterodera daverti, H. zeae, and Meloidogyne incognita on rice

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The reactions of five rice cultivars to the cyst nematodes Heterodera daverti and H. zeae and the root-knot nematode Meloidogyne incognita were determined in the greenhouse. The results showed that both H. daverti and H. zeae infected and reproduced successfully on some of the tested rice cultivars....