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Sample records for leaf roll virus

  1. Characterization of R genes involved in resistance to Cherry leaf roll virus in paradox hybrids

    USDA-ARS?s Scientific Manuscript database

    A single dominant ‘R’ gene (clrvR), in black walnuts (Juglans hindsii) or ‘paradox’ hybrids (J. hindsii x J. regia) confers resistance to Cherry leaf roll virus (CLRV), the causal agent of blackline disease. The identification and cloning of the ‘R’ gene is expected to aid the walnut breeding progra...

  2. 40 CFR 174.513 - Potato Leaf Roll Virus Resistance Gene (also known as orf1/orf2 gene); exemption from the...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 25 2013-07-01 2013-07-01 false Potato Leaf Roll Virus Resistance Gene... Virus Resistance Gene (also known as orf1/orf2 gene); exemption from the requirement of a tolerance. An... protectant Potato Leaf Roll Virus Resistance Gene (also known as orf1/orf2 gene) in or on all food...

  3. 40 CFR 174.513 - Potato Leaf Roll Virus Resistance Gene (also known as orf1/orf2 gene); exemption from the...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 24 2014-07-01 2014-07-01 false Potato Leaf Roll Virus Resistance Gene... Virus Resistance Gene (also known as orf1/orf2 gene); exemption from the requirement of a tolerance. An... protectant Potato Leaf Roll Virus Resistance Gene (also known as orf1/orf2 gene) in or on all food...

  4. 40 CFR 174.513 - Potato Leaf Roll Virus Resistance Gene (also known as orf1/orf2 gene); exemption from the...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Potato Leaf Roll Virus Resistance Gene... Virus Resistance Gene (also known as orf1/orf2 gene); exemption from the requirement of a tolerance. An... protectant Potato Leaf Roll Virus Resistance Gene (also known as orf1/orf2 gene) in or on all food...

  5. 40 CFR 174.513 - Potato Leaf Roll Virus Resistance Gene (also known as orf1/orf2 gene); exemption from the...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 25 2012-07-01 2012-07-01 false Potato Leaf Roll Virus Resistance Gene... Virus Resistance Gene (also known as orf1/orf2 gene); exemption from the requirement of a tolerance. An... protectant Potato Leaf Roll Virus Resistance Gene (also known as orf1/orf2 gene) in or on all food...

  6. 40 CFR 174.513 - Potato Leaf Roll Virus Resistance Gene (also known as orf1/orf2 gene); exemption from the...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 24 2011-07-01 2011-07-01 false Potato Leaf Roll Virus Resistance Gene... Virus Resistance Gene (also known as orf1/orf2 gene); exemption from the requirement of a tolerance. An... protectant Potato Leaf Roll Virus Resistance Gene (also known as orf1/orf2 gene) in or on all food...

  7. Host Species-Dependent Population Structure of a Pollen-Borne Plant Virus, Cherry Leaf Roll Virus

    PubMed Central

    Rebenstorf, Kathrin; Candresse, Thierry; Dulucq, Marie Josée; Büttner, Carmen; Obermeier, Christian

    2006-01-01

    Cherry leaf roll virus (CLRV) belongs to the Nepovirus genus within the family Comoviridae. It has a host range which includes a number of wild tree and shrub species. The serological and molecular diversity of CLRV was assessed using a collection of isolates and samples recovered from woody and herbaceous host plants from different geographical origins. Molecular diversity was assessed by sequencing a short (375-bp) region of the 3′ noncoding region (NCR) of the genomic RNAs while serological diversity was assessed using a panel of seven monoclonal antibodies raised initially against a walnut isolate of CLRV. The genomic region analyzed was shown to exhibit a significant degree of molecular variability with an average pairwise divergence of 8.5% (nucleotide identity). Similarly, serological variability proved to be high, with no single monoclonal antibody being able to recognize all isolates analyzed. Serological and molecular phylogenetic reconstructions showed a strong correlation. Remarkably, the diversity of CLRV populations is to a large extent defined by the host plant from which the viral samples are originally obtained. There are relatively few reports of plant viruses for which the genetic diversity is structured by the host plant. In the case of CLRV, we hypothesize that this situation may reflect the exclusive mode of transmission in natural plant populations by pollen and by seeds. These modes of transmission are likely to impose barriers to host change by the virus, leading to rapid biological and genetic separation of CLRV variants coevolving with different plant host species. PMID:16474152

  8. Comparison of diagnostic techniques for the detection and differentiation of Cherry leaf roll virus strains for quarantine purposes.

    PubMed

    Lebas, B S M; Veerakone, S; Liefting, L W; Tang, J; Perez-Egusquiza, Z; von Bargen, S; Ward, L

    2016-08-01

    Some strains of Cherry leaf roll virus (CLRV) are considered as quarantine pests in New Zealand. CLRV was detected in seven plant host species: Actinidia chinensis, Hydrangea macrophylla, Malus domestica, Plantago major, Ribes rubrum, Rubus idaeus and Rumex sp. collected from New Zealand between 2005 and 2012. Biological, serological and molecular techniques were compared for the detection and differentiation of CLRV isolates. The biological analysis revealed differences in symptomatology and disease severity among the isolates. The five isolates tested by ELISA were serologically related to each other using polyclonal antisera with only one out of four commercially-available antisera successfully detecting all of them. The phylogenetic analysis of sequences obtained from parts of the coat protein, polymerase and 3'-untranslated regions revealed that the New Zealand CLRV isolates clustered into two closely related but distinct phylogenetic groups with some isolates grouping differently depending on the gene studied. The New Zealand CLRV isolates were clearly distinct to overseas isolates found in phylogenetic groups A, D and E. The conventional RT-PCR using primers targeting the CLRV coat protein coding region is recommended for determining sequence differences between strains. These findings will be useful in making regulatory decisions with regard to the testing requirements and the CLRV strains to be regulated in New Zealand. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Cherry leaf roll virus

    USDA-ARS?s Scientific Manuscript database

    In the blackberry cultivar Himalaya Giant naturally infected in England, CLRV is reported to cause chlorotic mottling and line patterning in leaves. Infected plants are stunted and may be killed. In three red raspberry cultivars naturally infected by CLRV in New Zealand, infected plants had stunted,...

  10. Transgenic resistance in potato plants expressing potato leaf roll virus (PLRV) replicase gene sequences is RNA-mediated and suggests the involvement of post-transcriptional gene silencing.

    PubMed

    Vazquez Rovere, C; Asurmendi, S; Hopp, H E

    2001-07-01

    Genetically engineered expression of replicase encoding sequences has been proposed as an efficient system to confer protection against virus diseases by eliciting protection mechanisms in the plant. Potato leaf-roll was one of the first diseases for which this kind of protection was engineered in potato plants. However, details of the protecting mechanism were not reported, so far. The ORF2b of an Argentinean strain of PLRV was cloned and sequenced finding 94% and 97% of homology with Australian and Dutch strains, respectively. To elucidate the mechanism of protection against PLRV infection, three versions of ORF2b (non-translatable sense, translatable sense with an engineered ATG and antisense) were constructed under the control of the 35S CaMV promoter and the nos terminator and introduced in potato plants (cv. Kennebec) by Agrobacterium tumefaciens-mediated transformation. Grafting infection experiments showed that resistant transgenic plants could be obtained with any of the constructs, suggesting that the mechanism of protection is independent of the expression of protein and is RNA mediated. Field trial infection confirmed that resistant transgenic events were obtained. Biolistic transient transformation experiments of leaves derived from transgenic plants using a gene coding for the fusion protein GUS-ORF2b, followed by scoring of the number of GUS expressing leaf spots, supported that the protection is mediated by a post-transcriptional gene silencing mechanism.

  11. The silver lining of a viral agent: increasing seed yield and harvest index in Arabidopsis by ectopic expression of the potato leaf roll virus movement protein.

    PubMed

    Kronberg, Kristin; Vogel, Florian; Rutten, Twan; Hajirezaei, Mohammed-Reza; Sonnewald, Uwe; Hofius, Daniel

    2007-11-01

    Ectopic expression of viral movement proteins (MPs) has previously been shown to alter plasmodesmata (PD) function and carbon partitioning in transgenic plants, giving rise to the view of PD being dynamic and highly regulated structures that allow resource allocation to be adapted to environmental and developmental needs. However, most work has been restricted to solanaceous species and the potential use of MP expression to improve biomass and yield parameters has not been addressed in detail. Here we demonstrate that MP-mediated modification of PD function can substantially alter assimilate allocation, biomass production, and reproductive growth in Arabidopsis (Arabidopsis thaliana). These effects were achieved by constitutive expression of the potato leaf roll virus 17-kD MP (MP17) fused to green fluorescent protein (GFP) in different Arabidopsis ecotypes. The resulting transgenic plants were analyzed for PD localization of the MP17:GFP fusion protein and different lines with low to high expression levels were selected for further analysis. Low-level accumulation of MP17 resulted in enhanced sucrose efflux from source leaves and a considerably increased vegetative biomass production. In contrast, high MP17 levels impaired sucrose export, resulting in source leaf-specific carbohydrate accumulation and a strongly reduced vegetative growth. Surprisingly, later during development the MP17-mediated inhibition of resource allocation was reversed, and final seed yield increased in average up to 30% in different transgenic lines as compared to wild-type plants. This resulted in a strongly improved harvest index. The release of the assimilate export block was paralleled by a reduced PD binding of MP17 in senescing leaves, indicating major structural changes of PD during leaf senescence.

  12. Semi-Rolled Leaf2 modulates rice leaf rolling by regulating abaxial side cell differentiation.

    PubMed

    Liu, Xiaofei; Li, Ming; Liu, Kai; Tang, Ding; Sun, Mingfa; Li, Yafei; Shen, Yi; Du, Guijie; Cheng, Zhukuan

    2016-04-01

    Moderate leaf rolling maintains the erectness of leaves and minimizes the shadowing between leaves which is helpful to establish ideal plant architecture. Here, we describe asrl2(semi-rolled leaf2) rice mutant, which has incurved leaves due to the presence of defective sclerenchymatous cells on the abaxial side of the leaf and displays narrow leaves and reduced plant height. Map-based cloning revealed that SRL2 encodes a novel plant-specific protein of unknown biochemical function.SRL2 was mainly expressed in the vascular bundles of leaf blades, leaf sheaths, and roots, especially in their sclerenchymatous cells. The transcriptional activities of several leaf development-related YABBY genes were significantly altered in the srl2 mutant. Double mutant analysis suggested that SRL2 and SHALLOT-LIKE1(SLL1)/ROLLED LEAF9(RL9) function in distinct pathways that regulate abaxial-side leaf development. Hence, SRL2 plays an important role in regulating leaf development, particularly during sclerenchymatous cell differentiation.

  13. Semi-Rolled Leaf2 modulates rice leaf rolling by regulating abaxial side cell differentiation

    PubMed Central

    Liu, Xiaofei; Li, Ming; Liu, Kai; Tang, Ding; Sun, Mingfa; Li, Yafei; Shen, Yi; Du, Guijie; Cheng, Zhukuan

    2016-01-01

    Moderate leaf rolling maintains the erectness of leaves and minimizes the shadowing between leaves which is helpful to establish ideal plant architecture. Here, we describe a srl2 (semi-rolled leaf2) rice mutant, which has incurved leaves due to the presence of defective sclerenchymatous cells on the abaxial side of the leaf and displays narrow leaves and reduced plant height. Map-based cloning revealed that SRL2 encodes a novel plant-specific protein of unknown biochemical function. SRL2 was mainly expressed in the vascular bundles of leaf blades, leaf sheaths, and roots, especially in their sclerenchymatous cells. The transcriptional activities of several leaf development-related YABBY genes were significantly altered in the srl2 mutant. Double mutant analysis suggested that SRL2 and SHALLOT-LIKE1 (SLL1)/ROLLED LEAF9 (RL9) function in distinct pathways that regulate abaxial-side leaf development. Hence, SRL2 plays an important role in regulating leaf development, particularly during sclerenchymatous cell differentiation. PMID:26873975

  14. Cucumber leaf spot virus

    USDA-ARS?s Scientific Manuscript database

    Cucumber leaf spot virus (CLSV) was originally identified from cucumber (Cucumis sativus) in Germany, but has since been found in various parts of Europe, the UK, and the Middle East, including Jordan, Saudi Arabia, Bulgaria, Poland, and Spain. CLSV is known to cause symptoms ranging from chloroti...

  15. First report of seed-borne cherry leaf roll virus in wild potato, Solanum acaule, from South America

    USDA-ARS?s Scientific Manuscript database

    A virus, designated JCM-79, was isolated from wild potato (Solanum acaule Bitt.) plants grown from true seed received at USDA-APHIS Potato Quarantine Program from Peru. JCM-79 was mechanically transmissible to Nicotiana clevelandii, N.tabacum cv. Samsun NN, and Chenopodium quinoa. Symptoms in the ...

  16. Molecular cytogenetic characterization of a new leaf rolling triticale.

    PubMed

    Yang, E N; Yang, Z J; Zhang, J F; Zou, Y C; Ren, Z L

    2011-11-29

    Leaf rolling occurs in some cereal genotypes in response to drought. We identified and made a phenotypic, cytological and physiological analysis of a leaf-rolling genotype (CMH83) of hexaploid triticale (X Triticosecale Wittmack) that exhibited reduced plant height, rolled and narrow leaves. Gliadin electrophoresis of seed protein showed that CMH83 was genetically stable. Sequential Giemsa-C-banding and genomic in situ hybridization showed that CMH83 contains 12 rye chromosomes; two pairs of these chromosomes have reduced telomeric heterochromatin bands. Tests of relative water content and water loss rate of leaves of CMH83 compared with those of wheat cultivars indicated that rapid water loss after drought stress in CMH83 is associated with the leaf rolling phenotypes. Leaf rolling in CMH83 was a dominant trait in our inheritance studies. Triticale line CMH83 could be used to study drought resistance mechanisms in triticale.

  17. Development of molecular resistance in potato against potato leaf roll virus and potato virus Y through Agrobacterium-mediated double transgenesis

    USDA-ARS?s Scientific Manuscript database

    Potato leafroll virus (PLRV) and potato virus Y (PVY) are the two major viral problems for the potato production all over the world. Transgenic approaches involving the expression of viral genes are being developed to provide protection for plants against viral diseases. The purpose of this study w...

  18. SEMI-ROLLED LEAF1 Encodes a Putative Glycosylphosphatidylinositol-Anchored Protein and Modulates Rice Leaf Rolling by Regulating the Formation of Bulliform Cells1[W][OA

    PubMed Central

    Xiang, Jing-Jing; Zhang, Guang-Heng; Qian, Qian; Xue, Hong-Wei

    2012-01-01

    Leaf rolling is an important agronomic trait in rice (Oryza sativa) breeding and moderate leaf rolling maintains the erectness of leaves and minimizes shadowing between leaves, leading to improved photosynthetic efficiency and grain yields. Although a few rolled-leaf mutants have been identified and some genes controlling leaf rolling have been isolated, the molecular mechanisms of leaf rolling still need to be elucidated. Here we report the isolation and characterization of SEMI-ROLLED LEAF1 (SRL1), a gene involved in the regulation of leaf rolling. Mutants srl1-1 (point mutation) and srl1-2 (transferred DNA insertion) exhibit adaxially rolled leaves due to the increased numbers of bulliform cells at the adaxial cell layers, which could be rescued by complementary expression of SRL1. SRL1 is expressed in various tissues and is expressed at low levels in bulliform cells. SRL1 protein is located at the plasma membrane and predicted to be a putative glycosylphosphatidylinositol-anchored protein. Moreover, analysis of the gene expression profile of cells that will become epidermal cells in wild type but probably bulliform cells in srl1-1 by laser-captured microdissection revealed that the expression of genes encoding vacuolar H+-ATPase (subunits A, B, C, and D) and H+-pyrophosphatase, which are increased during the formation of bulliform cells, were up-regulated in srl1-1. These results provide the transcript profile of rice leaf cells that will become bulliform cells and demonstrate that SRL1 regulates leaf rolling through inhibiting the formation of bulliform cells by negatively regulating the expression of genes encoding vacuolar H+-ATPase subunits and H+-pyrophosphatase, which will help to understand the mechanism regulating leaf rolling. PMID:22715111

  19. Cotton leaf curl virus disease.

    PubMed

    Briddon, R W; Markham, P G

    2000-11-01

    Cotton is one of the most important crops of Pakistan, accounting for over 60% of foreign exchange earnings. The present epidemic of cotton leaf curl disease (CLCuD) originated in the Punjab region near the city of Multan and was first reported in 1985, although it was noted in this region as early as 1967. By the early 1990s, CLCuD had become the major limitation to cotton production in Pakistan and it has now spread into India and, more recently, south and west into other provinces of Pakistan. The very characteristic symptoms include leaf curling, darkened veins, vein swelling and enations that frequently develop into cup-shaped, leaf-like structures on the undersides of leaves. Identification of the vector of CLCuD as the whitefly Bemisia tabaci (Genn.) quickly led to the suggestion that the causative agent of the disease is a geminivirus. Researchers soon confirmed the presence of such a virus that is currently ascribed to the genus Begomovirus of the family Geminiviridae, However, in 1999, the aetiology of the disease was shown to be more complex than was originally assumed. Despite the identification of both a begomovirus and a so-called nanovirus-like component, the precise causal agent of CLCuD remains uncertain.

  20. SHALLOT-LIKE1 is a KANADI transcription factor that modulates rice leaf rolling by regulating leaf abaxial cell development.

    PubMed

    Zhang, Guang-Heng; Xu, Qian; Zhu, Xu-Dong; Qian, Qian; Xue, Hong-Wei

    2009-03-01

    As an important agronomic trait, rice (Oryza sativa L.) leaf rolling has attracted much attention from plant biologists and breeders. Moderate leaf rolling increases the photosynthesis of cultivars and hence raises grain yield. However, the relevant molecular mechanism remains unclear. Here, we show the isolation and functional characterization of SHALLOT-LIKE1 (SLL1), a key gene controlling rice leaf rolling. sll1 mutant plants have extremely incurved leaves due to the defective development of sclerenchymatous cells on the abaxial side. Defective development can be functionally rescued by expression of SLL1. SLL1 is transcribed in various tissues and accumulates in the abaxial epidermis throughout leaf development. SLL1 encodes a SHAQKYF class MYB family transcription factor belonging to the KANADI family. SLL1 deficiency leads to defective programmed cell death of abaxial mesophyll cells and suppresses the development of abaxial features. By contrast, enhanced SLL1 expression stimulates phloem development on the abaxial side and suppresses bulliform cell and sclerenchyma development on the adaxial side. Additionally, SLL1 deficiency results in increased chlorophyll and photosynthesis. Our findings identify the role of SLL1 in the modulation of leaf abaxial cell development and in sustaining abaxial characteristics during leaf development. These results should facilitate attempts to use molecular breeding to increase the photosynthetic capacity of rice, as well as other crops, by modulating leaf development and rolling.

  1. Analysis of the differential gene and protein expression profile of the rolled leaf mutant of transgenic rice (Oryza sativa L.).

    PubMed

    Zhu, Qiuqiang; Yu, Shuguang; Chen, Guanshui; Ke, Lanlan; Pan, Daren

    2017-01-01

    The importance of leaf rolling in rice (Oryza sativa L.) has been widely recognized. Although several studies have investigated rice leaf rolling and identified some related genes, knowledge of the molecular mechanism underlying rice leaf rolling, especially outward leaf rolling, is limited. Therefore, in this study, differential proteomics and gene expression profiling were used to analyze rolled leaf mutant of transgenic rice in order to investigate differentially expressed genes and proteins related to rice leaf rolling. To this end, 28 differentially expressed proteins related to rolling leaf traits were isolated and identified. Digital expression profiling detected 10 genes related to rice leaf rolling. Some of the proteins and genes detected are involved in lipid metabolism, which is related to the development of bulliform cells, such as phosphoinositide phospholipase C, Mgll gene, and At4g26790 gene. The "omics"-level techniques were useful for simultaneously isolating several proteins and genes related to rice leaf rolling. In addition, the results of the analysis of differentially expressed proteins and genes were closely consistent with those from a corresponding functional analysis of cellular mechanisms; our study findings might form the basis for further research on the molecular mechanisms underlying rice leaf rolling.

  2. Complete genome sequence of bean leaf crumple virus, a novel begomovirus infecting common bean in Colombia.

    PubMed

    Carvajal-Yepes, Monica; Zambrano, Leidy; Bueno, Juan M; Raatz, Bodo; Cuellar, Wilmer J

    2017-02-10

    A copy of the complete genome of a novel bipartite begomovirus infecting common bean (Phaseolus vulgaris L.) in Colombia was obtained by rolling-circle amplification (RCA), cloned, and sequenced. The virus is associated with leaf crumple symptoms and significant yield losses in Andean and Mesoamerican beans. Such symptoms have been reported increasingly in Colombia since at least 2002, and we detected the virus in leaf material collected since 2008. Sequence analysis showed that the virus is a member of a distinct species, sharing 81% and 76% nucleotide (nt) sequence identity (in DNA-A and DNA-B, respectively) to other begomoviruses infecting common bean in the Americas. The data obtained support the taxonomic status of this virus (putatively named 'bean leaf crumple virus', BLCrV) as a member of a novel species in the genus Begomovirus.

  3. Characterization of Rolled and Erect Leaf 1 in regulating leave morphology in rice

    PubMed Central

    Chen, Qiaoling; Xie, Qingjun; Gao, Ju; Wang, Wenyi; Sun, Bo; Liu, Bohan; Zhu, Haitao; Peng, Haifeng; Zhao, Haibing; Liu, Changhong; Wang, Jiang; Zhang, Jingliu; Zhang, Guiquan; Zhang, Zemin

    2015-01-01

    Leaf morphology, particularly in crop, is one of the most important agronomic traits because it influences the yield through the manipulation of photosynthetic capacity and transpiration. To understand the regulatory mechanism of leaf morphogenesis, an Oryza sativa dominant mutant, rolled and erect leaf 1 (rel1) has been characterized. This mutant has a predominant rolled leaf, increased leaf angle, and reduced plant height phenotype that results in a reduction in grain yield. Electron microscope observations indicated that the leaf incurvations of rel1 dominant mutants result from the alteration of the size and number of bulliform cells. Molecular cloning revealed that the rel1 dominant mutant phenotype is caused by the activation of the REL1 gene, which encodes a novel unknown protein, despite its high degree of conservation among monocot plants. Moreover, the downregulation of the REL1 gene in the rel1 dominant mutant restored the phenotype of this dominant mutant. Alternatively, overexpression of REL1 in wild-type plants induced a phenotype similar to that of the dominant rel1 mutant, indicating that REL1 plays a positive role in leaf rolling and bending. Consistent with the observed rel1 phenotype, the REL1 gene was predominantly expressed in the meristem of various tissues during plant growth and development. Nevertheless, the responsiveness of both rel1 dominant mutants and REL1-overexpressing plants to exogenous brassinosteroid (BR) was reduced. Moreover, transcript levels of BR response genes in the rel1 dominant mutants and REL1-overexpressing lines were significantly altered. Additionally, seven REL1-interacting proteins were also identified from a yeast two-hybrid screen. Taken together, these findings suggest that REL1 regulates leaf morphology, particularly in leaf rolling and bending, through the coordination of BR signalling transduction. PMID:26142419

  4. Replicative intermediates of maize streak virus found during leaf development.

    PubMed

    Erdmann, Julia B; Shepherd, Dionne N; Martin, Darren P; Varsani, Arvind; Rybicki, Edward P; Jeske, Holger

    2010-04-01

    Geminiviruses of the genera Begomovirus and Curtovirus utilize three replication modes: complementary-strand replication (CSR), rolling-circle replication (RCR) and recombination-dependent replication (RDR). Using two-dimensional gel electrophoresis, we now show for the first time that maize streak virus (MSV), the type member of the most divergent geminivirus genus, Mastrevirus, does the same. Although mastreviruses have fewer regulatory genes than other geminiviruses and uniquely express their replication-associated protein (Rep) from a spliced transcript, the replicative intermediates of CSR, RCR and RDR could be detected unequivocally within infected maize tissues. All replicative intermediates accumulated early and, to varying degrees, were already present in the shoot apex and leaves at different maturation stages. Relative to other replicative intermediates, those associated with RCR increased in prevalence during leaf maturation. Interestingly, in addition to RCR-associated DNA forms seen in other geminiviruses, MSV also apparently uses dimeric open circular DNA as a template for RCR.

  5. Stachytarpheta leaf curl virus is a novel monopartite begomovirus species.

    PubMed

    Xiong, Q; Fan, S; Guo, X; Zhou, X

    2005-11-01

    Begomovirus isolates were obtained from Stachytarpheta jamaicensis plants showing leaf curl and chlorosis symptoms collected in the Hainan province of China. The complete sequences of isolates Hn5-4, Hn6-1, Hn30 and Hn34 were determined to be 2748, 2751, 2748 and 2748 nucleotides long, respectively. The complete sequences of the four isolates share more than 94.9% nucleotide sequence identity, but all of them have less than 86% nucleotide sequence identity with other reported begomoviruses. The molecular data show that Hn5-4, Hn6-1, Hn30 and Hn34 are isolates of a distinct begomovirus species, for which the name Stachytarpheta leaf curl virus (StaLCV) is proposed. PCR and Southern blot analyses demonstrate that all the collected field samples are not associated with DNAbeta or DNA-B components. An infectious clone of StaLCV isolate Hn5-4 was constructed, and could efficiently infect Nicotiana benthamiana, N. tabacum Samsun, N. glutinosa, Lycopersicon esculentum and Petunia hybrida plants, inducing upward leaf roll and vein swelling symptoms. In addition, we illustrate that StaLCV can functionally interact with distinct DNAbeta molecules in plants.

  6. Notes on the ecology of rolled-leaf hispines (Chrysomelidae, Cassidinae) at La Gamba (Costa Rica).

    PubMed

    Schmitt, Michael; Frank, Meike

    2013-01-01

    A total of 301 adult hispine beetles of the genera Cephaloleia and Chelobasis were found in rolled leaves of plants of 17 species of Zingiberales (families Costaceae, Heliconiaceae, Maranthaceae, Musaceae, and Zingiberaceae) during a field study at La Gamba, Golfito region, Costa Rica. Of these beetles, Cephaloleia belti was recorded from 12 potential host plant species, C. distincta from 7, C. dilaticollis from 5, C., Chelobasis bicolor, C. championi, and C. histrionica from 3, Chelobasis perplexa and C. instabilis from 2, whereas C. trivittata from only one. Of the plant species, Heliconia latispatha had 7 beetle species in its leaf rolls, Calathea lutea had 5, H. imbricata and H. rostrata had 4, H. stricta and Musa paradisiaca had 3, H. wagneriana had 2, while on H. vaginalis, H. danielsiana, H. densiflora, H. longiflora, Calathea crotalifera, C. platystachya, Goeppertia lasiophylla, Alpinia purpurata, Costus pulverulentus and Costus barbatus, H. densiflora, H. vaginalis, and H. danielsana only hispines of one species were found. Cephaloleia belti occurred together with beetles of six other hispine species, whereas Cephaloleia trivittata never shared a leaf roll with another hispine species. The remaining beetle species aggregated with one to four other hispines. Adults of C. belti and C. championi were frequently seen, occasionally also with C. dilaticollis, C. histrionica, and Chelobasis perplexa, to co-occur with the carabid Calophaena ligata in the same leaf roll without any sign of interspecific aggression. A comparison of host choices and the phylogeny of the hispines and of their host plants revealed no signs that beetles used species level phylogenetic relationships within the Zingiberales to select food plants. Obviously, within this plant order, rolled-leaf hispines choose their plant hosts in a nearly opportunistic manner. Seemingly, they use differences among plants at higher taxonomic levels but within the Zingiberales, the availability of young

  7. Photosystem II functionality and antioxidant system changes during leaf rolling in post-stress emerging Ctenanthe setosa exposed to drought.

    PubMed

    Terzi, Rabiye; Saruhan, Neslihan; Sağlam, A; Nar, Hatice; Kadioğlu, A

    2009-12-01

    We studied the changes in antioxidant system and chlorophyll fluorescence parameters in post-stress emerging Ctenanthe setosa (Rosc.) Eichler (Marantaceae) plants (PSE plants) having reduced leaf area under drought stress causing leaf rolling and re-watering. PSE plants were compared to primary stressed plants (PS) in previous studies. The parameters were measured at different visual leaf rolling scores from 1 to 4 (1 is unrolled, 4 is tightly rolled and the others is intermediate form). Water potentials and stomatal conductance of leaves were gradually decreased during leaf rolling. Similarly, maximum quantum efficiency of open PS II center and quantum yield of PS II decreased during the rolling period. Non-photochemical quenching of chlorophyll fluorescence decreased at score 2 then increased while photochemical quenching did not change during leaf rolling. Electron transport rate decreased only at score 4 but approximately reached to score 1 level after re-watering. Superoxide dismutase activity was not constant at all leaf rolling scores. Ascorbate peroxidase, catalase and glutathione reductase activities generally tended to increase during leaf rolling. Lipid peroxidation and H 2 O 2 content increased at score 2 but decreased at the later scores. On the other hand, O 2 .- production increased during the rolling period. After re-watering of the plants having score 4 of leaf rolling, antioxidant enzyme activities were lower than those of score 1. Other physiological parameters also tended to reach the value of score 1. The results indicated that PSE plants gained drought tolerance by reducing leaf area effectively induced their antioxidant systems and protected the photosynthesis under drought stress similar to PS plants.

  8. Whitefly transmission of the Sweet potato leaf curl virus

    USDA-ARS?s Scientific Manuscript database

    The sweetpotato whitefly, Bemisia tabaci (Gennadius) is highly adaptive and polyphagous on taxonomically diverse species of plants on a global scale. This whitefly transmits numerous plant viruses, including Begomoviruses (Geminiviridae). We recently found the Sweet Potato Leaf Curl Virus (SPLCV) ...

  9. Tomato yellow leaf curl viruses: ménage à trois between the virus complex, the plant and the whitefly vector.

    PubMed

    Díaz-Pendón, Juan Antonio; Cañizares, M Carmen; Moriones, Enrique; Bejarano, Eduardo R; Czosnek, Henryk; Navas-Castillo, Jesús

    2010-07-01

    Tomato yellow leaf curl disease (TYLCD) is one of the most devastating viral diseases affecting tomato crops in tropical, subtropical and temperate regions of the world. Here, we focus on the interactions through recombination between the different begomovirus species causing TYLCD, provide an overview of the interactions with the cellular genes involved in viral replication, and highlight recent progress on the relationships between these viruses and their vector, the whitefly Bemisia tabaci. The tomato yellow leaf curl virus-like viruses (TYLCVs) are a complex of begomoviruses (family Geminiviridae, genus Begomovirus) including 10 accepted species: Tomato yellow leaf curl Axarquia virus (TYLCAxV), Tomato yellow leaf curl China virus (TYLCCNV), Tomato yellow leaf curl Guangdong virus (TYLCGuV), Tomato yellow leaf curl Indonesia virus (TYLCIDV), Tomato yellow leaf curl Kanchanaburi virus (TYLVKaV), Tomato yellow leaf curl Malaga virus (TYLCMalV), Tomato yellow leaf curl Mali virus (TYLCMLV), Tomato yellow leaf curl Sardinia virus (TYLCSV), Tomato yellow leaf curl Thailand virus (TYLCTHV), Tomato yellow leaf curl Vietnam virus (TYLCVNV) and Tomato yellow leaf curl virus(TYLCV). We follow the species demarcation criteria of the International Committee on Taxonomy of Viruses (ICTV), the most important of which is an 89% nucleotide identity threshold between full-length DNA-A component nucleotide sequences for begomovirus species. Strains of a species are defined by a 93% nucleotide identity threshold. The primary host of TYLCVs is tomato (Solanum lycopersicum), but they can also naturally infect other crops [common bean (Phaseolus vulgaris), sweet pepper (Capsicum annuum), chilli pepper (C. chinense) and tobacco (Nicotiana tabacum)], a number of ornamentals [petunia (Petuniaxhybrida) and lisianthus (Eustoma grandiflora)], as well as common weeds (Solanum nigrum and Datura stramonium). TYLCVs also infect the experimental host Nicotiana benthamiana. Infected tomato

  10. Leaf Rolling Controlled by the Homeodomain Leucine Zipper Class IV Gene Roc5 in Rice1[W

    PubMed Central

    Zou, Liang-ping; Sun, Xue-hui; Zhang, Zhi-guo; Liu, Peng; Wu, Jin-xia; Tian, Cai-juan; Qiu, Jin-long; Lu, Tie-gang

    2011-01-01

    Leaf rolling is considered an important agronomic trait in rice (Oryza sativa) breeding. To understand the molecular mechanism controlling leaf rolling, we screened a rice T-DNA insertion population and isolated the outcurved leaf1 (oul1) mutant showing abaxial leaf rolling. The phenotypes were caused by knockout of Rice outermost cell-specific gene5 (Roc5), an ortholog of the Arabidopsis (Arabidopsis thaliana) homeodomain leucine zipper class IV gene GLABRA2. Interestingly, overexpression of Roc5 led to adaxially rolled leaves, whereas cosuppression of Roc5 resulted in abaxial leaf rolling. Bulliform cell number and size increased in oul1 and Roc5 cosuppression plants but were reduced in Roc5-overexpressing lines. The data indicate that Roc5 negatively regulates bulliform cell fate and development. Gene expression profiling, quantitative polymerase chain reaction, and RNA interference (RNAi) analyses revealed that Protodermal Factor Like (PFL) was probably down-regulated in oul1. The mRNA level of PFL was increased in Roc5-overexpressing lines, and PFL-RNAi transgenic plants exhibit reversely rolling leaves by reason of increases of bulliform cell number and size, indicating that Roc5 may have a conserved function. These are, to our knowledge, the first functional data for a gene encoding a homeodomain leucine zipper class IV transcriptional factor in rice that modulates leaf rolling. PMID:21596949

  11. Rolling-leaf14 is a 2OG-Fe (II) oxygenase family protein that modulates rice leaf rolling by affecting secondary cell wall formation in leaves.

    PubMed

    Fang, Likui; Zhao, Fangming; Cong, Yunfei; Sang, Xianchun; Du, Qing; Wang, Dezhong; Li, Yunfeng; Ling, Yinghua; Yang, Zhenglin; He, Guanghua

    2012-06-01

    As an important agronomic trait, leaf rolling in rice (Oryza sativa L.) has attracted much attention from plant biologists and breeders. Moderate leaf rolling increases the amount of photosynthesis in cultivars and hence raises grain yield. Here, we describe the map-based cloning of the gene RL14, which was found to encode a 2OG-Fe (II) oxygenase of unknown function. rl14 mutant plants had incurved leaves because of the shrinkage of bulliform cells on the adaxial side. In addition, rl14 mutant plants displayed smaller stomatal complexes and decreased transpiration rates, as compared with the wild type. Defective development could be rescued functionally by the expression of wild-type RL14. RL14 was transcribed in sclerenchymatous cells in leaves that remained wrapped inside the sheath. In mature leaves, RL14 accumulated mainly in the mesophyll cells that surround the vasculature. Expression of genes related to secondary cell wall formation was affected in rl14-1 mutants, and cellulose and lignin content were altered in rl14-1 leaves. These results reveal that the RL14 gene affects water transport in leaves by affecting the composition of the secondary cell wall. This change in water transport results in water deficiency, which is the major reason for the abnormal shape of the bulliform cells.

  12. Association of tomato leaf curl Sudan virus with leaf curl disease of tomato in Jeddah, Saudi Arabia.

    PubMed

    Sohrab, Sayed Sartaj; Yasir, Muhammad; El-Kafrawy, Sherif Ali; Abbas, Ayman T; Mousa, Magdi Ali Ahmed; Bakhashwain, Ahmed A

    2016-06-01

    Tomato is an important vegetable crop and its production is adversely affected by leaf curl disease caused by begomovirus. Leaf curl disease is a serious concern for tomato crops caused by begomovirus in Jeddah, Kingdom of Saudi Arabia. Tomato leaf curl disease has been shown to be mainly caused either by tomato leaf curl Sudan virus or tomato yellow leaf curl virus as well as tomato leaf curl Oman virus. Many tomato plants infected with monopartite begomoviruses were also found to harbor a symptom enhancing betasatellites. Here we report the association of tomato leaf curl Sudan virus causing leaf curl disease of tomato in Jeddah, Kingdom of Saudi Arabia. The complete genome sequence analysis showed highest (99.9 %) identity with tomato leaf curl Sudan virus causing leaf curl disease in Arabian Peninsula. In phylogenetic relationships analysis, the identified virus formed closest cluster with tomato leaf curl Sudan virus. In recombination analysis study, the major parent was identified as tomato leaf curl Sudan virus. Findings of this study strongly supports the associated virus is a variant of tomato leaf curl Sudan virus causing disease in Sudan, Yemen and Arabian Peninsula. The betasatellites sequence analysis showed highest identity (99.8 %) with tomato leaf curl betasatellites-Amaranthus-Jeddah. The phylogenetic analysis result based on betasatellites formed closed cluster with tomato yellow leaf curl Oman betasatellites. The importance of these findings and occurrence of begomovirus in new geographic regions causing leaf curl disease of tomato in Jeddah, Kingdom of Saudi Arabia are discussed.

  13. SHALLOT-LIKE1 Is a KANADI Transcription Factor That Modulates Rice Leaf Rolling by Regulating Leaf Abaxial Cell Development[W][OA

    PubMed Central

    Zhang, Guang-Heng; Xu, Qian; Zhu, Xu-Dong; Qian, Qian; Xue, Hong-Wei

    2009-01-01

    As an important agronomic trait, rice (Oryza sativa L.) leaf rolling has attracted much attention from plant biologists and breeders. Moderate leaf rolling increases the photosynthesis of cultivars and hence raises grain yield. However, the relevant molecular mechanism remains unclear. Here, we show the isolation and functional characterization of SHALLOT-LIKE1 (SLL1), a key gene controlling rice leaf rolling. sll1 mutant plants have extremely incurved leaves due to the defective development of sclerenchymatous cells on the abaxial side. Defective development can be functionally rescued by expression of SLL1. SLL1 is transcribed in various tissues and accumulates in the abaxial epidermis throughout leaf development. SLL1 encodes a SHAQKYF class MYB family transcription factor belonging to the KANADI family. SLL1 deficiency leads to defective programmed cell death of abaxial mesophyll cells and suppresses the development of abaxial features. By contrast, enhanced SLL1 expression stimulates phloem development on the abaxial side and suppresses bulliform cell and sclerenchyma development on the adaxial side. Additionally, SLL1 deficiency results in increased chlorophyll and photosynthesis. Our findings identify the role of SLL1 in the modulation of leaf abaxial cell development and in sustaining abaxial characteristics during leaf development. These results should facilitate attempts to use molecular breeding to increase the photosynthetic capacity of rice, as well as other crops, by modulating leaf development and rolling. PMID:19304938

  14. Transmission of Sweet Potato Leaf Curl Virus by Bemisia tabaci

    USDA-ARS?s Scientific Manuscript database

    Sweetpotato, Ipomoea batatas (L.) Lam. (Solanales: Convolvulaceae), is an important world food crop, and Asia is the focal production region. Because it is vegetatively propagated, sweetpotato is especially prone to accumulate infections by several viruses. Sweet potato leaf curl virus (SPLCV) (ss...

  15. Mining cotton germplasm resources to fight Cotton Leaf Curl Virus

    USDA-ARS?s Scientific Manuscript database

    CLCuV (Cotton Leaf Curl Virus) is a major threat to cotton production in Pakistan and parts of India and has been reported in cotton producing countries in Africa, as well as China and Uzbekistan. Identifying sources of resistance to CLCuV helps not only countries such as Pakistan where the virus is...

  16. DNA forms indicate rolling circle and recombination-dependent replication of Abutilon mosaic virus.

    PubMed

    Jeske, H; Lütgemeier, M; Preiss, W

    2001-11-01

    Geminiviruses have spread worldwide and have become increasingly important in crop plants during recent decades. Recombination among geminiviruses was one major source of new variants. Geminiviruses replicate via rolling circles, confirmed here by electron microscopic visualization and two-dimensional gel analysis of Abutilon mosaic virus (AbMV) DNA. However, only a minority of DNA intermediates are consistent with this model. The majority are compatible with recombination-dependent replication (RDR). During development of naturally infected leaves, viral intermediates compatible with both models appeared simultaneously, whereas agro-infection of leaf discs with AbMV led to an early appearance of RDR forms but no RCR intermediates. Inactivation of viral genes ac2 and ac3 delayed replication, but produced the same DNA types as after wild-type infection, indicating that these genes were not essential for RDR in leaf discs. In conclusion, host factors alone or in combination with the viral AC1 protein are necessary and sufficient for the production of RDR intermediates. The consequences of an inherent geminiviral recombination activity for the use of pathogen-derived resistance traits are discussed.

  17. Joint Mapping and Allele Mining of the Rolled Leaf Trait in Rice (Oryza sativa L.)

    PubMed Central

    Wang, Chunchao; Nafisah; Joseph, Charles; Zhang, Wenzhong; Xu, Jianlong; Li, Zhikang

    2016-01-01

    The rolled leaf trait, long considered to be a key component of plant architecture, represents an important target trait for improving plant architecture at the population level. We therefore performed linkage mapping using a set of 262 highly variable RILs from two rice cultivars (Minghui 63 and 02428) with minor differences in leaf rolling index (LRI) in conjunction with GWAS mapping of a random subset of the 1127 germplasms from the 3K Rice Genomes Project (3K Rice). A total of seven main-effect loci were found to underlie the transgressive segregation of progenies from parents with minor differences in LRI. Five of these loci were previously identified and two (qRl7b and qRl9b) are newly reported with additional evidence from GWAS mapping for qRl7b. A total of 18 QTLs were identified by GWAS, including four newly identified QTLs. Six QTLs were confirmed by linkage mapping with the above RIL population, and 83.3% were found to be consistent with previously reported loci based on comparative mapping. We also performed allele mining with representative SNPs and identified the elite germplasms for the improvement of rolled leaf trait. Most favorable alleles at the detected loci were contributed by various 3K Rice germplasms. By a re-scanning of the candidate region with more saturated SNP markers, we dissected the region harboring gRl4-2 into three subregions, in which the average effect on LRI was 3.5% with a range from 2.4 to 4.1% in the third subregion, suggesting the presence of a new locus or loci within this region. The representative SNPs for favorable alleles in the reliable QTLs which were consistently identified in both bi-parental mapping and GWAS, such as qRl4, qRl5, qRl6, qRl7a, and qRl7b will be useful for future molecular breeding programs for ideal plant type in rice. PMID:27441398

  18. The complete nucleotide sequence of pelargonium leaf curl virus.

    PubMed

    McGavin, Wendy J; MacFarlane, Stuart A

    2016-05-01

    Investigation of a tombusvirus isolated from tulip plants in Scotland revealed that it was pelargonium leaf curl virus (PLCV) rather than the originally suggested tomato bushy stunt virus. The complete sequence of the PLCV genome was determined for the first time, revealing it to be 4789 nucleotides in size and to have an organization similar to that of the other, previously described tombusviruses. Primers derived from the sequence were used to construct a full-length infectious clone of PLCV that recapitulates the disease symptoms of leaf curling in systemically infected pelargonium plants.

  19. Notes on the ecology of rolled-leaf hispines (Chrysomelidae, Cassidinae) at La Gamba (Costa Rica)1

    PubMed Central

    Schmitt, Michael; Frank, Meike

    2013-01-01

    Abstract A total of 301 adult hispine beetles of the genera Cephaloleia and Chelobasis were found in rolled leaves of plants of 17 species of Zingiberales (families Costaceae, Heliconiaceae, Maranthaceae, Musaceae, and Zingiberaceae) during a field study at La Gamba, Golfito region, Costa Rica. Of these beetles, Cephaloleia belti was recorded from 12 potential host plant species, C. distincta from 7, C. dilaticollis from 5, C., Chelobasis bicolor, C. championi, and C. histrionica from 3, Chelobasis perplexa and C. instabilis from 2, whereas C. trivittata from only one. Of the plant species, Heliconia latispatha had 7 beetle species in its leaf rolls, Calathea lutea had 5, H. imbricata and H. rostrata had 4, H. stricta and Musa paradisiaca had 3, H. wagneriana had 2, while on H. vaginalis, H. danielsiana, H. densiflora, H. longiflora, Calathea crotalifera, C. platystachya, Goeppertia lasiophylla, Alpinia purpurata, Costus pulverulentus and Costus barbatus, H. densiflora, H. vaginalis, and H. danielsana only hispines of one species were found. Cephaloleia belti occurred together with beetles of six other hispine species, whereas Cephaloleia trivittata never shared a leaf roll with another hispine species. The remaining beetle species aggregated with one to four other hispines. Adults of C. belti and C. championi were frequently seen, occasionally also with C. dilaticollis, C. histrionica, and Chelobasis perplexa, to co-occur with the carabid Calophaena ligata in the same leaf roll without any sign of interspecific aggression. A comparison of host choices and the phylogeny of the hispines and of their host plants revealed no signs that beetles used species level phylogenetic relationships within the Zingiberales to select food plants. Obviously, within this plant order, rolled-leaf hispines choose their plant hosts in a nearly opportunistic manner. Seemingly, they use differences among plants at higher taxonomic levels but within the Zingiberales, the availability of

  20. Carrot yellow leaf virus Is Associated with Carrot Internal Necrosis

    PubMed Central

    Adams, Ian P.; Skelton, Anna; Macarthur, Roy; Hodges, Tobias; Hinds, Howard; Flint, Laura; Nath, Palash Deb; Boonham, Neil; Fox, Adrian

    2014-01-01

    Internal necrosis of carrot has been observed in UK carrots for at least 10 years, and has been anecdotally linked to virus infection. In the 2009 growing season some growers had up to 10% of yield with these symptoms. Traditional diagnostic methods are targeted towards specific pathogens. By using a metagenomic approach with high throughput sequencing technology, other, as yet unidentified causes of root necrosis were investigated. Additionally a statistical analysis has shown which viruses are most closely associated with disease symptoms. Carrot samples were collected from a crop exhibiting root necrosis (102 Affected: 99 Unaffected) and tested for the presence of the established carrot viruses: Carrot red leaf virus (CtRLV), Carrot mottle virus (CMoV), Carrot red leaf associated viral RNA (CtRLVaRNA) and Parsnip yellow fleck virus (PYFV). The presence of these viruses was not associated with symptomatic carrot roots either as single viruses or in combinations. A sub-sample of carrots of mixed symptom status was subjected to MiSeq sequencing. The results from these tests suggested Carrot yellow leaf virus (CYLV) was associated with symptomatic roots. Additionally a novel Torradovirus, a novel Closterovirus and two novel Betaflexiviradae related plant viruses were detected. A specific diagnostic test was designed for CYLV. Of the 102 affected carrots, 98% were positive for CYLV compared to 22% of the unaffected carrots. From these data we conclude that although we have yet to practically demonstrate a causal link, CYLV appears to be strongly associated with the presence of necrosis of carrots. PMID:25365290

  1. A detailed analysis of the leaf rolling mutant sll2 reveals complex nature in regulation of bulliform cell development in rice (Oryza sativa L.).

    PubMed

    Zhang, J-J; Wu, S-Y; Jiang, L; Wang, J-L; Zhang, X; Guo, X-P; Wu, C-Y; Wan, J-M

    2015-03-01

    Bulliform cells are large, thin-walled and highly vacuolated cells, and play an important role in controlling leaf rolling in response to drought and high temperature. However, the molecular mechanisms regulating bulliform cell development have not been well documented. Here, we report isolation and characterisation of a rice leaf-rolling mutant, named shallot-like 2 (sll2). The sll2 plants exhibit adaxially rolled leaves, starting from the sixth leaf stage, accompanied by increased photosynthesis and reduced plant height and tiller number. Histological analyses showed shrinkage of bulliform cells, resulting in inward-curved leaves. The mutant is recessive and revertible at a rate of 9%. The leaf rolling is caused by a T-DNA insertion. Cloning of the insertion using TAIL-PCR revealed that the T-DNA was inserted in the promoter region of LOC_Os07 g38664. Unexpectedly, the enhanced expression of LOC_Os07 g38664 by the 35S enhancer in the T-DNA is not responsible for the leaf rolling phenotype. Further, the enhancer also exerted a long-distance effect, including up-regulation of several bulliform cell-related genes. sll2 suppressed the outward leaf rolling of oul1 in the sll2oul1 double mutant. We conclude that leaf rolling in sll2 could be a result of the combined effect of multi-genes, implying a complex network in regulation of bulliform cell development.

  2. Anti-influenza virus activity of Ginkgo biloba leaf extracts.

    PubMed

    Haruyama, Takahiro; Nagata, Kyosuke

    2013-07-01

    We examined the influence of Ginkgo biloba leaf extract (EGb) on the infectivity of influenza viruses in Madin-Darby canine kidney (MDCK) cells. Plaque assays demonstrated that multiplication of influenza viruses after adsorption to host cells was not affected in the agarose overlay containing EGb. However, when the viruses were treated with EGb before exposure to cells, their infectivity was markedly reduced. In contrast, the inhibitory effect was not observed when MDCK cells were treated with EGb before infection with influenza viruses. Hemagglutination inhibition assays revealed that EGb interferes with the interaction between influenza viruses and erythrocytes. The inhibitory effect of EGb was observed against influenza A (H1N1 and H3N2) and influenza B viruses. These results suggest that EGb contains an anti-influenza virus substance(s) that directly affects influenza virus particles and disrupts the function of hemagglutinin in adsorption to host cells. In addition to the finding of the anti-influenza virus activity of EGb, our results demonstrated interesting and important insights into the screening system for anti-influenza virus activity. In general, the plaque assay using drug-containing agarose overlays is one of the most reliable methods for detection of antiviral activity. However, our results showed that EGb had no effects either on the number of plaques or on their sizes in the plaque assay. These findings suggest the existence of inhibitory activities against the influenza virus that were overlooked in past studies.

  3. Natural association of two different betasatellites with Sweet potato leaf curl virus in wild morning glory (Ipomoea purpurea) in India.

    PubMed

    Swapna Geetanjali, A; Shilpi, S; Mandal, Bikash

    2013-08-01

    Wild morning glory (Ipomoea purpurea) was observed to be affected by leaf curl and yellow vein diseases during summer-rainy season of 2009 in New Delhi, India. The virus was experimentally transmitted through whitefly, Bemisia tabaci to I. purpurea that reproduced the two distinct symptoms. Sequence analysis of multiple full-length clones obtained through rolling circle amplification from the leaf curl and yellow vein samples showed 91.8-95.3% sequence identity with Sweet potato leaf curl virus (SPLCV) and the isolates were phylogenetically distinct from those reported from Brazil, China, Japan and USA. Interestingly, two different betasatellites, croton yellow vein mosaic betasatellite and papaya leaf curl betasatellite were found with SPLCV in leaf curl and yellow vein diseases of I. purpurea, respectively. This study is the first report of occurrence of SPLCV in wild morning glory in India. SPLCV was known to infect other species of morning glory; our study revealed that I. purpurea, a new species of morning glory was a natural host of SPLCV. To date, betasatellite associated with SPLCV in Ipomoea spp. is not known. Our study provides evidence of natural association of two different betasatellites with SPLCV in leaf curl and yellow vein diseases of I. purpurea.

  4. Virus Hunting: Discovery of New Episomal Circular Viruses by Rolling Circle Techniques.

    PubMed

    Vanmechelen, Bert; Rector, Annabel; Maes, Piet

    2017-02-06

    Many methods for the discovery of novel viruses are based on amplification of the virus using consensus or degenerate PCR primers. A downside of this approach is that it requires prior knowledge of the viral nucleotide sequence to be applicable. Presented in this unit is a method for the sequence-independent amplification of circular viral genomes that is based on the rolling-circle mechanism used by certain viruses in their natural replication cycle. The amplification of the virus of interest is coupled to the isolation of the viral genome by gel extraction following a restriction digestion. Once isolated, the sequence of the viral genome can be determined by nanopore sequencing, a rapid and inexpensive next-generation sequencing technology that generates long reads in real time. The method described in this unit was originally developed for the discovery of papillomaviruses, but can be used for the identification of all types of circular DNA viruses. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

  5. Effects of Clinacanthus siamensis leaf extract on influenza virus infection.

    PubMed

    Wirotesangthong, Mali; Nagai, Takayuki; Yamada, Haruki; Amnuoypol, Surattana; Mungmee, Chutichot

    2009-02-01

    Ethanolic extracts of 20 medicinal plants were screened for influenza virus NA inhibition and in vitro antiviral activities using MDCK cells in an MTT assay. The vaccine proteins of influenza virus A/New Caledonia/20/99 (H1N1), mouse-adapted influenza virus A/Guizhou/54/89 (A/G)(H3N2) and mouse-adapted influenza virus B/Ibaraki/2/85 (B/I) were used in the NA inhibition assay, and mouse-adapted influenza viruses A/PR/8/34 (H1N1), A/G and B/I were used in the in vitro antiviral assay. The results of the in vitro antiviral assay indicated that the A/G virus was the most susceptible and an extract of the leaf of CS possessed the highest in vitro anti-A/G virus activity (41.98%). Therefore, the A/G virus and the CS extract were selected for studying in vivo anti-influenza virus activity. BALB/c mice were treated with CS extract (100 mg/kg per day, 5 times) orally from 4 hr before to 4 days after infection. CS extract elicited significant production of anti-influenza virus IgG(1) antibody in BAW and increased mouse weight compared to oseltamivir (0.1 mg/kg per day) on day 19 or water on days 17-19 of infection. Moreover, CS extract produced a higher anti-influenza virus IgA antibody level in BAW compared to oseltamivir, and a tendency towards an increase in anti-influenza virus IgA compared to water was shown. The results suggest that CS extract has a protective effect against influenza virus infection.

  6. The relationship between leaf rolling and ascorbate-glutathione cycle enzymes in apoplastic and symplastic areas of Ctenanthe setosa subjected to drought stress.

    PubMed

    Saruhan, Neslihan; Terzi, Rabiye; Saglam, Aykut; Kadioglu, Asim

    2009-01-01

    The ascorbate-glutathione (ASC-GSH) cycle has an important role in defensive processes against oxidative damage generated by drought stress. In this study, the changes that take place in apoplastic and symplastic ASC-GSH cycle enzymes of the leaf and petiole were investigated under drought stress causing leaf rolling in Ctenanthe setosa (Rose.) Eichler (Marantaceae). Apoplastic and symplastic extractions of leaf and petiole were performed at different visual leaf rolling scores from 1 to 4 (1 is unrolled, 4 is tightly rolled and the others are intermediate forms). Glutathione reductase (GR), a key enzyme in the GSH regeneration cycle, and ascorbate (ASC) were present in apoplastic spaces of the leaf and petiole, whereas dehydroascorbate reductase (DHAR), which uses glutathione as reductant, monodehydroascorbate reductase (MDHAR), which uses NAD(P)H as reductant, and glutathione were absent. GR, DHAR and MDHAR activities increased in the symplastic and apoplastic areas of the leaf. Apoplastic and symplastic ASC and dehydroascorbate (DHA), the oxidized form of ascorbate, rose at all scores except score 4 of symplastic ASC in the leaf. On the other hand, while reduced glutathione (GSH) content was enhanced, oxidized glutathione (GSSG) content decreased in the leaf during rolling. As for the petiole, GR activity increased in the apoplastic area but decreased in the symplastic area. DHAR and MDHAR activities increased throughout all scores, but decreased to the score 1 level at score 4. The ASC content of the apoplast increased during leaf rolling. Conversely, symplastic ASC content increased at score 2, however decreased at the later scores. While the apoplastic DHA content declined, symplastic DHA rose at score 2, but later was down to the level of score 1. While GSH content enhanced during leaf rolling, GSSG content did not change except at score 2. As well, there were good correlations between leaf rolling and ASC-GSH cycle enzyme activities in the leaf (GR and DHAR

  7. Tête à Tête of Tomato Yellow Leaf Curl Virus and Tomato Yellow Leaf Curl Sardinia Virus in Single Nuclei

    PubMed Central

    Morilla, Gabriel; Krenz, Björn; Jeske, Holger; Bejarano, Eduardo R.; Wege, Christina

    2004-01-01

    Since 1997 two distinct geminivirus species, Tomato yellow leaf curl Sardinia virus (TYLCSV) and Tomato yellow leaf curl virus (TYLCV), have caused a similar yellow leaf curl disease in tomato, coexisted in the fields of southern Spain, and very frequently doubly infected single plants. Tomatoes as well as experimental test plants (e.g., Nicotiana benthamiana) showed enhanced symptoms upon mixed infections under greenhouse conditions. Viral DNA accumulated to a similar extent in singly and doubly infected plants. In situ tissue hybridization showed TYLCSV and TYLCV DNAs to be confined to the phloem in both hosts, irrespective of whether they were inoculated individually or in combination. The number of infected nuclei in singly or doubly infected plants was determined by in situ hybridization of purified nuclei. The percentage of nuclei containing viral DNA (i.e., 1.4% in tomato or 6% in N. benthamiana) was the same in plants infected with either TYLCSV, TYLCV, or both. In situ hybridization of doubly infected plants, with probes that discriminate between both DNAs, revealed that at least one-fifth of infected nuclei harbored DNAs from both virus species. Such a high number of coinfected nuclei may explain why recombination between different geminivirus DNAs occurs frequently. The impact of these findings for epidemiology and for resistance breeding concerning tomato yellow leaf curl diseases is discussed. PMID:15367638

  8. Leaf-rolling sawflies (Hymenoptera, Pamphiliidae, Pamphiliinae) of Tianmushan Mountains, Zhejiang Province, China.

    PubMed

    Shinohara, Akihiko; Wei, Mei-Cai

    2016-02-02

    Two species of Neurotoma, six species of Onycholyda, and five species of Pamphilius are recorded from Tianmushan, Zhejiang Province, China, and a key is given to these three genera and 13 species. Three new species, Onycholyda atra Shinohara & Wei, sp. nov. from Zhejiang Province, O. fulvicornis Shinohara, sp. nov. from Shaanxi Province and Zhejiang Province, and Pamphilius padus Shinohara, sp. nov. from Zhejiang Province, are described. New distribution records are: Onycholyda shaanxiana Shinohara, 1999, from Hubei Province, Zhejiang Province and Guangxi Zhuang Autonomous Region, O. subquadrata (Maa, 1944) from Zhejiang Province, O. tianmushana Shinohara & Xiao, 2006, from Hunan Province and Jiangxi Province, Pamphilius palliceps Shinohara & Xiao, 2006, from Anhui Province, P. lizejiani Shinohara in Shinohara & Wei, 2012, from Zhejiang Province, P. shengi Wei in Wei & Xiao, 1999, from Hubei Province and Zhejiang Province, and P. qinlingicus Wei in Wu & Wei, 2010, from Zhejiang Province. The larva of P. padus feeds on Padus obtusata (Rosaceae) and that of P. palliceps feeds on Rosa multiflora (Rosaceae), both singly making a leaf-roll. Rubus peltatus (Rosaceae) is recorded as a host plant of O. atra based on the observation of oviposition.

  9. Alfalfa Leaf Curl Virus: an Aphid-Transmitted Geminivirus.

    PubMed

    Roumagnac, Philippe; Granier, Martine; Bernardo, Pauline; Deshoux, Maëlle; Ferdinand, Romain; Galzi, Serge; Fernandez, Emmanuel; Julian, Charlotte; Abt, Isabelle; Filloux, Denis; Mesléard, François; Varsani, Arvind; Blanc, Stéphane; Martin, Darren P; Peterschmitt, Michel

    2015-09-01

    The family Geminiviridae comprises seven genera differentiated by genome organization, sequence similarity, and insect vector. Capulavirus, an eighth genus, has been proposed to accommodate two newly discovered highly divergent geminiviruses that presently have no known vector. Alfalfa leaf curl virus, identified here as a third capulavirus, is shown to be transmitted by Aphis craccivora. This is the first report of an aphid-transmitted geminivirus.

  10. Alfalfa Leaf Curl Virus: an Aphid-Transmitted Geminivirus

    PubMed Central

    Roumagnac, Philippe; Granier, Martine; Bernardo, Pauline; Deshoux, Maëlle; Ferdinand, Romain; Galzi, Serge; Fernandez, Emmanuel; Julian, Charlotte; Abt, Isabelle; Filloux, Denis; Mesléard, François; Varsani, Arvind; Blanc, Stéphane; Martin, Darren P.

    2015-01-01

    The family Geminiviridae comprises seven genera differentiated by genome organization, sequence similarity, and insect vector. Capulavirus, an eighth genus, has been proposed to accommodate two newly discovered highly divergent geminiviruses that presently have no known vector. Alfalfa leaf curl virus, identified here as a third capulavirus, is shown to be transmitted by Aphis craccivora. This is the first report of an aphid-transmitted geminivirus. PMID:26109720

  11. [Molecular detection of tomato yellow leaf curl virus (TYLCV)].

    PubMed

    Li, Chang-Bao; Cui, Yan-Ling; Zhang, Li-Ying; Li, Chuan-You

    2012-03-01

    Tomato yellow leaf curl virus (TYLCV) is currently considered as one of the most devastating viruses in cultivated tomatoes (Solanum lycopersicum) worldwide. We reported here the development of a PCR-based method to quickly detect TYLCV using the primer pairs (TYLCV-F: 5'-ACG CAT GCC TCT AAT CCA GTG TA-3' and TYLCV-R: 5'-CCA ATA AGG CGT AAG CGT GTA GAC-3'), which was designed based on the genome sequence of TYLCV. A TYLCV-specific band of 543 bp was amplified from infected tomato plants. This protocol provides a rapid, reliable, and sensitive tool for molecular detection and identification of TYLCV in the industrial seedling and virus resistance breeding to facilitate safe and sustainable production of tomato.

  12. Pepper leaf curl Lahore virus requires the DNA B component of Tomato leaf curl New Delhi virus to cause leaf curl symptoms

    PubMed Central

    2010-01-01

    Background Begomoviruses are whitefly-transmitted geminiviruses with genomes that consist of either two components (known as DNA A and DNA B) or a single component (homologous to the DNA A component of bipartite begomoviruses). Monopartite begomoviruses are often associated with a symptom-modulating DNA satellite (collectively known as betasatellites). Both bipartite and monopartite begomoviruses with associated satellites have previously been identified in chillies showing leaf curl symptoms in Pakistan. Results A chilli plant (Capsicum annum) with chilli leaf curl disease symptoms was found to contain a begomovirus, a betasatellite and the DNA B component of Tomato leaf curl New Delhi virus (ToLCNDV). The begomovirus consisted of 2747 nucleotides and had the highest sequence identity (99%) with Pepper leaf curl Lahore virus (PepLCLV-[PK: Lah:04], acc. no. AM404179). Agrobacterium-mediated inoculation of the clone to Nicotiana benthamiana, induced very mild symptoms and low levels of viral DNA, detected in systemically infected leaves by PCR. No symptoms were induced in Nicotiana tabacum or chillies either in the presence or absence of a betasatellite. However, inoculation of PepLCLV with the DNA B component of ToLCNDV induced leaf curl symptoms in N. benthamiana, N. tabacum and chillies and viral DNA accumulated to higher levels in comparison to plants infected with just PepLCLV. Conclusions Based on our previous efforts aimed at understanding of diversity of begomoviruses associated with chillies, we propose that PepLCLV was recently mobilized into chillies upon its interaction with DNA B of ToLCNDV. Interestingly, the putative rep-binding iterons found on PepLCLV (GGGGAC) differ at two base positions from those of ToLCNDV (GGTGTC). This is the first experimental demonstration of the infectivity for a bipartite begomovirus causing chilli leaf curl disease in chillies from Pakistan and suggests that component capture is contributing to the emerging complexity of

  13. Sweet Potato Leaf Curl Virus: Virus Reservoir in Species of Wild Morning Glory

    USDA-ARS?s Scientific Manuscript database

    Recent increases in populations of the Sweetpotato leaf curl virus (SPLCV) vector, the sweetpotato whitefly, Bemisia tabaci (Gennadius), led to a dramatic increase in the disease in sweetpotato (Ipomoea batatas). Knowledge of crop or weed species that occur in sweetpotato growing areas and can serv...

  14. Association of a recombinant Cotton leaf curl Bangalore virus with yellow vein and leaf curl disease of okra in India.

    PubMed

    Venkataravanappa, V; Lakshminarayana Reddy, C N; Devaraju, A; Jalali, Salil; Krishna Reddy, M

    2013-09-01

    A begomovirus isolate (OY136A) collected from okra plants showing upward leaf curling, vein clearing, vein thickening and yellowing symptoms from Bangalore rural district, Karnataka, India was characterized. The sequence comparisons revealed that, this virus isolate share highest nucleotide identity with isolates of Cotton leaf curl Bangalore virus (CLCuBV) (AY705380) (92.8 %) and Okra enation leaf curl virus (81.1-86.2 %). This is well supported by phylogentic analysis showing, close clustering of the virus isolate with CLCuBV. With this data, based on the current taxonomic criteria for the genus Begomovirus, the present virus isolate is classified as a new strain of CLCuBV, for which CLCuBV-[India: Bangalore: okra: 2006] additional descriptor is proposed. The betasatellite (KC608158) associated with the virus is having more than 95 % sequence similarity with the cotton leaf curl betasatellites (CLCuB) available in the GenBank.The recombination analysis suggested, emergence of this new strain of okra infecting begomovirus might have been from the exchange of genetic material between BYVMV and CLCuMuV. The virus was successfully transmitted by whitefly and grafting. The host range of the virus was shown to be very narrow and limited to two species in the family Malvaceae, okra (Abelmoschus esculentus) and hollyhock (Althaea rosea), and four in the family Solanaceae.

  15. Molecular characterization and phylogenetic analysis of sugarcane yellow leaf virus isolates from China

    USDA-ARS?s Scientific Manuscript database

    Sugarcane yellow leaf virus (SCYLV), the causal agent of sugarcane yellow leaf disease (YLD), was first reported in China in 2006. In order to determine the distribution existence of SCYLV in major sugarcane-growing provinces in China, leaf samples were collected from 22 sugarcane clones (Saccharum ...

  16. Evaluating Weeds as Hosts of Tomato yellow leaf curl virus.

    PubMed

    Smith, Hugh A; Seijo, Teresa E; Vallad, Gary E; Peres, Natalia A; Druffel, Keri L

    2015-08-01

    Bemisia tabaci (Gennadius) biotype B transmits Tomato yellow leaf curl virus (TYLCV), which affects tomato production globally. Prompt destruction of virus reservoirs is a key component of virus management. Identification of weed hosts of TYLCV will be useful for reducing such reservoirs. The status of weeds as alternate hosts of TYLCV in Florida remains unclear. In greenhouse studies, B. tabaci adults from a colony reared on TYLCV-infected tomato were established in cages containing one of four weeds common to horticultural fields in central and south Florida. Cages containing tomato and cotton were also infested with viruliferous whiteflies as a positive control and negative control, respectively. Whitefly adults and plant tissue were tested periodically over 10 wk for the presence of TYLCV using PCR. After 10 wk, virus-susceptible tomato plants were placed in each cage to determine if whiteflies descended from the original adults were still infective. Results indicate that Bidens alba, Emilia fosbergii, and Raphanus raphanistrum are not hosts of TYLCV, and that Amaranthus retroflexus is a host.

  17. First report of Sugarcane yellow leaf virus infecting Columbus Grass (Sorghum almum) in Florida

    USDA-ARS?s Scientific Manuscript database

    Sugarcane yellow leaf virus (SCYLV) [genus Polerovirus, family Luteoviridae] is the causal agent of sugarcane yellow leaf disease. SCYLV is widespread in Florida where sugarcane was the only known natural host of this virus. During spring 2015, we collected (leaves or stalks) and tested several gras...

  18. Chili leaf curl betasatellite is associated with a distinct recombinant begomovirus, Pepper leaf curl Lahore virus, in Capsicum in Pakistan.

    PubMed

    Tahir, Muhammad; Haider, Muhammad Saleem; Briddon, Rob W

    2010-04-01

    Capsium spp. are an important vegetable crop cultivated through Pakistan. Leaf curl disease is the major disease of Capsicum spp. in Pakistan caused by viruses. The disease has previously been shown to be associated with begomoviruses and betasatellites. We have cloned and sequenced a begomovirus and its associated betasatellite from Capsicum originating from central Pakistan. The begomovirus isolated was distinct from all previously characterised viruses and we propose the name Pepper leaf curl Lahore virus (PepLCLV) for this new species. Comparison of the sequence of PepLCLV with previously characterised begomoviruses shows it likely to have resulted from recombination between Papaya leaf curl virus and Chili leaf curl virus (ChiLCV), two species that have previously been identified in Pakistan. The betasatellite associated with PepLCLV in Capsicum was identified as Chili leaf curl betasatellite (ChLCB). This is the first identification of a cognate begomovirus for ChLCB infecting Capsicum, although this betasatellite has been shown in association with ChiLCV infecting potato in Pakistan. PepLCLV is one of an increasing number of monopartite begomoviruses shown to be associated with a betasatellite and one of the numerous species that affect Capsicum. In view of their only having been identified in Pakistan, PepLCLV and ChLCB likely represent a geographically distinct, Capsicum adapted, begomovirus-betasatellite complex.

  19. Ongoing geographical spread of Tomato yellow leaf curl virus.

    PubMed

    Mabvakure, Batsirai; Martin, Darren P; Kraberger, Simona; Cloete, Leendert; van Brunschot, Sharon; Geering, Andrew D W; Thomas, John E; Bananej, Kaveh; Lett, Jean-Michel; Lefeuvre, Pierre; Varsani, Arvind; Harkins, Gordon W

    2016-11-01

    Tomato yellow leaf curl virus (TYLCV) seriously impacts tomato production throughout tropical and sub-tropical regions of the world. It has a broad geographical distribution and continues to spread to new regions in the Indian and Pacific Oceans including Australia, New Caledonia and Mauritius. We undertook a temporally-scaled, phylogeographic analysis of all publicly available, full genome sequences of TYLCV, together with 70 new genome sequences from Australia, Iran and Mauritius. This revealed that whereas epidemics in Australia and China likely originated through multiple independent viral introductions from the East-Asian region around Japan and Korea, the New Caledonian epidemic was seeded by a variant from the Western Mediterranean region and the Mauritian epidemic by a variant from the neighbouring island of Reunion. Finally, we show that inter-continental scale movements of TYLCV to East Asia have, at least temporarily, ceased, whereas long-distance movements to the Americas and Australia are probably still ongoing.

  20. A leaf-rolling weevil benefits from general saprophytic fungi in polysaccharide degradation

    USDA-ARS?s Scientific Manuscript database

    Insects form symbiosis with fungi widely, especially those feeding on leaf litter. As dead plant tissues provide poor quality diets which contain relatively high levels of indigestible lignin and cellulose, saprophytic fungi may increase nutrient availability by polysaccharide degradation. Although ...

  1. Molecular Characterization of Tomato leaf curl Palampur virus and Pepper leaf curl betasatellite Naturally Infecting Pumpkin (Cucurbita moschata) in India.

    PubMed

    Namrata, Jaiswal; Saritha, R K; Datta, D; Singh, M; Dubey, R S; Rai, A B; Rai, M

    2010-10-01

    Pumpkin cultivation in India is affected by severe incidence of a yellow vein mosaic disease. Tomato leaf curl New Delhi virus and Squash leaf curl China virus are known to be associated with this disease in India. We were able to identify a third begomovirus-Tomato leaf curl Palampur virus (ToLCPMV), from pumpkin showing typical symptoms of the disease at Varanasi based on the sequence of complete DNA-A genome of the virus. The complete DNA-A sequence of the virus shared more than 99% sequence identity with other ToLCPMV isolates available in the GenBank and clustered with them in the phylogenetic analysis. This betasatellite amplified from the same infected sample has been identified as Pepper leaf curl betasatellite (PepLCB) which also infects chilli in India. There was 92% sequence identity between the two isolates. This is the first report of natural infection of ToLCPMV on pumpkin and association of PepLCB with yellow vein mosaic disease of pumpkin in India.

  2. First report of citrus leaf blotch virus in lemon in China

    USDA-ARS?s Scientific Manuscript database

    Citrus leaf blotch virus (CLBV) is a species of genus Citrivirus in the family Betaflexiviridae. The virus infects several species of the genus Citrus spp., but has not been previously reported from Lemon [Citrus limon (L.)]. The virus was identified in a lemon tree displaying yellow vein clearing i...

  3. Cotton leaf curl Burewala virus with intact or mutant transcriptional activator proteins: complexity of cotton leaf curl disease.

    PubMed

    Kumar, Jitendra; Gunapati, Samatha; Alok, Anshu; Lalit, Adarsh; Gadre, Rekha; Sharma, Naresh C; Roy, Joy K; Singh, Sudhir P

    2015-05-01

    Cotton leaf curl disease (CLCuD) is a serious disease of cotton on the Indian subcontinent. In the present study, three cotton leaf curl viruses, cotton leaf curl Burewala virus (CLCuBuV), cotton leaf curl Kokhran virus (CLCuKoV) and cotton leaf curl Multan virus (CLCuMV), and their associated satellites, cotton leaf curl Multan betasatellite (CLCuMB) and cotton leaf curl Multan alphasatellite (CLCuMA), were detected. CLCuBuV with either intact (CLCuBuV-1) or mutant (CLCuBuV-2) transcriptional activator protein (TrAP) were detected in different plants. Agroinoculation with CLCuBuV-1 or CLCuBuV-2 together with CLCuMB and CLCuMA, resulted in typical leaf curling and stunting of tobacco plants. Inoculation with CLCuKoV or an isolate of CLCuMV (CLCuMV-2), together with CLCuMB and CLCuMA, induced severe leaf curling, while the other isolate of CLCuMV (CLCuMV-1), which was recombinant in origin, showed mild leaf curling in tobacco. To investigate the effect of intact or mutant TrAP and also the recombination events, CLCuBuV-1, CLCuBuV-2, CLCuMV-1 or CLCuMV-2 together with the satellites (CLCuMA and CLCuMB) were transferred to cotton via whitefly-mediated transmission. Cotton plants containing CLCuBuV-1, CLCuBuV-2 or CLCuMV-2 together with satellites showed curling and stunting, whereas the plants having CLCuMV-1 and the satellites showed only mild and indistinguishable symptoms. CLCuBuV-1 (intact TrAP) showed severe symptoms in comparison to CLCuBuV-2 (mutant TrAP). The present study reveals that two types of CLCuBuV, one with an intact TrAP and the other with a mutant TrAP, exist in natural infection of cotton in India. Additionally, CLCuMuV-1, which has a recombinant origin, induces mild symptoms in comparison to the other CLCuMV isolates.

  4. Frequent Occurrence of Tomato Leaf Curl New Delhi Virus in Cotton Leaf Curl Disease Affected Cotton in Pakistan

    PubMed Central

    Zaidi, Syed Shan-e-Ali; Shafiq, Muhammad; Amin, Imran; Scheffler, Brian E.; Scheffler, Jodi A.; Briddon, Rob W.; Mansoor, Shahid

    2016-01-01

    Cotton leaf curl disease (CLCuD) is the major biotic constraint to cotton production on the Indian subcontinent, and is caused by monopartite begomoviruses accompanied by a specific DNA satellite, Cotton leaf curl Multan betasatellite (CLCuMB). Since the breakdown of resistance against CLCuD in 2001/2002, only one virus, the “Burewala” strain of Cotton leaf curl Kokhran virus (CLCuKoV-Bur), and a recombinant form of CLCuMB have consistently been identified in cotton across the major cotton growing areas of Pakistan. Unusually a bipartite isolate of the begomovirus Tomato leaf curl virus was identified in CLCuD-affected cotton recently. In the study described here we isolated the bipartite begomovirus Tomato leaf curl New Delhi virus (ToLCNDV) from CLCuD-affected cotton. To assess the frequency and geographic occurrence of ToLCNDV in cotton, CLCuD-symptomatic cotton plants were collected from across the Punjab and Sindh provinces between 2013 and 2015. Analysis of the plants by diagnostic PCR showed the presence of CLCuKoV-Bur in all 31 plants examined and ToLCNDV in 20 of the samples. Additionally, a quantitative real-time PCR analysis of the levels of the two viruses in co-infected plants suggests that coinfection of ToLCNDV with the CLCuKoV-Bur/CLCuMB complex leads to an increase in the levels of CLCuMB, which encodes the major pathogenicity (symptom) determinant of the complex. The significance of these results are discussed. PMID:27213535

  5. Standardized RT-PCR conditions for detection and identification of eleven viruses of potato and Potato spindle tuber viroid

    USDA-ARS?s Scientific Manuscript database

    Standardized RT-PCR procedures were developed and validated for detection of Alfalfa mosaic virus (AMV), Impatiens necrotic spot virus (INSV), Tobacco rattle virus (TRV), Tomato spotted wilt virus (TSWV), Potato leaf roll virus (PLRV), Potato mop top virus (PMTV), Potato virus A (PVA), Potato viru...

  6. Status of Sugarcane yellow leaf virus and its impact in different progenies

    USDA-ARS?s Scientific Manuscript database

    Yellow leaf disease caused by Sugarcane yellow leaf virus (SCYLV) a Polerovirus is an important disease for sugarcane industries worldwide. High yield losses up to 50% were reported in susceptible varieties. Most of the commercial cultivars in Florida are infected with SCYLV; therefore, there is a ...

  7. First report of an alphasatellite associated with Okra enation leaf curl virus.

    PubMed

    Chandran, S A; Packialakshmi, R M; Subhalakshmi, K; Prakash, C; Poovannan, K; Nixon Prabu, A; Gopal, P; Usha, R

    2013-06-01

    An alphasatellite DNA associated with Okra enation leaf curl virus (OELCuV) which causes enation and leaf curling in okra (Abelmoschus esculentus) plants was characterized. The full-length DNA comprises 1,350 nucleotides and shows typical genome organization of an alphasatellite. It shows the highest nucleotide sequence identity (79.7 %) to Hollyhock yellow vein virus-associated symptomless alphasatellite (HoYVSLA). This is the first report of the association of an alphasatellite with OELCuV from India.

  8. Novel Synthetic Promoters from the Cestrum Yellow Leaf Curling Virus.

    PubMed

    Sahoo, Dipak Kumar; Sarkar, Shayan; Maiti, Indu B; Dey, Nrisingha

    2016-01-01

    Constitutive promoters direct gene expression uniformly in most tissues and cells at all stages of plant growth and development; they confer steady levels of transgene expression in plant cells and hence their demand is high in plant biology. The gene silencing due to promoter homology can be avoided by either using diverse promoters isolated from different plant and viral genomes or by designing synthetic promoters. The aim of this chapter was to describe the basic protocols needed to develop and analyze novel, synthetic, nearly constitutive promoters from Cestrum yellow leaf curling virus (CmYLCV) through promoter/leader deletion and activating cis-sequence analysis. We also describe the methods to evaluate the strength of the promoters efficiently in various transient expression systems like agroinfiltration assay, gene-gun method, and assay in tobacco protoplasts. Besides, the detailed methods for developing transgenic plants (tobacco and Arabidopsis) for evaluation of the promoter using the GUS reporter gene are also described. The detailed procedure for electrophoretic mobility shift assay (EMSA) coupled with super-shift EMSA analysis are also described for showing the binding of tobacco transcription factor, TGA1a to cis-elements in the CmYLCV distal promoter region.

  9. Tomato yellow leaf curl virus C4 protein is a determinant of disease phenotype in tomato

    USDA-ARS?s Scientific Manuscript database

    Tomato yellow leaf curl virus (TYLCV) is a monopartite begomovirus. Its genome contains six open reading frames, with V1 and V2 in sense, and C1 to C4 in complementary orientation. The functions of V1 and V2 are for coat protein and pre-coat, respectively. C1 is for virus replication, C2 for trans-a...

  10. Biological characterization and complete genomic sequence of Carrot thin leaf virus

    USDA-ARS?s Scientific Manuscript database

    The host range of a cilantro isolate of Carrot thin leaf virus (CTLV-Cs) was determined to include 15 plant species. The virus was also transmitted to 9 of 11 tested apiaceous species by aphids. Complete genomic sequences of CTLV-Cs and a carrot isolate of CTLV were determined. Their genomic sequenc...

  11. Whitefly transmission of Sweet potato leaf curl virus in sweetpotato germplasm

    USDA-ARS?s Scientific Manuscript database

    Sweetpotato, Ipomoea batatas (L.) Lam., is among an extensive number of plant species attacked by Bemisia tabaci (Gennadius). Because this important world food crop is vegetatively propagated, it can conveniently accumulate infections by several viruses. Sweet potato leaf curl virus (SPLCV) (ssDNA...

  12. Management of sweet potato leaf curl virus in sweetpotatoes using insecticides

    USDA-ARS?s Scientific Manuscript database

    Sweetpotato leaf curl virus (SPLCV), which is transmitted by the sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), can severely affect yields of commercial sweetpotatoes, Ipomoea batatas (L.) Lam. (Convolvulaceae). This virus occurs every year at the U.S. Vegetable Laborato...

  13. Sweet Potato Leaf Curl Virus: Efficiency of Acquisition, Retention and Transmission by Bemisia tabaci (Hemiptera: Aleyrodidae)

    USDA-ARS?s Scientific Manuscript database

    The sweetpotato whitefly, Bemisia tabaci (Gennadius), is a global pest which damages plants directly by feeding on leaves. Moreover, the problem is compounded because B. tabaci also vectors numerous plant viruses, including Begomoviruses (Geminiviridae) such as the Sweet Potato Leaf Curl Virus (SPL...

  14. Tomato Leaf Curl New Delhi Virus: An Emerging Virus Complex Threatening Vegetable and Fiber Crops.

    PubMed

    Moriones, Enrique; Praveen, Shelly; Chakraborty, Supriya

    2017-09-21

    The tomato leaf curl New Delhi virus (ToLCNDV) (genus Begomovirus, family Geminiviridae) represents an important constraint to tomato production, as it causes the most predominant and economically important disease affecting tomato in the Indian sub-continent. However, in recent years, ToLCNDV has been fast extending its host range and spreading to new geographical regions, including the Middle East and the western Mediterranean Basin. Extensive research on the genome structure, protein functions, molecular biology, and plant-virus interactions of ToLCNDV has been conducted in the last decade. Special emphasis has been given to gene silencing suppression ability in order to counteract host plant defense responses. The importance of the interaction with DNA alphasatellites and betasatellites in the biology of the virus has been demonstrated. ToLCNDV genetic variability has been analyzed, providing new insights into the taxonomy, host adaptation, and evolution of this virus. Recombination and pseudorecombination have been shown as motors of diversification and adaptive evolution. Important progress has also been made in control strategies to reduce disease damage. This review highlights these various achievements in the context of the previous knowledge of begomoviruses and their interactions with plants.

  15. Phylogenetic lineage of Tobacco leaf curl virus in Korea and estimation of recombination events implicated in their sequence variation.

    PubMed

    Park, Jungan; Lee, Hyejung; Kim, Mi-Kyung; Kwak, Hae-Ryun; Auh, Chung-Kyoon; Lee, Kyeong-Yeoll; Kim, Sunghan; Choi, Hong-Soo; Lee, Sukchan

    2011-08-01

    New strains of Tobacco leaf curl virus (TbLCV) were isolated from tomato plants in four different local communities of Korea, and hence were designated TbLCV-Kr. Phylogenetic analysis of the sequences of the whole genome and of individual ORFs of these viruses indicated that they are closely related to the Tobacco leaf curl Japan virus (TbLCJV) cluster, which includes Honeysuckle yellow vein virus (HYVV), Honeysuckle yellow vein mosaic virus (HYVMV), and TbLCJV isolates. Four putative recombination events were recognized within these virus sequences, suggesting that the sequence variations observed in these viruses may be attributable to intraspecific and interspecific recombination events involving some TbLCV-Kr isolates, Papaya leaf curl virus (PaLCV), and a local isolate of Tomato yellow leaf curl virus (TYLCV). Copyright © 2011 Elsevier B.V. All rights reserved.

  16. Significant increase in titer of Raspberry bushy dwarf virus when present in combination with Raspberry leaf mottle virus and its effect on raspberry plants

    USDA-ARS?s Scientific Manuscript database

    Raspberry crumbly fruit is a virus-induced disease widespread in the Pacific Northwest (PNW). Raspberry bushy dwarf virus (RBDV) has been attributed as the causal agent of the disease. Recently, the identification of two new viruses: Raspberry leaf mottle virus (RLMV) and Raspberry latent virus (RpL...

  17. Host range and genetic diversity of croton yellow vein mosaic virus, a weed-infecting monopartite begomovirus causing leaf curl disease in tomato.

    PubMed

    Pramesh, D; Mandal, Bikash; Phaneendra, Chigurupati; Muniyappa, V

    2013-03-01

    Croton yellow vein mosaic virus (CYVMV) is a widely occurring begomovirus in Croton bonplandianum, a common weed in the Indian subcontinent. In this study, CYVMV (genus Begomovirus, family Geminiviridae) was transmitted by whiteflies (Bemisia tabaci) to as many as 35 plant species belonging to 11 families, including many vegetables, tobacco varieties, ornamentals and weeds. CYVMV produced bright yellow vein symptoms in croton, whereas in all the other host species, the virus produced leaf curl symptoms. CYVMV produced leaf curl in 13 tobacco species and 22 cultivars of Nicotiana tabacum and resembled tobacco leaf curl virus (TobLCV) in host reactions. However, CYVMV was distinguished from TobLCV in four differential hosts, Ageratum conyzoides, C. bonplandianum, Euphorbia geniculata and Sonchus bracyotis. The complete genome sequences of four isolates originating from northern, eastern and southern India revealed that a single species of DNA-A and a betasatellite, croton yellow vein mosaic betasatellite (CroYVMB) were associated with the yellow vein mosaic disease of croton. The sequence identity among the isolates of CYVMV DNA-A and CroYVMB occurring in diverse plant species was 91.8-97.9 % and 83.3-100 %, respectively. The CYVMV DNA-A and CroYVMB generated through rolling-circle amplification of the cloned DNAs produced typical symptoms of yellow vein mosaic and leaf curling in croton and tomato, respectively. The progeny virus from both the croton and tomato plants was transmitted successfully by B. tabaci. The present study establishes the etiology of yellow vein mosaic disease of C. bonplandianum and provides molecular evidence that a weed-infecting monopartite begomovirus causes leaf curl in tomato.

  18. Characterization and distribution of tomato yellow margin leaf curl virus, a begomovirus from Venezuela.

    PubMed

    Nava, A; Londoño, A; Polston, J E

    2013-02-01

    A begomovirus causing mottling and leaf deformation in tomato from the State of Mérida was cloned and sequenced. The virus has a bipartite genome comprised of a DNA-A (2,572 nucleotides) and a DNA-B (2,543 nucleotides) with a genome organization typical of New World begomoviruses. Both components share a common region of 115 nucleotides with 98 % sequence identity. Phylogenetic analysis indicated that while no virus sequences were closely related, the A component was distantly related to those of two other tomato-infecting viruses, tomato leaf deformation virus and Merremia mosaic virus; and the DNA-B, to those of pepper huasteco yellow vein virus and Rhynchosia golden mosaic Yucatan virus. The DNA-A and DNA-B sequences were submitted to GenBank (accession no. AY508993 and AY508994, respectively) and later accepted by the International Committee on Taxonomy of Viruses as the genome of a member of a unique virus species with the name Tomato yellow margin leaf curl virus (TYMLCV). Tomato (Solanum lycopersicum L. 'Fl. Lanai') plants inoculated with cloned TYMLCV DNA-A and DNA-B became systemically infected and showed chlorotic margins and leaf curling. The distribution of TYMLCV in tomato-producing states in Venezuela was determined by nucleic acid spot hybridization analysis of 334 tomato leaf samples collected from ten states using a TYMLCV-specific probe and confirmed by PCR and sequencing of the PCR fragment. TYMLCV was detected in samples from the states of Aragua, Guárico, and Mérida, suggesting that TYMLCV is widely distributed in Venezuela.

  19. Tomato leaf curl Karnataka virus from Bangalore, India, Appears to be a Recombinant Begomovirus.

    PubMed

    Chatchawankanphanich, Orawan; Maxwell, Douglas P

    2002-06-01

    ABSTRACT The genome of Tomato leaf curl virus (ToLCV) from Bangalore, India, a whitefly-transmitted geminivirus, was cloned (pIND9) and sequenced. The circular DNA of 2,759 nucleotides (U38239) is organized similarly to that of other begomoviruses with monopartite genomes. Comparison of the nucleotide sequence of pIND9 with other tomato-associated begomoviruses from India (Tomato leaf curl Bangalore virus [ToLCBV, Z48182]) and Tomato leaf curl New Delhi virus-Severe (ToLCNdV-Svr, U15015) showed moderate DNA sequence identities (82 to 87%) between capsid protein (CP) genes but low identities (66 to 67%) for the intergenic regions and the replication-associated protein (Rep) genes (75 to 81% identity). Phylogenetic trees generated with nucleotide sequences of the Rep and CP genes of 26 begomoviruses indicated that this ToLCV is distinct from other begomoviruses and that it may be a recombinant virus derived from at least three different viral lineages. Tomatoes (Lycopersicon esculentum) inoculated with the cloned DNA monomer of ToLCV (pIND9) via particle bombardment developed leaf curling and yellowing symptoms. The virus was transmitted by Bemisia tabaci biotype B from tomatoes infected via particle bombardment to healthy tomatoes and by sap inoculation from infected tomatoes to tomato, Nicotiana benthamiana and N. tabacum. This ToLCV is a distinct member of the genus Begomovirus from India that differs from the previously characterized Tomato leaf curl Sadasivanagar virus isolate Bangalore 1 (L12739), ToLCBV (Z48182), ToLCBV isolate Bangalore 4 (AF165098), and the bipartite ToLCNdV (U15015, U15016). Thus, this ToLCV is named Tomato leaf curl Karnataka virus (ToLCKV).

  20. Anti-influenza virus activity of Myrica rubra leaf ethanol extract evaluated using Madino-Darby canine kidney (MDCK) cells.

    PubMed

    Mochida, Kyo

    2008-11-01

    Myrica rubra leaf ethanol extract was added to culture medium of Madino-Darby canine kidney (MDCK) cells inoculated with influenza virus, and the inhibition of influenza virus replication was measured. Myrica rubra leaf ethanol extract showed anti-influenza virus activity irrespective of the hemagglutinin antigen type in the influenza virus type A (H1N1), its subtype (H3N2), and type B.

  1. Relationship between Tomato yellow leaf curl viruses and the whitefly vector.

    PubMed

    Ssekyewa, Charles; Van Damme, Patrick L J; Hofte, Monica

    2007-01-01

    Tomato Yellow Leaf Curl Virus (TYLCV) and Tomato Leaf Curl Virus (ToLCV) are the currently known begomoviruses in Uganda. The relationship with their whiteflies (Bemisia tabaci) vector and its management were not known in Uganda. A direct relationship was expected between these begomoviruses and whiteflies at Buwama in Mpigi district of Uganda. Farmer practices were expected to have limited efficacy. To investigate this, a completely randomised block design was used for all trials with six treatments replicated three times. Treatments evaluated were farmer whitefly pest management practices. These included both chemical pesticide and non-pesticide applications. Data on whitefly population and tomato yellow leaf curl virus disease incidence was recorded weekly, and analysed using SAS and SPSS statistical programmes for ANOVA, and correlations. Ranked means, coefficients of variation and standard errors were noted. Virus-vector relationship field studies established that virus occurrence varied in space and time, and with management practices, crop development stage, and weather conditions. A negative relationship (R = -0.14, p 0.04) was established between number of plants infected with TYLCV (sensu lato) and percentage marketable tomato yield. Tomato maturity was inversely propotional to whitefly infestation (R = -0.5, p 0.0001). Uprooting and application of dimethoate was most effective of the six treatments. Tomato leaf curl virus diseases and whitefly management options were established in Uganda.

  2. Functional Analysis of Cotton Leaf Curl Kokhran Virus/Cotton Leaf Curl Multan Betasatellite RNA Silencing Suppressors

    PubMed Central

    Saeed, Muhammad; Briddon, Rob W.; Dalakouras, Athanasios; Krczal, Gabi; Wassenegger, Michael

    2015-01-01

    In South Asia, Cotton leaf curl disease (CLCuD) is caused by a complex of phylogenetically-related begomovirus species and a specific betasatellite, Cotton leaf curl Multan betasatellite (CLCuMuB). The post-transcriptional gene silencing (PTGS) suppression activities of the transcriptional activator protein (TrAP), C4, V2 and βC1 proteins encoded by Cotton leaf curl Kokhran virus (CLCuKoV)/CLCuMuB were assessed in Nicotiana benthamiana. A variable degree of local silencing suppression was observed for each viral protein tested, with V2 protein exhibiting the strongest suppression activity and only the C4 protein preventing the spread of systemic silencing. The CLCuKoV-encoded TrAP, C4, V2 and CLCuMuB-encoded βC1 proteins were expressed in Escherichia coli and purified. TrAP was shown to bind various small and long nucleic acids including single-stranded (ss) and double-stranded (ds) RNA and DNA molecules. C4, V2, and βC1 bound ssDNA and dsDNA with varying affinities. Transgenic expression of C4 under the constitutive 35S Cauliflower mosaic virus promoter and βC1 under a dexamethasone inducible promoter induced severe developmental abnormalities in N. benthamiana. The results indicate that homologous proteins from even quite closely related begomoviruses may differ in their suppressor activity and mechanism of action. The significance of these findings is discussed. PMID:26512705

  3. Brugmansia suaveolens mottle virus, a novel potyvirus causing leaf mottling of Brugmansia suaveolens in Brazil.

    PubMed

    Lucinda, N; Nagata, T; Inoue-Nagata, A K; Salaroli, R B; Kitajima, E W

    2008-01-01

    A potyvirus was isolated from Brugmansia suaveolens showing leaf mottling and tentatively named Brugmansia suaveolens mottle virus (BsMoV). The virus (isolate Bs-Campinas) could infect some solanaceous plants and two Chenopodium species, and was transmitted by aphids. Symptomatic leaves contained flexuous particles and cylindrical inclusions. RT-PCR amplification using potyvirus universal primers produced a DNA fragment of 1851 nt (3' terminal genomic region), which shared 71% nucleotide identity with Pepper mottle virus, the best-matched potyvirus sequence. Since this identity value is below the threshold currently used to discriminate Potyvirus species, Brugmansia suaveolens mottle virus most likely represents a new Potyvirus species.

  4. Molecular characterization of Tobacco leaf curl Pusa virus, a new monopartite Begomovirus associated with tobacco leaf curl disease in India.

    PubMed

    Singh, Manoj K; Singh, K; Haq, Q M R; Mandal, B; Varma, A

    2011-10-01

    Leaf curl disease of tobacco (TbLCD) is endemic in India. A monopartite Begomovirus, a betasatellite and an alphasatellite were found associated with the disease in Pusa, Bihar. The DNA-A of the Begomovirus associated with TbLCD in Pusa, Bihar was found to comprise of 2707 nt with a typical Old World begomovirus-like genome organization. The full-length sequence of DNA-A [HQ180391] showed that the Pusa isolate is a newly described member of the genus Begomovirus, as it had <89% sequence homology with DNA-A of all the known begomoviruses. The isolate is tentatively named as Tobacco leaf curl Pusa virus [India:Pusa:2010]. The betasatellite (HQ180395) associated with TbLCD in Pusa was identified as a variant of Tomato leaf curl Bangladesh betasatellite [IN:Raj:03], with which it shared 90.4% sequence identity. The alphasatellite (HQ180392) associated with the disease had highest 87% nucleotide sequence identity with Tomato leaf curl alphasatellite. The Begomovirus, betasatellite, and alphasatellite associated with TbLCD in Pusa, Bihar, India were found to be recombinants of extant begomoviruses, betasatellites and alphasatellites spreading in the Indian sub-continent and South-East Asia.

  5. Development of Cotton leaf curl virus resistant transgenic cotton using antisense ßC1 gene.

    PubMed

    Sohrab, Sayed Sartaj; Kamal, Mohammad A; Ilah, Abdul; Husen, Azamal; Bhattacharya, P S; Rana, D

    2016-05-01

    Cotton leaf curl virus (CLCuV) is a serious pathogen causing leaf curl disease and affecting the cotton production in major growing areas. The transgenic cotton (Gossypium hirsutum cv. Coker 310) plants were developed by using βC1 gene in antisense orientation gene driven by Cauliflower mosaic virus-35S promoter and nos (nopaline synthase) terminator and mediated by Agrobacterium tumefaciens transformation and somatic embryogenesis system. Molecular confirmation of the transformants was carried out by polymerase chain reaction (PCR) and Southern blot hybridization. The developed transgenic and inoculated plants remained symptomless till their growth period. In conclusion, the plants were observed as resistant to CLCuV.

  6. Infectious clones of Tomato leaf curl Palampur virus with a defective DNA B and their pseudo-recombination with Tomato leaf curl New Delhi virus

    PubMed Central

    2011-01-01

    Background Tomato leaf curl Palampur virus (ToLCPMV) is a bipartite begomovirus which has been reported from India and Iran but infectious clones have not been obtained. We have previously shown the association of Zucchini yellow mosaic virus (ZYMV), a potyvirus, with severe leaf curl disease of muskmelon in Pakistan. However, the severity of symptoms in the field and yield losses led us to believe that some other agent, such as a begomovirus, could be associated with the disease. Results A bipartite begomovirus associated with a severe yellow leaf curl disease on muskmelon in Pakistan has been characterized. Analysis of the complete nucleotide sequence of the DNA A and DNA B components of the begomovirus showed that it has the highest DNA sequence identity with ToLCPMV. However, the gene encoding the nuclear shuttle protein (NSP) was truncated in comparison to previously characterised isolates. Agrobacterium-mediated inoculation of Nicotiana benthamiana with the ToLCPMV clones obtained here did not result in symptoms. However, inoculation of plants with the DNA A component of ToLCPMV and the DNA B component of Tomato leaf curl New Delhi virus (ToLCNDV) lead to systemic infection with leaf curl symptoms. This suggested that the lack of infectivity of the ToLCPMV clones was due to the defect in DNA B. The DNA B of ToLCPMV was able to move systemically when inoculated with DNA A of the either virus. Agro-infiltration of muskmelon with the DNA A and DNA B components of ToLCPMV did not lead to symptomatic infection whereas inoculation with the DNA A with the DNA B of ToLCNDV resulted in a hypersensitive response (HR) along the veins. Additionally, agro-infiltration of muskmelon with a construct for the expression of the NSP gene of ToLCNDV under the control of the cauliflower mosaic virus 35S promoter induced a HR, suggesting that this is the gene causing the HR. Conclusions Both ToLCPMV and ZYMV are associated with muskmelon leaf curl disease in Pakistan. However, the

  7. Molecular diversity of Cotton leaf curl Gezira virus isolates and their satellite DNAs associated with okra leaf curl disease in Burkina Faso

    PubMed Central

    2010-01-01

    Okra leaf curl disease (OLCD) is a major constraint on okra (Abelmoschus esculentus) production and is widespread in Africa. Using a large number of samples representative of the major growing regions in Burkina Faso (BF), we show that the disease is associated with a monopartite begomovirus and satellite DNA complexes. Twenty-three complete genomic sequences of Cotton leaf curl Gezira virus (CLCuGV) isolates associated with OLCD, sharing 95 to 99% nucleotide identity, were cloned and sequenced. Six betasatellite and four alphasatellite (DNA-1) molecules were also characterized. The six isolates of betasatellite associated with CLCuGV isolates correspond to Cotton leaf curl Gezira betasatellite (CLCuGB) (88 to 98% nucleotide identity). One isolate of alphasatellite is a variant of Cotton leaf curl Gezira alphasatellite (CLCuGA) (89% nucleotide identity), whereas the three others isolates appear to correspond to a new species of alphasatellite (CLCuGA most similar sequence present 52 to 60% nucleotide identity), provisionally named Okra leaf curl Burkina Faso alphasatellite (OLCBFA). Recombination analysis of the viruses demonstrated the interspecies recombinant origin of all CLCuGV isolates, with parents being close to Hollyhock leaf crumple virus (AY036009) and Tomato leaf curl Diana virus (AM701765). Combined with the presence of satellites DNA, these results highlight the complexity of begomoviruses associated with OLCD. PMID:20178575

  8. Properties of a virus causing mosaic and leaf curl disease of Celosia argentea L. in Nigeria.

    PubMed

    Owolabi, T A; Taiwo, M A; Thottappilly, G A; Shoyinka, S A; Proll, E; Rabenstein, F

    1998-06-01

    A sap transmissible virus, causing mosaic and leaf curl disease of Celosia argentea, was isolated at vegetable farms in Amuwo Odofin, Tejuoso, and Abule Ado, Lagos, Nigeria. The virus had a restricted host range confined to a few species of the Amaranthaceae, Chenopodiaceae and Solanaceae families. It failed to infect several other species of the Aizoaceae, Brassicaceae, Cucurbitaceae, Fabaceae, Lamiaceae, Malvaceae, Poaceae and Tiliaceae families. The virus was transmitted in a non-persistent manner by Aphis spiraecola and Toxoptera citricidus but not by eight other aphid species tested. There was no evidence of transmission by seeds of C. argentae varieties. The viral coat protein had a relative molecular mass (M(r)) of about 30.2 K. Electron microscopy of purified virus preparations revealed flexuous rod shaped particles of about 750 nm in length. Serological studies were performed using the enzyme-linked immunosorbent assay (ELISA), immunosorbent electron microscopy (ISEM) and Western blot analysis. The virus reacted positively with an universal potyvirus group monoclonal antibody (MoAb) and MoAb P-3-3H8 raised against peanut stripe potyvirus. It also reacted with polyclonal antibodies raised against several potyviruses including asparagus virus-1 (AV-1), turnip mosaic virus (TuMV), maize dwarf mosaic virus (MDMV), watermelon mosaic virus (WMV-2), plum pox virus (PPV), soybean mosaic virus (SoyMV), lettuce mosaic virus (LMV), bean common mosaic virus (BCMV) and beet mosaic virus (BMV) in at least one of the serological assays used. On the basis of host range, mode of transmission, and available literature data, the celosia virus seems to be different from potyviruses previously reported to infect vegetables in Nigeria. The name celosia mosaic virus (CIMV) has been proposed for this virus.

  9. Unravel the genetic basis of Sugarcane Yellow Leaf Virus (SCYLV) resistance in Saccharum spp. hybrid

    USDA-ARS?s Scientific Manuscript database

    Sugarcane (Saccharum Spp.) produces 80% of the world’s table sugar along with several other byproducts. The production of sugarcane is vulnerable due to infestation of sugarcane yellow leaf virus (SCYLV) worldwide. A genetic mapping study was conducted using an F1 segregating population derived from...

  10. Papaya is not a host for Tomato Yellow Leaf Curl Virus

    USDA-ARS?s Scientific Manuscript database

    The economic value of tomato production is threatened by tomato yellow leaf-curl virus TYLCV and its vector, the silverleaf whitefly Bemisia tabaci biotype B (Gennadius) (Hemiptera: Aleyrodidae). Use of papaya Carica papaya L. as a banker plant for a whitefly parasitoid shows promise as a whitefly m...

  11. Multiple Introductions of the Old World Begomovirus Tomato yellow leaf curl virus into the New World▿

    PubMed Central

    Duffy, Siobain; Holmes, Edward C.

    2007-01-01

    A phylogenetic analysis of three genomic regions revealed that Tomato yellow leaf curl virus (TYLCV) from western North America is distinct from TYLCV isolated in eastern North America and the Caribbean. This analysis supports a second introduction of this Old World begomovirus into the New World, most likely from Asia. PMID:17827315

  12. Complete nucleotide sequence of Rose yellow leaf virus, a new member of the family Tombusviridae

    USDA-ARS?s Scientific Manuscript database

    The genome of the Rose yellow leaf virus (RYLV) has been determined to be 3918 nucleotides containing seven open reading frames (ORFs). ORF1 encodes a 27 kDa peptide (p27). ORF2 shares a common start codon with ORF1 and continues through the amber stop codon of p27 to encode a 87 kDa (p87) protein t...

  13. Is the begomovirus, sweet potato leaf curl virus, really seed transmitted in sweetpotato?

    USDA-ARS?s Scientific Manuscript database

    Sweetpotato is one of the major root crops in the world and is also widely grown in the southern United States. Sweet potato leaf curl virus (SPLCV) is a begomovirus posing a serious threat to sweetpotato production worldwide and is primarily transmitted by whitefly (Bemisia tabaci) or through veget...

  14. Replication of Tomato Yellow Leaf Curl Virus in Its Whitefly Vector, Bemisia tabaci.

    PubMed

    Pakkianathan, Britto Cathrin; Kontsedalov, Svetlana; Lebedev, Galina; Mahadav, Assaf; Zeidan, Muhammad; Czosnek, Henryk; Ghanim, Murad

    2015-10-01

    Tomato yellow leaf curl virus (TYLCV) is a begomovirus transmitted exclusively by the whitefly Bemisia tabaci in a persistent, circulative manner. Replication of TYLCV in its vector remains controversial, and thus far, the virus has been considered to be nonpropagative. Following 8 h of acquisition on TYLCV-infected tomato plants or purified virions and then transfer to non-TYLCV-host cotton plants, the amounts of virus inside whitefly adults significantly increased (>2-fold) during the first few days and then continuously decreased, as measured by the amounts of genes on both virus DNA strands. Reported alterations in insect immune and defense responses upon virus retention led us to hypothesize a role for the immune response in suppressing virus replication. After virus acquisition, stress conditions were imposed on whiteflies, and the levels of three viral gene sequences were measured over time. When whiteflies were exposed to TYLCV and treatment with two different pesticides, the virus levels continuously increased. Upon exposure to heat stress, the virus levels gradually decreased, without any initial accumulation. Switching of whiteflies between pesticide, heat stress, and control treatments caused fluctuating increases and decreases in virus levels. Fluorescence in situ hybridization analysis confirmed these results and showed virus signals inside midgut epithelial cell nuclei. Combining the pesticide and heat treatments with virus acquisition had significant effects on fecundity. Altogether, our results demonstrate for the first time that a single-stranded DNA plant virus can replicate in its hemipteran vector. Plant viruses in agricultural crops are of great concern worldwide. Many of them are transmitted from infected to healthy plants by insects. Persistently transmitted viruses often have a complex association with their vectors; however, most are believed not to replicate within these vectors. Such replication is important, as it contributes to the

  15. Interaction of tomato yellow leaf curl virus with diverse betasatellites enhances symptom severity.

    PubMed

    Ito, Takuya; Kimbara, Junji; Sharma, Pradeep; Ikegami, Masato

    2009-01-01

    The complete nucleotide sequence was determined for a begomovirus isolated from tomato exhibiting leaf curling and yellowing symptoms in Tochigi Prefecture in Japan. The genome organization of this virus was similar to those of other Old World monopartite begomoviruses. Neither a DNA betasatellite nor a DNA-B component was detected. It had the highest total nucleotide sequence identity (99%) with tomato yellow leaf curl virus-Israel[Japan:Tosa:2005] (TYLCV-IL[JR:Tos:05]) and TYLCV-Israel[Japan:Haruno:2005] (TYLCV-IL[JR:Han:05]). Its coat protein V1 also showed an identical amino acid sequence with those of TYLCV-IL[JR:Tos:05] and TYLCV-IL[JR:Han:05]. Thus, the begomovirus was determined to be an isolate of TYLCV-IL designated as TYLCV-Israel[Japan:Tochigi:2007] (TYLCV-IL[JR:Toc:07]). We investigated the interaction of TYLCV-IL[JR:Toc:07] with two known satellites associated with tomato yellow dwarf disease in Japan, tobacco leaf curl Japan betasatellite [Japan:Ibaraki:2006] and honeysuckle yellow vein mosaic betasatellite [Japan:Nara:2006], as well as with tomato leaf curl Philippines betasatellite [Philippines:Laguna1:2008], in tomato and Nicotiana benthamiana plants. TYLCV-IL[JR:Toc:07] trans-replicated these betasatellites, inducing more severe tomato yellow leaf curl disease-related symptoms than TYLCV-IL[JR:Toc:07] alone.

  16. Molecular characterization and pathogenicity of tomato yellow leaf curl virus in China.

    PubMed

    Zhang, Hui; Gong, Huanran; Zhou, Xueping

    2009-10-01

    Several tomato production regions in China were surveyed for tomato yellow leaf curl disease (TYLCD), and 31 tomato leaf samples showing TYLCD-like symptoms were collected. The partial or full-length genomes of these isolates were sequenced and tomato yellow leaf curl virus (TYLCV) was detected in Shanghai, Zhejiang, Jiangsu Shandong and Hebei provinces of China. The TYLCV isolates found in China share high sequence identity ([98%) and have more than 97% sequence identity with TYLCVIL[ IL:Reo] (X15656). Phylogenetic relationship analysis reveals that although with little genetic variability, they can form two groups and all the TYLCV isolates in China belong to the group I. An infectious clone of TYLCV-[CN:SH2] (AM282874) was constructed and agro-inoculated into Nicotiana benthamiana, N. tabacum Samsun, N. glutinosa, Solanum lycopersicum, Petunia hybrida, Cucumis sativus, Gossypium hirsutum, S. melongena, and Capsicum annuum. TYLCV-[CN:SH2] can induce severe leaf curling and stunting symptoms in these plants except C. sativus, G. hirsutum, S. melongena and C. annuum.We verified that TYLCV can trans-replicate tomato yellow leaf curl China virus DNA-b in N. benthamiana and S. lycopersicum and induced more severe symptoms with distortion and yellow vein.

  17. Real-time RT-PCR for detection of Raspberry bushy dwarf virus, Raspberry leaf mottle virus and characterizing synergistic interactions in mixed infections

    USDA-ARS?s Scientific Manuscript database

    Two TaqMan-based real-time One-Step RT-PCR assays were developed for the rapid and efficient detection of Raspberry bushy dwarf virus (RBDV) and Raspberry leaf mottle virus (RLMV), two of the most common raspberry viruses in North America and Europe. The primers and probes were designed from conser...

  18. Transcriptome analysis to identify differentially expressed genes associated with ty-5 against tomato yellow leaf curl virus and Sw-7 against tomato spotted wilt virus in tomato

    USDA-ARS?s Scientific Manuscript database

    Tomato yellow leaf curl virus (TYLCV) and Tomato spotted wilt virus (TSWV) are two of the most economically important viruses on tomato (Solanum lycopersicum) worldwide. Developing a cultivar with resistance would be the most effective and economical means for viral disease management. However, id...

  19. Survey of apple chlorotic leaf spot virus and apple stem grooving virus occurrence in Korea and frequency of mixed infections in apple

    USDA-ARS?s Scientific Manuscript database

    Due to the absence of knowledge of the distribution of Apple stem grooving virus (ASGV) and Apple chlorotic leaf spot virus (ACLSV) in apples in Korea, we carried out a survey for these viruses in Gyeongsang and Chungcheong provinces in 2014. A total of 65 samples were collected and tested by RT-PCR...

  20. Quantification and Localization of Watermelon Chlorotic Stunt Virus and Tomato Yellow Leaf Curl Virus (Geminiviridae) in Populations of Bemisia tabaci (Hemiptera, Aleyrodidae) with Differential Virus Transmission Characteristics

    PubMed Central

    Kollenberg, Mario; Winter, Stephan; Götz, Monika

    2014-01-01

    Bemisia tabaci (Gennadius) is one of the economically most damaging insects to crops in tropical and subtropical regions. Severe damage is caused by feeding and more seriously by transmitting viruses. Those of the genus begomovirus (Geminiviridae) cause the most significant crop diseases and are transmitted by B. tabaci in a persistent circulative mode, a process which is largely unknown. To analyze the translocation and to identify critical determinants for transmission, two populations of B. tabaci MEAM1 were compared for transmitting Watermelon chlorotic stunt virus (WmCSV) and Tomato yellow leaf curl virus (TYLCV). Insect populations were chosen because of their high and respectively low virus transmission efficiency to compare uptake and translocation of virus through insects. Both populations harbored Rickettsia, Hamiltonella and Wolbachia in comparable ratios indicating that endosymbionts might not contribute to the different transmission rates. Quantification by qPCR revealed that WmCSV uptake and virus concentrations in midguts and primary salivary glands were generally higher than TYLCV due to higher virus contents of the source plants. Both viruses accumulated higher in insects from the efficiently compared to the poorly transmitting population. In the latter, virus translocation into the hemolymph was delayed and virus passage was impeded with limited numbers of viruses translocated. FISH analysis confirmed these results with similar virus distribution found in excised organs of both populations. No virus accumulation was found in the midgut lumen of the poor transmitter because of a restrained virus translocation. Results suggest that the poorly transmitting population comprised insects that lacked transmission competence. Those were selected to develop a population that lacks virus transmission. Investigations with insects lacking transmission showed that virus concentrations in midguts were reduced and only negligible virus amounts were found at the

  1. Quantification and localization of Watermelon chlorotic stunt virus and Tomato yellow leaf curl virus (Geminiviridae) in populations of Bemisia tabaci (Hemiptera, Aleyrodidae) with differential virus transmission characteristics.

    PubMed

    Kollenberg, Mario; Winter, Stephan; Götz, Monika

    2014-01-01

    Bemisia tabaci (Gennadius) is one of the economically most damaging insects to crops in tropical and subtropical regions. Severe damage is caused by feeding and more seriously by transmitting viruses. Those of the genus begomovirus (Geminiviridae) cause the most significant crop diseases and are transmitted by B. tabaci in a persistent circulative mode, a process which is largely unknown. To analyze the translocation and to identify critical determinants for transmission, two populations of B. tabaci MEAM1 were compared for transmitting Watermelon chlorotic stunt virus (WmCSV) and Tomato yellow leaf curl virus (TYLCV). Insect populations were chosen because of their high and respectively low virus transmission efficiency to compare uptake and translocation of virus through insects. Both populations harbored Rickettsia, Hamiltonella and Wolbachia in comparable ratios indicating that endosymbionts might not contribute to the different transmission rates. Quantification by qPCR revealed that WmCSV uptake and virus concentrations in midguts and primary salivary glands were generally higher than TYLCV due to higher virus contents of the source plants. Both viruses accumulated higher in insects from the efficiently compared to the poorly transmitting population. In the latter, virus translocation into the hemolymph was delayed and virus passage was impeded with limited numbers of viruses translocated. FISH analysis confirmed these results with similar virus distribution found in excised organs of both populations. No virus accumulation was found in the midgut lumen of the poor transmitter because of a restrained virus translocation. Results suggest that the poorly transmitting population comprised insects that lacked transmission competence. Those were selected to develop a population that lacks virus transmission. Investigations with insects lacking transmission showed that virus concentrations in midguts were reduced and only negligible virus amounts were found at the

  2. A Rapid and Efficient Method for Construction of an Infectious Clone of Tomato yellow leaf curl virus.

    PubMed

    Bang, Bongjun; Lee, Jongyun; Kim, Sunyoung; Park, Jungwook; Nguyen, Thao Thi; Seo, Young-Su

    2014-09-01

    Tomato yellow leaf curl virus (TYLCV), a member of the genus Begomovirus, is responsible for one of the most devastating viral diseases in tomato-growing countries and is becoming a serious problem in many subtropical and tropical countries. The climate in Korea is getting warmer and developing subtropical features in response to global warming. These changes are being accompanied by TYLCV, which is now becoming a large problem in the Korean tomato industry. The most effective way to reduce damage caused by TYLCV is to breed resistant varieties of tomatoes. To accomplish this, it is necessary to establish a simple inoculation technique for the efficient evaluation of resistance to TYLCV. Here, we present the rolling circle amplification (RCA) method, which employs a bacteriophage using phi-29 DNA polymerase for construction of infectious TYLCV clones. The RCA method is simple, does not require sequence information for cloning, and is less expensive and time consuming than conventional PCR based-methods. Furthermore, RCA-based construction of an infectious clone can be very useful to other emerging and unknown geminiviruses in Korea.

  3. Characterization and occurrence of squash chlorotic leaf spot virus, a tentative new torradovirus infecting cucurbits in Sudan.

    PubMed

    Lecoq, H; Verdin, E; Tepfer, M; Wipf-Scheibel, C; Millot, P; Dafalla, G; Desbiez, C

    2016-06-01

    During a survey conducted in Sudan in 2012, a virus with spherical particles was isolated from a squash plant showing chlorotic leaf spots. The virus was transmitted mechanically and by two whitefly species, but not by aphids. RT-PCR with generic torradovirus primers yielded a band of expected size from total RNA of a symptomatic plant. Next-generation sequencing confirmed that this is tentatively a new torradovirus, for which we propose the name 'squash chlorotic leaf spot virus'. Using specific RT-PCR primers, the virus was detected in cucurbit samples collected since 1992 at different locations in Sudan.

  4. Antiviral effect of flavonol glycosides isolated from the leaf of Zanthoxylum piperitum on influenza virus.

    PubMed

    Ha, Song-Yi; Youn, Hana; Song, Chang-Seon; Kang, Se Chan; Bae, Jong Jin; Kim, Hee Tae; Lee, Kwang Min; Eom, Tae Hoon; Kim, In Su; Kwak, Jong Hwan

    2014-04-01

    The ethanol extract of Zanthoxylum piperitum (L.) DC. showed in vitro antiviral activity against influenza A virus. Three flavonol glycosides were isolated from the EtOAc fraction of Z. piperitum leaf by means of activity-guided chromatographic separation. Structures of isolated compounds were identified as quercetin 3-O-β-D-galactopyranoside (1), quercetin 3-O-α-L-rhamnopyranoside (2), kaempferol 3-O-α-L-rhamnopyranoside (3) by comparing their spectral data with literature values. The anti-influenza viral activity of isolates was evaluated using a plaque reduction assay against influenza A/NWS/33 (H1N1) virus. The compounds also were subjected to neuraminidase inhibition assay in influenza A/NWS/33 virus. Compounds 1-3 exhibited antiviral activity against an influenza A virus in vitro, and inhibited the neuraminidase activity at relatively high concentrations.

  5. Transcript mapping of Cotton leaf curl Burewala virus and its cognate betasatellite, Cotton leaf curl Multan betasatellite

    PubMed Central

    2012-01-01

    Background Whitefly-transmitted geminiviruses (family Geminiviridae, genus Begomovirus) are major limiting factors for the production of numerous dicotyledonous crops throughout the warmer regions of the world. In the Old World a small number of begomoviruses have genomes consisting of two components whereas the majority have single-component genomes. Most of the monopartite begomoviruses associate with satellite DNA molecules, the most important of which are the betasatellites. Cotton leaf curl disease (CLCuD) is one of the major problems for cotton production on the Indian sub-continent. Across Pakistan, CLCuD is currently associated with a single begomovirus (Cotton leaf curl Burewala virus [CLCuBuV]) and the cotton-specific betasatellite Cotton leaf curl Multan betasatellite (CLCuMuB), both of which have recombinant origins. Surprisingly, CLCuBuV lacks C2, one of the genes present in all previously characterized begomoviruses. Virus-specific transcripts have only been mapped for few begomoviruses, including one monopartite begomovirus that does not associate with betasatellites. Similarly, the transcripts of only two betasatellites have been mapped so far. The study described has investigated whether the recombination/mutation events involved in the evolution of CLCuBuV and its associated CLCuMuB have affected their transcription strategies. Results The major transcripts of CLCuBuV and its associated betasatellite (CLCuMuB) from infected Nicotiana benthamiana plants have been determined. Two complementary-sense transcripts of ~1.7 and ~0.7 kb were identified for CLCuBuV. The ~1.7 kb transcript appears similar in position and size to that of several begomoviruses and likely directs the translation of C1 and C4 proteins. Both complementary-sense transcripts can potentially direct the translation of C2 and C3 proteins. A single virion-sense transcript of ~1 kb, suitable for translation of the V1 and V2 genes was identified. A predominant complementary

  6. Sequence characterization of tomato leaf curl Sinaloa virus and tomato severe leaf curl virus: phylogeny of New World begomoviruses and detection of recombination.

    PubMed

    Rojas, A; Kvarnheden, A; Marcenaro, D; Valkonen, J P T

    2005-07-01

    Diseases caused by begomoviruses (family Geminiviridae, genus Begomovirus) constitute a serious constraint to tomato production in Nicaragua. In this study, the complete nucleotide (nt) sequences of the DNA-A and DNA-B components were determined for the first time for Tomato leaf curl Sinaloa virus (ToLCSinV). In addition, the complete nt sequence was determined for the DNA-A component of two isolates of Tomato severe leaf curl virus (ToSLCV). The genome organization of ToLCSinV and ToSLCV was identical to the bipartite genomes of other begomoviruses described from the Americas. A phylogenetic analysis of DNA-A including 45 begomovirus species showed that the indigenous begomoviruses of the New World can be divided into three major clades and an intermediate group: AbMV clade, SLCV clade, "Brazil clade", and BGYMV group. Phylogenetic analyses of the DNA-A and DNA-B components and their open reading frames indicated that ToLCSinV and ToSLCV belong to different clades: ToLCSinV to the AbMV clade, and ToSLCV to the SLCV clade. The two Nicaraguan isolates of ToSLCV showed a close relationship with ToSLCV from Guatemala (ToSLCV-[GT96-1]) and Tomato chino La Paz virus (ToChLPV), but differed significantly in the AV1 and AC1 regions, respectively. Computer-based predictions indicated that recombination with another begomovirus had taken place within AV1 of ToSLCV dividing this species into two strains. A high probability was also found that ToChLPV is involved in the evolution of ToSLCV.

  7. Newly identified RNAs of raspberry leaf blotch virus encoding a related group of proteins.

    PubMed

    Lu, Yuwen; McGavin, Wendy; Cock, Peter J A; Schnettler, Esther; Yan, Fei; Chen, Jianping; MacFarlane, Stuart

    2015-11-01

    Members of the genus Emaravirus, including Raspberry leaf blotch virus (RLBV), are enveloped plant viruses with segmented genomes of negative-strand RNA, although the complete genome complement for any of these viruses is not yet clear. Currently, wheat mosaic virus has the largest emaravirus genome comprising eight RNAs. Previously, we identified five genomic RNAs for RLBV; here, we identify a further three RNAs (RNA6-8). RNA6-8 encode proteins that have clear homologies to one another, but not to any other emaravirus proteins. The proteins self-interacted in yeast two-hybrid and bimolecular fluorescence complementation (BiFC) experiments, and the P8 protein interacted with the virus nucleocapsid protein (P3) using BiFC. Expression of two of the proteins (P6 and P7) using potato virus X led to an increase in virus titre and symptom severity, suggesting that these proteins may play a role in RLBV pathogenicity; however, using two different tests, RNA silencing suppression activity was not detected for any of the RLBV proteins encoded by RNA2-8.

  8. Complete nucleotide sequence of rose yellow leaf virus, a new member of the family Tombusviridae.

    PubMed

    Mollov, Dimitre; Lockhart, Ben; Zlesak, David C

    2014-10-01

    The genome of the rose yellow leaf virus (RYLV) has been determined to be 3918 nucleotides long and to contain seven open reading frames (ORFs). ORF1 encodes a 27-kDa peptide (p27). ORF2 shares a common start codon with ORF1 and continues through the amber stop codon of p27 to encode an 87-kDa (p87) protein that has amino acid similarity to the RNA-dependent RNA polymerase (RdRp) of members of the family Tombusviridae. ORFs 3 and 4 have no significant amino acid similarity to known functional viral ORFs. ORF5 encodes a 6-kDa (p6) protein that has similarity to movement proteins of members of the Tombusviridae. ORF5A has no conventional start codon and overlaps with p6. A putative +1 frameshift mechanism allows p6 translation to continue through the stop codon and results in a 12-kDa protein that has high homology to the carmovirus p13 movement protein. The 37-kDa protein encoded by ORF6 has amino acid sequence similarity to coat proteins (CP) of members of the Tombusviridae. ORF7 has no significant amino acid similarity to known viral ORFs. Phylogenetic analysis of the RdRp amino acid sequences grouped RYLV together with the unclassified Rosa rugosa leaf distortion virus (RrLDV), pelargonium line pattern virus (PLPV), and pelargonium chlorotic ring pattern virus (PCRPV) in a distinct subgroup of the family Tombusviridae.

  9. Seed Transmission of Tomato yellow leaf curl virus in White Soybean (Glycine max).

    PubMed

    Kil, Eui-Joon; Park, Jungho; Choi, Hong-Soo; Kim, Chang-Seok; Lee, Sukchan

    2017-08-01

    Tomato yellow leaf curl virus (TYLCV) infection of the common bean (Phaseolus vulgaris) has been reported, but soybean (Glycine max) has not previously been identified as a TYLCV host. Five cultivars of white soybean were agro-inoculated using an infectious TYLCV clone. At 30 days post-inoculation, they showed infection rates of 25% to 100%. Typical TYLCV symptoms were not observed in any inoculated plants. To examine whether TYLCV was transmitted in soybean seeds, DNA was isolated from bundles of five randomly selected seeds from TYLCV-inoculated soybean plants and amplified with a TYLCV-specific primer set. With the exception of one bundle, all bundles of seeds were verified to be TYLCV-infected. Virus dissemination was also confirmed in three of the 14 bunches. Viral replication was also identified in seeds and seedlings. This is the first report demonstrating that soybean is a TYLCV host, and that TYLCV is a seed-transmissible virus in white soybean.

  10. Tomato yellow leaf curl virus: No evidence for replication in the insect vector Bemisia tabaci

    PubMed Central

    Sánchez-Campos, Sonia; Rodríguez-Negrete, Edgar A.; Cruzado, Lucía; Grande-Pérez, Ana; Bejarano, Eduardo R.; Navas-Castillo, Jesús; Moriones, Enrique

    2016-01-01

    Begomovirus ssDNA plant virus (family Geminiviridae) replication within the Bemisia tabaci vector is controversial. Transovarial transmission, alteration to whitefly biology, or detection of viral transcripts in the vector are proposed as indirect evidence of replication of tomato yellow leaf curl virus (TYLCV). Recently, contrasting direct evidence has been reported regarding the capacity of TYLCV to replicate within individuals of B. tabaci based on quantitave PCR approaches. Time-course experiments to quantify complementary and virion sense viral nucleic acid accumulation within B. tabaci using a recently implemented two step qPCR procedure revealed that viral DNA quantities did not increase for time points up to 96 hours after acquisition of the virus. Our findings do not support a recent report claiming TYLCV replication in individuals of B. tabaci. PMID:27476582

  11. Efficient Regeneration and Selection of Virus-free Sweetpotato Plants from Sweet Potato Leaf Curl Virus Infected Materials and Their Effects on Yields in Field Trials

    USDA-ARS?s Scientific Manuscript database

    Sweet potato leaf curl virus (SPLCV) is an emerging virus disease in sweetpotato (Ipomoea batata) in the U.S. The incidence of SPLCV infection on sweetpotato increased dramatically in recent years due to the explosion of whitefly, Bemisia tabaci (Gennadius) populations. Among several sweetpotato v...

  12. Natural Occurrence of Tomato leaf curl New Delhi virus in Iranian Cucurbit Crops

    PubMed Central

    Yazdani-Khameneh, Sara; Aboutorabi, Samaneh; Shoori, Majid; Aghazadeh, Azin; Jahanshahi, Parastoo; Golnaraghi, Alireza; Maleki, Mojdeh

    2016-01-01

    The main areas for field-grown vegetable production in Iran were surveyed during the years of 2012–2014 to determine the occurrence of begomoviruses infecting these crops. A total of 787 leaf samples were collected from vegetables and some other host plants showing virus-like symptoms and tested by an enzyme-linked immunosorbent assay (ELISA) using polyclonal antibodies produced against Tomato yellow leaf curl virus (TYLCV). According to the ELISA results, 81 samples (10.3%) positively reacted with the virus antibodies. Begomovirus infections were confirmed by polymerase chain reaction (PCR) using previously described TYLCV-specific primer pair TYLCV-Sar/TYLCV-Isr or universal primer pair Begomo-F/Begomo-R. The PCR tests using the primer pair TYLCV-Sar/TYLCV-Isr resulted in the amplification of the expected fragments of ca. 0.67-kb in size for ELISA-positive samples tested from alfalfa, pepper, spinach and tomato plants, confirming the presence of TYLCV. For one melon sample, having a week reaction in ELISA and no reaction in PCR using TYLCV-specific primers, the PCR reaction using the primer pair Begomo-F/Begomo-R resulted in the amplification fragments of the expected size of ca. 2.8 kb. The nucleotide sequences of the DNA amplicons derived from the isolate, Kz-Me198, were determined and compared with other sequences available in GenBank. BLASTN analysis confirmed the begomovirus infection of the sample and showed 99% identities with Tomato leaf curl New Delhi virus (ToLCNDV); phylogenetic analysis supported the results of the database searches. This study reports the natural occurrence of TYLCV in different hosts in Iran. Our results also reveal the emergence of ToLCNDV in Iranian cucurbit crops. PMID:27298595

  13. Potential threat of a new pathotype of Papaya leaf distortion mosaic virus infecting transgenic papaya resistant to Papaya ringspot virus.

    PubMed

    Bau, H-J; Kung, Y-J; Raja, J A J; Chan, S-J; Chen, K-C; Chen, Y-K; Wu, H-W; Yeh, S-D

    2008-07-01

    A virus identified as a new pathotype of Papaya leaf distortion mosaic virus (PLDMV, P-TW-WF) was isolated from diseased papaya in an isolated test-field in central Taiwan, where transgenic papaya lines resistant to Papaya ringspot virus (PRSV) were evaluated. The infected plants displayed severe mosaic, distortion and shoe-stringing on leaves; stunting in apex; and water-soaking on petioles and stems. This virus, which did not react in enzyme-linked immunosorbent assay with the antiserum to the PRSV coat protein, infected only papaya, but not the other 18 plant species tested. Virions studied under electron microscope exhibited morphology and dimensions of potyvirus particles. Reverse transcription-polymerase chain reaction conducted using potyvirus-specific primers generated a 1,927-nucleotide product corresponding to the 3' region of a potyvirus, showing high sequence identity to the CP gene and 3' noncoding region of PLDMV. Search for similar isolates with the antiserum against CP of P-TW-WF revealed scattered occurrence of PLDMV in Taiwan. Phylogenetic analysis of PLDMV isolates of Taiwan and Japan indicated that the Taiwan isolates belong to a separate genetic cluster. Since all the Taiwan isolates infected only papaya, unlike the cucurbit-infecting Japanese P type isolates, the Taiwan isolates are considered a new pathotype of PLDMV. Susceptibility of all our PRSV-resistant transgenic papaya lines to PLDMV indicates that the virus is an emerging threat for the application of PRSV-resistant transgenic papaya in Taiwan and elsewhere.

  14. Identification and characterization of privet leaf blotch-associated virus, a novel idaeovirus.

    PubMed

    Navarro, Beatriz; Loconsole, Giuliana; Giampetruzzi, Annalisa; Aboughanem-Sabanadzovic, Nina; Ragozzino, Antonio; Ragozzino, Ester; Di Serio, Francesco

    2017-09-01

    A novel virus has been identified by next-generation sequencing (NGS) in privet (Ligustrum japonicum L.) affected by a graft-transmissible disease characterized by leaf blotch symptoms resembling infectious variegation, a virus-like privet disease with an unclear aetiology. This virus, which has been tentatively named 'privet leaf blotch-associated virus' (PrLBaV), was absent in non-symptomatic privet plants, as revealed by NGS and reverse transcription-polymerase chain reaction (RT-PCR). Molecular characterization of PrLBaV showed that it has a segmented genome composed of two positive single-stranded RNAs, one of which (RNA1) is monocistronic and codes for the viral replicase, whereas the other (RNA2) contains two open reading frames (ORFs), ORF2a and ORF2b, coding for the putative movement (p38) and coat (p30) proteins, respectively. ORF2b is very probably expressed through a subgenomic RNA starting with six nucleotides (AUAUCU) that closely resemble those found in the 5'-terminal end of genomic RNA1 and RNA2 (AUAUUU and AUAUAU, respectively). The molecular signatures identified in the PrLBaV RNAs and proteins resemble those of Raspberry bushy dwarf virus (RBDV), currently the only member of the genus Idaeovirus. These data, together with phylogenetic analyses, are consistent with the proposal of considering PrLBaV as a representative of the second species in the genus Idaeovirus. Transient expression of a recombinant PrLBaV p38 fused to green fluorescent protein in leaves of Nicotiana benthamiana, coupled with confocal laser scanning microscopy assays, showed that it localizes at cell plasmodesmata, strongly supporting its involvement in viral movement/trafficking and providing the first functional characterization of an idaeovirus encoded protein. © 2016 BSPP AND JOHN WILEY & SONS LTD.

  15. Engineered Disease Resistance in Cotton Using RNA-Interference to Knock down Cotton leaf curl Kokhran virus-Burewala and Cotton leaf curl Multan betasatellite Expression.

    PubMed

    Ahmad, Aftab; Zia-Ur-Rehman, Muhammad; Hameed, Usman; Qayyum Rao, Abdul; Ahad, Ammara; Yasmeen, Aneela; Akram, Faheem; Bajwa, Kamran Shahzad; Scheffler, Jodi; Nasir, Idrees Ahmad; Shahid, Ahmad Ali; Iqbal, Muhammad Javed; Husnain, Tayyab; Haider, Muhammad Saleem; Brown, Judith K

    2017-09-14

    Cotton leaf curl virus disease (CLCuD) is caused by a suite of whitefly-transmitted begomovirus species and strains, resulting in extensive losses annually in India and Pakistan. RNA-interference (RNAi) is a proven technology used for knockdown of gene expression in higher organisms and viruses. In this study, a small interfering RNA (siRNA) construct was designed to target the AC1 gene of Cotton leaf curl Kokhran virus-Burewala (CLCuKoV-Bu) and the βC1 gene and satellite conserved region of the Cotton leaf curl Multan betasatellite (CLCuMB). The AC1 gene and CLCuMB coding and non-coding regions function in replication initiation and suppression of the plant host defense pathway, respectively. The construct, Vβ, was transformed into cotton plants using the Agrobacterium-mediated embryo shoot apex cut method. Results from fluorescence in situ hybridization and karyotyping assays indicated that six of the 11 T₁ plants harbored a single copy of the Vβ transgene. Transgenic cotton plants and non-transgenic (susceptible) test plants included as the positive control were challenge-inoculated using the viruliferous whitefly vector to transmit the CLCuKoV-Bu/CLCuMB complex. Among the test plants, plant Vβ-6 was asymptomatic, had the lowest amount of detectable virus, and harbored a single copy of the transgene on chromosome six. Absence of characteristic leaf curl symptom development in transgenic Vβ-6 cotton plants, and significantly reduced begomoviral-betasatellite accumulation based on real-time polymerase chain reaction, indicated the successful knockdown of CLCuKoV-Bu and CLCuMB expression, resulting in leaf curl resistant plants.

  16. Detection of tomato yellow leaf curl Thailand virus by PCR without DNA extraction.

    PubMed

    Ieamkhang, Supaporn; Riangwong, Lumpueng; Chatchawankanphanich, Orawan

    2005-11-01

    We report the simple and rapid method for detection of tomato yellow leaf curl Thailand virus (TYLCTHV) based on the direct capture of virus particles to the surface of a polymerase chain reaction (PCR) tube. This method allowed PCR without the time-consuming procedures of DNA extraction from infected plant tissue. A small amount of tomato tissue (approximately 10 mg) was ground in extraction buffer to release viruses from plant tissues. The constituents of the plant extract that might inhibit PCR activity were discarded by washing the tube with PBST buffer before adding the PCR mixture to the tube. This method was used for detection of TYLCTHV with plant sap solution diluted up to 1:20,000 and was more sensitive than an enzyme-linked immunosorbent assay (ELISA) method. In addition, this method can be used for detection of TYLCTHV in viruliferous whiteflies. The PCR tubes with captured TYLCTHV could be used for PCR, after storage at 4 degrees C for 4 wk. The method presented here was used for detection of begomoviruses in cucurbit and pepper. In addition, this method was effectively used to detect papaya ringspot virus in papaya and zucchini yellow mosaic virus in cucumber by reverse transcriptase (RT)-PCR.

  17. Multiple forms of vector manipulation by a plant-infecting virus: Bemisia tabaci and tomato yellow leaf curl virus.

    PubMed

    Liu, Baiming; Preisser, Evan L; Chu, Dong; Pan, Huipeng; Xie, Wen; Wang, Shaoli; Wu, Qingjun; Zhou, Xuguo; Zhang, Youjun

    2013-05-01

    For many insect-vectored plant viruses, the relationship between feeding behavior and vector competence may prove integral to an understanding of the epidemiology of the resulting plant disease. While plant-infecting viruses are well known to change host plant physiology in a way that makes them more attractive to vectors, viral manipulation of the vectors themselves has only recently been reported. Previous research suggested that the rapid spread of Tomato yellow leaf curl virus (TYLCV) throughout China has been facilitated by its primary vector, the whitefly Bemisia tabaci. We conducted two experiments testing the impact of TYLCV infection of the host plant (tomato) and vector (B. tabaci biotypes B and Q) on whitefly feeding behavior. Whiteflies of biotypes B and Q both appeared to find TYLCV-infected plants more attractive, probing them more quickly and having a greater number of feeding bouts; this did not, however, alter the total time spent feeding. Viruliferous whiteflies fed more readily than uninfected whiteflies and spent more time salivating into sieve tube elements. Because vector salivation is essential for viral transmission, this virally mediated alteration of behavior should provide TYLCV a direct fitness benefit. This is the first report of such manipulation by a nonpropagative virus that belongs to an exclusively plant-infecting family of viruses (Geminiviridae). In the context of previous research showing that feeding on TYLCV-infected plants harms biotype B but helps biotype Q, the fact that both biotypes were equally affected by TYLCV also suggests that the virus may alter the biotype B-biotype Q competitive interaction in favor of biotype Q.

  18. The autophagy pathway participates in resistance to tomato yellow leaf curl virus infection in whiteflies.

    PubMed

    Wang, Lan-Lan; Wang, Xin-Ru; Wei, Xue-Mei; Huang, Huang; Wu, Jian-Xiang; Chen, Xue-Xin; Liu, Shu-Sheng; Wang, Xiao-Wei

    2016-09-01

    Macroautophagy/autophagy plays an important role against pathogen infection in mammals and plants. However, little has been known about the role of autophagy in the interactions of insect vectors with the plant viruses, which they transmit. Begomoviruses are a group of single-stranded DNA viruses and are exclusively transmitted by the whitefly Bemisia tabaci in a circulative manner. In this study, we found that the infection of a begomovirus, tomato yellow leaf curl virus (TYLCV) could activate the autophagy pathway in the Middle East Asia Minor 1 (MEAM1) species of the B. tabaci complex as evidenced by the formation of autophagosomes and ATG8-II. Interestingly, the activation of autophagy led to the subsequent degradation of TYLCV coat protein (CP) and genomic DNA. While feeding the whitefly with 2 autophagy inhibitors (3-methyladenine and bafilomycin A1) and silencing the expression of Atg3 and Atg9 increased the viral load; autophagy activation via feeding of rapamycin notably decreased the amount of viral CP and DNA in the whitefly. Furthermore, we found that activation of whitefly autophagy could inhibit the efficiency of virus transmission; whereas inhibiting autophagy facilitated virus transmission. Taken together, these results indicate that TYLCV infection can activate the whitefly autophagy pathway, which leads to the subsequent degradation of virus. Furthermore, our report proves that an insect vector uses autophagy as an intrinsic antiviral program to repress the infection of a circulative-transmitted plant virus. Our data also demonstrate that TYLCV may replicate and trigger complex interactions with the insect vector.

  19. A Novel Strain of Tomato Leaf Curl New Delhi Virus Has Spread to the Mediterranean Basin

    PubMed Central

    Fortes, Isabel M.; Sánchez-Campos, Sonia; Fiallo-Olivé, Elvira; Díaz-Pendón, Juan A.; Navas-Castillo, Jesús; Moriones, Enrique

    2016-01-01

    Tomato leaf curl New Delhi virus (ToLCNDV) is a whitefly-transmitted bipartite begomovirus (genus Begomovirus, family Geminiviridae) that causes damage to multiple cultivated plant species mainly belonging to the Solanaceae and Cucurbitaceae families. ToLCNDV was limited to Asian countries until 2012, when it was first reported in Spain, causing severe epidemics in cucurbit crops. Here, we show that a genetically-uniform ToLCNDV population is present in Spain, compatible with a recent introduction. Analyses of ToLCNDV isolates reported from other parts of the world indicated that this virus has a highly heterogeneous population genetically with no evident geographical, plant host or year-based phylogenetic groups observed. Isolates emerging in Spain belong to a strain that seems to have evolved by recombination. Isolates of this strain seem adapted to infecting cucurbits, but poorly infect tomatoes. PMID:27834936

  20. Purification of particles of subterranean clover red leaf virus using an industrial-grade cellulase.

    PubMed

    Waterhouse, P M; Helms, K

    1984-07-01

    Particles of two isolates of subterranean clover red leaf virus were purified by a method in which infected plant tissue was digested with an industrial-grade cellulase, Celluclast 2.0 L type X. The yields of virus particles using this enzyme were comparable with those obtained using either of two laboratory-grade cellulases, Cellulase type 1 (Sigma) and Driselase. However, the specific infectivity or aphid transmissibility of the particles purified using Celluclast was 10-100 times greater than those of preparations obtained using laboratory-grade cellulases or no enzyme. The main advantage of using Celluclast is that at present in Australia its cost is only ca. 1% of laboratory-grade cellulases.

  1. Detection and molecular characterization of tomato yellow leaf curl virus naturally infecting Lycopersicon esculentum in Egypt.

    PubMed

    Rabie, M; Ratti, C; Abdel Aleem, E; Fattouh, F

    2017-01-01

    Tomato yellow leaf curl virus (TYLCV) infections of tomato crops in Egypt were widely spread in 2014. Infected symptomatic tomato plants from different governorates were sampled. TYLCV strains Israel and Mild (TYLCV-IL, TYLCV-Mild) were identified by multiplex and real-time PCR. In addition, nucleotide sequence analysis of the V1 and V2 protein genes, revealed ten TYLCV Egyptian isolates (TYLCV from TY1 to 10). Phylogenetic analysis showed their high degree of relatedness with TYLCV-IL Jordan isolate (98%). Here we have showed the complete nucleotide sequence of the TYLCV Egyptian isolate TY10, sampled from El Beheira. A high degree of similarity to other previously reported Egyptian isolates and isolates from Jordan and Japan reflect the importance of phylogenetic analysis in monitoring virus genetic diversity and possibilities for divergence of more virulent strains or genotypes.

  2. A new betasatellite associated with cotton leaf curl Burewala virus infecting tomato in India: influence on symptoms and viral accumulation.

    PubMed

    Kumar, Jitendra; Gunapati, Samatha; Singh, Sudhir P; Kumar, Abhinav; Lalit, Adarsh; Sharma, Naresh C; Puranik, Rekha; Tuli, Rakesh

    2013-06-01

    A begomovirus and its associated alpha- and betasatellite were detected in tomato plants affected with leaf curl disease. Based on a nucleotide sequence identity of 99 %, this begomovirus was designated an isolate of cotton leaf curl Burewala virus (CLCuBuV). The alphasatellite exhibited 93 % sequence identity to cotton leaf curl Burewala alphasatellite (CLCuBuA) and is hence referred to here as a variant of CLCuBuA. The detected betasatellite was recombinant in nature and showed 70 % sequence identity to the known betasatellites. Inoculation of healthy tomato with CLCuBuV plus betasatellite, either in the presence or the absence of alphasatellite, led to typical leaf curling, while inoculation with CLCuBuV in the absence of betasatellite resulted in mild symptoms. This confirmed the role of the betasatellite in expression of disease symptoms. We propose to name the newly detected betasatellite tomato leaf curl Hajipur betasatellite (ToLCHJB).

  3. Use of Tomato leaf curl virus (TYLCV) truncated Rep gene sequence to engineer TYLCV resistance in tomato plants.

    PubMed

    Ben Tamarzizt, H; Gharsallah Chouchane, S; Lengliz, R; Maxwell, D P; Marrakchi, M; Fakhfakh, H; Gorsane, F

    2009-01-01

    Tomato yellow leaf curl disease causes severe losses in tomato production throughout Mediterranean countries including Tunisia. In order to generate engineered resistance to this disease, an intron-hairpin RNA construct harboring a Tomato yellow leaf curl Sardinia virus (TYLCSV) truncated replication-associated protein (Rep) gene was used to transform genotype of tomato plants. Prepared transgenic plants were agro-inoculated with Tunisian infectious strain of TYLCSV and screened for the resistance to infection. The infected transgenic plants were divided into 3 different groups according to their specific symptoms. Only one of them contained transgenic plants fully resistant to the tomato yellow leaf curl disease.

  4. Molecular characterization and infectivity of Papaya leaf curl China virus infecting tomato in China.

    PubMed

    Zhang, Hui; Ma, Xin-ying; Qian, Ya-juan; Zhou, Xue-ping

    2010-02-01

    Papaya leaf curl China virus (PaLCuCNV) was previously reported as a distinct begomovirus infecting papaya in southern China. Based on molecular diagnostic survey, 13 PaLCuCNV isolates were obtained from tomato plants showing leaf curl symptoms in Henan and Guangxi Provinces of China. Complete nucleotide sequences of 5 representative isolates (AJ558116, AJ558117, AJ704604, FN256260, and FN297834) were determined to be 2738-2751 nucleotides, which share 91.7%-97.9% sequence identities with PaLCuCNV isolate G2 (AJ558123). DNA-beta was not found to be associated with PaLCuCNV isolates. To investigate the infectivity of PaLCuCNV, an infectious clone of PaLCuCNV-[CN:HeNZM1] was constructed and agro-inoculated into Nicotiana benthamiana, N. tabacum Samsun, N. glutinosa, Solanum lycopersicum and Petunia hybrida plants, which induced severe leaf curling and crinkling symptoms in these plants. Southern blot analysis and polymerase chain reaction (PCR) indicated a systemic infection of test plants by the agro-infectious clone.

  5. Rapid fluorescent reporter quantification by leaf disc analysis and its application in plant-virus studies

    PubMed Central

    2014-01-01

    Background Fluorescent proteins are extraordinary tools for biology studies due to their versatility; they are used extensively to improve comprehension of plant-microbe interactions. The viral infection process can easily be tracked and imaged in a plant with fluorescent protein-tagged viruses. In plants, fluorescent protein genes are among the most commonly used reporters in transient RNA silencing and heterologous protein expression assays. Fluorescence intensity is used to quantify fluorescent protein accumulation by image analysis or spectroscopy of protein extracts; however, these methods might not be suitable for medium- to large-scale comparisons. Results We report that laser scanners, used routinely in proteomic studies, are suitable for quantitative imaging of plant leaves that express different fluorescent protein pairs. We developed a microtiter plate fluorescence spectroscopy method for direct quantitative comparison of fluorescent protein accumulation in intact leaf discs. We used this technique to measure a fluorescent reporter in a transient RNA silencing suppression assay, and also to monitor early amplification dynamics of a fluorescent protein-labeled potyvirus. Conclusions Laser scanners allow dual-color fluorescence imaging of leaf samples, which might not be acquired in standard stereomicroscope devices. Fluorescence microtiter plate analysis of intact leaf discs can be used for rapid, accurate quantitative comparison of fluorescent protein accumulation. PMID:25053970

  6. Citrus leaf blotch virus invades meristematic regions in Nicotiana benthamiana and citrus.

    PubMed

    Agüero, Jesús; Vives, María Carmen; Velázquez, Karelia; Ruiz-Ruiz, Susana; Juárez, Jose; Navarro, Luis; Moreno, Pedro; Guerri, José

    2013-08-01

    To invade systemically host plants, viruses need to replicate in the infected cells, spread to neighbouring cells through plasmodesmata and move to distal parts of the plant via sieve tubes to start new infection foci. To monitor the infection of Nicotiana benthamiana plants by Citrus leaf blotch virus (CLBV), leaves were agroinoculated with an infectious cDNA clone of the CLBV genomic RNA expressing green fluorescent protein (GFP) under the transcriptional control of a duplicate promoter of the coat protein subgenomic RNA. Fluorescent spots first appeared in agroinfiltrated leaves 11-12 days after infiltration, indicating CLBV replication. Then, after entering the phloem vascular system, CLBV was unloaded in the upper parts of the plant and invaded all tissues, including flower organs and meristems. GFP fluorescence was not visible in citrus plants infected with CLBV-GFP. Therefore, to detect CLBV in meristematic regions, Mexican lime (Citrus aurantifolia) plants were graft inoculated with CLBV, with Citrus tristeza virus (CTV), a virus readily eliminated by shoot-tip grafting in vitro, or with both simultaneously. Although CLBV was detected by hybridization and real-time reverse transcription-polymerase chain reaction (RT-PCR) in 0.2-mm shoot tips in all CLBV-inoculated plants, CTV was not detected. These results explain the difficulty in eliminating CLBV by shoot-tip grafting in vitro. © 2013 BSPP AND JOHN WILEY & SONS LTD.

  7. The Involvement of Heat Shock Proteins in the Establishment of Tomato Yellow Leaf Curl Virus Infection

    PubMed Central

    Gorovits, Rena; Czosnek, Henryk

    2017-01-01

    Tomato yellow leaf curl virus (TYLCV), a begomovirus, induces protein aggregation in infected tomatoes and in its whitefly vector Bemisia tabaci. The interactions between TYLCV and HSP70 and HSP90 in plants and vectors are necessity for virus infection to proceed. In infected host cells, HSP70 and HSP90 are redistributed from a soluble to an aggregated state. These aggregates contain, together with viral DNA/proteins and virions, HSPs and components of the protein quality control system such as ubiquitin, 26S proteasome subunits, and the autophagy protein ATG8. TYLCV CP can form complexes with HSPs in tomato and whitefly. Nonetheless, HSP70 and HSP90 play different roles in the viral cell cycle in the plant host. In the infected host cell, HSP70, but not HSP90, participates in the translocation of CP from the cytoplasm into the nucleus. Viral amounts decrease when HSP70 is inhibited, but increase when HSP90 is downregulated. In the whitefly vector, HSP70 impairs the circulative transmission of TYLCV; its inhibition increases transmission. Hence, the efficiency of virus acquisition by whiteflies depends on the functionality of both plant chaperones and their cross-talk with other protein mechanisms controlling virus-induced aggregation. PMID:28360921

  8. Characterization of Cestrum yellow leaf curling virus: a new member of the family Caulimoviridae.

    PubMed

    Stavolone, Livia; Ragozzino, Antonio; Hohn, Thomas

    2003-12-01

    Cestrum yellow leaf curling virus (CmYLCV) has been characterized as the aetiological agent of the Cestrum parqui mosaic disease. The virus genome was cloned and the clone was proven to be infectious to C. parqui. The presence of typical viroplasms in virus-infected plant tissue and the information obtained from the complete genomic sequence confirmed CmYLCV as a member of the Caulimoviridae family. All characteristic domains conserved in plant pararetroviruses were found in CmYLCV. Its genome is 8253 bp long and contains seven open reading frames (ORFs). Phylogenetic analysis of the relationships with other members of the Caulimoviridae revealed that CmYLCV is closely related to the Soybean chlorotic mottle virus (SbCMV)-like genus and particularly to SbCMV. However, in contrast to the other members of this genus, the primer-binding site is located in the intercistronic region following ORF Ib rather than within this ORF, and an ORF corresponding to ORF VII is missing.

  9. Molecular phylogenetics and evolution of host plant use in the Neotropical rolled leaf 'hispine' beetle genus Cephaloleia (Chevrolat) (Chrysomelidae: Cassidinae).

    PubMed

    McKenna, Duane D; Farrell, Brian D

    2005-10-01

    Here, we report the results of a species level phylogenetic study of Cephaloleia beetles designed to clarify relationships and patterns of host plant taxon and tissue use among species. Our study is based on up to 2088bp of mtDNA sequence data. Maximum parsimony, maximum likelihood, and Bayesian methods of phylogenetic inference consistently recover a monophyletic Cephaloleia outside of a basal clade of primarily palm feeding species (the 'Arecaceae-feeding clade'), and C. irregularis. In all three analyses, the 'Arecaceae-feeding clade' includes Cephaloleia spp. with unusual morphological features, and a few species currently placed in other cassidine genera and tribes. All three analyses also recover a clade that includes all Zingiberales feeding Cephaloleia and most Cephaloleia species (the 'Zingiberales-feeding clade'). Two notable clades are found within the 'Zingiberales-feeding clade.' One is comprised of beetles that normally feed only on the young rolled leaves of plants in the families Heliconiaceae and Marantaceae (the 'Heliconiaceae & Marantaceae-feeding clade'). The other is comprised of relative host tissue generalist, primarily Zingiberales feeding species (the 'generalist-feeding clade'). A few species in the 'generalist-feeding clade' utilize Cyperaceae or Poaceae as hosts. Overall, relatively basal Cephaloleia (e.g., the 'Arecaceae clade') feed on relatively basal monocots (e.g., Cyclanthaceae and Arecaceae), and relatively derived Cephaloleia (e.g., the 'Zingiberales-feeding clade') feed on relatively derived monocots (mostly in the order Zingiberales). Zingiberales feeding and specialization on young rolled Zingiberales leaves have each apparently evolved just once in Cephaloleia.

  10. The begomoviruses Honeysuckle yellow vein mosaic virus and Tobacco leaf curl Japan virus with DNAbeta satellites cause yellow dwarf disease of tomato.

    PubMed

    Ogawa, T; Sharma, P; Ikegami, M

    2008-11-01

    The complete nucleotide sequences of two begomoviruses (Nara virus-1 and Nara virus-2), a satellite DNA (DNAbeta-Nara) and defective DNAs were obtained from honeysuckle (Lonicera japonica) showing characteristic yellow vein mosaic symptoms in Nara Prefecture, Japan. One begomovirus (Ibaraki virus) and a satellite DNA (DNAbeta-Ibaraki) was isolated and cloned from honeysuckle plants exhibited typical yellowing of veins and small elliptical shaped enations along veins on the under side of the leaves in Ibaraki Prefecture, Japan. The genome organization of the three viruses is the same as those of other Old World monopartite begomoviruses. Nara virus-1 had overall nucleotide sequence identity with Nara virus-2 of 94% and Ibaraki virus of 90%. DNAbeta-Nara had overall nucleotide sequence identity with DNAbeta-Ibaraki of 83%. Comparison of the nucleotide sequences with other begomoviruses revealed that Nara virus-1 and Nara virus-2 are strains of Honeysuckle yellow vein mosaic virus (HYVMV), hence named as HYVMV-Nara1 and HYVMV-Nara2, whereas Ibaraki virus was a strain of Tobacco leaf curl Japan virus (TbLCJV), designated as TbLCJV-Hs[Iba]. HYVMV-Nara1 and HYVMV-Nara2 have hybrid genomes, which are likely to have formed recombination between HYVMV and TbLCJV. TbLCJV-Hs[Iba] or HYVMV-Nara2 could infect and cause yellowing, leaf crinkling and stunting symptoms when partial tandem dimeric constructs were agroinoculated on tomato plants. However, in the presence of DNAbeta, both TbLCJV-Hs[Iba] or HYVMV-Nara2 produced more severe stunting symptoms in tomato plants. Therefore, these viruses along with their satellites are causal agents of tomato yellow dwarf disease in Japan, and honeysuckle acts as a potential reservoir host. Previously available evidence indicated that DNAbeta elements do not contain iteron sequences of their helper viruses; hence this is the first evidence that DNAbeta satellites have the iteron of their helper virus.

  11. Generation of transgenic papaya with double resistance to Papaya ringspot virus and Papaya leaf-distortion mosaic virus.

    PubMed

    Kung, Yi-Jung; Bau, Huey-Jiunn; Wu, Yi-Ling; Huang, Chiung-Huei; Chen, Tsui-Miao; Yeh, Shyi-Dong

    2009-11-01

    During the field tests of coat protein (CP)-transgenic papaya lines resistant to Papaya ringspot virus (PRSV), another Potyvirus sp., Papaya leaf-distortion mosaic virus (PLDMV), appeared as an emerging threat to the transgenic papaya. In this investigation, an untranslatable chimeric construct containing the truncated CP coding region of the PLDMV P-TW-WF isolate and the truncated CP coding region with the complete 3' untranslated region of PRSV YK isolate was transferred into papaya (Carica papaya cv. Thailand) via Agrobacterium-mediated transformation to generate transgenic plants with resistance to PLDMV and PRSV. Seventy-five transgenic lines were obtained and challenged with PRSV YK or PLDMV P-TW-WF by mechanical inoculation under greenhouse conditions. Thirty-eight transgenic lines showing no symptoms 1 month after inoculation were regarded as highly resistant lines. Southern and Northern analyses revealed that four weakly resistant lines have one or two inserts of the construct and accumulate detectable amounts of transgene transcript, whereas nine resistant lines contain two or three inserts without significant accumulation of transgene transcript. The results indicated that double virus resistance in transgenic lines resulted from double or more copies of the insert through the mechanism of RNA-mediated posttranscriptional gene silencing. Furthermore, three of nine resistant lines showed high levels of resistance to heterologous PRSV strains originating from Hawaii, Thailand, and Mexico. Our transgenic lines have great potential for controlling a number of PRSV strains and PLDMV in Taiwan and elsewhere.

  12. The nucleotide sequence and genomic organization of Citrus leaf blotch virus: candidate type species for a new virus genus.

    PubMed

    Vives, M C; Galipienso, L; Navarro, L; Moreno, P; Guerri, J

    2001-08-15

    The complete nucleotide sequence of Citrus leaf blotch virus (CLBV) was determined. CLBV genomic RNA (gRNA) has 8747 nt, excluding the 3'-terminal poly(A) tail, and contains three open reading frames (ORFs) and untranslated regions (UTR) of 73 and 541 nucleotides at the 5' and 3' termini, respectively. ORF1 potentially encodes a 227.4-kDa polypeptide, which has methyltransferase, papain-like protease, helicase, and RNA-dependent RNA polymerase motifs. ORF2 encodes a 40.2-kDa polypeptide containing a motif characteristic of cell-to-cell movement proteins. The 40.7-kDa polypeptide encoded by ORF3 was identified as the coat protein. The genome organization of CLBV resembles that of viruses in the genus Trichovirus, but they differ in various aspects: (i) in trichoviruses ORF2 overlaps ORFs 1 and 3, whereas in CLBV, ORFs 2 and 3 are separated and ORFs 1 and 2 overlap in one nucleotide; (ii) CLBV gRNA and CP are larger than those of trichoviruses; and (iii) the CLBV 3' UTR is larger than that of trichoviruses. Phylogenetic comparisons based on CP amino acid signatures clearly separates CLBV from trichoviruses. Also contrasting with trichoviruses, CLBV could not be transmitted to Chenopodium quinoa Willd. Considering these singularities, we propose that CLBV should be included in a new virus genus. Copyright 2001 Academic Press.

  13. Molecular analysis of new isolates of Tomato leaf curl Philippines virus and an associated betasatellite occurring in the Philippines.

    PubMed

    Sharma, Pradeep; Matsuda, N; Bajet, N B; Ikegami, M

    2011-02-01

    Three new begomovirus isolates and one betasatellite were obtained from a tomato plant exhibiting leaf curl symptom in Laguna, the Philippines. Typical begomovirus DNA components representing the three isolates (PH01, PH02 and PH03) were cloned, and their full-length sequences were determined to be 2754 to 2746 nucleotides. The genome organizations of these isolates were similar to those of other Old World monopartite begomoviruses. The sequence data indicated that PH01 and PH02 were variants of strain B of the species Tomato leaf curl Philippines virus, while PH03 was a variant of strain A of the species Tomato leaf curl Philippines virus. These isolates were designated ToLCPV-B[PH:Lag1:06], ToLCPV-B[PH:Lag2:06], and ToLCPV-A[PH:Lag3:06], respectively. Phylogenetic analysis revealed that the present isolates form a separate monophyletic cluster with indigenous begomoviruses reported earlier in the Philippines. A betasatellite isolated from same sample belongs to the betasatellite species Tomato leaf curl Philippines betasatellite and designated Tomato leaf curl Philippines betasatellite-[Philippines:Laguna1:2006], ToLCPHB-[PH:Lag1:06]. When co-inoculated with this betasatellite, tomato leaf curl Philippines virus induced severe symptoms in N. benthamiana and Solanum lycopersicum plants. Using a PVX-mediated transient assay, we found that the C4 and C2 proteins of tomato leaf curl Philippines virus and the βC1 protein of ToLCPHB-[PH:Lag1:06] function as a suppressor of RNA silencing.

  14. Melon chlorotic leaf curl virus: characterization and differential reassortment with closest relatives reveal adaptive virulence in the squash leaf curl virus clade and host shifting by the host-restricted bean calico mosaic virus.

    PubMed

    Idris, A M; Mills-Lujan, K; Martin, K; Brown, J K

    2008-02-01

    The genome components of the Melon chlorotic leaf curl virus (MCLCuV) were cloned from symptomatic cantaloupe leaves collected in Guatemala during 2002. The MCLCuV DNA-A and DNA-B components shared their closest nucleotide identities among begomoviruses, at approximately 90 and 81%, respectively, with a papaya isolate of MCLCuV from Costa Rica. The closest relatives at the species level were other members of the Squash leaf curl virus (SLCV) clade, which is endemic in the southwestern United States and Mexico. Biolistic inoculation of cantaloupe seedlings with the MCLCuV DNA-A and -B components resulted in the development of characteristic disease symptoms, providing definitive evidence of causality. MCLCuV experimentally infected species within the Cucurbitaceae, Fabaceae, and Solanaceae. The potential for interspecific reassortment was examined for MCLCuV and its closest relatives, including the bean-restricted Bean calico mosaic virus (BCaMV), and three other cucurbit-infecting species, Cucurbit leaf crumple virus (CuLCrV), SLCV, and SMLCV. The cucurbit viruses have distinct but overlapping host ranges. All possible reassortants were established using heterologous combinations of the DNA-A or DNA-B components. Surprisingly, only certain reassortants arising from MCLCuV and BCaMV, or MCLCuV and CuLCrV, were viable in bean, even though it is a host of all of the "wild-type" (parent) viruses. The bean-restricted BCaMV was differentially assisted in systemically infecting the cucurbit test species by the components of the four cucurbit-adapted begomoviruses. In certain heterologous combinations, the BCaMV DNA-A or -B component was able to infect one or more cucurbit species. Generally, the reassortants were less virulent in the test hosts than the respective wild-type (parent) viruses, strongly implicating adaptive modulation of virulence. This is the first illustration of reassortment resulting in the host range expansion of a host-restricted begomovirus.

  15. Molecular Evidence for Occurrence of Tomato leaf curl New Delhi virus in Ash Gourd (Benincasa hispida) Germplasm Showing a Severe Yellow Stunt Disease in India.

    PubMed

    Roy, Anirban; Spoorthi, P; Panwar, G; Bag, Manas Kumar; Prasad, T V; Kumar, Gunjeet; Gangopadhyay, K K; Dutta, M

    2013-06-01

    An evaluation of 70 accessions of ash gourd germplasm grown at National Bureau of Plant Genetic Resources, New Delhi, India during Kharif season (2010) showed natural occurrence of a yellow stunt disease in three accessions (IC554690, IC036330 and Pusa Ujjwal). A set of begomovirus specific primers used in PCR gave expected amplicon from all the symptomatic plants; however no betasatellite was detected. Complete genome of the begomovirus (DNA-A and DNA-B), amplified through rolling circle amplification, was cloned and sequenced. The begomovirus under study shared high sequence identities to different isolates of Tomato leaf curl New Delhi virus (ToLCNDV) and clustered with them. Among those isolates, the DNA-A and DNA-B of the present begomovirus isolate showed highest 99.6 and 96.8 % sequence identities, respectively with an isolate reported on pumpkin from India (DNA-A: AM286433, DNA-B: AM286435). Based on the sequence analysis, the begomovirus obtained from ash gourd was considered as an isolate of ToLCNDV. Thus, the present findings constitute the first report of occurrence of a new yellow stunt disease in ash gourd from India and demonstrated the association of ToLCNDV with the symptomatic samples. Occurrence of ToLCNDV in ash gourd germplasm not only adds up a new cucurbitaceous host of this virus but also raises the concern about the perpetuation of this virus in absence of its main host tomato and thus has an epidemiological relevance for understanding the rapid spread of this virus in tomato and other hosts in Indian sub-continent.

  16. The Complete Nucleotide Sequence and Biotype Variability of Papaya leaf distortion mosaic virus.

    PubMed

    Maoka, Tetsuo; Hataya, Tatsuji

    2005-02-01

    ABSTRACT The complete nucleotide sequence of the genome of Papaya leaf distortion mosaic virus (PLDMV) was determined. The viral RNA genome of strain LDM (leaf distortion mosaic) comprised 10,153 nucleotides, excluding the poly(A) tail, and contained one long open reading frame encoding a polyprotein of 3,269 amino acids (molecular weight 373,347). The polyprotein contained nine putative proteolytic cleavage sites and some motifs conserved in other potyviral polyproteins with 44 to 50% identities, indicating that PLDMV is a distinct species in the genus Potyvirus. Like the W biotype of Papaya ringspot virus (PRSV), the non-papaya-infecting biotype of PLDMV (PLDMV-C) was found in plants of the family Cucurbitaceae. The coat protein (CP) sequence of PLDMV-C in naturally infected-Trichosanthes bracteata was compared with those of three strains of the P biotype (PLDMV-P), LDM and two additional strains M (mosaic) and YM (yellow mosaic), which are biologically different from each other. The CP sequences of three strains of PLDMV-P share high identities of 95 to 97%, while they share lower identities of 88 to 89% with that of PLDMV-C. Significant changes in hydrophobicity and a deletion of two amino acids at the N-terminal region of the CP of PLDMV-C were observed. The finding of two biotypes of PLDMV implies the possibility that the papaya-infecting biotype evolved from the cucurbitaceae-infecting potyvirus, as has been previously suggested for PRSV. In addition, a similar evolutionary event acquiring infectivity to papaya may arise frequently in viruses in the family Cucurbitaceae.

  17. Rolling-circle replication of viroids, viroid-like satellite RNAs and hepatitis delta virus: variations on a theme.

    PubMed

    Flores, Ricardo; Grubb, Douglas; Elleuch, Amine; Nohales, María-Ángeles; Delgado, Sonia; Gago, Selma

    2011-01-01

    Viroids and viroid-like satellite RNAs from plants, and the human hepatitis delta virus (HDV) RNA share some properties that include small size, circularity and replication through a rolling-circle mechanism. Replication occurs in different cell compartments (nucleus, chloroplast and membrane-associated cytoplasmatic vesicles) and has three steps: RNA polymerization, cleavage and ligation. The first step generates oligomeric RNAs that result from the reiterative transcription of the circular templates of one or both polarities, and is catalyzed by either the RNA-dependent RNA polymerase of the helper virus on which viroid-like satellite RNAs are functionally dependent, or by host DNA-dependent RNA polymerases that, remarkably, viroids and HDV redirect to transcribe RNA templates. Cleavage is mediated by host enzymes in certain viroids and viroid-like satellite RNAs, while in others and in HDV is mediated by cis-acting ribozymes of three classes. Ligation appears to be catalyzed mainly by host enzymes. Replication most likely also involves many other non-catalytic proteins of host origin and, in HDV, the single virus-encoded protein.

  18. Detection of Papaya leaf distortion mosaic virus by reverse-transcription loop-mediated isothermal amplification.

    PubMed

    Shen, Wentao; Tuo, Decai; Yan, Pu; Li, Xiaoying; Zhou, Peng

    2014-01-01

    Papaya leaf distortion mosaic virus (PLDMV) can infect transgenic papaya resistant to a related pathogen, Papaya ringspot virus (PRSV), posing a substantial threat to papaya production in China. Current detection methods, however, are unable to be used for rapid detection in the field. Here, a reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of PLDMV, using a set of four RT-LAMP primers designed based on the conserved sequence of PLDMV CP. The RT-LAMP method detected specifically PLDMV and was highly sensitive, with a detection limit of 1.32×10(-6) μg of total RNA per reaction. Indeed, the reaction was 10 times more sensitive than one-step RT-PCR, while also requiring significantly less time and equipment. The effectiveness of RT-LAMP and one-step RT-PCR in detecting the virus were compared using 90 field samples of non-transgenic papaya and 90 field samples of commercialized PRSV-resistant transgenic papaya from Hainan Island. None of the non-transgenic papaya tested positive for PLDMV using either method. In contrast, 19 of the commercialized PRSV-resistant transgenic papaya samples tested positive by RT-LAMP assay, and 6 of those tested negative by RT-PCR. Therefore, the PLDMV-specific RT-LAMP is a simple, rapid, sensitive, and cost-effective tool in the field diagnosis and control of PLDMV.

  19. Tomato yellow leaf curl virus (TYLCV-IL): a seed-transmissible geminivirus in tomatoes.

    PubMed

    Kil, Eui-Joon; Kim, Sunhoo; Lee, Ye-Ji; Byun, Hee-Seong; Park, Jungho; Seo, Haneul; Kim, Chang-Seok; Shim, Jae-Kyoung; Lee, Jung-Hwan; Kim, Ji-Kwang; Lee, Kyeong-Yeoll; Choi, Hong-Soo; Lee, Sukchan

    2016-01-08

    Tomato yellow leaf curl virus (TYLCV) is one of the most well-known tomato-infecting begomoviruses and transmitted by Bemisia tabaci. Seed transmission has previously been reported for some RNA viruses, but TYLCV has not previously been described as a seed-borne virus. In 2013 and 2014, without whitefly-mediated transmission, TYLCV was detected in young tomato plants germinated from fallen fruits produced from TYLCV-infected tomato plants in the previous cultivation season. In addition, TYLCV-Israel (TYLCV-IL) was also detected in seeds and their seedlings of TYLCV-infected tomato plants that were infected by both viruliferous whitefly-mediated transmission and agro-inoculation. The seed infectivity was 20-100%, respectively, and the average transmission rate to seedlings was also 84.62% and 80.77%, respectively. TYLCV-tolerant tomatoes also produced TYLCV-infected seeds, but the amount of viral genome was less than seen in TYLCV-susceptible tomato plants. When tomato plants germinated from TYLCV-infected seeds, non-viruliferous whiteflies and healthy tomato plants were placed in an insect cage together, TYLCV was detected from whiteflies as well as receiver tomato plants six weeks later. Taken together, TYLCV-IL can be transmitted via seeds, and tomato plants germinated from TYLCV-infected seeds can be an inoculum source of TYLCV. This is the first report about TYLCV seed transmission in tomato.

  20. Method: low-cost delivery of the cotton leaf crumple virus-induced gene silencing system

    PubMed Central

    2012-01-01

    Background We previously developed a virus-induced gene silencing (VIGS) vector for cotton from the bipartite geminivirusCotton leaf crumple virus (CLCrV). The original CLCrV VIGS vector was designed for biolistic delivery by a gene gun. This prerequisite limited the use of the system to labs with access to biolistic equipment. Here we describe the adaptation of this system for delivery by Agrobacterium (Agrobacterium tumefaciens). We also describe the construction of two low-cost particle inflow guns. Results The biolistic CLCrV vector was transferred into two Agrobacterium binary plasmids. Agroinoculation of the binary plasmids into cotton resulted in silencing and GFP expression comparable to the biolistic vector. Two homemade low-cost gene guns were used to successfully inoculate cotton (G. hirsutum) and N. benthamiana with either the CLCrV VIGS vector or the Tomato golden mosaic virus (TGMV) VIGS vector respectively. Conclusions These innovations extend the versatility of CLCrV-based VIGS for analyzing gene function in cotton. The two low-cost gene guns make VIGS experiments affordable for both research and teaching labs by providing a working alternative to expensive commercial gene guns. PMID:22853641

  1. Female-Biased Symbionts and Tomato Yellow Leaf Curl Virus Infections in Bemisia tabaci

    PubMed Central

    Guo, Huifang; Qu, Yufeng; Liu, Xiangdong; Zhong, Wanfang; Fang, Jichao

    2014-01-01

    The female-biased infection of facultative symbionts has been found in Bemisia tabaci; however, whether there are any differences in tomato yellow leaf curl virus (TYLCV) and obligate symbiont infection rates between females and males is unknown. Determining whether such differences exist would be very important for understanding the spread of the plant virus and of the symbionts. We compared both symbiont infection types, including obligate and facultative symbionts, and the rates of TYLCV infection in both sexes in five field populations from Jiangsu Province, China. The obligate symbiont Portiera aleyrodidarum was not found in every whitefly tested. In all tested populations, more females than males were found to harbor P. aleyrodidarum; and more females than males also harbored Hamiltonella defense, the most common facultative symbiont as well as Cardinium. In addition to female-biased symbiont infections, there were also female-biased TYLCV infections, and the infection frequencies of this plant virus in females were higher than those in males. Taken together, these results suggested that both the female-biased symbiont infections and female-biased TYLCV infections promoted the rapid spread of TYLCV in China. PMID:24465416

  2. Association of an Alphasatellite with Tomato Yellow Leaf Curl Virus and Ageratum Yellow Vein Virus in Japan is Suggestive of a Recent Introduction

    PubMed Central

    Shahid, Muhammad Shafiq; Ikegami, Masato; Waheed, Abdul; Briddon, Rob W.; Natsuaki, Keiko T.

    2014-01-01

    Samples were collected in 2011 from tomato plants exhibiting typical tomato leaf curl disease symptoms in the vicinity of Komae, Japan. PCR mediated amplification, cloning and sequencing of all begomovirus components from two plants from different fields showed the plants to be infected by Tomato yellow leaf curl virus (TYLCV) and Ageratum yellow vein virus (AYVV). Both viruses have previously been shown to be present in Japan, although this is the first identification of AYVV on mainland Japan; the virus previously having been shown to be present on the Okinawa Islands. The plant harboring AYVV was also shown to contain the betasatellite Tomato leaf curl Java betasatellite (ToLCJaB), a satellite not previously shown to be present in Japan. No betasatellite was associated with the TYLCV infected tomato plants analyzed here, consistent with earlier findings for this virus in Japan. Surprisingly both plants were also found to harbor an alphasatellite; no alphasatellites having previously been reported from Japan. The alphasatellite associated with both viruses was shown to be Sida yellow vein China alphasatellite which has previously only been identified in the Yunnan Province of China and Nepal. The results suggest that further begomoviruses, and their associated satellites, are being introduced to Japan. The significance of these findings is discussed. PMID:24424499

  3. Association of an alphasatellite with tomato yellow leaf curl virus and ageratum yellow vein virus in Japan is suggestive of a recent introduction.

    PubMed

    Shahid, Muhammad Shafiq; Ikegami, Masato; Waheed, Abdul; Briddon, Rob W; Natsuaki, Keiko T

    2014-01-14

    Samples were collected in 2011 from tomato plants exhibiting typical tomato leaf curl disease symptoms in the vicinity of Komae, Japan. PCR mediated amplification, cloning and sequencing of all begomovirus components from two plants from different fields showed the plants to be infected by Tomato yellow leaf curl virus (TYLCV) and Ageratum yellow vein virus (AYVV). Both viruses have previously been shown to be present in Japan, although this is the first identification of AYVV on mainland Japan; the virus previously having been shown to be present on the Okinawa Islands. The plant harboring AYVV was also shown to contain the betasatellite Tomato leaf curl Java betasatellite (ToLCJaB), a satellite not previously shown to be present in Japan. No betasatellite was associated with the TYLCV infected tomato plants analyzed here, consistent with earlier findings for this virus in Japan. Surprisingly both plants were also found to harbor an alphasatellite; no alphasatellites having previously been reported from Japan. The alphasatellite associated with both viruses was shown to be Sida yellow vein China alphasatellite which has previously only been identified in the Yunnan Province of China and Nepal. The results suggest that further begomoviruses, and their associated satellites, are being introduced to Japan. The significance of these findings is discussed.

  4. Leaf rolling and stem fasciation in grass pea (Lathyrus sativus L.) mutant are mediated through glutathione-dependent cellular and metabolic changes and associated with a metabolic diversion through cysteine during phenotypic reversal.

    PubMed

    Talukdar, Dibyendu; Talukdar, Tulika

    2014-01-01

    A Lathyrus sativus L. mutant isolated in ethylmethane sulfonate-treated M2 progeny of mother variety BioL-212 and designated as rlfL-1 was characterized by inwardly rolled-leaf and stem and bud fasciations. The mutant exhibited karyomorphological peculiarities in both mitosis and meiosis with origin of aneuploidy. The mitosis was vigorous with high frequency of divisional cells and their quick turnover presumably steered cell proliferations. Significant transcriptional upregulations of cysteine and glutathione synthesis and concomitant stimulations of glutathione-mediated antioxidant defense helped rlfL-1 mutant to maintain balanced reactive oxygen species (ROS) metabolisms, as deduced by ROS-imaging study. Glutathione synthesis was shut down in buthionine sulfoximine- (BSO-) treated mother plant and mutant, and leaf-rolling and stems/buds fasciations in the mutant were reversed, accompanied by normalization of mitotic cell division process. Antioxidant defense was downregulated under low glutathione-redox but cysteine-desulfurations and photorespiratory glycolate oxidase transcripts were markedly overexpressed, preventing cysteine overaccumulation but resulted in excess H2O2 in BSO-treated mutant. This led to oxidative damage in proliferating cells, manifested by severe necrosis in rolled-leaf and fasciated stems. Results indicated vital role of glutathione in maintaining abnormal proliferations in plant organs, and its deficiency triggered phenotypic reversal through metabolic diversions of cysteine and concomitant cellular and metabolic modulations.

  5. Leaf Rolling and Stem Fasciation in Grass Pea (Lathyrus sativus L.) Mutant Are Mediated through Glutathione-Dependent Cellular and Metabolic Changes and Associated with a Metabolic Diversion through Cysteine during Phenotypic Reversal

    PubMed Central

    Talukdar, Dibyendu; Talukdar, Tulika

    2014-01-01

    A Lathyrus sativus L. mutant isolated in ethylmethane sulfonate-treated M2 progeny of mother variety BioL-212 and designated as rlfL-1 was characterized by inwardly rolled-leaf and stem and bud fasciations. The mutant exhibited karyomorphological peculiarities in both mitosis and meiosis with origin of aneuploidy. The mitosis was vigorous with high frequency of divisional cells and their quick turnover presumably steered cell proliferations. Significant transcriptional upregulations of cysteine and glutathione synthesis and concomitant stimulations of glutathione-mediated antioxidant defense helped rlfL-1 mutant to maintain balanced reactive oxygen species (ROS) metabolisms, as deduced by ROS-imaging study. Glutathione synthesis was shut down in buthionine sulfoximine- (BSO-) treated mother plant and mutant, and leaf-rolling and stems/buds fasciations in the mutant were reversed, accompanied by normalization of mitotic cell division process. Antioxidant defense was downregulated under low glutathione-redox but cysteine-desulfurations and photorespiratory glycolate oxidase transcripts were markedly overexpressed, preventing cysteine overaccumulation but resulted in excess H2O2 in BSO-treated mutant. This led to oxidative damage in proliferating cells, manifested by severe necrosis in rolled-leaf and fasciated stems. Results indicated vital role of glutathione in maintaining abnormal proliferations in plant organs, and its deficiency triggered phenotypic reversal through metabolic diversions of cysteine and concomitant cellular and metabolic modulations. PMID:24987684

  6. Genetic dissection of novel QTLs for resistance to leaf spots and Tomato spotted wilt virus in peanut (Arachis hypogaea L.)

    USDA-ARS?s Scientific Manuscript database

    Peanut is an import crop, economically and nutritiously, but high production cost is a serious challenge to peanut farmers as exemplified by chemical spray to control foliar diseases such as leaf spots and thrips, the vectors of tomato spotted wilt virus (TSWV). The objective of this research was to...

  7. Comparing the natural spread of Sugarcane yellow leaf virus in Florida to its natural spread in Louisiana

    USDA-ARS?s Scientific Manuscript database

    Among reports sugarcane infected with Sugarcane yellow leaf virus (SCYLV), the incidence of infections has been higher in Florida than in Louisiana. An experiment was planted in 2003 at two USDA, ARS sugarcane research locations, the Sugarcane Field Station, Canal Point, Florida and the Sugarcane R...

  8. One-step reverse transcription loop mediated isothermal amplification assay for detection of Apple chlorotic leaf spot virus

    USDA-ARS?s Scientific Manuscript database

    A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the detection of Apple chlorotic leaf spot virus (ACLSV) was developed. In this method, a set of four primers was designed based on the conserved regions in the coat protein gene of ACLSV, and was synthesized for the ...

  9. Identification of differentially expressed genes in tomato associated with R-lines Ty-5 against tomato yellow leaf curl virus (TYLCV) and Sw-7 against tomato spotted wilt virus (TSWV)

    USDA-ARS?s Scientific Manuscript database

    Tomato yellow leaf curl virus (TYLCV) and tomato spotted wilt virus (TSWV) are two most economically important viruses affecting tomato productions worldwide. Developing a cultivar with resistance to these viruses would be the most effective and economical means of disease management. Comparative ...

  10. Implications Of Host Plant Resistance Against Whitefly-Transmitted Tomato Yellow Leaf Curl Virus In Tomato For Virus Epidemics And Management

    USDA-ARS?s Scientific Manuscript database

    Whitefly-transmitted Tomato yellow leaf curl virus (TYLCV) severely impacts tomato production in southeastern USA. Growers typically spray insecticides against whiteflies and plant TYLCV-resistant genotypes. Semi-dominant genes such as TY-1 and TY-2 confer resistance to TYLCV. Resistant genotypes ar...

  11. Conservation of major and minor jelly-roll capsid proteins in Polinton (Maverick) transposons suggests that they are bona fide viruses

    PubMed Central

    2014-01-01

    Reviewers This article was reviewed by Lakshminarayan M. Iyer and I. King Jordan. For complete reviews, see the Reviewers’ Reports section. Polintons (also known as Mavericks) and Tlr elements of Tetrahymena thermophila represent two families of large DNA transposons widespread in eukaryotes. Here, we show that both Polintons and Tlr elements encode two key virion proteins, the major capsid protein with the double jelly-roll fold and the minor capsid protein, known as the penton, with the single jelly-roll topology. This observation along with the previously noted conservation of the genes for viral genome packaging ATPase and adenovirus-like protease strongly suggests that Polintons and Tlr elements combine features of bona fide viruses and transposons. We propose the name ‘Polintoviruses’ to denote these putative viruses that could have played a central role in the evolution of several groups of DNA viruses of eukaryotes. PMID:24773695

  12. Conservation of major and minor jelly-roll capsid proteins in Polinton (Maverick) transposons suggests that they are bona fide viruses.

    PubMed

    Krupovic, Mart; Bamford, Dennis H; Koonin, Eugene V

    2014-04-29

    Polintons (also known as Mavericks) and Tlr elements of Tetrahymena thermophila represent two families of large DNA transposons widespread in eukaryotes. Here, we show that both Polintons and Tlr elements encode two key virion proteins, the major capsid protein with the double jelly-roll fold and the minor capsid protein, known as the penton, with the single jelly-roll topology. This observation along with the previously noted conservation of the genes for viral genome packaging ATPase and adenovirus-like protease strongly suggests that Polintons and Tlr elements combine features of bona fide viruses and transposons. We propose the name 'Polintoviruses' to denote these putative viruses that could have played a central role in the evolution of several groups of DNA viruses of eukaryotes.

  13. Tomato leaf curl New Delhi virus: a widespread bipartite begomovirus in the territory of monopartite begomoviruses.

    PubMed

    Zaidi, Syed Shan-E-Ali; Martin, Darren P; Amin, Imran; Farooq, Muhammad; Mansoor, Shahid

    2016-08-23

    Tomato leaf curl New Delhi virus (ToLCNDV) is an exceptional Old World bipartite begomovirus. On the Indian subcontinent, a region in which monopartite DNA satellite-associated begomoviruses with mostly narrow geographical ranges predominate, it is widespread, with a geographical range also including the Far East, Middle East, North Africa and Europe. The success of ToLCNDV probably derives from its broad host range and highly flexible genomic configuration: its DNA-A component is capable of productively interacting with, and trans-replicating, diverse DNA-B components and betasatellites. An understanding of the capacity of ToLCNDV to infect a variety of hosts and spread across a broad and ecologically variable geographical range could illuminate the potential economic threats associated with similar begomoviral invasions. Towards this end, we used available ToLCNDV sequences to reconstruct the history of ToLCNDV spread.

  14. Lamium amplexicaule (Lamiaceae): a weed reservoir for tomato yellow leaf curl virus (TYLCV) in Korea.

    PubMed

    Kil, Eui-Joon; Park, Jungan; Lee, Hyejung; Kim, Jaedeok; Choi, Hong-Soo; Lee, Kyeong-Yeoll; Kim, Chang-Seok; Lee, Sukchan

    2014-06-01

    After the first identification of tomato yellow leaf curl virus (TYLCV) in the southern part of Korea in 2008, TYLCV has rapidly spread to tomato farms in most regions of Korea. From 2008 to 2010, a survey of natural weed hosts that could be reservoirs of TYLCV was performed in major tomato production areas of Korea. About 530 samples were collected and identified as belonging to 25 species from 11 families. PCR and Southern hybridization were used to detect TYLCV in samples, and replicating forms of TYLCV DNA were detected in three species (Achyranthes bidentata, Lamium amplexicaule, and Veronica persica) by Southern hybridization. TYLCV transmission mediated by Bemisia tabaci from TYLCV-infected tomato plants to L. amplexicaule was confirmed, and TYLCV-infected L. amplexicaule showed symptoms such as yellowing, stunting, and leaf curling. TYLCV from infected L. amplexicaule was also transmitted to healthy tomato and L. amplexicaule plants by B. tabaci. The rate of infection of L. amplexicaule by TYLCV was similar to that of tomato. This report is the first to show that L. amplexicaule is a reservoir weed host for TYLCV.

  15. Genetic diversity, host range, and distribution of tomato yellow leaf curl virus in Iran.

    PubMed

    Shirazi, M; Mozafari, J; Rakhshandehroo, F; Shams-Bakhsh, M

    2014-01-01

    Tomato yellow leaf curl virus (TYLCV) is considered one of the most important tomato pathogens in tropical and subtropical regions including Iran. During the years 2007 to 2009, a total number of 510 symptomatic and asymptomatic vegetable, ornamental and weed samples were collected from fields and greenhouses in ten provinces of Iran. Symptoms included stunting, yellowing, leaf curl and flower senescence. PCR with specific primers showed TYLCV infection in 184 samples (36%) such as cucumber, pepper, tomato and several weeds from seven provinces. Based on the geographical origin, host range and symptoms, twenty three representative isolates were selected for phylogenetic analysis. An amplicon with a size about 608 base pair (bp) comprising partial sequence of the coat (CP) and movement protein (MP) coding regions of the viral genome was sequenced and compared with the corresponding selected sequences available in GenBank for Iran and worldwide. Phylogenetic analyses on the basis of the nucleotide sequences indicated two geographically separated clades. Isolates collected from Hormozgan, Khuzestan and Kerman provinces were grouped together with other Iranian isolates including TYLCV-Ir2, TYLCV-Kahnooj, and an isolate from Oman. It was also revealed that isolates collected from Boushehr, Fars, Tehran, and Isfahan placed close to the Iranian isolate TYLCV-Abadeh and isolates from Israel and Egypt. No correlation was found between the genetic variation and the host species, but selected Iranian isolates were grouped on the basis of the geographical origins. Results of this study indicated a high genetic diversity among Iranian TYLCV isolates.

  16. Recessive Resistance Derived from Tomato cv. Tyking-Limits Drastically the Spread of Tomato Yellow Leaf Curl Virus

    PubMed Central

    Pereira-Carvalho, Rita C.; Díaz-Pendón, Juan A.; Fonseca, Maria Esther N.; Boiteux, Leonardo S.; Fernández-Muñoz, Rafael; Moriones, Enrique; Resende, Renato O.

    2015-01-01

    The tomato yellow leaf curl disease (TYLCD) causes severe damage to tomato (Solanum lycopersicum L.) crops throughout tropical and subtropical regions of the world. TYLCD is associated with a complex of single-stranded circular DNA plant viruses of the genus Begomovirus (family Geminiviridae) transmitted by the whitefy Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae). The tomato inbred line TX 468-RG is a source of monogenic recessive resistance to begomoviruses derived from the hybrid cv. Tyking F1. A detailed analysis of this germplasm source against tomato yellow leaf curl virus-Israel (TYLCV-IL), a widespread TYLCD-associated virus, showed a significant restriction to systemic virus accumulation even under continuous virus supply. The resistance was effective in limiting the onset of TYLCV-IL in tomato, as significantly lower primary spread of the virus occurred in resistant plants. Also, even if a limited number of resistant plants could result infected, they were less efficient virus sources for secondary spread owing to the impaired TYLCV-IL accumulation. Therefore, the incorporation of this resistance into breeding programs might help TYLCD management by drastically limiting TYLCV-IL spread. PMID:26008699

  17. A sensitive and specific hyperbranched rolling circle amplification assay and test strip for white spot syndrome virus.

    PubMed

    Zhao, Yu-Ran; Yin, Wei-Li; Yue, Zhi-Qin; Li, Ba-Fang

    2014-01-01

    White spot syndrome virus (WSSV) is a global threat to the prawn industry, and there is no simple method for field-based testing of this virus. We designed a padlock probe and primers to the capsid protein gene VP28 of WSSV, and established a hyperbranched rolling circle amplification (HRCA) assay and a corresponding strip-based test. The assay and the test strip both had similar high accuracy and specificity, and their sensitivity was about 10 copies/μL, which is 100 times higher than conventional PCR. In this study, 68 batches of prawns were tested for WSSV with the HRCA assay and test strip, and the results were compared with the PCR assay. The results indicated that both the assay and test strip had accuracy similar to each other and to the PCR results. However, the assay and strip were more sensitive and user-friendly than PCR. Establishment of this method will provide a rapid detection of WSSV and also a basis for field-based detection of animal disease.

  18. Molecular characterization and an infectious clone construction of sweet potato leaf curl virus (SPLCV) isolated from Korea.

    PubMed

    Choi, E; Lee, G; Park, J; Lee, T K; Choi, H S; Lee, S

    2012-01-01

    Sweet potato leaf curl disease (SPLCD) was primarily identified in sweet potato fields in Korea in 2003, and the complete genomic sequence of sweet potato leaf curl virus (SPLCV) has been cloned. The genome of the Korean SPLCV isolate (SPLCV-KR) comprises 2,828 nucleotides with six open reading frames in DNA-A, similar to a monopartite begomovirus. Additionally, neither the genome B genomic component nor the DNA beta sequence was detected. The results of phylogenetic analysis using the maximum parsimony method showed that SPLCV-KR is more closely related to SPLCV-US (US) than SPLCV-CN (China) and SPLCV-JP (Japan). A tandem repeat dimer of SPLCV-KR was cloned and found to be infectious in sweet potatoes (Ipomoea batatas) via biolistic inoculation. The SPLCV-infected sweet potatoes exhibited mild leaf curl symptoms of SPLCD, and the newly-replicated viral DNA was detected via Southern blot analysis. Results of biotic, molecular, and phylogenetic characterization suggest that SPLCV-KR is a new strain of SPLCV and is importantly placed in the evolutionary progression from curtoviruses to begomoviruses. sweet potato leaf curl virus; sweet potato leaf curl disease; phylogenetic analysis; infectious clone; biolistic infection.

  19. The role of corchorus in spreading of tomato yellow leaf curl virus on tomato in Jeddah, Saudi Arabia.

    PubMed

    Sohrab, Sayed Sartaj

    2016-03-01

    Corchorus (Corchorus capsularis L. and Corchorus olitorius L.) is one of the most important fiber crops grown in tropical and subtropical regions throughout the world. Field survey was conducted and naturally infected leaf samples were collected from corchorus and tomato plants in Jeddah, Saudi Arabia. The causal virus was transmitted by whiteflies to tomato plants and begomovirus infection was confirmed by Polymerase chain reaction. The complete viral genome and associated betasatellites were amplified, cloned and sequenced from both corchorus and tomato samples. The genetic variability and phylogenetic relationships were determined for both isolates (corchorus and tomato). The complete genome sequences showed highest (99.5 % nt) similarity with tomato yellow leaf curl virus (TYLCV) and formed closest cluster with TYLCV-Tomato reported from Jizan and Al-Qasim, Saudi Arabia and betasatellites sequences showed highest similarity (99.8 % nt) with Tomato yellow leaf curl betasatellites-Jeddah followed by Tomato yellow leaf curl Oman betasatellites and formed closed cluster with TYLCV-Tomato. On the basis of results obtained from whiteflies transmission, sequence similarity and phylogenetic relationships; it is concluded that the identified virus could be a variant of TYLCV circulating in the Kingdom. The significance of this study demonstrated that the corchorus is serving as reservoir and alternative host and playing an important role in spreading the begomovirus associated disease in the Kingdom of Saudi Arabia.

  20. Assessment of the genetic diversity of tomato yellow leaf curl virus.

    PubMed

    Wan, H J; Yuan, W; Wang, R Q; Ye, Q J; Ruan, M Y; Li, Z M; Zhou, G Z; Yao, Z P; Yang, Y J

    2015-01-26

    The objective of the present study was to analyze the genetic diversity of tomato yellow leaf curl virus (TYLCV). Representative TYLCV sequences were searched in the National Center for Biotechnology Information database. Comprehensive analysis of TYLCV was performed using bioinformatics by examining gene structure, sequence alignments, phylogeny, GC content, and homology. Forty-eight representative TYLCV sequences were selected from 48 regions in 29 countries. The results showed that all TYLCV sequences were 2752-2794 nucleotides in length, which encoded 6 open reading frames (AV1, AV2, AC1, AC2, AC3, and AC4). GC content ranged from 0.41-0.42. Sequence alignment showed a number of insertions and deletions within these TYLCV sequences. Phylogenetic tree results revealed that the sequences were divided into 10 classes; homology of the sequences ranged from 72.8 to 98.6%. All 48 sequences contained the typical structure of TYLCV, including open reading frames and intergenic regions. These results provide a theoretical basis for the identification and evolution of the virus in the future.

  1. A new virus in Luteoviridae is associated with raspberry leaf curl disease

    USDA-ARS?s Scientific Manuscript database

    To determine the etiology of Raspberry Leaf Curl Disease (RLCD), which causes leaf curling, leaf distortion, leaf chlorosis, shoot dwarfing, shoot proliferation in raspberries and can kill plants within three years, a next generation sequences approach was applied. Two red raspberry plants collected...

  2. Effects of the mutation of selected genes of cotton leaf curl Kokhran virus on infectivity, symptoms and the maintenance of cotton leaf curl Multan betasatellite.

    PubMed

    Iqbal, Zafar; Sattar, M Naeem; Kvarnheden, Anders; Mansoor, Shahid; Briddon, Rob W

    2012-10-01

    Cotton leaf curl Kokhran virus (CLCuKoV) is a cotton-infecting monopartite begomovirus (family Geminiviridae). The effects of mutation of the coat protein (CP), V2, C2 and C4 genes of CLCuKoV on infectivity and symptoms in Nicotiana benthamiana were investigated. Each mutation introduced a premature stop codon which would lead to premature termination of translation of the gene. Mutation of the CP gene abolished infectivity. However, transient expression of the CLCuKoV CP gene under the control of the Cauliflower mosaic virus 35S promoter (35S-Ko(CP)), at the point of inoculation, led to a small number of plants in which viral DNA could be detected by PCR in tissues distal to the inoculation site. Mutations of the V2, C2 and C4 genes reduced infectivity. The V2 and C2 mutants did not induce symptoms, whereas the C4 mutation was associated with attenuated symptoms. Infections of plants with the C4 mutant were associated with viral DNA levels equivalent to the wild-type virus, whereas viral DNA levels for the V2 mutant were low, detectable by Southern blot hybridisation, and for the C2 mutant were detectable only by PCR. Significantly, transient expression of the CLCuKoV C2 gene at the point of inoculation, raised virus DNA levels in tissues distal to the inoculation site such that they could be detected by Southern hybridisation, although they remained at well below the levels seen for the wild-type virus, but reduced the infectivity of the virus. These findings are consistent with earlier mutation studies of monopartite begomoviruses and our present knowledge concerning the functions of the four genes suggesting that the CP is essential for long distance spread of the virus in plants, the C4 is involved in modulating symptoms, the C2 interferes with host defence and the V2 is involved in virus movement. The results also suggest that the V2, C2 and C4 may be pathogenicity determinants. Additionally the effects of the mutations of CLCuKoV genes on infections of the

  3. Sweet pepper confirmed as a reservoir host for tomato yellow leaf curl virus by both agro-inoculation and whitefly-mediated inoculation.

    PubMed

    Kil, Eui-Joon; Byun, Hee-Seong; Kim, Sunhoo; Kim, Jaedeok; Park, Jungan; Cho, Seungchan; Yang, Dong-Cheol; Lee, Kyeong-Yeoll; Choi, Hong-Soo; Kim, Ji-Kwang; Lee, Sukchan

    2014-09-01

    Tomato yellow leaf curl virus (TYLCV), a member of the genus Begomovirus, has a single-stranded DNA genome. TYLCV can induce severe disease symptoms on tomato plants, but other hosts plants such as cucurbits and peppers are asymptomatic. A full-length DNA clone of a Korean TYLCV isolate was constructed by rolling-circle amplification from TYLCV-infected tomatoes in Korea. To assess relative susceptibility of sweet pepper varieties to TYLCV, 19 cultivars were inoculated with cloned TYLCV by agro-inoculation. All TYLCV-infected sweet peppers were asymptomatic, even though Southern hybridization and polymerase chain reaction analysis showed TYLCV genomic DNA accumulation in roots, stems, and newly produced shoots. Southern hybridization indicated that TYLCV replicated and moved systemically from agro-inoculated apical shoot tips to roots or newly produced shoots of sweet peppers. Whitefly-mediated inoculation experiments showed that TYLCV can be transmitted to tomatoes from TYLCV-infected sweet peppers. Taken together, these results indicate that sweet pepper can be a reservoir for TYLCV in nature.

  4. Simultaneous detection of papaya ringspot virus, papaya leaf distortion mosaic virus, and papaya mosaic virus by multiplex real-time reverse transcription PCR.

    PubMed

    Huo, P; Shen, W T; Yan, P; Tuo, D C; Li, X Y; Zhou, P

    2015-12-01

    Both the single infection of papaya ringspot virus (PRSV), papaya leaf distortion mosaic virus (PLDMV) or papaya mosaic virus (PapMV) and double infection of PRSV and PLDMV or PapMV which cause indistinguishable symptoms, threaten the papaya industry in Hainan Island, China. In this study, a multiplex real-time reverse transcription PCR (RT-PCR) was developed to detect simultaneously the three viruses based on their distinctive melting temperatures (Tms): 81.0±0.8°C for PRSV, 84.7±0.6°C for PLDMV, and 88.7±0.4°C for PapMV. The multiplex real-time RT-PCR method was specific and sensitive in detecting the three viruses, with a detection limit of 1.0×10(1), 1.0×10(2), and 1.0×10(2) copies for PRSV, PLDMV, and PapMV, respectively. Indeed, the reaction was 100 times more sensitive than the multiplex RT-PCR for PRSV, and 10 times more sensitive than multiplex RT-PCR for PLDMV. Field application of the multiplex real-time RT-PCR demonstrated that some non-symptomatic samples were positive for PLDMV by multiplex real-time RT-PCR but negative by multiplex RT-PCR, whereas some samples were positive for both PRSV and PLDMV by multiplex real-time RT-PCR assay but only positive for PLDMV by multiplex RT-PCR. Therefore, this multiplex real-time RT-PCR assay provides a more rapid, sensitive and reliable method for simultaneous detection of PRSV, PLDMV, PapMV and their mixed infections in papaya.

  5. Frequent occurrence of tomato leaf curl New Delhi virus in cotton leaf curl disease affected cotton in Pakistan

    USDA-ARS?s Scientific Manuscript database

    Cotton leaf curl disease (CLCuD) in the Indian subcontinent is associated with several distinct monopartite begomoviruses and DNA satellites. However, only a single begomovirus was associated with breakdown of resistance against CLCuD in previously resistant cotton varieties. The monopartite begomov...

  6. Narrow Bottlenecks Affect Pea Seedborne Mosaic Virus Populations during Vertical Seed Transmission but not during Leaf Colonization

    PubMed Central

    Johansen, Elisabeth Ida; Simon, Vincent; Jacquemond, Mireille; Senoussi, Rachid

    2014-01-01

    The effective size of populations (Ne) determines whether selection or genetic drift is the predominant force shaping their genetic structure and evolution. Populations having high Ne adapt faster, as selection acts more intensely, than populations having low Ne, where random effects of genetic drift dominate. Estimating Ne for various steps of plant virus life cycle has been the focus of several studies in the last decade, but no estimates are available for the vertical transmission of plant viruses, although virus seed transmission is economically significant in at least 18% of plant viruses in at least one plant species. Here we study the co-dynamics of two variants of Pea seedborne mosaic virus (PSbMV) colonizing leaves of pea plants (Pisum sativum L.) during the whole flowering period, and their subsequent transmission to plant progeny through seeds. Whereas classical estimators of Ne could be used for leaf infection at the systemic level, as virus variants were equally competitive, dedicated stochastic models were needed to estimate Ne during vertical transmission. Very little genetic drift was observed during the infection of apical leaves, with Ne values ranging from 59 to 216. In contrast, a very drastic genetic drift was observed during vertical transmission, with an average number of infectious virus particles contributing to the infection of a seedling from an infected mother plant close to one. A simple model of vertical transmission, assuming a cumulative action of virus infectious particles and a virus density threshold required for vertical transmission to occur fitted the experimental data very satisfactorily. This study reveals that vertically-transmitted viruses endure bottlenecks as narrow as those imposed by horizontal transmission. These bottlenecks are likely to slow down virus adaptation and could decrease virus fitness and virulence. PMID:24415934

  7. Complete nucleotide sequences of okra isolates of Cotton leaf curl Gezira virus and their associated DNA-beta from Niger.

    PubMed

    Shih, S L; Kumar, S; Tsai, W S; Lee, L M; Green, S K

    2009-01-01

    Okra (Abelmoschus esculentus) is a major crop in Niger. In the fall of 2007, okra leaf curl disease was observed in Niger and the begomovirus and DNA-beta satellite were found associated with the disease. The complete nucleotide sequences of DNA-A (FJ469626 and FJ469627) and associated DNA-beta satellites (FJ469628 and FJ469629) were determined from two samples. This is the first report of molecular characterization of okra-infecting begomovirus and their associated DNA-beta from Niger. The begomovirus and DNA-beta have been identified as Cotton leaf curl Gezira virus and Cotton leaf curl Gezira betasatellite, respectively, which are reported to also infect okra in Egypt, Mali and Sudan.

  8. Complete genome sequence and analysis of blackcurrant leaf chlorosis associated virus, a new member of the genus Idaeovirus.

    PubMed

    James, Delano; Phelan, James

    2017-02-11

    Blackcurrant leaf chlorosis associated virus (BCLCaV) was isolated from symptomatic blackcurrants (Ribes nigrum cv. Baldwin). The virus has a genome organization similar to that of raspberry bushy dwarf virus (RBDV), the type member of the genus Idaeovirus. The RNA-1of this virus encodes the replicase complex (ORF1, Mr 197 kDa), while RNA-2 encodes a putative movement protein (ORF2a, Mr 38.8 kDa) and the putative coat protein (ORF2b, Mr 30 kDa). A concatenated form of BCLCaV RNA-2 was detected by next-generation sequencing and confirmed by RT-PCR. BCLCaV is a new member of the genus Idaeovirus.

  9. Bean leaf beetle (Coleoptera: Chrysomelidae) management for reduction of bean pod mottle virus.

    PubMed

    Krell, Rayda K; Pedigo, Larry P; Hill, John H; Rice, Marlin E

    2004-04-01

    Bean pod mottle virus (BPMV) is a management concern for soybean, Glycine max (L.), producers in the North Central states because it can cause yield loss and reduce seed quality by induction of seed coat mottling. The main vector of BPMV is the bean leaf beetle, Cerotoma trifurcata (Forster). An experiment was conducted in 2000 and 2001 at two locations in northwestern and central Iowa to test three insecticide treatments for suppression of bean leaf beetles, and subsequently, BPMV. Treatments of insecticide applications with lambda-cyhalothrin were 1) a single early-season application (23 g [AI] /ha) (2.5 oz/acre) at the VE-VC soybean developmental stage; 2) two early-season applications, the first the same as treatment 1 and a second at the same rate 9-13 d later; 3) a single early-season application the same as treatment 1, followed by a mid-season application (28 g [AI] /ha (3.2 oz/acre) at approximately R2 (flowering, near 15 July); and 4) an unsprayed control. Application of lambda-cyhalothrin after soybean emergence and again as first-generation bean leaf beetles emerged in northwestern Iowa in 2000 (treatment 3) significantly reduced beetle densities through mid-season, BPMV field incidence by 31.5%, and seed coat mottling by 31.2%, compared with the unsprayed control. Similar effects were measured at the same location when insecticide was applied twice at early season (treatment 2). Yield was 453.7 kg/ha (6.74 bu/acre) greater in treatment 2 and 525.20 kg/ha (7.80 bu/acre) greater in treatment 3 than in the unsprayed control at the northwestern site in 2000. At both locations in 2001 fewer treatment effects were observed, which was likely related to lower beetle populations in that year. Early-season insecticide sprays targeted at overwintered beetles on VC-VE reduced the initial population of vector insects and may have contributed to a lower first-generation population because of reduced overwintered beetle oviposition. In 1 year at one location there

  10. Flupyradifurone effectively manages whitefly Bemisia tabaci MED (Hemiptera: Aleyrodidae) and tomato yellow leaf curl virus in tomato.

    PubMed

    Roditakis, Emmanouil; Stavrakaki, Marianna; Grispou, Maria; Achimastou, Aikaterini; Van Waetermeulen, Xavier; Nauen, Ralf; Tsagkarakou, Anastasia

    2017-08-01

    The cotton whitefly Bemisia tabaci (Gennadius) is among the most important pests of numerous crops and a vector of more than 100 plant viruses, causing significant crop losses worldwide. Managing this pest as well as inhibiting the transmission of major viruses such as tomato yellow leaf curl virus (TYLCV) are of utmost importance for sustainable yields. The efficacy against both whitefly and virus transmission of the novel systemic butenolide insecticide flupyradifurone was investigated in this study. The inhibition of TYLCV transmission by flupyradifurone was compared to that by thiamethoxam, a neonicotinoid insecticide reported to inhibit virus transmission. The experiment was performed under high virus pressure conditions (10 viruliferous insects per plant for 48 h) using a fully characterized field strain of B. tabaci. The insecticides were foliarly applied at recommended label rates under greenhouse conditions. Flupyradifurone suppressed virus transmission by 85% while levels of suppression after thiamethoxam treatments were just 25% and significantly lower. In untreated control plots, 100% of plants were infected by TYLCV. The observed difference in the potential to suppress virus transmission is linked to a strong knockdown effect as well as prolonged feeding inhibition in flupyradifurone treatments. Flupyradifurone is shown to be an extremely useful, fast-acting, new chemical tool in integrated crop management offering simultaneous control of whiteflies and strong suppression of viral infections via its rapid knockdown action and good residual activity. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  11. Tomato chlorotic leaf distortion virus, a new bipartite begomovirus infecting Solanum lycopersicum and Capsicum chinense in Venezuela.

    PubMed

    Zambrano, Karla; Geraud-Pouey, Francis; Chirinos, Doris; Romay, Gustavo; Marys, Edgloris

    2011-12-01

    Virus isolate T217L was obtained from a diseased tomato (Solanum lycopersicum) plant showing leaf deformation and chlorotic mottle symptoms near Maracaibo in the state of Zulia, Venezuela. Full-length DNA-A and DNA-B molecules of T217L were cloned and sequenced. The genome organization of T217L was identical to the bipartite genomes of other begomoviruses described from the Americas. Characteristic disease symptoms were reproduced in S. lycopersicum and Capsicum annum plants inoculated using the cloned viral DNA-A and DNA-B components, confirming disease aetiology. A sequence analysis of DNA-A showed that the T217L isolate has the highest sequence identity (84%) with sida yellow mosaic Yucatan virus (SiYMYuV), sida golden mosaic Honduras virus (SiGMHV) and bean dwarf mosaic virus (BDMV) isolates. This is less than the 89% identity in the DNA-A component that has been defined as the threshold value for the demarcation of species in the genus Begomovirus. The molecular data show that isolate T217L belongs to a novel tentative begomovirus species, for which the name tomato chlorotic leaf distortion virus is proposed. TCLDV was also detected in symptomatic C. chinense plants growing near the T217L-infected plant.

  12. Monitoring the dynamics of emergence of a non-canonical recombinant of Tomato yellow leaf curl virus and displacement of its parental viruses in tomato.

    PubMed

    Belabess, Z; Dallot, S; El-Montaser, S; Granier, M; Majde, M; Tahiri, A; Blenzar, A; Urbino, C; Peterschmitt, M

    2015-12-01

    Recombinant viruses are increasingly being reported but the dynamics of their emergence is rarely documented. A new recombinant Tomato yellow leaf curl virus (TYLCV-IS76) was detected for the first time in 2010 in Southern Morocco (Souss). An original diagnostic tool was needed to fit its unusual recombination profile. Although IS76 was detected following the appearance of Tylc symptoms on tolerant tomato plants, symptoms could not be associated to IS76 or to a synergy with criniviruses. According to infection profiles of Tylc-associated viruses determined on 879 plant samples collected between 1998 and 2014 and a Bayesian inference applied to genomic sequences of representatives of TYLCV, IS76 emerged in Southern Morocco at the end of the 1990s, replaced the parental viruses between 2004 and 2012 in Souss and is spreading towards the North of Morocco. The emergence of IS76 coincides with the increasing use of tolerant cultivars in the 2000s.

  13. In Vitro Inhibition of Cytopathic Effect of Influenza Virus and Human Immunodeficiency Virus by Bamboo Leaf Extract Solution and Sodium Copper Chlorophyllin

    PubMed Central

    Ito, Akiko; Tsuneki, Akeno; Yoshida, Yu; Ryoke, Kazuo; Kaidoh, Toshiyuki; Kageyama, Seiji

    2016-01-01

    Background Although the link between oral and oropharyngeal health status and susceptibility to infection has long been recognized, there is a limit to the selection of antiseptics for oral care. Methods Madin-Darby canine kidney (MDCK) cells were exposed to influenza virus and cultured in the presence or absence of test reagents: bamboo leaf extract solution and sodium copper chrolophyllin. MDCK cells were pre-incubated with the reagents to assess the inhibitory activity at adsorption (viral attachment). Similarly, anti-HIV activity and the inhibitory mechanism at adsorption were assessed by MT-2 cell culture system. Mixture of HIV and bamboo leaf extract solution was fixed and examined by transmission electron microscopy. Results The 50% inhibitory concentration (IC50) of bamboo leaf extract solution against influenza virus and the 50% cytotoxic concentration (CC50) in MDCK cells of the solution lay between 0.0313–0.0625% and 0.5–1.0%. The solution inhibited the influenza virus adsorption at the concentration of 0.5% (P < 0.05). The values of IC50 and CC50 of sodium copper chlorophyllin lay between 50–100 µM and 200–400 µM, respectively. This inhibited the virus adsorption at 200 µM (P < 0.05). The bamboo leaf extract solution showed values of IC50 against HIV and CC50 in MT-2 cells at around 0.0313% and between 0.25–0.5%, respectively. This solution inhibited HIV adsorption at 1.25% (P < 0.05). The IC50 and CC50 of sodium copper chlorophyllin lay between 50–100 µM and 200–400 µM, respectively. Sodium copper chlorophyllin inhibited HIV adsorption at 2.5 mM (P < 0.05). HIV particles survived after the exposure to 0.5% bamboo leaf extract solution. Conclusion Sodium copper chlorophyllin exerted antiviral activities against influenza virus and HIV as the major ingredient of bamboo leaf extract solution by blocking adsorption. This mechanism of action is different completely from the one of povidone-iodine. PMID:27046952

  14. In Vitro Inhibition of Cytopathic Effect of Influenza Virus and Human Immunodeficiency Virus by Bamboo Leaf Extract Solution and Sodium Copper Chlorophyllin.

    PubMed

    Ito, Akiko; Tsuneki, Akeno; Yoshida, Yu; Ryoke, Kazuo; Kaidoh, Toshiyuki; Kageyama, Seiji

    2016-03-01

    Although the link between oral and oropharyngeal health status and susceptibility to infection has long been recognized, there is a limit to the selection of antiseptics for oral care. Madin-Darby canine kidney (MDCK) cells were exposed to influenza virus and cultured in the presence or absence of test reagents: bamboo leaf extract solution and sodium copper chrolophyllin. MDCK cells were pre-incubated with the reagents to assess the inhibitory activity at adsorption (viral attachment). Similarly, anti-HIV activity and the inhibitory mechanism at adsorption were assessed by MT-2 cell culture system. Mixture of HIV and bamboo leaf extract solution was fixed and examined by transmission electron microscopy. The 50% inhibitory concentration (IC50) of bamboo leaf extract solution against influenza virus and the 50% cytotoxic concentration (CC50) in MDCK cells of the solution lay between 0.0313-0.0625% and 0.5-1.0%. The solution inhibited the influenza virus adsorption at the concentration of 0.5% (P < 0.05). The values of IC50 and CC50 of sodium copper chlorophyllin lay between 50-100 µM and 200-400 µM, respectively. This inhibited the virus adsorption at 200 µM (P < 0.05). The bamboo leaf extract solution showed values of IC50 against HIV and CC50 in MT-2 cells at around 0.0313% and between 0.25-0.5%, respectively. This solution inhibited HIV adsorption at 1.25% (P < 0.05). The IC50 and CC50 of sodium copper chlorophyllin lay between 50-100 µM and 200-400 µM, respectively. Sodium copper chlorophyllin inhibited HIV adsorption at 2.5 mM (P < 0.05). HIV particles survived after the exposure to 0.5% bamboo leaf extract solution. Sodium copper chlorophyllin exerted antiviral activities against influenza virus and HIV as the major ingredient of bamboo leaf extract solution by blocking adsorption. This mechanism of action is different completely from the one of povidone-iodine.

  15. The spread of tomato yellow leaf curl virus from the Middle East to the world.

    PubMed

    Lefeuvre, Pierre; Martin, Darren P; Harkins, Gordon; Lemey, Philippe; Gray, Alistair J A; Meredith, Sandra; Lakay, Francisco; Monjane, Adérito; Lett, Jean-Michel; Varsani, Arvind; Heydarnejad, Jahangir

    2010-10-28

    The ongoing global spread of Tomato yellow leaf curl virus (TYLCV; Genus Begomovirus, Family Geminiviridae) represents a serious looming threat to tomato production in all temperate parts of the world. Whereas determining where and when TYLCV movements have occurred could help curtail its spread and prevent future movements of related viruses, determining the consequences of past TYLCV movements could reveal the ecological and economic risks associated with similar viral invasions. Towards this end we applied Bayesian phylogeographic inference and recombination analyses to available TYLCV sequences (including those of 15 new Iranian full TYLCV genomes) and reconstructed a plausible history of TYLCV's diversification and movements throughout the world. In agreement with historical accounts, our results suggest that the first TYLCVs most probably arose somewhere in the Middle East between the 1930s and 1950s (with 95% highest probability density intervals 1905-1972) and that the global spread of TYLCV only began in the 1980s after the evolution of the TYLCV-Mld and -IL strains. Despite the global distribution of TYLCV we found no convincing evidence anywhere other than the Middle East and the Western Mediterranean of epidemiologically relevant TYLCV variants arising through recombination. Although the region around Iran is both the center of present day TYLCV diversity and the site of the most intensive ongoing TYLCV evolution, the evidence indicates that the region is epidemiologically isolated, which suggests that novel TYLCV variants found there are probably not direct global threats. We instead identify the Mediterranean basin as the main launch-pad of global TYLCV movements.

  16. Yoctomole electrochemical genosensing of Ebola virus cDNA by rolling circle and circle to circle amplification.

    PubMed

    Carinelli, S; Kühnemund, M; Nilsson, M; Pividori, M I

    2017-07-15

    This work addresses the design of an Ebola diagnostic test involving a simple, rapid, specific and highly sensitive procedure based on isothermal amplification on magnetic particles with electrochemical readout. Ebola padlock probes were designed to detect a specific L-gene sequence present in the five most common Ebola species. Ebola cDNA was amplified by rolling circle amplification (RCA) on magnetic particles. Further re-amplification was performed by circle-to-circle amplification (C2CA) and the products were detected in a double-tagging approach using a biotinylated capture probe for immobilization on magnetic particles and a readout probe for electrochemical detection by square-wave voltammetry on commercial screen-printed electrodes. The electrochemical genosensor was able to detect as low as 200 ymol, corresponding to 120 cDNA molecules of L-gene Ebola virus with a limit of detection of 33 cDNA molecules. The isothermal double-amplification procedure by C2CA combined with the electrochemical readout and the magnetic actuation enables the high sensitivity, resulting in a rapid, inexpensive, robust and user-friendly sensing strategy that offers a promising approach for the primary care in low resource settings, especially in less developed countries. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. In vitro rearing of Perkinsiella saccharicida and the use of leaf segments to assay Fiji disease virus transmission.

    PubMed

    Hughes, G L; Allsopp, P G; Brumbley, S M; Johnson, K N; O'Neill, S L

    2008-07-01

    Fiji leaf gall (FLG) is caused by the Reovirus, Fiji disease virus (FDV), which is transmitted to sugarcane by planthoppers of the genus Perkinsiella. Low vector transmission rates and slow disease symptom development make experimentation within the FDV-Perkinsiella-sugarcane system inherently difficult. A laboratory-based technique was devised to rear the vector using sugarcane leaves as a food source. Planthoppers were reared on sugarcane leaf segments embedded in agarose enclosed within plastic containers. To provide a nondestructive assay for determination of the inoculation potential of planthoppers, FDV was detected by reverse transcription-polymerase chain reaction (RT-PCR) in newly infected sugarcane leaf segments following exposure to viruliferous planthoppers. Leaf segment inoculation correlated with development of FLG symptoms in whole plants that were fed on by the same planthoppers. Analysis of FDV RNAs within the planthopper, measured by quantitative RT-PCR (qRT-PCR), indicated that FDV RNA concentration was associated with successful inoculation of the leaf segment, transmission of FDV to sugarcane and subsequent development of FLG in plants. Quantification of FDV RNA within planthoppers provided an additional measure to assess vector competence in individuals.

  18. Immunity to tomato yellow leaf curl virus in transgenic tomato is associated with accumulation of transgene small RNA.

    PubMed

    Leibman, Diana; Prakash, Shanmugam; Wolf, Dalia; Zelcer, Aaron; Anfoka, Ghandi; Haviv, Sabrina; Brumin, Marina; Gaba, Victor; Arazi, Tzahi; Lapidot, Moshe; Gal-On, Amit

    2015-11-01

    Gene silencing is a natural defense response of plants against invading RNA and DNA viruses. The RNA post-transcriptional silencing system has been commonly utilized to generate transgenic crop plants that are "immune" to plant virus infection. Here, we applied this approach against the devastating DNA virus tomato yellow leaf curl virus (TYLCV) in its host tomato (Solanum lycopersicum L.). To generate broad resistance to a number of different TYLCV viruses, three conserved sequences (the intergenic region [NCR], V1-V2 and C1-C2 genes) from the genome of the severe virus (TYLCV) were synthesized as a single insert and cloned into a hairpin configuration in a binary vector, which was used to transform TYLCV-susceptible tomato plants. Eight of 28 independent transgenic tomato lines exhibited immunity to TYLCV-Is and to TYLCV-Mld, but not to tomato yellow leaf curl Sardinia virus, which shares relatively low sequence homology with the transgene. In addition, a marker-free (nptII-deleted) transgenic tomato line was generated for the first time by Agrobacterium-mediated transformation without antibiotic selection, followed by screening of 1180 regenerated shoots by whitefly-mediated TYLCV inoculation. Resistant lines showed a high level of transgene-siRNA (t-siRNA) accumulation (22% of total small RNA) with dominant sizes of 21 nt (73%) and 22 nt (22%). The t-siRNA displayed hot-spot distribution ("peaks") along the transgene, with different distribution patterns than the viral-siRNA peaks observed in TYLCV-infected tomato. A grafting experiment demonstrated the mobility of 0.04% of the t-siRNA from transgenic rootstock to non-transformed scion, even though scion resistance against TYLCV was not achieved.

  19. Characterization of Apricot pseudo-chlorotic leaf spot virus, A Novel Trichovirus Isolated from Stone Fruit Trees.

    PubMed

    Liberti, D; Marais, A; Svanella-Dumas, L; Dulucq, M J; Alioto, D; Ragozzino, A; Rodoni, B; Candresse, T

    2005-04-01

    ABSTRACT A trichovirus closely related to Apple chlorotic leaf spot virus (ACLSV) was detected in symptomatic apricot and Japanese plum from Italy. The Sus2 isolate of this agent cross-reacted with anti-ACLSV polyclonal reagents but was not detected by broad-specificity anti- ACLSV monoclonal antibodies. It had particles with typical trichovirus morphology but, contrary to ACLSV, was unable to infect Chenopodium quinoa and C. amaranticolor. The sequence of its genome (7,494 nucleotides [nt], missing only approximately 30 to 40 nt of the 5' terminal sequence) and the partial sequence of another isolate were determined. The new virus has a genomic organization similar to that of ACLSV, with three open reading frames coding for a replication-associated protein (RNA-dependent RNA polymerase), a movement protein, and a capsid protein, respectively. However, it had only approximately 65 to 67% nucleotide identity with sequenced isolates of ACLSV. The differences in serology, host range, genome sequence, and phylogenetic reconstructions for all viral proteins support the idea that this agent should be considered a new virus, for which the name Apricot pseudo-chlorotic leaf spot virus (APCLSV) is proposed. APCLSV shows substantial sequence variability and has been recovered from various Prunus sources coming from seven countries, an indication that it is likely to have a wide geographical distribution.

  20. Amplicon based RNA interference targeting V2 gene of cotton leaf curl Kokhran virus-Burewala strain can provide resistance in transgenic cotton plants

    USDA-ARS?s Scientific Manuscript database

    An RNAi based gene construct designated “C2” was used to target the V2 region of the cotton leaf curl virus (CLCuV) genome which is responsible for virus movement. The construct was transformed into two elite cotton varieties MNH-786 and VH-289. A shoot apex method of plant transformation using Agr...

  1. Aphid transmission of Lettuce necrotic leaf curl virus, a member of a tentative new subgroup within the genus Torradovirus.

    PubMed

    Verbeek, Martin; Dullemans, Annette M; van der Vlugt, René A A

    2017-02-20

    Lettuce necrotic leaf curl virus (LNLCV) was described as the first non-tomato-infecting member of the genus Torradovirus. Until today, the virus was found only in The Netherlands in two different areas in open field crops of lettuce. In 2015, LNLCV was accepted by the ICTV as a new member of the genus Torradovirus. The tomato-infecting (TI) torradoviruses Tomato torrado virus (ToTV), Tomato marchitez virus (ToMarV) and Tomato chocolàte virus (ToChV) are transmitted by at least three whitefly species in a semi-persistent and stylet-borne manner. As LNLCV was transmitted in open fields in The Netherlands, where whiteflies are present only in low incidence, transmission studies were set up to identify the natural vector of LNLCV. Whitefly species which survive Dutch open field conditions during summer, as well as lettuce colonizing aphid species, were tested for their ability to transmit LNLCV. Lengths of acquisition and inoculation periods were chosen in accordance with the conditions for TI torradoviruses. Transmission experiments involving whiteflies were never successful. Transmission with aphids was only successful in case of the lettuce-currant aphid, Nasonovia ribisnigri. Localization of LNLCV virions in N. ribisnigri with a nested RT-PCR indicated the stylets as possible retention sites. The willow-carrot aphid Cavariella aegopodii did not transmit LNLCV in our transmission experiment but the virus could be detected in the stylets of this aphid, leaving C. aegopodii as a possible vector for LNLCV.

  2. The hypersensitive response to tomato leaf curl New Delhi virus nuclear shuttle protein is inhibited by transcriptional activator protein.

    PubMed

    Hussain, Mazhar; Mansoor, Shahid; Iram, Shazia; Zafar, Yusuf; Briddon, Rob W

    2007-12-01

    The hypersensitive response (HR) is a common feature of plant disease resistance reactions and a type of programmed cell death (PCD). Many pathogens are able to modulate pathways involved in cell death. In contrast to animal viruses, inhibitors of PCD activity have not been identified for plant-infecting viruses. Previously, we have reported that the nuclear shuttle protein (NSP) of Tomato leaf curl New Delhi virus (ToLCNDV) induces an HR in Nicotiana tabacum and Lycopersicon esculentum plants when expressed under the control of the Cauliflower mosaic virus 35S promoter. However, HR is not evident in plants infected with ToLCNDV, suggesting that the virus encodes a factor (or factors) that counters this response. Analysis of all ToLCNDV-encoded genes pinpointed the transcriptional activator protein (TrAP) as the factor mediating the anti-HR effect. Deletion mutagenesis showed the central region of TrAP, containing a zinc finger domain and nuclear localization signal, to be important in inhibiting the HR. These results demonstrate that TrAP counters HR-induced cell death, the first such activity identified for a plant-infecting virus.

  3. Detection and characterization of a new betasatellite: variation in disease symptoms of tomato leaf curl Pakistan virus-India due to associated betasatellite.

    PubMed

    Kumar, Jitendra; Singh, Sudhir P; Kumar, Abhinav; Khan, Jawaid A; Tuli, Rakesh

    2013-01-01

    A begomovirus and its associated betasatellites were amplified and sequenced from tobacco plants affected with leaf curl disease. The begomovirus was identified as a new strain of tomato leaf curl Pakistan virus (ToLCPKV), which is referred to here as ToLCPKV-India. A previously known betasatellite [tomato leaf curl Patna betasatellite (ToLCPaB)] and a new betasatellite were also found in leaf-curl-affected samples. The use of infectious clones of ToLCPKV-IN plus ToLCPaB for agroinoculation led to typical leaf curl, while ToLCPKV-IN together with the new betasatellite resulted in curling and chlorosis of leaves. Based on these disease symptoms, we propose to name the new betasatellite tobacco leaf chlorosis betasatellite (TbLChB).

  4. Genetic diversity of tomato-infecting Tomato yellow leaf curl virus (TYLCV) isolates in Korea.

    PubMed

    Kim, Sue Hoon; Oh, Sung; Oh, Tae-Kyun; Park, Jae Sung; Kim, Sei Chang; Kim, Seong Hwan; Kim, Young Shik; Hong, Jeum Kyu; Sim, Sang-Yun; Park, Kwon Seo; Lee, Hwan Gu; Kim, Kyung Jae; Choi, Chang Won

    2011-02-01

    Epidemic outbreaks of Tomato yellow leaf curl virus (TYLCV) diseases occurred in greenhouse grown tomato (Solanum lycopersicum) plants of Busan (TYLCV-Bus), Boseong (TYLCV-Bos), Hwaseong (TYLCV-Hwas), Jeju Island (TYLCV-Jeju), and Nonsan (TYLCV-Nons) in Korea during 2008-2009. Tomato disease by TYLCV has never occurred in Korea before. We synthesized the full-length genomes of each TYLCV isolate from the tomato plants collected at each area and determined their nucleotides (nt) sequences and deduced the amino acids of six open reading frames in the genomes. TYLCV-Bus and -Bos genomes shared higher nt identities with four Japanese isolates -Ng, -Omu, -Mis, and -Miy. On the other hand, TYLCV-Hwas, -Jeju, and -Nons genomes shared higher nt identities with five Chinese isolates TYLCV-AH1, -ZJ3, -ZJHZ12, -SH2, -Sh10, and two Japanese isolates -Han and -Tosa. On the basis of a neighbor-joining tree, five Korean TYLCV isolates were separated into three clades. TYLCV-Bus and -Bos formed the first clade, clustering with four Japanese isolates TYLCV-Mis, -Omu, -Ng, and -Miy. TYLCV-Jeju and -Nons formed the second clade, clustering with two Chinese isolates -ZJHZ212 and -Sh10. TYLCV-Hwas was clustered with two Japanese isolates -Han and -Tosa and three Chinese isolates -AH1, -ZJ3, and -SH2. Two fragments that had a potentially recombinant origin were identified using the RDP, GENECONV, BootScan, MaxChi, Chimaera, SiScan, and 3Seq methods implemented in RDP3.41. On the basis of RDP analysis, all TYLCV isolates could originated from the interspecies recombination between TYLCV-Mld[PT] isolated from Portugal as a major parent and TYLCTHV-MM isolated from Myanmar as a minor parent.

  5. Highly sensitive serological methods for detecting tomato yellow leaf curl virus in tomato plants and whiteflies.

    PubMed

    Xie, Yan; Jiao, Xiaoyang; Zhou, Xueping; Liu, Huan; Ni, Yuequn; Wu, Jianxiang

    2013-05-06

    Tomato yellow leaf curl virus (TYLCV) is a member of the genus Begomovirus in the family Geminiviridae, which causes severe losses in tomato production in tropic and subtropic regions. The purified TYLCV virions were used as the immunogen to produce monoclonal antibodies (MAbs) using the hybridoma technology. MAb-based dot enzyme-linked immunosorbent assay (dot-ELISA) and direct tissue blot immunoassay (DTBIA) were developed for sensitive, simple, and rapid detection of TYLCV in field tomato and whitefly (Bemisia tabaci) samples collected from TYLCV prevalent provinces in China. Using the hybridoma technology, six murine MAbs (1C4, 8D10, 6E3, 2F2, 3F4 and 4G3) against TYLCV were prepared. Using the MAb 1C4, dot-ELISA and DTBIA were then established for detecting TYLCV in field tomato and whitefly samples collected from TYLCV prevalent provinces in China. The dot-ELISA could detect TYLCV in infected tissue crude extract diluted at 1:5,120 (w/v, g mL-1), and in viruliferous whitefly homogenate diluted at 1:128 (individual whitefly/μL), respectively. Field tomato samples (n=487) and whitefly samples (n=110) from TYLCV prevalent districts in China were screened for the presence of TYLCV using the two developed methods, and the results were further confirmed by PCR and nucleotide sequencing. The survey revealed that TYLCV is widespread on tomato plants in Zhejiang, Shandong and Henan provinces in China. The developed dot-ELISA is very suitable for the routine detection of TYLCV in field tomato and whitefly samples, and the DTBIA is more suitable for the routine detection of TYLCV in large-scale tomato plant samples collected from TYLCV prevalent areas.

  6. Highly sensitive serological methods for detecting tomato yellow leaf curl virus in tomato plants and whiteflies

    PubMed Central

    2013-01-01

    Background Tomato yellow leaf curl virus (TYLCV) is a member of the genus Begomovirus in the family Geminiviridae, which causes severe losses in tomato production in tropic and subtropic regions. Methods The purified TYLCV virions were used as the immunogen to produce monoclonal antibodies (MAbs) using the hybridoma technology. MAb-based dot enzyme-linked immunosorbent assay (dot-ELISA) and direct tissue blot immunoassay (DTBIA) were developed for sensitive, simple, and rapid detection of TYLCV in field tomato and whitefly (Bemisia tabaci) samples collected from TYLCV prevalent provinces in China. Results Using the hybridoma technology, six murine MAbs (1C4, 8D10, 6E3, 2F2, 3F4 and 4G3) against TYLCV were prepared. Using the MAb 1C4, dot-ELISA and DTBIA were then established for detecting TYLCV in field tomato and whitefly samples collected from TYLCV prevalent provinces in China. The dot-ELISA could detect TYLCV in infected tissue crude extract diluted at 1:5,120 (w/v, g mL-1), and in viruliferous whitefly homogenate diluted at 1:128 (individual whitefly/μL), respectively. Field tomato samples (n=487) and whitefly samples (n=110) from TYLCV prevalent districts in China were screened for the presence of TYLCV using the two developed methods, and the results were further confirmed by PCR and nucleotide sequencing. The survey revealed that TYLCV is widespread on tomato plants in Zhejiang, Shandong and Henan provinces in China. Conclusions The developed dot-ELISA is very suitable for the routine detection of TYLCV in field tomato and whitefly samples, and the DTBIA is more suitable for the routine detection of TYLCV in large-scale tomato plant samples collected from TYLCV prevalent areas. PMID:23647724

  7. Difference of physiological characters in dark green islands and yellow leaf tissue of Cucumber mosaic virus (CMV)-infected Nicotiana tabacum leaves.

    PubMed

    Shang, Jing; Xi, De-Hui; Yuan, Shu; Xu, Fei; Xu, Mo-Yun; Qi, Hai-Long; Wang, Shao-Dong; Huang, Qing-Rong; Wen, Lin; Lin, Hong-Hui

    2010-01-01

    Dark green islands (DGIs) are a common symptom of plants systemically infected with the mosaic virus. DGIs are clusters of green leaf cells that are free of virus but surrounded by yellow leaf tissue that is full of virus particles. In Cucumber mosaic virus (CMV)-infected Nicotiana tabacum leaves, the respiration and photosynthesis capabilities of DGIs and yellow leaf tissues were measured. The results showed that the cyanide-resistant respiration was enhanced in yellow leaf tissue and the photosynthesis was declined, while in DGIs they were less affected. The activities of the oxygen-scavenging enzymes catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) in infected leaves were significantly higher than those in the healthy leaves, and the enzyme activities in DGIs were always lower than in the yellow leaf tissues. Reactive oxygen species (ROS) staining showed that the hydrogen peroxide content in yellow leaf tissues was apparently higher than that in DGIs, while the superoxide content was on the contrary. Formation of DGIs may be a strategy of the host plants resistance to the CMV infection.

  8. Molecular and biological characterization of a new Tomato mild yellow leaf curl Aragua virus strain producing severe symptoms in tomato.

    PubMed

    Romay, Gustavo; Chirinos, Dorys T; Geraud-Pouey, Francis; Gillis, Annika; Mahillon, Jacques; Desbiez, Cécile; Bragard, Claude

    2017-06-20

    Tomato mild yellow leaf curl Aragua virus (ToMYLCV) is a begomovirus first reported infecting tomato (Solanum lycopersicum) and milkweed (Euphorbia heterophylla) in Venezuela. In this study, a ToMYLCV isolate (Zulia-219) was completely sequenced and its host range was evaluated. The DNA-A and DNA-B components of isolate Zulia-219 showed 93 and 85% nucleotide sequence identity with the respective counterparts of the ToMYLCV type strain. According to current demarcation criteria for begomovirus species, Zulia-219 is a new strain of ToMYLCV. Interestingly, tomato plants inoculated with ToMYLCV Zulia-219 displayed severe symptoms, including severe chlorotic leaf curling, in contrast to mild symptoms associated with the type strain of this begomovirus. These results indicate potential risks associated with this new ToMYLCV strain for tomato production in Venezuela.

  9. Recruitment of the Host Plant Heat Shock Protein 70 by Tomato Yellow Leaf Curl Virus Coat Protein Is Required for Virus Infection

    PubMed Central

    Gorovits, Rena; Moshe, Adi; Ghanim, Murad; Czosnek, Henryk

    2013-01-01

    A functional capsid protein (CP) is essential for host plant infection and insect transmission of Tomato yellow leaf curl virus (TYLCV) and other monopartite begomoviruses. We have previously shown that TYLCV CP specifically interacts with the heat shock protein 70 (HSP70) of the virus insect vector, Bemisia tabaci. Here we demonstrate that during the development of tomato plant infection with TYLCV, a significant amount of HSP70 shifts from a soluble form into insoluble aggregates. CP and HSP70 co-localize in these aggregates, first in the cytoplasm, then in the nucleus of cells associated with the vascular system. CP-HSP70 interaction was demonstrated by co-immunopreciptation in cytoplasmic - but not in nuclear extracts from leaf and stem. Inhibition of HSP70 expression by quercetin caused a decrease in the amount of nuclear CP aggregates and a re-localization of a GFP-CP fusion protein from the nucleus to the cytoplasm. HSP70 inactivation resulted in a decrease of TYLCV DNA levels, demonstrating the role of HSP70 in TYLCV multiplication in planta. The current study reveals for the first time the involvement of plant HSP70 in TYLCV CP intracellular movement. As described earlier, nuclear aggregates contained TYLCV DNA-CP complexes and infectious virions. Showing that HSP70 localizes in these large nuclear aggregates infers that these structures operate as nuclear virus factories. PMID:23894631

  10. New Insecticides for Management of Tomato Yellow Leaf Curl, a Virus Vectored by the Silverleaf Whitefly, Bemisia tabaci

    PubMed Central

    Smith, H. A.; Giurcanu, M. C.

    2014-01-01

    Greenhouse studies using a randomized complete block design were carried out to evaluate the effect of six insecticides on transmission of Tomato yellow leaf curl virus (TYLCV) by the silverleaf whitefly, Bemisia tabaci biotype B Gennadius (Hemiptera: Aleyrodidae) to tomato, Lycopersicon esculentum (Miller) (Solanales: Solanaceae), seedlings that were inoculated with whiteflies from a TYLCV colony in cages 3, 7, or 14 d after treatment with insecticide. The purpose was to reveal differences in residual efficacy of four materials that are nearing registration for use on tomato—cyazypyr, flupyradifurone, pyrafluquinazon, and sulfoxaflor—and to compare them with two established insecticides, pymetrozine and a zeta-cypermethrin/bifenthrin combination. Differences in efficacy were expected because these six materials represent five distinct modes of action and both contact and systemic materials. Percentage of tomato seedlings expressing virus symptoms tended to be lowest in seedlings treated with flupyradifurone. The zeta-cypermethrin/bifenthrin insecticide demonstrated comparable efficacy to flupyradifurone in some trials at 3 and 7 d after treatment inoculations, but not the 14 d after treatment inoculation. Pyrafluquinazon was not statistically different from cyazypyr or sulfoxaflor in percentage of plants with virus symptoms in any trial. Percentage virus in the cyazypyr and sulfoxaflor treatments was not statistically different in the 3 and 7 d after treatment inoculations. Among seedlings treated with insecticide, percentage with virus symptoms tended to be highest in the seedlings treated with pymetrozine. PMID:25368089

  11. New insecticides for management of tomato yellow leaf curl, a virus vectored by the silverleaf whitefly, Bemisia tabaci.

    PubMed

    Smith, H A; Giurcanu, M C

    2014-01-01

    Greenhouse studies using a randomized complete block design were carried out to evaluate the effect of six insecticides on transmission of Tomato yellow leaf curl virus (TYLCV) by the silverleaf whitefly, Bemisia tabaci biotype B Gennadius (Hemiptera: Aleyrodidae) to tomato, Lycopersicon esculentum (Miller) (Solanales: Solanaceae), seedlings that were inoculated with whiteflies from a TYLCV colony in cages 3, 7, or 14 d after treatment with insecticide. The purpose was to reveal differences in residual efficacy of four materials that are nearing registration for use on tomato-cyazypyr, flupyradifurone, pyrafluquinazon, and sulfoxaflor-and to compare them with two established insecticides, pymetrozine and a zeta-cypermethrin/bifenthrin combination. Differences in efficacy were expected because these six materials represent five distinct modes of action and both contact and systemic materials. Percentage of tomato seedlings expressing virus symptoms tended to be lowest in seedlings treated with flupyradifurone. The zeta-cypermethrin/bifenthrin insecticide demonstrated comparable efficacy to flupyradifurone in some trials at 3 and 7 d after treatment inoculations, but not the 14 d after treatment inoculation. Pyrafluquinazon was not statistically different from cyazypyr or sulfoxaflor in percentage of plants with virus symptoms in any trial. Percentage virus in the cyazypyr and sulfoxaflor treatments was not statistically different in the 3 and 7 d after treatment inoculations. Among seedlings treated with insecticide, percentage with virus symptoms tended to be highest in the seedlings treated with pymetrozine. © The Author 2014. Published by Oxford University Press on behalf of the Entomological Society of America.

  12. Peptide Aptamers That Bind to Geminivirus Replication Proteins Confer a Resistance Phenotype to Tomato Yellow Leaf Curl Virus and Tomato Mottle Virus Infection in Tomato

    PubMed Central

    Reyes, Maria Ines; Nash, Tara E.; Dallas, Mary M.; Ascencio-Ibáñez, J. Trinidad

    2013-01-01

    Geminiviruses constitute a large family of single-stranded DNA viruses that cause serious losses in important crops worldwide. They often exist in disease complexes and have high recombination and mutation rates, allowing them to adapt rapidly to new hosts and environments. Thus, an effective resistance strategy must be general in character and able to target multiple viruses. The geminivirus replication protein (Rep) is a good target for broad-based disease control because it is highly conserved and required for viral replication. In an earlier study, we identified a set of peptide aptamers that bind to Rep and reduce viral replication in cultured plant cells. In this study, we selected 16 of the peptide aptamers for further analysis in yeast two-hybrid assays. The results of these experiments showed that all 16 peptide aptamers interact with all or most of the Rep proteins from nine viruses representing the three major Geminiviridae genera and identified two peptide aptamers (A22 and A64) that interact strongly with different regions in the Rep N terminus. Transgenic tomato lines expressing A22 or A64 and inoculated with Tomato yellow leaf curl virus or Tomato mottle virus exhibited delayed viral DNA accumulation and often contained lower levels of viral DNA. Strikingly, the effect on symptoms was stronger, with many of the plants showing no symptoms or strongly attenuated symptoms. Together, these results established the efficacy of using Rep-binding peptide aptamers to develop crops that are resistant to diverse geminiviruses. PMID:23824791

  13. Implication of the bacterial endosymbiont Rickettsia spp. in interactions of the whitefly Bemisia tabaci with tomato yellow leaf curl virus.

    PubMed

    Kliot, Adi; Cilia, Michelle; Czosnek, Henryk; Ghanim, Murad

    2014-05-01

    Numerous animal and plant viruses are transmitted by arthropod vectors in a persistent, circulative manner. Tomato yellow leaf curl virus (TYLCV) is transmitted by the sweet potato whitefly Bemisia tabaci. We report here that infection with Rickettsia spp., a facultative endosymbiont of whiteflies, altered TYLCV-B. tabaci interactions. A B. tabaci strain infected with Rickettsia acquired more TYLCV from infected plants, retained the virus longer, and exhibited nearly double the transmission efficiency compared to an uninfected B. tabaci strain with the same genetic background. Temporal and spatial antagonistic relationships were discovered between Rickettsia and TYLCV within the whitefly. In different time course experiments, the levels of virus and Rickettsia within the insect were inversely correlated. Fluorescence in situ hybridization analysis of Rickettsia-infected midguts provided evidence for niche exclusion between Rickettsia and TYLCV. In particular, high levels of the bacterium in the midgut resulted in higher virus concentrations in the filter chamber, a favored site for virus translocation along the transmission pathway, whereas low levels of Rickettsia in the midgut resulted in an even distribution of the virus. Taken together, these results indicate that Rickettsia, by infecting the midgut, increases TYLCV transmission efficacy, adding further insights into the complex association between persistent plant viruses, their insect vectors, and microorganism tenants that reside within these insects. Interest in bacterial endosymbionts in arthropods and many aspects of their host biology in agricultural and human health systems has been increasing. A recent and relevant studied example is the influence of Wolbachia on dengue virus transmission by mosquitoes. In parallel with our recently studied whitefly-Rickettsia-TYLCV system, other studies have shown that dengue virus levels in the mosquito vector are inversely correlated with bacterial load. Our work

  14. Severe outbreak of tomato yellow leaf curl Sardinia virus on pepper in southern Italy.

    PubMed

    Comes, Soccorsa; Fanigliulo, Angela; Pacella, Rosa; Crescenzi, Aniello

    2009-01-01

    During summer and autumn 2008 a severe outbreak of pepper leaf curl disease (PLCD) was observed in pepper crops under plastic tunnels in the ionic coast of Basilicata region. Its incidence reached, in some cases, values close to 50%. The beginning of infections was recorded along the perimeter of the tunnels, where it reached a percentage of almost 100%. The infection then progressively spread towards the central areas of the greenhouses. Large populations of whiteflies, identificated as Bemisia tabaci, were observed on the infected crops. Detection assays for TYLCSV and TYLCV were performed in order to ascertain the etiologic agent: 190 symptomatic samples were collected from different fields and assayed in DAS-ELISA using a broad-spectrum reagent combination (distributed by Bioreba AG) detecting TYLCV, TYLCSV and other Begamoviruses: of these, 176 samples resulted positive. In order to discriminate between TYLCSV, TYLCV or any other Begamovirus, 15 positive samples were analyzed by PCR using a couple of synthetic oligonucleotides allowing the amplification of the whole coat protein (CP) gene. RFLP analysis performed on the PCR product, 1008 bp long, showed the presence of only TYLCSV in all assayed samples. The molecular characterization performed by phylogenetic analysis of the sequenced coat protein gene revealed that the isolate shares a similarity of about 97% with the corresponding sequence of a tomato TYLCSV isolate from Sicily (Z28390) and is almost identical with the pepper isolate CAB-It recovered in the same area in 2007 (TYLCSV was first recorded on pepper in Italy in 2007 in Policoro-MT, Fanigliulo et al., 2008. Comm. Appl. Biol. Sci, Ghent University, 73/2, 2008), indicating that there is a very low variability in TYLCSV population in the surveyed area. The further diffusion of PLCD and its hazard has to be connected with the presence of wide tomato cultivations, of weed hosts alternative to pepper (Solanum nigrum, Datura stramonium, Sonchus asper

  15. 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.

  16. Comparison of transmission of Papaya leaf curl China virus among four cryptic species of the whitefly Bemisia tabaci complex

    PubMed Central

    Guo, Tao; Guo, Qi; Cui, Xi-Yun; Liu, Yin-Quan; Hu, Jian; Liu, Shu-Sheng

    2015-01-01

    Begomoviruses are transmitted by cryptic species of the whitefly Bemisia tabaci complex, often in a species-specific manner. Papaya leaf curl China virus (PaLCuCNV) has been recorded to infect several crops including papaya, tomato and tobacco in China. To help assess the risks of spread of this virus, we compared the acquisition, retention and transmission of PaLCuCNV among four species of whiteflies, Middle East-Asia Minor 1 (MEAM1), Mediterranean (MED), Asia 1 and Asia II 7. All four species of whiteflies are able to acquire, retain and transmit the virus, but with different levels of efficiency. Transmission tests using tomato as the host plant showed that MEAM1 transmitted PaLCuCNV with substantially higher efficiency than did MED, Asia 1 and Asia II 7. Furthermore, accumulation of PaLCuCNV in the whiteflies was positively associated with its efficiency of transmitting the virus. Altogether, these findings indicate that MEAM1 is the most efficient vector for PaLCuCNV in the four species of whiteflies, and suggest that risks of PaLCuCNV pandemics are high in regions where MEAM1 occurs. PMID:26486606

  17. Identification of a Severe Acute Respiratory Syndrome Coronavirus-Like Virus in a Leaf-Nosed Bat in Nigeria

    PubMed Central

    Quan, Phenix-Lan; Firth, Cadhla; Street, Craig; Henriquez, Jose A.; Petrosov, Alexandra; Tashmukhamedova, Alla; Hutchison, Stephen K.; Egholm, Michael; Osinubi, Modupe O. V.; Niezgoda, Michael; Ogunkoya, Albert B.; Briese, Thomas; Rupprecht, Charles E.; Lipkin, W. Ian

    2010-01-01

    Bats are reservoirs for emerging zoonotic viruses that can have a profound impact on human and animal health, including lyssaviruses, filoviruses, paramyxoviruses, and severe acute respiratory syndrome coronaviruses (SARS-CoVs). In the course of a project focused on pathogen discovery in contexts where human-bat contact might facilitate more efficient interspecies transmission of viruses, we surveyed gastrointestinal tissue obtained from bats collected in caves in Nigeria that are frequented by humans. Coronavirus consensus PCR and unbiased high-throughput pyrosequencing revealed the presence of coronavirus sequences related to those of SARS-CoV in a Commerson’s leaf-nosed bat (Hipposideros commersoni). Additional genomic sequencing indicated that this virus, unlike subgroup 2b CoVs, which includes SARS-CoV, is unique, comprising three overlapping open reading frames between the M and N genes and two conserved stem-loop II motifs. Phylogenetic analyses in conjunction with these features suggest that this virus represents a new subgroup within group 2 CoVs. PMID:21063474

  18. Comparison of transmission of Papaya leaf curl China virus among four cryptic species of the whitefly Bemisia tabaci complex.

    PubMed

    Guo, Tao; Guo, Qi; Cui, Xi-Yun; Liu, Yin-Quan; Hu, Jian; Liu, Shu-Sheng

    2015-10-21

    Begomoviruses are transmitted by cryptic species of the whitefly Bemisia tabaci complex, often in a species-specific manner. Papaya leaf curl China virus (PaLCuCNV) has been recorded to infect several crops including papaya, tomato and tobacco in China. To help assess the risks of spread of this virus, we compared the acquisition, retention and transmission of PaLCuCNV among four species of whiteflies, Middle East-Asia Minor 1 (MEAM1), Mediterranean (MED), Asia 1 and Asia II 7. All four species of whiteflies are able to acquire, retain and transmit the virus, but with different levels of efficiency. Transmission tests using tomato as the host plant showed that MEAM1 transmitted PaLCuCNV with substantially higher efficiency than did MED, Asia 1 and Asia II 7. Furthermore, accumulation of PaLCuCNV in the whiteflies was positively associated with its efficiency of transmitting the virus. Altogether, these findings indicate that MEAM1 is the most efficient vector for PaLCuCNV in the four species of whiteflies, and suggest that risks of PaLCuCNV pandemics are high in regions where MEAM1 occurs.

  19. A Sequence-Independent Strategy for Detection and Cloning of Circular DNA Virus Genomes by Using Multiply Primed Rolling-Circle Amplification

    PubMed Central

    Rector, Annabel; Tachezy, Ruth; Van Ranst, Marc

    2004-01-01

    The discovery of novel viruses has often been accomplished by using hybridization-based methods that necessitate the availability of a previously characterized virus genome probe or knowledge of the viral nucleotide sequence to construct consensus or degenerate PCR primers. In their natural replication cycle, certain viruses employ a rolling-circle mechanism to propagate their circular genomes, and multiply primed rolling-circle amplification (RCA) with φ29 DNA polymerase has recently been applied in the amplification of circular plasmid vectors used in cloning. We employed an isothermal RCA protocol that uses random hexamer primers to amplify the complete genomes of papillomaviruses without the need for prior knowledge of their DNA sequences. We optimized this RCA technique with extracted human papillomavirus type 16 (HPV-16) DNA from W12 cells, using a real-time quantitative PCR assay to determine amplification efficiency, and obtained a 2.4 × 104-fold increase in HPV-16 DNA concentration. We were able to clone the complete HPV-16 genome from this multiply primed RCA product. The optimized protocol was subsequently applied to a bovine fibropapillomatous wart tissue sample. Whereas no papillomavirus DNA could be detected by restriction enzyme digestion of the original sample, multiply primed RCA enabled us to obtain a sufficient amount of papillomavirus DNA for restriction enzyme analysis, cloning, and subsequent sequencing of a novel variant of bovine papillomavirus type 1. The multiply primed RCA method allows the discovery of previously unknown papillomaviruses, and possibly also other circular DNA viruses, without a priori sequence information. PMID:15113879

  20. Use of Posttranscription Gene Silencing in Squash to Induce Resistance against the Egyptian Isolate of the Squash Leaf Curl Virus

    PubMed Central

    Taha, Omnia; Farouk, Inas; Abdallah, Abdelhadi

    2016-01-01

    Squash leaf curl virus (SqLCV) is a bipartite begomovirus affecting squash plants. It is transmitted by whitefly Bemisia tabaci biotype B causing severe leaf curling, vein banding, and molting ending by stunting. In this study full-length genomic clone of SqLCV Egyptian isolated and posttranscriptional gene silencing (PTGS) has been induced to develop virus resistance. The Noubaria SqLCV has more than 95% homology with Jordon, Israel, Lebanon, Palestine, and Cairo isolates. Two genes fragment from SqLCV introduced in sense and antisense orientations using pFGC5049 vector to be expressed as hairpin RNA. The first fragment was 348 bp from replication associated protein gene (Rep). The second fragment was 879 bp representing the full sequence of the movement protein gene (BC1). Using real-time PCR, a silencing record of 97% has been recorded to Rep/TrAP construct; as a result it has prevented the appearance of viral symptoms in most tested plants up to two months after infection, while construct containing the BC1 gene scored a reduction in the accumulation of viral genome expression as appearing in real-time PCR results 4.6-fold giving a silencing of 79%, which had a positive effect on symptoms development in most tested plants. PMID:27034922

  1. Genome organization of Tobacco leaf curl Zimbabwe virus, a new, distinct monopartite begomovirus associated with subgenomic defective DNA molecules.

    PubMed

    Paximadis, M; Rey, M E

    2001-12-01

    The complete DNA A of the begomovirus Tobacco leaf curl Zimbabwe virus (TbLCZWV) was sequenced: it comprises 2767 nucleotides with six major open reading frames encoding proteins with molecular masses greater than 9 kDa. Full-length TbLCZWV DNA A tandem dimers, cloned in binary vectors (pBin19 and pBI121) and transformed into Agrobacterium tumefaciens, were systemically infectious upon agroinoculation of tobacco and tomato. Efforts to identify a DNA B component were unsuccessful. These findings suggest that TbLCZWV is a new member of the monopartite group of begomoviruses. Phylogenetic analysis identified TbLCZWV as a distinct begomovirus with its closest relative being Chayote mosaic virus. Abutting primer PCR amplified ca. 1300 bp molecules, and cloning and sequencing of two of these molecules revealed them to be subgenomic defective DNA molecules originating from TbLCZWV DNA A. Variable symptom severity associated with tobacco leaf curl disease and TbLCZWV is discussed.

  2. Insecticidal Effects on the Spatial Progression of Tomato Yellow Leaf Curl Virus and Movement of Its Whitefly Vector in Tomato.

    PubMed

    Dempsey, M; Riley, D G; Srinivasan, R

    2017-03-08

    Commercial management of whitefly-transmitted Tomato yellow leaf curl virus (TYLCV) typically relies on insecticide control of whitefly vectors as a first line of defense. We quantified this effect in crop tunnel studies, with validation in a tomato field setting. Tomato yellow leaf curl virus-infected and Bemisia tabaci (Gennadius)-infested source plants were planted at the beginning of tunneled rows to serve as inoculum source, so that movement of whiteflies and TYLCV symptoms could be tracked down the length of the tunnel over time. Tunnel study results showed that proximity to the source plant was a more important factor than insecticide treatments. Insecticide-treated tomato transplants did tend to suppress whitefly incidence and slowed TYLCV movement in comparison with the untreated check; however, tomato plants planted closer to the source plant had higher incidence of whiteflies and TYLCV infection, regardless of treatment. In a large tomato plot study with a controlled inoculum source, insecticide treatments significantly reduced the spread of TYLCV. When uninhibited by insecticide treatment, 80% of the TYLCV spread was restricted to <15 m from the source plant (<11 m in the validation study), with insecticide treatment generally reducing the distance and magnitude of this spread.

  3. Molecular characterization and prevalence of two capulaviruses: Alfalfa leaf curl virus from France and Euphorbia caput-medusae latent virus from South Africa.

    PubMed

    Bernardo, Pauline; Muhire, Brejnev; François, Sarah; Deshoux, Maëlle; Hartnady, Penelope; Farkas, Kata; Kraberger, Simona; Filloux, Denis; Fernandez, Emmanuel; Galzi, Serge; Ferdinand, Romain; Granier, Martine; Marais, Armelle; Monge Blasco, Pablo; Candresse, Thierry; Escriu, Fernando; Varsani, Arvind; Harkins, Gordon W; Martin, Darren P; Roumagnac, Philippe

    2016-06-01

    Little is known about the prevalence, diversity, evolutionary processes, genomic structures and population dynamics of viruses in the divergent geminivirus lineage known as the capulaviruses. We determined and analyzed full genome sequences of 13 Euphorbia caput-medusae latent virus (EcmLV) and 26 Alfalfa leaf curl virus (ALCV) isolates, and partial genome sequences of 23 EcmLV and 37 ALCV isolates. While EcmLV was asymptomatic in uncultivated southern African Euphorbia caput-medusae, severe alfalfa disease symptoms were associated with ALCV in southern France. The prevalence of both viruses exceeded 10% in their respective hosts. Besides using patterns of detectable negative selection to identify ORFs that are probably functionally expressed, we show that ALCV and EcmLV both display evidence of inter-species recombination and biologically functional genomic secondary structures. Finally, we show that whereas the EcmLV populations likely experience restricted geographical dispersion, ALCV is probably freely moving across the French Mediterranean region. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. Suppression of Resistance-breaking Beet Necrotic Yellow Vein Virus Isolates by Beet Oak-leaf Virus in Sugar Beet.

    USDA-ARS?s Scientific Manuscript database

    Rhizomania, a very serious disease of sugar beet (Beta vulgaris L.), is caused by Beet necrotic yellow vein virus (BNYVV). Resistance allele Rz1 has been widely incorporated into commercial cultivars. Recently, resistance-breaking strains of Beet necrotic yellow vein virus (RB-BNYVV) were identified...

  5. Molecular Characterization of Tomato Leaf Curl China Virus, Infecting Tomato Plants in China, and Functional Analyses of Its Associated Betasatellite▿†

    PubMed Central

    Yang, Xiuling; Guo, Wei; Ma, Xinying; An, Qianli; Zhou, Xueping

    2011-01-01

    A novel tomato-infecting begomovirus from Guangxi province, China, was identified and characterized, for which the name Tomato leaf curl China virus (ToLCCNV) was proposed. Phylogenetic and recombination analyses of the virus genomic sequences suggested that ToLCCNV may have arisen by recombination among Tomato leaf curl Vietnam virus (ToLCVV), Tomato leaf curl Gujarat virus (ToLCGV), and an unknown virus. A betasatellite molecule was found to be associated with ToLCCNV (ToLCCNB), and its complete nucleotide sequences were determined. Infectious clones of ToLCCNV and ToLCCNB were constructed and then used for agro-inoculation of plants; ToLCCNV alone infected Nicotiana benthamiana, Nicotiana glutinosa, Petunia hybrida, and Solanum lycopersicum plants, but no symptoms were induced. ToLCCNB was required for induction of leaf curl disease in these hosts. The βC1 protein of ToLCCNB was identified as a suppressor of RNA silencing and accumulated primarily in the nucleus. Deletion mutagenesis of βC1 showed that the central part of βC1 (amino acids 44 to 74) was responsible for both the suppressor activity and nuclear localization. PMID:21378048

  6. Cotton Leaf Curl Multan Virus-Derived Viral Small RNAs Can Target Cotton Genes to Promote Viral Infection

    PubMed Central

    Wang, Jinyan; Tang, Yafei; Yang, Yuwen; Ma, Na; Ling, Xitie; Kan, Jialiang; He, Zifu; Zhang, Baolong

    2016-01-01

    RNA silencing is a conserved mechanism in plants that targets viruses. Viral small RNAs (vsiRNAs) can be generated from viral double-stranded RNA replicative intermediates within the infected host, or from host RNA-dependent RNA polymerases activity on viral templates. The abundance and profile of vsiRNAs in viral infections have been reported previously. However, the involvement of vsiRNAs during infection of the Geminiviridae family member cotton leaf curl virus (CLCuD), which causes significant economic losses in cotton growing regions, remains largely uncharacterized. Cotton leaf curl Multan virus (CLCuMuV) associated with a betasatellite called Cotton leaf curl Multan betasatellite (CLCuMuB) is a major constraint to cotton production in South Asia and is now established in Southern China. In this study, we obtained the profiles of vsiRNAs from CLCuMV and CLCuMB in infected upland cotton (Gossypium hirsutum) plants by deep sequencing. Our data showed that vsiRNA that were derived almost equally from sense and antisense CLCuD DNA strands accumulated preferentially as 21- and 22-nucleotide (nt) small RNA population and had a cytosine bias at the 5′-terminus. Polarity distribution revealed that vsiRNAs were almost continuously present along the CLCuD genome and hotspots of sense and antisense strands were mainly distributed in the Rep proteins region of CLCuMuV and in the C1 protein of CLCuMuB. In addition, hundreds of host transcripts targeted by vsiRNAs were predicted, many of which encode transcription factors associated with biotic and abiotic stresses. Quantitative real-time polymerase chain reaction analysis of selected potential vsiRNA targets showed that some targets were significantly down-regulated in CLCuD-infected cotton plants. We also verified the potential function of vsiRNA targets that may be involved in CLCuD infection by virus-induced gene silencing (VIGS) and 5′-rapid amplification of cDNA end (5′-RACE). Here, we provide the first report

  7. Examination of the leaf-drop symptom of virus-infected potato using anther culture-derived haploids.

    PubMed

    Valkonen, J P; Rokka, V M; Watanabe, K N

    1998-10-01

    ABSTRACT Necrotic lesions and vein necrosis characteristic of the hypersensitive response (HR) controlled by the dominant resistance gene Ny develop in potato cv. Pito after infection with potato virus Y ordinary strain (PVY masculine) at a low temperature (16/18 degrees C night/day). In contrast, at high temperatures (19/24 degrees C night/day), large coalesced lesions develop in the lower infected leaves, which wither and remain hanging from stems forming the leaf-drop symptom; mosaic symptoms with no necrosis also develop in the top leaves. The genetic basis of the leaf-drop symptom and its dependence on temperature were examined using a novel approach involving 58 haploids (2n = 24) derived from 'Pito' (2n = 48) through anther culture. These haploids and 'Pito' were graft-inoculated with PVY(O) at 19/24 to 25 degrees C (night/day). Necrotic symptoms were expressed in 28 haploids, of which 18 haploids (phenotype class N) developed top necrosis, vein necrosis, or both and necrotic lesions that are characteristic of HR. Ten haploids showed leaf drop similar to 'Pito' (phenotype class LD). Thirty haploids were susceptible and showed only mosaic symptoms (phenotype class S). These data indicated that necrosis was induced by a single dominant gene, Ny, in the simplex condition. However, the three distinct phenotypic classes (N, LD, and S) among the haploids grown under the same environmental conditions showed that another locus (gene) was involved in modifying the HR triggered by Ny. Data suggested that this locus contains a dominant temperature-dependent modifier (Tdm) gene that alters the expression of PVY-induced HR at higher temperatures, resulting in leaf drop.

  8. Genetic Dissection of Novel QTLs for Resistance to Leaf Spots and Tomato Spotted Wilt Virus in Peanut (Arachis hypogaea L.)

    PubMed Central

    Pandey, Manish K.; Wang, Hui; Khera, Pawan; Vishwakarma, Manish K.; Kale, Sandip M.; Culbreath, Albert K.; Holbrook, C. Corley; Wang, Xingjun; Varshney, Rajeev K.; Guo, Baozhu

    2017-01-01

    Peanut is an important crop, economically and nutritiously, but high production cost is a serious challenge to peanut farmers as exemplified by chemical spray to control foliar diseases such as leaf spots and thrips, the vectors of tomato spotted wilt virus (TSWV). The objective of this research was to map the quantitative trait loci (QTLs) for resistance to leaf spots and TSWV in one recombinant inbred line (RIL) mapping population of “Tifrunner × GT-C20” for identification of linked markers for marker-assisted breeding. Here, we report the improved genetic linkage map with 418 marker loci with a marker density of 5.3 cM/loci and QTLs associated with multi-year (2010–2013) field phenotypes of foliar disease traits, including early leaf spot (ELS), late leaf spot (LLS), and TSWV. A total of 42 QTLs were identified with phenotypic variation explained (PVE) from 6.36 to 15.6%. There were nine QTLs for resistance to ELS, 22 QTLs for LLS, and 11 QTLs for TSWV, including six, five, and one major QTLs with PVE higher than 10% for resistance to each disease, respectively. Of the total 42 QTLs, 34 were mapped on the A sub-genome and eight mapped on the B sub-genome suggesting that the A sub-genome harbors more resistance genes than the B sub-genome. This genetic linkage map was also compared with two diploid peanut physical maps, and the overall co-linearity was 48.4% with an average co-linearity of 51.7% for the A sub-genome and 46.4% for the B sub-genome. The identified QTLs associated markers and potential candidate genes will be studied further for possible application in molecular breeding in peanut genetic improvement for disease resistance. PMID:28197153

  9. Genetic Dissection of Novel QTLs for Resistance to Leaf Spots and Tomato Spotted Wilt Virus in Peanut (Arachis hypogaea L.).

    PubMed

    Pandey, Manish K; Wang, Hui; Khera, Pawan; Vishwakarma, Manish K; Kale, Sandip M; Culbreath, Albert K; Holbrook, C Corley; Wang, Xingjun; Varshney, Rajeev K; Guo, Baozhu

    2017-01-01

    Peanut is an important crop, economically and nutritiously, but high production cost is a serious challenge to peanut farmers as exemplified by chemical spray to control foliar diseases such as leaf spots and thrips, the vectors of tomato spotted wilt virus (TSWV). The objective of this research was to map the quantitative trait loci (QTLs) for resistance to leaf spots and TSWV in one recombinant inbred line (RIL) mapping population of "Tifrunner × GT-C20" for identification of linked markers for marker-assisted breeding. Here, we report the improved genetic linkage map with 418 marker loci with a marker density of 5.3 cM/loci and QTLs associated with multi-year (2010-2013) field phenotypes of foliar disease traits, including early leaf spot (ELS), late leaf spot (LLS), and TSWV. A total of 42 QTLs were identified with phenotypic variation explained (PVE) from 6.36 to 15.6%. There were nine QTLs for resistance to ELS, 22 QTLs for LLS, and 11 QTLs for TSWV, including six, five, and one major QTLs with PVE higher than 10% for resistance to each disease, respectively. Of the total 42 QTLs, 34 were mapped on the A sub-genome and eight mapped on the B sub-genome suggesting that the A sub-genome harbors more resistance genes than the B sub-genome. This genetic linkage map was also compared with two diploid peanut physical maps, and the overall co-linearity was 48.4% with an average co-linearity of 51.7% for the A sub-genome and 46.4% for the B sub-genome. The identified QTLs associated markers and potential candidate genes will be studied further for possible application in molecular breeding in peanut genetic improvement for disease resistance.

  10. A severe symptom phenotype in tomato in Mali is caused by a reassortant between a novel recombinant begomovirus (Tomato yellow leaf curl Mali virus) and a betasatellite.

    PubMed

    Chen, Li-Fang; Rojas, Maria; Kon, Tatsuya; Gamby, Kadiatou; Xoconostle-Cazares, Beatriz; Gilbertson, Robert L

    2009-05-01

    Tomato production in West Africa has been severely affected by begomovirus diseases, including yellow leaf curl and a severe symptom phenotype, characterized by extremely stunted and distorted growth and small deformed leaves. Here, a novel recombinant begomovirus from Mali, Tomato yellow leaf curl Mali virus (TYLCMLV), is described that, alone, causes tomato yellow leaf curl disease or, in combination with a betasatellite, causes the severe symptom phenotype. TYLCMLV is an Old World monopartite begomovirus with a hybrid genome composed of sequences from Tomato yellow leaf curl virus-Mild (TYLCV-Mld) and Hollyhock leaf crumple virus (HoLCrV). A TYLCMLV infectious clone induced leaf curl and yellowing in tomato, leaf curl, crumpling and yellowing in Nicotiana benthamiana and common bean, mild symptoms in N. glutinosa, and a symptomless infection in Datura stramonium. In a field-collected sample from a tomato plant showing the severe symptom phenotype in Mali, TYLCMLV was detected together with a betasatellite, identified as Cotton leaf curl Gezira betasatellite (CLCuGB). Tomato plants co-agroinoculated with TYLCMLV and CLCuGB developed severely stunted and distorted growth and small crumpled leaves. These symptoms were more severe than those induced by TYLCMLV alone, and were similar to the severe symptom phenotype observed in the field in Mali and in other West African countries. TYLCMLV and CLCuGB also induced more severe symptoms than TYLCMLV in the other solanaceous hosts, but not in common bean. The increased symptom severity was associated with hyperplasia of phloem-associated cells, but relatively little increase in TYLCMLV DNA levels. In surveys of tomato virus diseases in West Africa, TYLCMLV was commonly detected in plants with leaf curl and yellow leaf curl symptoms, whereas CLCuGB was infrequently detected and always in association with the severe symptom phenotype. Together, these results indicate that TYLCMLV causes tomato yellow leaf curl disease

  11. Development of plants resistant to Papaya leaf distortion mosaic virus by intergeneric hybridization between Carica papaya and Vasconcellea cundinamarcensis

    PubMed Central

    Tarora, Kazuhiko; Shudo, Ayano; Kawano, Shinji; Yasuda, Keiji; Ueno, Hiroki; Matsumura, Hideo; Urasaki, Naoya

    2016-01-01

    In this study, we confirmed that Vasconcellea cundinamarcensis resists Papaya leaf distortion mosaic virus (PLDMV), and used it to produce intergeneric hybrids with Carica papaya. From the cross between C. papaya and V. cundinamarcensis, we obtained 147 seeds with embryos. Though C. papaya is a monoembryonic plant, multiple embryos were observed in all 147 seeds. We produced 218 plants from 28 seeds by means of embryo-rescue culture. All plants had pubescence on their petioles and stems characteristic of V. cundinamarcensis. Flow cytometry and PCR of 28 plants confirmed they were intergeneric hybrids. To evaluate virus resistance, mechanical inoculation of PLDMV was carried out. The test showed that 41 of 134 intergeneric hybrid plants showed no symptoms and were resistant. The remaining 93 hybrids showed necrotic lesions on the younger leaves than the inoculated leaves. In most of the 93 hybrids, the necrotic lesions enclosed the virus and prevented further spread. These results suggest that the intergeneric hybrids will be valuable material for PLDMV-resistant papaya breeding. PMID:28163589

  12. Chilli leaf curl virus-based vector for phloem-specific silencing of endogenous genes and overexpression of foreign genes.

    PubMed

    Kushwaha, Nirbhay Kumar; Chakraborty, Supriya

    2017-03-01

    Geminiviruses are the largest and most devastating group of plant viruses which contain ssDNA as a genetic material. Geminivirus-derived virus-induced gene silencing (VIGS) vectors have emerged as an efficient and simple tool to study functional genomics in various plants. However, previously developed VIGS vectors have certain limitations, owing to their inability to be used in tissue-specific functional study. In the present study, we developed a Chilli leaf curl virus (ChiLCV)-based VIGS vector for its tissue-specific utilization by replacing the coat protein gene (open reading frame (ORF) AV1) with the gene of interest for phytoene desaturase (PDS) of Nicotiana benthamiana. Functional validation of ChiLCV-based VIGS in N. benthamiana resulted in systemic silencing of PDS exclusively in the phloem region of inoculated plants. Furthermore, expression of enhanced green fluorescence protein (EGFP) using the same ChiLCV vector was verified in the phloem region of the inoculated plants. Our results also suggested that, during the early phase of infection, ChiLCV was associated with the phloem region, but at later stage of pathogenesis, it can spread into the adjoining non-vascular tissues. Taken together, the newly developed ChiLCV-based vector provides an efficient and versatile tool, which can be exploited to unveil the unknown functions of several phloem-specific genes.

  13. Development of plants resistant to Papaya leaf distortion mosaic virus by intergeneric hybridization between Carica papaya and Vasconcellea cundinamarcensis.

    PubMed

    Tarora, Kazuhiko; Shudo, Ayano; Kawano, Shinji; Yasuda, Keiji; Ueno, Hiroki; Matsumura, Hideo; Urasaki, Naoya

    2016-12-01

    In this study, we confirmed that Vasconcellea cundinamarcensis resists Papaya leaf distortion mosaic virus (PLDMV), and used it to produce intergeneric hybrids with Carica papaya. From the cross between C. papaya and V. cundinamarcensis, we obtained 147 seeds with embryos. Though C. papaya is a monoembryonic plant, multiple embryos were observed in all 147 seeds. We produced 218 plants from 28 seeds by means of embryo-rescue culture. All plants had pubescence on their petioles and stems characteristic of V. cundinamarcensis. Flow cytometry and PCR of 28 plants confirmed they were intergeneric hybrids. To evaluate virus resistance, mechanical inoculation of PLDMV was carried out. The test showed that 41 of 134 intergeneric hybrid plants showed no symptoms and were resistant. The remaining 93 hybrids showed necrotic lesions on the younger leaves than the inoculated leaves. In most of the 93 hybrids, the necrotic lesions enclosed the virus and prevented further spread. These results suggest that the intergeneric hybrids will be valuable material for PLDMV-resistant papaya breeding.

  14. The P0 gene of Sugarcane yellow leaf virus encodes an RNA silencing suppressor with unique activities

    SciTech Connect

    Mangwende, Tichaona Wang Mingli Borth, Wayne Hu, John Moore, Paul H. Mirkov, T. Erik Albert, Henrik H.

    2009-02-05

    The Sugarcane yellow leaf virus (SCYLV) P0, a member of the highly heterologous proteins of poleroviruses, is a suppressor of posttranscriptional gene silencing (PTGS) and has additional activities not seen in other P0 proteins. The P0 protein in previously tested poleroviruses (Beet western yellows virus and Cucurbit aphid-borne yellows virus), suppresses local, but not systemic, PTGS induced by both sense GFP and inverted repeat GF using its F-box-like domain to mediate destabilization of the Argonaute1 protein. We now report that the SCYLV P0 protein not only suppressed local PTGS induced by sense GFP and inverted repeat GF in Nicotiana benthamiana, but also triggered a dosage dependent cell death phenotype in infiltrated leaves and suppressed systemic sense GFP-PTGS. Deletion of the first 15 N-terminal amino acid residues of SCYLV P0 abolished suppression of both local and systemic PTGS and the induction of cell death. In contrast, only systemic PTGS and cell death were lost when the 15 C-terminal amino acid residues were deleted. We conclude that the 15 C-terminal amino acid residue region of SCYLV P0 is necessary for suppressing systemic PTGS and inducing cell death, but is not required for suppression of local PTGS.

  15. Field evaluation of yield effects on the U.S.A. heirloom sweet potato cultivars infected by sweet potato leaf curl virus

    USDA-ARS?s Scientific Manuscript database

    The incidence of Sweet potato leaf curl virus (SPLCV), a Begomovirus, infection of sweetpotato Ipomoea batatas (L.) Lam. (Convolvulaceae) in South Carolina, USA has increased rapidly in recent years. This is likely due to the use of infected propagating materials and the increasing population of it...

  16. Transcriptome profiling to discover defense-related genes associated with resistance line ty-5 against Tomato yellow leaf curl virus in tomato

    USDA-ARS?s Scientific Manuscript database

    Tomato yellow leaf curl virus (TYLCV), a whitefly-transmitted begomovirus, has caused serious economic losses to tomato crops in the U.S. and around the world. The most effective management would be the use of a TYLCV-resistant tomato cultivar. Several sources of TYLCV resistance genes have been ide...

  17. Molecular Characterization of the Meyer Lemon Isolate of Citrus Tatter Leaf Virus: Complete Genome Sequence and Development of Biologically Active In Vitro Transcripts

    USDA-ARS?s Scientific Manuscript database

    Citrus tatter leaf virus isolated from Meyer lemon trees (CTLV-ML) from California and Florida induces bud union incompatibility of citrus trees grafted on the widely used trifoliate and trifoliate hybrid rootstocks. The complete genome sequence of CTLV-ML was determined to be 6,495 nucleotides (nts...

  18. A begomovirus associated with Ageratum yellow vein disease in Indonesia: evidence for natural recombination between tomato leaf curl Java virus and Ageratum yellow vein virus-[Java].

    PubMed

    Kon, T; Kuwabara, K; Hidayat, S H; Ikegami, M

    2007-01-01

    A begomovirus (2747 nucleotides) and a satellite DNA beta component (1360 nucleotides) have been isolated from Ageratum conyzoides L. plants with yellow vein symptoms growing in Java, Indonesia. The begomovirus is most closely related to Tomato leaf curl Java virus (ToLCJV) (91 and 98% in the total nucleotide and coat protein amino acid sequences, respectively), although the products of ORFs C1 and C4 are more closely related to those of Ageratum yellow vein virus-[Java] (91 and 95% identity, respectively). For this reason, the begomovirus it is considered to be a strain of ToLCJV and is referred to as ToLCJV-Ageratum. The virus probably derives from a recombination event in which nucleotides 2389-2692 of ToLCJV have been replaced with the corresponding region of the AYVV-[Java] genome, which includes the 5' part of the intergenic region and the C1 and C4 ORFs. Infection of A. conyzoides with ToLCJV-Ageratum alone produced no symptoms, but co-infection with DNAbeta induced yellow vein symptoms. Symptoms induced in Nicotiana benthamiana by ToLCJV-Ageratum, ToLCJV and AYVV-[Java] are consistent with the exchange of pathogenicity determinant ORF C4 during recombination.

  19. Altered gene expression changes in Arabidopsis leaf tissues and protoplasts in response to Plum pox virus infection

    PubMed Central

    Babu, Mohan; Griffiths, Jonathan S; Huang, Tyng-Shyan; Wang, Aiming

    2008-01-01

    Background Virus infection induces the activation and suppression of global gene expression in the host. Profiling gene expression changes in the host may provide insights into the molecular mechanisms that underlie host physiological and phenotypic responses to virus infection. In this study, the Arabidopsis Affymetrix ATH1 array was used to assess global gene expression changes in Arabidopsis thaliana plants infected with Plum pox virus (PPV). To identify early genes in response to PPV infection, an Arabidopsis synchronized single-cell transformation system was developed. Arabidopsis protoplasts were transfected with a PPV infectious clone and global gene expression changes in the transfected protoplasts were profiled. Results Microarray analysis of PPV-infected Arabidopsis leaf tissues identified 2013 and 1457 genes that were significantly (Q ≤ 0.05) up- (≥ 2.5 fold) and downregulated (≤ -2.5 fold), respectively. Genes associated with soluble sugar, starch and amino acid, intracellular membrane/membrane-bound organelles, chloroplast, and protein fate were upregulated, while genes related to development/storage proteins, protein synthesis and translation, and cell wall-associated components were downregulated. These gene expression changes were associated with PPV infection and symptom development. Further transcriptional profiling of protoplasts transfected with a PPV infectious clone revealed the upregulation of defence and cellular signalling genes as early as 6 hours post transfection. A cross sequence comparison analysis of genes differentially regulated by PPV-infected Arabidopsis leaves against uniEST sequences derived from PPV-infected leaves of Prunus persica, a natural host of PPV, identified orthologs related to defence, metabolism and protein synthesis. The cross comparison of genes differentially regulated by PPV infection and by the infections of other positive sense RNA viruses revealed a common set of 416 genes. These identified genes

  20. Molecular dissection of Tomato leaf curl virus resistance in tomato line TY172 derived from Solanum peruvianum.

    PubMed

    Anbinder, Ilana; Reuveni, Moshe; Azari, Raviv; Paran, Ilan; Nahon, Sahadia; Shlomo, Haviva; Chen, Lea; Lapidot, Moshe; Levin, Ilan

    2009-08-01

    Tomato yellow leaf curl virus (TYLCV) is devastating to tomato (Solanum lycopersicum) crops and resistant cultivars are highly effective in controlling the disease. The breeding line TY172, originating from Solanum peruvianum, is highly resistant to TYLCV. To map quantitative trait loci (QTLs) controlling TYLCV resistance in TY172, appropriate segregating populations were analyzed using 69 polymorphic DNA markers spanning the entire tomato genome. Results show that TYLCV resistance in TY172 is controlled by a previously unknown major QTL, originating from the resistant line, and four additional minor QTLs. The major QTL, we term Ty-5, maps to chromosome 4 and accounts for 39.7-46.6% of the variation in symptom severity among segregating plants (LOD score 33-35). The minor QTLs, originated either from the resistant or susceptible parents, were mapped to chromosomes 1, 7, 9 and 11, and contributed 12% to the variation in symptom severity in addition to Ty-5.

  1. Implication of the Whitefly Bemisia tabaci Cyclophilin B Protein in the Transmission of Tomato yellow leaf curl virus

    PubMed Central

    Kanakala, Surapathrudu; Ghanim, Murad

    2016-01-01

    Tomato yellow leaf curl virus (TYLCV) is a single-stranded (ssDNA) begomoviruses that causes severe damage to tomato and several other crops worldwide. TYLCV is exclusively transmitted by the sweetpotato whitefly, Bemisia tabaci in a persistent circulative and propagative manner. Previous studies have shown that the transmission, retention, and circulation of TYLCV in its vector involves interaction with insect and endosymbiont proteins, which aid in the transmission of the virus, or have a protective role in response to the presence of the virus in the insect body. However, only a low number of such proteins have been identified. Here, the role of B. tabaci Cyclophilin B (CypB) in the transmission of TYLCV protein was investigated. Cyclophilins are a large family of cellular prolyl isomerases that have many molecular roles including facilitating protein-protein interactions in the cell. One cyclophilin protein has been implicated in aphid-luteovirus interactions. We demonstrate that the expression of CypB from B. tabaci is altered upon TYLCV acquisition and retention. Further experiments used immunocapture-PCR and co-immunolocalization and demonstrated a specific interaction and colocalization between CypB and TYLCV in the the midgut, eggs, and salivary glands. Membrane feeding of anti-CypB antibodies and TYLCV-infected plants showed a decrease in TYLCV transmission, suggesting a critical role that CypB plays in TYLCV transmission. Further experiments, which used membrane feeding with the CypB inhibitor Cyclosporin A showed decrease in CypB-TYLCV colocalization in the midgut and virus transmission. Altogether, our results indicate that CypB plays an important role in TYLCV transmission by B. tabaci. PMID:27895657

  2. Virus-induced gene silencing of P23k in barley leaf reveals morphological changes involved in secondary wall formation.

    PubMed

    Oikawa, Ai; Rahman, Abidur; Yamashita, Tetsuro; Taira, Hideharu; Kidou, Shin-Ichiro

    2007-01-01

    P23k is a monocot-unique protein that is highly expressed in the scutellum of germinating barley seed. Previous expression analyses suggested that P23k is involved in sugar translocation and/or sugar metabolism. However, the role of P23k in barley physiology remains unclear. Here, to elucidate its physiological function, BSMV-based virus-induced gene silencing (VIGS) of P23k in barley leaves was performed. Expression and localization analyses of P23k mRNA in barley leaves showed up-regulation of P23k transcript with increased photosynthetic activity and the localization of these transcripts to the vascular bundles and sclerenchyma, where secondary wall formation is most active. VIGS of the P23k gene led to abnormal leaf development, asymmetric orientation of main veins, and cracked leaf edges caused by mechanical weakness. In addition, histochemical analyses indicated that the distribution of P23k in leaves coincides with the distribution of cell wall polysaccharides. Considering these results together, it is proposed that P23k is involved in the synthesis of cell wall polysaccharides and contributes to secondary wall formation in barley leaves.

  3. Binding of Human GII.4 Norovirus Virus-Like Particles to Carbohydrates of Romaine Lettuce Leaf Cell Wall Materials

    PubMed Central

    Esseili, Malak A.

    2012-01-01

    Norovirus (NoV) genogroup II genotype 4 (GII.4) strains are the dominant cause of the majority of food-borne outbreaks, including those that involve leafy greens, such as lettuce. Since human NoVs use carbohydrates of histo-blood group antigens as receptors/coreceptors, we examined the role of carbohydrates in the attachment of NoV to lettuce leaves by using virus-like particles (VLPs) of a human NoV/GII.4 strain. Immunofluorescence analysis showed that the VLPs attached to the leaf surface, especially to cut edges, stomata, and along minor veins. Binding was quantified using enzyme-linked immunosorbent assay (ELISA) performed on cell wall materials (CWM) from innermost younger leaves and outermost lamina of older leaves. The binding to CWM of older leaves was significantly (P < 0.05) higher (1.5- to 2-fold) than that to CWM of younger leaves. Disrupting the carbohydrates of CWM or porcine gastric mucin (PGM) (a carbohydrate control) using 100 mM sodium periodate (NaIO4) significantly decreased the binding an average of 17% in younger leaves, 43% in older leaves, and 92% for PGM. In addition, lectins recognizing GalNAc, GlcNAc, and sialic acid at 100 μg/ml significantly decreased the binding an average of 41%, 33%, and 20% on CWM of older leaves but had no effect on younger leaves. Lectins recognizing α-d-Gal, α-d-Man/α-d-Glc, and α-l-Fuc showed significant inhibition on CWM of older leaves as well as that of younger leaves. All lectins, except for the lectin recognizing α-d-Gal, significantly inhibited NoV VLP binding to PGM. Collectively, our results indicate that NoV VLPs bind to lettuce CWM by utilizing multiple carbohydrate moieties. This binding may enhance virus persistence on the leaf surface and prevent effective decontamination. PMID:22138991

  4. Complete genome sequence of an isolate of papaya leaf distortion mosaic virus from commercialized PRSV-resistant transgenic papaya in China.

    PubMed

    Tuo, D; Shen, W; Yan, P; Li, Ch; Gao, L; Li, X; Li, H; Zhou, P

    2013-01-01

    Papaya leaf distortion mosaic virus is highly destructive to commercial papaya production. Here, the complete genome sequence was determined for an isolate of papaya leaf distortion mosaic virus, designated PLDMV-DF, infecting the commercialized papaya ringspot virus (PRSV)-resistant transgenic papaya from China. Excluding the 3'-poly (A) tail, the sequence shares high sequence identity to several PLDMV isolates from Taiwan and Japan and is phylogenetically most closely related to the isolate from Japan. Infection of PLDMV-DF in transgenic PRSV-resistant papaya may indicate emergence of this disease in genetically engineered plants. The reported sequence for this isolate may help generate bi-transgenic papaya resistant to PRSV and PLDMV.

  5. Real-time quantitative PCR assay for the quantification of virus and satellites causing leaf curl disease in cotton in Pakistan.

    PubMed

    Shafiq, Muhammad; Iqbal, Zafar; Ali, Irfan; Abbas, Qamar; Mansoor, Shahid; Briddon, Rob W; Amin, Imran

    2017-10-01

    Cotton leaf curl disease (CLCuD) is the major biotic constraint to cotton production in Pakistan and northwestern India. The disease is caused by monopartite begomoviruses in association with a specific DNA satellite, Cotton leaf curl Multan betasatellite. The virus-betasatellite complex is also frequently associated with another DNA satellite-like molecule; an alphasatellite. A quantitative real-time PCR (qPCR) assay to detect all three components of the monopartite begomovirus/betasatellite/alphasatellite complex which causes CLCuD was established. This was used to investigate the relationship between symptoms and virus/satellite titre. Not surprisingly the analysis showed that, overall, there was a reasonable correlation between symptom severity and virus/satellite titre - more severe symptoms usually being associated with more virus/satellite. However, cotton plants were identified with no or very mild symptoms with relatively high virus/satellite titres and plants with severe symptoms but relatively low virus/satellite titres. This may be attributed to the resistance/susceptibility of the cotton variety - tolerant plants being able to sustain a relatively high virus/satellite titre whilst exhibiting mild symptoms. The usefulness of this qPCR procedure in the screening for resistance in cotton against CLCuD is discussed. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Rolling-circle amplification of Torque teno virus (TTV) complete genomes from human and swine sera and identification of a novel swine TTV genogroup.

    PubMed

    Niel, Christian; Diniz-Mendes, Leonardo; Devalle, Sylvie

    2005-05-01

    Multiply primed rolling-circle amplification is a novel technology that uses bacteriophage phi29 DNA polymerase to amplify circular DNA molecules, without the need for prior knowledge of their sequences. In an attempt to detect Torque teno virus (TTV), rolling-circle amplification was used to amplify DNA extracted from eight human and four pig serum samples. All samples gave high molecular weight (>30 kb) amplification products. By restriction endonuclease digestion, these products generated DNA fragments whose sizes were consistent with those of human TTV (3.8 kb) and swine TTV (Sd-TTV; 2.9 kb) genomes. Two TTV isolates derived from a single AIDS patient, as well as two Sd-TTV isolates derived from a single pig, were characterized by complete nucleotide sequencing. One of the Sd-TTV isolates showed very low (43-45 %) nucleotide sequence similarity to the other Sd-TTV isolate and to the prototype isolate Sd-TTV31, and could be considered the prototype of a novel genogroup.

  7. Association of tomato leaf curl New Delhi virus DNA-B with bhendi yellow vein mosaic virus in okra showing yellow vein mosaic disease symptoms.

    PubMed

    Venkataravanappa, V; Lakshminarayana Reddy, C N; Jalali, S; Krishna Reddy, M

    2015-06-01

    Okra samples showing yellow vein mosaic, vein twisting and bushy appearance were collected from different locations of India during the surveys conducted between years 2005-2009. The dot blot and PCR detection revealed that 75.14% of the samples were associated with monopartite begomovirus and remaining samples with bipartite virus. Whitefly transmission was established for three samples representing widely separated geographical locations which are negative to betasatellites and associated with DNA-B. Genome components of these three representative isolates were cloned and sequenced. The analysis of DNA-A-like sequence revealed that three begomovirus isolates shared more than 93% nucleotide sequence identity with bhendi yellow vein mosaic virus from India (BYVMV), a monopartite begomovirus species that was reported previously as causative agent of bhendi yellow mosaic disease in association of bhendi yellow vein mosaic betasatellite. Further, the DNA-B-like sequences associated with the three virus isolates shared no more than 90% sequence identity with tomato leaf curl New Delhi virus (ToLCNDV). Analyses of putative iteron-binding sequence required for trans-replication suggests that begomovirus sequences shared compatible rep-binding iterons with DNA-B of ToLCNDV. Our data suggest that the monopartite begomovirus associated with okra yellow vein disease has captured DNA-B of ToLCNDV to infect okra. Widespread distribution of the complex shows the increasing trend of the capturing of DNA-B of ToLCNDV by monopartite begomoviruses in the Indian subcontinent. The recombination analysis showed that the DNA-A might have been derived from the inter-specific recombination of begomoviruses, while DNA-B was derived from the ToLCNDV infecting different hosts.

  8. Rolling Reloaded

    ERIC Educational Resources Information Center

    Jones, Simon A.; Nieminen, John M.

    2008-01-01

    Not so long ago a new observation about rolling motion was described: for a rolling wheel, there is a set of points with instantaneous velocities directed at or away from the centre of the wheel; these points form a circle whose diameter connects the centre of the wheel to the wheel's point of contact with the ground (Sharma 1996 "Eur. J. Phys."…

  9. Genetic diversity and distribution of a distinct strain of Chili leaf curl virus and associated betasatellite infecting tomato and pepper in Oman.

    PubMed

    Khan, Akhtar J; Akhtar, Sohail; Al-Zaidi, Amal M; Singh, Achuit K; Briddon, Rob W

    2013-10-01

    Tomato and pepper are widely grown in Oman for local consumption. A countrywide survey was conducted during 2010-2011 to collect samples and assess the diversity of begomoviruses associated with leaf curl disease of tomato and pepper. A virus previously only identified on the Indian subcontinent, chili leaf curl virus (ChLCV), was found associated with tomato and pepper diseases in all vegetable grown areas of Oman. Some of the infected plant samples were also found to contain a betasatellite. A total of 19 potentially full-length begomovirus and eight betasatellite clones were sequenced. The begomovirus clones showed >96% nucleotide sequence identity, showing them to represent a single species. Comparisons to sequences available in the databases showed the highest levels of nucleotide sequence identity (88.0-91.1%) to isolates of the "Pakistan" strain of ChLCV (ChLCV-PK), indicating the virus from Oman to be a distinct strain, for which the name Oman strain (ChLCV-OM) is proposed. An analysis for recombination showed ChLCV-OM likely to have originated by recombination between ChLCV-PK (the major parent), pepper leaf curl Lahore virus and a third strain of ChLCV. The betasatellite sequences obtained were shown to have high levels of identity to isolates of tomato leaf curl betasatellite (ToLCB) previous shown to be present in Oman. For the disease in tomato Koch's postulates were satisfied by Agrobacterium-mediated inoculation of virus and betasatellites clones. This showed the symptoms induced by the virus in the presence of the betasatellite to be enhanced, although viral DNA levels were not affected. ChLCV-OM is the fourth begomovirus identified in tomato in Oman and the first in Capsicum. The significance of these findings is discussed.

  10. Inhibition of binding of tomato yellow leaf curl virus rep to its replication origin by artificial zinc-finger protein.

    PubMed

    Mori, Tomoaki; Takenaka, Kosuke; Domoto, Fumiya; Aoyama, Yasuhiro; Sera, Takashi

    2013-06-01

    Previously we demonstrated that inhibition of replication-associated protein (Rep) binding to its replication origin by artificial zinc-finger proteins (AZPs) is a powerful method to prevent plant virus infection in vivo. In the present study, we applied the AZP technology to Tomato yellow leaf curl virus (TYLCV), which is a limiting factor in tomato cultivation worldwide. First, we determined 5'-ATCGGTGT ATCGGTGT-3' in the 195-bp intergenic region of the TYLCV-Israel strain, a strain reported first among TYLCV strains, as the Rep-binding site by gel shift assays. We then constructed a 6-finger AZP that bound to a 19-bp DNA including the Rep-binding site. We demonstrated that the binding affinity of the AZP was >1,000-fold greater than that of Rep and that the AZP inhibited Rep binding completely in vitro. Because the binding capability of the AZP was same as that of the AZP previously designed for geminivirus-resistant Arabidopsis thaliana, we predict that the present AZP will prevent TYLCV infection in vivo.

  11. Rapid accumulation and low degradation: key parameters of Tomato yellow leaf curl virus persistence in its insect vector Bemisia tabaci.

    PubMed

    Becker, Nathalie; Rimbaud, Loup; Chiroleu, Frédéric; Reynaud, Bernard; Thébaud, Gaël; Lett, Jean-Michel

    2015-12-02

    Of worldwide economic importance, Tomato yellow leaf curl virus (TYLCV, Begomovirus) is responsible for one of the most devastating plant diseases in warm and temperate regions. The DNA begomoviruses (Geminiviridae) are transmitted by the whitefly species complex Bemisia tabaci. Although geminiviruses have long been described as circulative non-propagative viruses, observations such as long persistence of TYLCV in B. tabaci raised the question of their possible replication in the vector. We monitored two major TYLCV strains, Mild (Mld) and Israel (IL), in the invasive B. tabaci Middle East-Asia Minor 1 cryptic species, during and after the viral acquisition, within two timeframes (0-144 hours or 0-20 days). TYLCV DNA was quantified using real-time PCR, and the complementary DNA strand of TYLCV involved in viral replication was specifically quantified using anchored real-time PCR. The DNA of both TYLCV strains accumulated exponentially during acquisition but remained stable after viral acquisition had stopped. Neither replication nor vertical transmission were observed. In conclusion, our quantification of the viral loads and complementary strands of both Mld and IL strains of TYLCV in B. tabaci point to an efficient accumulation and preservation mechanism, rather than to a dynamic equilibrium between replication and degradation.

  12. Rapid accumulation and low degradation: key parameters of Tomato yellow leaf curl virus persistence in its insect vector Bemisia tabaci

    PubMed Central

    Becker, Nathalie; Rimbaud, Loup; Chiroleu, Frédéric; Reynaud, Bernard; Thébaud, Gaël; Lett, Jean-Michel

    2015-01-01

    Of worldwide economic importance, Tomato yellow leaf curl virus (TYLCV, Begomovirus) is responsible for one of the most devastating plant diseases in warm and temperate regions. The DNA begomoviruses (Geminiviridae) are transmitted by the whitefly species complex Bemisia tabaci. Although geminiviruses have long been described as circulative non-propagative viruses, observations such as long persistence of TYLCV in B. tabaci raised the question of their possible replication in the vector. We monitored two major TYLCV strains, Mild (Mld) and Israel (IL), in the invasive B. tabaci Middle East-Asia Minor 1 cryptic species, during and after the viral acquisition, within two timeframes (0–144 hours or 0–20 days). TYLCV DNA was quantified using real-time PCR, and the complementary DNA strand of TYLCV involved in viral replication was specifically quantified using anchored real-time PCR. The DNA of both TYLCV strains accumulated exponentially during acquisition but remained stable after viral acquisition had stopped. Neither replication nor vertical transmission were observed. In conclusion, our quantification of the viral loads and complementary strands of both Mld and IL strains of TYLCV in B. tabaci point to an efficient accumulation and preservation mechanism, rather than to a dynamic equilibrium between replication and degradation. PMID:26625871

  13. Functional Characterization of a Bidirectional Plant Promoter from Cotton Leaf Curl Burewala Virus Using an Agrobacterium-Mediated Transient Assay

    PubMed Central

    Ashraf, Muhammad Aleem; Shahid, Ahmad Ali; Rao, Abdul Qayyum; Bajwa, Kamran Shehzad; Husnain, Tayyab

    2014-01-01

    The C1 promoter expressing the AC1 gene, and V1 promoter expressing the AV1 gene are located in opposite orientations in the large intergenic region of the Cotton leaf curl Burewala virus (CLCuBuV) genome. Agro-infiltration was used to transiently express putative promoter constructs in Nicotiana tabacum and Gossypium hirsutum leaves, which was monitored by a GUS reporter gene, and revealed that the bidirectional promoter of CLCuBuV transcriptionally regulates both the AC1 and AV1 genes. The CLCuBuV C1 gene promoter showed a strong, consistent transient expression of the reporter gene (GUS) in N. tabacum and G. hirsutum leaves and exhibited GUS activity two- to three-fold higher than the CaMV 35S promoter. The CLCuBuV bidirectional genepromoter is a nearly constitutive promoter that contains basic conserved elements. Many cis-regulatory elements (CREs) were also analyzed within the bidirectional plant promoters of CLCuBuV and closely related geminiviruses, which may be helpful in understanding the transcriptional regulation of both the virus and host plant. PMID:24424501

  14. Functional Characterization of a Strong Bi-directional Constitutive Plant Promoter Isolated from Cotton Leaf Curl Burewala Virus

    PubMed Central

    Khan, Zainul A.; Abdin, Malik Z.; Khan, Jawaid A.

    2015-01-01

    Cotton leaf curl Burewala virus (CLCuBuV), belonging to the genus Begomovirus, possesses single-stranded monopartite DNA genome. The bidirectional promoters representing Rep and coat protein (CP) genes of CLCuBuV were characterized and their efficacy was assayed. Rep and CP promoters of CLCuBuV and 35S promoter of Cauliflower mosaic virus (CaMV) were fused with β-glucuronidase (GUS) and green fluorescent protein (GFP) reporter genes. GUS activity in individual plant cells driven by Rep, CP and 35S promoters was estimated using real-time PCR and fluorometric GUS assay. Histochemical staining of GUS in transformed tobacco (Nicotiana tabacum cv. Xanthi) leaves showed highest expression driven by Rep promoter followed by 35S promoter and CP promoter. The expression level of GUS driven by Rep promoter in transformed tobacco plants was shown to be two to four-fold higher than that of 35S promoter, while the expression by CP promoter was slightly lower. Further, the expression of GFP was monitored in agroinfiltrated leaves of N. benthamiana, N. tabacum and cotton (Gossypium hirsutum) plants using confocal laser scanning microscopy. Rep promoter showed strong consistent transient expression in tobacco and cotton leaves as compared to 35S promoter. The strong constitutive CLCuBuV Rep promoter developed in this study could be very useful for high level expression of transgenes in a wide variety of plant cells. PMID:25799504

  15. Tomato plant cell death induced by inhibition of HSP90 is alleviated by Tomato yellow leaf curl virus infection.

    PubMed

    Moshe, Adi; Gorovits, Rena; Liu, Yule; Czosnek, Henryk

    2016-02-01

    To ensure a successful long-term infection cycle, begomoviruses must restrain their destructive effect on host cells and prevent drastic plant responses, at least in the early stages of infection. The monopartite begomovirus Tomato yellow leaf curl virus (TYLCV) does not induce a hypersensitive response and cell death on whitefly-mediated infection of virus-susceptible tomato plants until diseased tomatoes become senescent. The way in which begomoviruses evade plant defences and interfere with cell death pathways is still poorly understood. We show that the chaperone HSP90 (heat shock protein 90) and its co-chaperone SGT1 (suppressor of the G2 allele of Skp1) are involved in the establishment of TYLCV infection. Inactivation of HSP90, as well as silencing of the Hsp90 and Sgt1 genes, leads to the accumulation of damaged ubiquitinated proteins and to a cell death phenotype. These effects are relieved under TYLCV infection. HSP90-dependent inactivation of 26S proteasome degradation and the transcriptional activation of the heat shock transcription factors HsfA2 and HsfB1 and of the downstream genes Hsp17 and Apx1/2 are suppressed in TYLCV-infected tomatoes. Following suppression of the plant stress response, TYLCV can replicate and accumulate in a permissive environment. © 2015 BSPP AND JOHN WILEY & SONS LTD.

  16. Effects of Isaria fumosorosea on TYLCV (Tomato Yellow Leaf Curl Virus) Accumulation and Transmitting Capacity of Bemisia tabaci

    PubMed Central

    2016-01-01

    Tomato yellow leaf curl virus (TYLCV) is transmitted by the Bemisia tabaci pest Middle East-Asia Minor 1 (MEAM1) in China. Isaria fumosorosea is a fungal pathogen of B. tabaci. However, the effects of fungal infection on TYLCV expression and transmission by MEAM1 are unclear. In this study, potted tomatoes containing second instar nymphs of MEAM1 were treated with I. fumosorosea IfB01 strain and the relationship between fungal infection in MEAM1 and its TYLCV transmission capacity was investigated. The results indicated that a significantly (p < 0.05) decreased incidence of transmission of TYLCV-infected plants (ITYPs) transmitted by second instar nymphs of MEAM1 infected with fungus. Further, we found a negative correlation between fungal conidial concentrations and eclosion rates of MEAM1, and a positive correlation between ITYPs and eclosion. In addition, when each plant was exposed to three adults treated with fungus, a significantly decreased transmission of TYLCV (TYTE) was observed in the infected group. However, the incidence of TYLCV-carrying MEAM1 adults (ITYAs) was not significantly different in the infected and control groups (p < 0.05). Nevertheless, a significant decrease in viral accumulation using TYLCV AC2 gene as a marker was observed in the fungus-infected MEAM1. In conclusion, the results suggested that I. fumosorosea infection decreases TYLCV accumulation in MEAM1 and subsequently reduces its transmission. Our study provides new insights into the relationship between host plant, plant virus, insect vector, and entomopathogenic fungus. PMID:27716852

  17. 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

  18. Fine mapping of the tomato yellow leaf curl virus resistance gene Ty-2 on chromosome 11 of tomato.

    PubMed

    Yang, Xiaohui; Caro, Myluska; Hutton, Samuel F; Scott, John W; Guo, Yanmei; Wang, Xiaoxuan; Rashid, Md Harunur; Szinay, Dora; de Jong, Hans; Visser, Richard G F; Bai, Yuling; Du, Yongchen

    2014-01-01

    Resistances to begomoviruses, including bipartite tomato mottle virus and monopartite tomato yellow leaf curl virus (TYLCV), have been introgressed to cultivated tomato (Solanum lycopersicum) from wild tomato accessions. A major gene, Ty-2 from S. habrochaites f. glabratum accession "B6013," that confers resistance to TYLCV was previously mapped to a 19-cM region on the long arm of chromosome 11. In the present study, approximately 11,000 plants were screened and nearly 157 recombination events were identified between the flanking markers C2_At1g07960 (82.5 cM, physical distance 51.387 Mb) and T0302 (89 cM, 51.878 Mb). Molecular marker analysis of recombinants and TYLCV evaluation of progeny from these recombinants localized Ty-2 to an approximately 300,000-bp interval between markers UP8 (51.344 Mb) and M1 (51.645 Mb). No recombinants were identified between TG36 and C2_At3g52090, a region of at least 115 kb, indicating severe recombination suppression in this region. Due to the small interval, fluorescence in situ hybridization analysis failed to clarify whether recombination suppression is caused by chromosomal rearrangements. Candidate genes predicted based on tomato genome annotation were analyzed by RT-PCR and virus-induced gene silencing. Results indicate that the NBS gene family present in the Ty-2 region is likely not responsible for the Ty-2-conferred resistance and that two candidate genes might play a role in the Ty-2-conferred resistance. Several markers very tightly linked to the Ty-2 locus are presented and useful for marker-assisted selection in breeding programs to introgress Ty-2 for begomovirus resistance.

  19. Virus-induced gene silencing (VIGS) of genes expressed in root, leaf, and meiotic tissues of wheat.

    PubMed

    Bennypaul, Harvinder S; Mutti, Jasdeep S; Rustgi, Sachin; Kumar, Neeraj; Okubara, Patricia A; Gill, Kulvinder S

    2012-03-01

    Barley stripe mosaic virus (BSMV)-based virus-induced gene silencing (VIGS) is an effective strategy for rapid functional analysis of genes in wheat leaves, but its utility to transiently express genes, and silencing in other tissues including root, flower, and developing grains, has not been demonstrated in monocots. We monitored green fluorescent protein (GFP) expression to demonstrate the utility of BSMV as a transient expression vector and silenced genes in various wheat tissues to expand VIGS utility to characterize tissue-specific genes. An antisense construct designed for coronatine insensitive1 (COI1) showed an 85% decrease in COI1 transcript level in roots accompanied by a 26% reduction in root length. Similarly, silencing of seed-specific granule-bound starch synthase by antisense and hairpin constructs resulted in up to 82% reduction in amylose content of the developing grains. VIGS of meiosis-specific genes demonstrated by silencing wheat homologue of disrupted meiosis cDNA1 (DMC1) by an antisense construct resulted in a 75-80% reduction in DMC1 transcript level accompanied by an average of 37.2 univalents at metaphase I. The virus-based transient GFP expression was observed in the leaf, phloem, and root cortex at 10-17 days post-inoculation. A novel observation was made that 8-11% of the first selfed generation progeny showed VIGS inheritance and that this proportion increased to 53-72% in the second and to 90-100% in the third generations. No viral symptoms were observed in the progeny, making it possible to study agronomic traits by VIGS. VIGS inheritance is particularly useful to study genes expressing during seed germination or other stages of early plant growth.

  20. Rolling Ribbons

    NASA Astrophysics Data System (ADS)

    Raux, P. S.; Reis, P. M.; Bush, J. W. M.; Clanet, C.

    2010-07-01

    We present the results of a combined experimental and theoretical investigation of rolling elastic ribbons. Particular attention is given to characterizing the steady shapes that arise in static and dynamic rolling configurations. In both cases, above a critical value of the forcing (either gravitational or centrifugal), the ribbon assumes a two-lobed, peanut shape similar to that assumed by rolling droplets. Our theoretical model allows us to rationalize the observed shapes through consideration of the ribbon’s bending and stretching in response to the applied forcing.

  1. Microsatellite Diversity of Soybean Genotypes Differing in Bean Pod Mottle Virus Leaf Symptom

    USDA-ARS?s Scientific Manuscript database

    Soybean, Glycine max (L.) Merr, is the most important source of vegetable oil and protein meal in the world. Bean pod mottle virus (BPMV) is a threat to soybean yield and productivity in most soybean growing states of the USA. In the absence of complete resistance to BPMV, partial resistance of so...

  2. How to pattern a leaf

    USDA-ARS?s Scientific Manuscript database

    Leaf development presents a tremendous resource for tackling the question of patterning in biology. Leaves can be simple or highly dissected. They may have elaborated parts such as the tendrils of a pea leaf or the rolled blade of a carnivorous pitcher plant. Despite the variation in size, shape, an...

  3. Colorimetric monitoring of rolling circle amplification for detection of H5N1 influenza virus using metal indicator.

    PubMed

    Hamidi, Seyed Vahid; Ghourchian, Hedayatollah

    2015-10-15

    A new colorimetric method for monitoring of rolling circle amplification was developed. At first H5N1 target hybrids with padlock probe (PLP) and then PLP is circularized upon the action of T4 ligase enzyme. Subsequently, the circular probe is served as a template for hyperbranched rolling circle amplification (HRCA) by utilizing Bst DNA polymerase enzyme. By improving the reaction, pyrophosphate is produced via DNA polymerization and chelates the Mg(2+) in the buffer solution. This causes change in solution color in the presence of hydroxy naphthol blue (HNB) as a metal indicator. By using pH shock instead of heat shock and isothermal RCA reaction not only the procedure becomes easier, but also application of HNB for colorimetric detection of RCA reaction further simplifies the assay. The responses of the biosensor toward H5N1 were linear in the concentration range from 0.16 to 1.20 pM with a detection limit of 28 fM.

  4. Effect of Raspberry bushy dwarf virus, Raspberry leaf mottle virus, and Raspberry latent virus on plant growth and fruit crumbliness in ‘Meeker’ red Raspberry

    USDA-ARS?s Scientific Manuscript database

    Raspberry crumbly fruit in red raspberry (Rubus idaeus L.), widespread in the Pacific Northwest of the United States and British Columbia, Canada, is most commonly caused by a virus infection. Raspberry bushy dwarf virus (RBDV) has long been attributed as the causal agent of the disease. Recently, t...

  5. Rolling Robot

    NASA Technical Reports Server (NTRS)

    Larimer, Stanley J.; Lisec, Thomas R.; Spiessbach, Andrew J.

    1990-01-01

    Proposed rolling robot routinely traverses rough terrain, clearing rocks as high as 1 m. Climbs steps 1 m high and spans ditches 2.3 m wide. Simple but rugged semiautonomous rover has large wheels and articulated body. With combined yaw, roll, and four-wheel drive, robot crawls slowly to pass over soft or sandy terrain. Senses terrain along corridor, chooses path to avoid insurmountable obstacles, and monitors state of vehicle for unexpected hazards.

  6. Nucleotide sequences of a Korean isolate of apple stem grooving virus associated with black necrotic leaf spot disease on pear (Pyrus pyrifolia).

    PubMed

    Shim, Hyekyung; Min, Yeonju; Hong, Sungyoul; Kwon, Moonsik; Kim, Daehyun; Kim, Hyunran; Choi, Yongmoon; Lee, Sukchan; Yang, Jaemyung

    2004-10-31

    Pear black necrotic leaf spot (PBNLS) is a disease of pears caused by capillovirus-like particles, which can be observed under the electron microscope. The disease was analyzed by Western blot analysis with antisera raised against apple stem grooving virus (ASGV) coat protein. cDNAs covering the entire genome were synthesized by RT-PCR and RACE using RNA isolated from Chenopodium quinoa infected with sap extracted from pear leaves carrying black necrotic spot disease. The complete genome sequence of the putative pear virus, 6497 nucleotides in length excluding the poly (A) tail, was determined and analyzed. It contains two overlapping open reading frames (ORFs). ORF1, spans from nucleotide position 37 to 6354, producing a putative protein of 241 kDa. ORF2, which is in a different reading frame within ORF1, begins at nucleotide 4788 and terminates at 5750, and produces a putative protein of 36 kDa. The 241 kDa protein contains sequences related to the NTP-binding motifs of helicases and RNA-dependent RNA polymerases. The 36-kDa protein contains the consensus sequence GDSG found in the active sites of several cellular and viral serine proteases. Morphological and serological analysis, and sequence comparison between the putative pear virus, ASGV, citrus tatter leaf virus and cherry virus A of the capillovirus suggest that PBNLS may be caused by a Korean isolate of ASGV.

  7. Improved rolling circle amplification (RCA) of hepatitis B virus (HBV) relaxed-circular serum DNA (RC-DNA).

    PubMed

    Martel, Nora; Gomes, Selma A; Chemin, Isabelle; Trépo, Christian; Kay, Alan

    2013-11-01

    For functional analysis of HBV isolates, epidemiological studies and correct identification of recombinant genomes, the amplification of complete genomes is necessary. A method for completely in vitro amplification of full-length HBV genomes starting from serum RC-DNA is described. This uses in vitro completion/ligation of plus-strand HBV RC-DNA and amplification using Rolling-Circle Amplification, eventually followed by a genomic PCR. The method can amplify complete HBV genomes from sera with viral loads ranging from >1.0E+8 IU/ml down to 1.0E+3 IU/ml. The method can be applied to archived sera that have undergone long-term storage or to archived DNA serum extracts. The genomes can easily be cloned. HBV genotypes A-G can all be amplified with no apparent problems. A recombinant subgenotype A3/genotype E genome was identified and fully sequenced.

  8. A developmentally regulated lipocalin-like gene is overexpressed in Tomato yellow leaf curl virus-resistant tomato plants upon virus inoculation, and its silencing abolishes resistance.

    PubMed

    Sade, Dagan; Eybishtz, Assaf; Gorovits, Rena; Sobol, Iris; Czosnek, Henryk

    2012-10-01

    To discover genes involved in tomato resistance to Tomato yellow leaf curl virus (TYLCV), we previously compared cDNA libraries from susceptible (S) and resistant (R) tomato lines. Among the genes preferentially expressed in R plants and upregulated by TYLCV infection was a gene encoding a lipocalin-like protein. This gene was termed Solanum lycopersicum virus resistant/susceptible lipocalin (SlVRSLip). The SlVRSLip structural gene sequence of R and S plants was identical. SlVRSLip was expressed in leaves during a 15-day window starting about 40 days after sowing (20 days after planting). SlVRSLip was upregulated by Bemisia tabaci (the TYLCV vector) feeding on R plant leaves, and even more strongly upregulated following whitefly-mediated TYLCV inoculation. Silencing of SlVRSLip in R plants led to the collapse of resistance upon TYLCV inoculation and to a necrotic response along the stem and petioles accompanied by ROS production. Contrary to previously identified tomato lipocalin gene DQ222981, SlVRSLip was not regulated by cold, nor was it regulated by heat or salt. The expression of SlVRSLip was inhibited in R plants in which the hexose transporter gene LeHT1 was silenced. In contrast, the expression of LeHT1 was not inhibited in SlVRSLip-silenced R plants. Hence, in the hierarchy of the gene network conferring TYLCV resistance, SlVRSLip is downstream of LeHT1. Silencing of another gene involved in resistance, a Permease-I like protein, did not affect the expression of SlVRSLip and LeHT1; expression of the Permease was not affected by silencing SlVRSLip or LeHT1, suggesting that it does not belong to the same network. The triple co-silencing of SlVRSLip, LeHT1 and Permease provoked an immediate cessation of growth of R plants upon infection and the accumulation of large amounts of virus. SlVRSLip is the first lipocalin-like gene shown to be involved in resistance to a plant virus.

  9. Phylogenetic analysis and inflow route of Tomato yellow leaf curl virus (TYLCV) and Bemisia tabaci in Korea.

    PubMed

    Lee, Hyejung; Song, Woogeun; Kwak, Hae-Ryun; Kim, Jae-Deok; Park, Jungan; Auh, Chung-Kyoon; Kim, Dae-Hyun; Lee, Kyeong-Yeoll; Lee, Sukchan; Choi, Hong-Soo

    2010-11-01

    Tomato yellow leaf curl virus (TYLCV) is a member of the genus Begomovirus of the family Geminiviridae, members of which are characterized by closed circular single-stranded DNA genomes of 2.7-2.8 kb in length, and include viruses transmitted by the Bemisia tabaci whitefly. No reports of TYLCV in Korea are available prior to 2008, after which TYLCV spread rapidly to most regions of the southern Korean peninsula (Gyeongsang-Do, Jeolla-Do and Jeju-Do). Fifty full sequences of TYLCV were analyzed in this study, and the AC1, AV1, IR, and full sequences were analyzed via the muscle program and bayesian analysis. Phylogenetic analysis demonstrated that the Korea TYLCVs were divided into two subgroups. The TYLCV Korea 1 group (Masan) originated from TYLCV Japan (Miyazaki) and the TYLCV Korea 2 group (Jeju/Jeonju) from TYLCV Japan (Tosa/Haruno). A B. tabaci phylogenetic tree was constructed with 16S rRNA and mitochondria cytochrome oxidase I (MtCOI) sequences using the muscle program and MEGA 4.0 in the neighbor-joining algorithm. The sequence data of 16S rRNA revealed that Korea B. tabaci was closely aligned to B. tabaci isolated in Iran and Nigeria. The Q type of B. tabaci, which was originally identified as a viruliferous insect in 2008, was initially isolated in Korea as a non-viruliferous insect in 2005. Therefore, we suggest that two TYLCV Japan isolates were introduced to Korea via different routes, and then transmitted by native B. tabaci.

  10. Resistance to tomato leaf curl New Delhi virus in melon is controlled by a major QTL located in chromosome 11.

    PubMed

    Sáez, Cristina; Esteras, Cristina; Martínez, Cecilia; Ferriol, María; Dhillon, Narinder P S; López, Carmelo; Picó, Belén

    2017-07-14

    Identification of three genomic regions and underlying candidate genes controlling the high level of resistance to ToLCNDV derived from a wild melon. SNP markers appropriated for MAS management of resistance. Tomato leaf curl New Delhi virus (ToLCNDV) is a bipartite begomovirus that severely affects melon crop (Cucumis melo) in the main production areas of Spain since 2012. In this work, we evaluated the degree of resistance of four accessions (two belonging to the subsp. agrestis var. momordica and two to the wild agrestis group) and their corresponding hybrids with a susceptible commercial melon belonging to the subsp. melo (Piel de Sapo, PS). The analysis using quantitative PCR (qPCR) allowed us to select one wild agrestis genotype (WM-7) with a high level of resistance and use it to construct segregating populations (F 2 and backcrosses). These populations were phenotyped for symptom severity and virus content using qPCR, and genotyped with different sets of SNP markers. Phenotyping and genotyping results in the F 2 and BC1s populations derived from the WM-7 × PS cross were used for QTL analysis. Three genomic regions controlling resistance to ToLCNDV were found, one major locus in chromosome 11 and two additional regions in chromosomes 12 and 2. The highest level of resistance (no or mild symptoms and very low viral titer) was obtained with the homozygous WM-7WM-7 genotype at the major QTL in chromosome 11, even with PSPS genotypes at the other two loci. The resistance derived from WM-7 is useful to develop new melon cultivars and the linked SNPs selected in this paper will be highly useful in marker-assisted breeding for ToLCNDV resistance in melon.

  11. Discovering host genes involved in the infection by the Tomato Yellow Leaf Curl Virus complex and in the establishment of resistance to the virus using Tobacco Rattle Virus-based post transcriptional gene silencing.

    PubMed

    Czosnek, Henryk; Eybishtz, Assaf; Sade, Dagan; Gorovits, Rena; Sobol, Iris; Bejarano, Eduardo; Rosas-Díaz, Tábata; Lozano-Durán, Rosa

    2013-03-22

    The development of high-throughput technologies allows for evaluating gene expression at the whole-genome level. Together with proteomic and metabolomic studies, these analyses have resulted in the identification of plant genes whose function or expression is altered as a consequence of pathogen attacks. Members of the Tomato yellow leaf curl virus (TYLCV) complex are among the most important pathogens impairing production of agricultural crops worldwide. To understand how these geminiviruses subjugate plant defenses, and to devise counter-measures, it is essential to identify the host genes affected by infection and to determine their role in susceptible and resistant plants. We have used a reverse genetics approach based on Tobacco rattle virus-induced gene silencing (TRV-VIGS) to uncover genes involved in viral infection of susceptible plants, and to identify genes underlying virus resistance. To identify host genes with a role in geminivirus infection, we have engineered a Nicotiana benthamiana line, coined 2IRGFP, which over-expresses GFP upon virus infection. With this system, we have achieved an accurate description of the dynamics of virus replication in space and time. Upon silencing selected N. benthamiana genes previously shown to be related to host response to geminivirus infection, we have identified eighteen genes involved in a wide array of cellular processes. Plant genes involved in geminivirus resistance were studied by comparing two tomato lines: one resistant (R), the other susceptible (S) to the virus. Sixty-nine genes preferentially expressed in R tomatoes were identified by screening cDNA libraries from infected and uninfected R and S genotypes. Out of the 25 genes studied so far, the silencing of five led to the total collapse of resistance, suggesting their involvement in the resistance gene network. This review of our results indicates that TRV-VIGS is an exquisite reverse genetics tool that may provide new insights into the molecular

  12. Discovering Host Genes Involved in the Infection by the Tomato Yellow Leaf Curl Virus Complex and in the Establishment of Resistance to the Virus Using Tobacco Rattle Virus-based Post Transcriptional Gene Silencing

    PubMed Central

    Czosnek, Henryk; Eybishtz, Assaf; Sade, Dagan; Gorovits, Rena; Sobol, Iris; Bejarano, Eduardo; Rosas-Díaz, Tábata; Lozano-Durán, Rosa

    2013-01-01

    The development of high-throughput technologies allows for evaluating gene expression at the whole-genome level. Together with proteomic and metabolomic studies, these analyses have resulted in the identification of plant genes whose function or expression is altered as a consequence of pathogen attacks. Members of the Tomato yellow leaf curl virus (TYLCV) complex are among the most important pathogens impairing production of agricultural crops worldwide. To understand how these geminiviruses subjugate plant defenses, and to devise counter-measures, it is essential to identify the host genes affected by infection and to determine their role in susceptible and resistant plants. We have used a reverse genetics approach based on Tobacco rattle virus-induced gene silencing (TRV-VIGS) to uncover genes involved in viral infection of susceptible plants, and to identify genes underlying virus resistance. To identify host genes with a role in geminivirus infection, we have engineered a Nicotiana benthamiana line, coined 2IRGFP, which over-expresses GFP upon virus infection. With this system, we have achieved an accurate description of the dynamics of virus replication in space and time. Upon silencing selected N. benthamiana genes previously shown to be related to host response to geminivirus infection, we have identified eighteen genes involved in a wide array of cellular processes. Plant genes involved in geminivirus resistance were studied by comparing two tomato lines: one resistant (R), the other susceptible (S) to the virus. Sixty-nine genes preferentially expressed in R tomatoes were identified by screening cDNA libraries from infected and uninfected R and S genotypes. Out of the 25 genes studied so far, the silencing of five led to the total collapse of resistance, suggesting their involvement in the resistance gene network. This review of our results indicates that TRV-VIGS is an exquisite reverse genetics tool that may provide new insights into the molecular

  13. Tomato yellow leaf curl virus infection mitigates the heat stress response of plants grown at high temperatures.

    PubMed

    Anfoka, Ghandi; Moshe, Adi; Fridman, Lilia; Amrani, Linoy; Rotem, Or; Kolot, Mikhail; Zeidan, Mouhammad; Czosnek, Henryk; Gorovits, Rena

    2016-01-21

    Cultured tomatoes are often exposed to a combination of extreme heat and infection with Tomato yellow leaf curl virus (TYLCV). This stress combination leads to intense disease symptoms and yield losses. The response of TYLCV-susceptible and resistant tomatoes to heat stress together with viral infection was compared. The plant heat-stress response was undermined in TYLCV infected plants. The decline correlated with the down-regulation of heat shock transcription factors (HSFs) HSFA2 and HSFB1, and consequently, of HSF-regulated genes Hsp17, Apx1, Apx2 and Hsp90. We proposed that the weakened heat stress response was due to the decreased capacity of HSFA2 to translocate into the nuclei of infected cells. All the six TYLCV proteins were able to interact with tomato HSFA2 in vitro, moreover, coat protein developed complexes with HSFA2 in nuclei. Capturing of HSFA2 by viral proteins could suppress the transcriptional activation of heat stress response genes. Application of both heat and TYLCV stresses was accompanied by the development of intracellular large protein aggregates containing TYLCV proteins and DNA. The maintenance of cellular chaperones in the aggregated state, even after recovery from heat stress, prevents the circulation of free soluble chaperones, causing an additional decrease in stress response efficiency.

  14. Correlation of meteorological parameters and remotely sensed normalized difference vegetation index (NDVI) with cotton leaf curl virus (CLCV) in Multan

    NASA Astrophysics Data System (ADS)

    Ahmed, A.; Akhtar, A.; Khalid, B.; Shamim, A.

    2013-06-01

    Climate change and weather has a profound effect on the spread of Cotton Leaf Curl Virus (CLCV) which is transmitted by whitefly. Climate change is altering temperature and precipitation patterns, resulting in the shift of some insect/pest from small population to large population thus effecting crops yield. To find out the relationship between the weather conditions, outburst of CLCV and changes in Normalized Difference Vegetation Index (NDVI) values due to the outburst of CLCV, a study was carried out for tehsil Multan. Data was acquired for the months of June, July, August and September for the year 2010. Regression analysis between CLCV and meteorological conditions as well as between CLCV and NDVI was performed. Meteorological parameters included temperature, humidity, precipitation, cloud cover, wind direction, pan evaporation and sunshine hours. NDVI values were calculated from SPOT satellite imagery (1km) using ArcMap10 and WinDisp v5.1. Correlation coefficients obtained in most of the cases were acceptable however the significance F and P-value were higher than their critical value at 95% level of significance. Therefore significant correlation was found only between CLCV and temperature and between CLCV and PAN evaporation during the month of July.

  15. Tomato yellow leaf curl virus infection mitigates the heat stress response of plants grown at high temperatures

    PubMed Central

    Ghandi, Anfoka; Adi, Moshe; Lilia, Fridman; Linoy, Amrani; Or, Rotem; Mikhail, Kolot; Mouhammad, Zeidan; Henryk, Czosnek; Rena, Gorovits

    2016-01-01

    Cultured tomatoes are often exposed to a combination of extreme heat and infection with Tomato yellow leaf curl virus (TYLCV). This stress combination leads to intense disease symptoms and yield losses. The response of TYLCV-susceptible and resistant tomatoes to heat stress together with viral infection was compared. The plant heat-stress response was undermined in TYLCV infected plants. The decline correlated with the down-regulation of heat shock transcription factors (HSFs) HSFA2 and HSFB1, and consequently, of HSF-regulated genes Hsp17, Apx1, Apx2 and Hsp90. We proposed that the weakened heat stress response was due to the decreased capacity of HSFA2 to translocate into the nuclei of infected cells. All the six TYLCV proteins were able to interact with tomato HSFA2 in vitro, moreover, coat protein developed complexes with HSFA2 in nuclei. Capturing of HSFA2 by viral proteins could suppress the transcriptional activation of heat stress response genes. Application of both heat and TYLCV stresses was accompanied by the development of intracellular large protein aggregates containing TYLCV proteins and DNA. The maintenance of cellular chaperones in the aggregated state, even after recovery from heat stress, prevents the circulation of free soluble chaperones, causing an additional decrease in stress response efficiency. PMID:26792235

  16. Synthesis and biological activity evaluation of novel amino acid derivatives as potential elicitors against Tomato yellow leaf curl virus.

    PubMed

    Deng, Yufang; He, Shun; Geng, Qianqian; Duan, Yongheng; Guo, Mingcheng; Li, Jianqiang; Cao, Yongsong

    2015-12-01

    Disease caused by Tomato yellow leaf curl virus (TYLCV) brings serious production losses of cultivated tomato worldwide. In our previous study, two novel amino acid derivatives exerted satisfactory antiviral activities against TYLCV. In this study, the variation of TYLCV, the transcriptional expression level of Ty-1 and the enzyme activities of POD and PPO in tomato were monitored after treatment with two amino acid derivatives to illustrate the antiviral mechanism. The results showed the symptom severity caused by TYLCV was reduced significantly by two compounds and was associated with the inhibition of viral DNA level at the early stage. Among three levels of concentration, the highest inhibition rate of CNBF-His was 40.66% at 1000 mg/L, for CNBF-Asn, the highest inhibition rate was 36.26% at 2000 mg/L 30 days post-inoculation. Two compounds could also enhance the activities of PPO and POD and the transcriptional expression level of Ty-1 which correlates with plant resistance in tomato. In the field test, two compounds increased the yields of tomato and the maximum increase of yield was 37.66%. This is the first report of novel amino acid derivatives inducing resistance in tomato plant against TYLCV. It is suggested that amino acid derivatives have the potential to be an effective approach against TYLCV in tomato plant.

  17. Tomato yellow leaf curl virus differentially influences plant defence responses to a vector and a non-vector herbivore.

    PubMed

    Su, Qi; Mescher, Mark C; Wang, Shaoli; Chen, Gong; Xie, Wen; Wu, Qingjun; Wang, Wenkai; Zhang, Youjun

    2016-03-01

    Plants frequently engage in simultaneous interactions with diverse classes of biotic antagonists. Differential induction of plant defence pathways by these antagonists, and interactions between pathways, can have important ecological implications; however, these effects are currently not well understood. We explored how Tomato yellow leaf curl virus (TYLCV) influenced the performance of its vector (Bemisia tabaci) and a non-vector herbivore (Tetranychus urticae) occurring separately or together on tomato plants (Solanum lycopersicum). TYLCV enhanced the performance of B. tabaci, although this effect was statistically significant only in the absence of T. urticae, which adversely affected B. tabaci performance regardless of infection status. In contrast, the performance of T. urticae was enhanced (only) by the combined presence of TYLCV and B. tabaci. Analyses of phytohormone levels and defence gene expression in wild-type tomatoes and various plant-defence mutants indicate that the enhancement of herbivore performance (for each species) entails the disruption of downstream defences in the jasmonic acid (JA) pathway. For T. urticae, this disruption appears to involve antagonistic effects of salicylic acid (SA), which is cumulatively induced to high levels by B. tabaci and TYLCV. In contrast, TYLCV was found to suppress JA-mediated responses to B. tabaci via mechanisms independent of SA.

  18. RNA interference-based resistance in transgenic tomato plants against Tomato yellow leaf curl virus-Oman (TYLCV-OM) and its associated betasatellite.

    PubMed

    Ammara, Um e; Mansoor, Shahid; Saeed, Muhammad; Amin, Imran; Briddon, Rob W; Al-Sadi, Abdullah Mohammed

    2015-03-04

    Tomato yellow leaf curl virus (TYLCV), a monopartite begomovirus (family Geminiviridae) is responsible for heavy yield losses for tomato production around the globe. In Oman at least five distinct begomoviruses cause disease in tomato, including TYLCV. Unusually, TYLCV infections in Oman are sometimes associated with a betasatellite (Tomato leaf curl betasatellite [ToLCB]; a symptom modulating satellite). RNA interference (RNAi) can be used to develop resistance against begomoviruses at either the transcriptional or post-transcriptional levels. A hairpin RNAi (hpRNAi) construct to express double-stranded RNA homologous to sequences of the intergenic region, coat protein gene, V2 gene and replication-associated gene of Tomato yellow leaf curl virus-Oman (TYLCV-OM) was produced. Initially, transient expression of the hpRNAi construct at the site of virus inoculation was shown to reduce the number of plants developing symptoms when inoculated with either TYLCV-OM or TYLCV-OM with ToLCB-OM to Nicotiana benthamiana or tomato. Solanum lycopersicum L. cv. Pusa Ruby was transformed with the hpRNAi construct and nine confirmed transgenic lines were obtained and challenged with TYLCV-OM and ToLCB-OM by Agrobacterium-mediated inoculation. For all but one line, for which all plants remained symptomless, inoculation with TYLCV-OM led to a proportion (≤25%) of tomato plants developing symptoms of infection. For inoculation with TYLCV-OM and ToLCB-OM all lines showed a proportion of plants (≤45%) symptomatic. However, for all infected transgenic plants the symptoms were milder and virus titre in plants was lower than in infected non-transgenic tomato plants. These results show that RNAi can be used to develop resistance against geminiviruses in tomato. The resistance in this case is not immunity but does reduce the severity of infections and virus titer. Also, the betasatellite may compromise resistance, increasing the proportion of plants which ultimately show symptoms.

  19. Expression of stress-response proteins upon whitefly-mediated inoculation of Tomato yellow leaf curl virus in susceptible and resistant tomato plants.

    PubMed

    Gorovits, Rena; Akad, Fouad; Beery, Hila; Vidavsky, Favi; Mahadav, Assaf; Czosnek, Henryk

    2007-11-01

    To better understand the nature of resistance of tomato to the whitefly (Bemisia tabaci, B biotype)-transmitted Tomato yellow leaf curl virus (TYLCV), whiteflies and TYLCV were considered as particular cases of biotic stresses and virus resistance as a particular case of successful response to these stresses. Two inbred tomato lines issued from the same breeding program that used Solanum habrochaites as a TYLCV resistance source, one susceptible and the other resistant, were used to compare the expression of key proteins involved at different stages of the plant response with stresses: mitogen-activated protein kinases (MAPKs), cellular heat shock proteins (HSPs, proteases), and pathogenesis-related (PR) proteins. The two biotic stresses-non-viruliferous whitefly feeding and virus infection with viruliferous insects--led to a slow decline in abundance of MAPKs, HSPs, and chloroplast protease FtsH (but not chloroplast protease ClpC), and induced the activities of the PR proteins, beta-1,3-glucanase, and peroxidase. This decline was less pronounced in virus-resistant than in virus-susceptible lines. Contrary to whitefly infestation and virus infection, inoculation with the fungus Sclerotinia sclerotiorum induced a rapid accumulation of the stress proteins studied, followed by a decline; the virus-susceptible and -resistant tomato lines behaved similarly in response to the fungus.

  20. Rapid spread of a recently introduced virus (tomato yellow leaf curl virus) and its vector Bemisia tabaci (Hemiptera: Aleyrodidae) in Liaoning Province, China.

    PubMed

    Zhang, Wan-Min; Fu, Hai-Bin; Wang, Wen-Hang; Piao, Chun-Shu; Tao, Yun-Li; Guo, Dong; Chu, Dong

    2014-02-01

    In Liaoning Province, China, tomato yellow leaf curl virus (TYLCV) was first detected in 2009 and in only four counties. To quantify the spread of TYLCV and to identify potential factors influencing its spread in Liaoning Province, we assayed for TYLCV within 1,055 whiteflies (Bemisia tabaci (Gennadius) complex) from 74 populations and 29 counties in 2011. The B. tabaci species of these individuals was determined based on molecular markers. TYLCV was found in 13 counties (Donggang, Liaoyang, Kazuo, Lingyuan, Heishan, Liaozhong, Kaiyuan, Taian, Dawa, Dashiqiao, Beizhen, Linghai, and Xingcheng) and was most frequently detected in the central plain. In addition, the percentage of whiteflies with TYLCV was significantly higher in B. tabaci Q than in B. tabaci B but was unrelated to the hosts (pepper, eggplant, tomato, cucumber, and kidney bean) on which the whiteflies had been collected. These results demonstrate that TYLCV has spread rapidly in Liaoning Province since its first detection and suggest that its spread is more closely associated with the introduction of B. tabaci Q than with the species of host plant. These findings also indicate that controls are now needed to reduce the further spread of TYLCV and that these controls should include the management of B. tabaci Q populations.

  1. Viruses Roll the Dice: The Stochastic Behavior of Viral Genome Molecules Accelerates Viral Adaptation at the Cell and Tissue Levels

    PubMed Central

    Miyashita, Shuhei; Ishibashi, Kazuhiro; Kishino, Hirohisa; Ishikawa, Masayuki

    2015-01-01

    Recent studies on evolutionarily distant viral groups have shown that the number of viral genomes that establish cell infection after cell-to-cell transmission is unexpectedly small (1–20 genomes). This aspect of viral infection appears to be important for the adaptation and survival of viruses. To clarify how the number of viral genomes that establish cell infection is determined, we developed a simulation model of cell infection for tomato mosaic virus (ToMV), a positive-strand RNA virus. The model showed that stochastic processes that govern the replication or degradation of individual genomes result in the infection by a small number of genomes, while a large number of infectious genomes are introduced in the cell. It also predicted two interesting characteristics regarding cell infection patterns: stochastic variation among cells in the number of viral genomes that establish infection and stochastic inequality in the accumulation of their progenies in each cell. Both characteristics were validated experimentally by inoculating tobacco cells with a library of nucleotide sequence–tagged ToMV and analyzing the viral genomes that accumulated in each cell using a high-throughput sequencer. An additional simulation model revealed that these two characteristics enhance selection during tissue infection. The cell infection model also predicted a mechanism that enhances selection at the cellular level: a small difference in the replication abilities of coinfected variants results in a large difference in individual accumulation via the multiple-round formation of the replication complex (i.e., the replication machinery). Importantly, this predicted effect was observed in vivo. The cell infection model was robust to changes in the parameter values, suggesting that other viruses could adopt similar adaptation mechanisms. Taken together, these data reveal a comprehensive picture of viral infection processes including replication, cell-to-cell transmission, and

  2. Viruses roll the dice: the stochastic behavior of viral genome molecules accelerates viral adaptation at the cell and tissue levels.

    PubMed

    Miyashita, Shuhei; Ishibashi, Kazuhiro; Kishino, Hirohisa; Ishikawa, Masayuki

    2015-03-01

    Recent studies on evolutionarily distant viral groups have shown that the number of viral genomes that establish cell infection after cell-to-cell transmission is unexpectedly small (1-20 genomes). This aspect of viral infection appears to be important for the adaptation and survival of viruses. To clarify how the number of viral genomes that establish cell infection is determined, we developed a simulation model of cell infection for tomato mosaic virus (ToMV), a positive-strand RNA virus. The model showed that stochastic processes that govern the replication or degradation of individual genomes result in the infection by a small number of genomes, while a large number of infectious genomes are introduced in the cell. It also predicted two interesting characteristics regarding cell infection patterns: stochastic variation among cells in the number of viral genomes that establish infection and stochastic inequality in the accumulation of their progenies in each cell. Both characteristics were validated experimentally by inoculating tobacco cells with a library of nucleotide sequence-tagged ToMV and analyzing the viral genomes that accumulated in each cell using a high-throughput sequencer. An additional simulation model revealed that these two characteristics enhance selection during tissue infection. The cell infection model also predicted a mechanism that enhances selection at the cellular level: a small difference in the replication abilities of coinfected variants results in a large difference in individual accumulation via the multiple-round formation of the replication complex (i.e., the replication machinery). Importantly, this predicted effect was observed in vivo. The cell infection model was robust to changes in the parameter values, suggesting that other viruses could adopt similar adaptation mechanisms. Taken together, these data reveal a comprehensive picture of viral infection processes including replication, cell-to-cell transmission, and evolution

  3. Development and application of triple antibody sandwich enzyme-linked immunosorbent assays for begomovirus detection using monoclonal antibodies against Tomato yellow leaf curl Thailand virus.

    PubMed

    Seepiban, Channarong; Charoenvilaisiri, Saengsoon; Warin, Nuchnard; Bhunchoth, Anjana; Phironrit, Namthip; Phuangrat, Bencharong; Chatchawankanphanich, Orawan; Attathom, Supat; Gajanandana, Oraprapai

    2017-05-30

    Tomato yellow leaf curl Thailand virus, TYLCTHV, is a begomovirus that causes severe losses of tomato crops in Thailand as well as several countries in Southeast and East Asia. The development of monoclonal antibodies (MAbs) and serological methods for detecting TYLCTHV is essential for epidemiological studies and screening for virus-resistant cultivars. The recombinant coat protein (CP) of TYLCTHV was expressed in Escherichia coli and used to generate MAbs against TYLCTHV through hybridoma technology. The MAbs were characterized and optimized to develop triple antibody sandwich enzyme-linked immunosorbent assays (TAS-ELISAs) for begomovirus detection. The efficiency of TAS-ELISAs for begomovirus detection was evaluated with tomato, pepper, eggplant, okra and cucurbit plants collected from several provinces in Thailand. Molecular identification of begomoviruses in these samples was also performed through PCR and DNA sequence analysis of the CP gene. Two MAbs (M1 and D2) were generated and used to develop TAS-ELISAs for begomovirus detection. The results of begomovirus detection in 147 field samples indicated that MAb M1 reacted with 2 begomovirus species, TYLCTHV and Tobacco leaf curl Yunnan virus (TbLCYnV), whereas MAb D2 reacted with 4 begomovirus species, TYLCTHV, TbLCYnV, Tomato leaf curl New Delhi virus (ToLCNDV) and Squash leaf curl China virus (SLCCNV). Phylogenetic analyses of CP amino acid sequences from these begomoviruses revealed that the CP sequences of begomoviruses recognized by the narrow-spectrum MAb M1 were highly conserved, sharing 93% identity with each other but only 72-81% identity with MAb M1-negative begomoviruses. The CP sequences of begomoviruses recognized by the broad-spectrum MAb D2 demonstrated a wider range of amino acid sequence identity, sharing 78-96% identity with each other and 72-91% identity with those that were not detected by MAb D2. TAS-ELISAs using the narrow-specificity MAb M1 proved highly efficient for the detection of

  4. Rolling Uphill

    ERIC Educational Resources Information Center

    Cross, Rod

    2017-01-01

    In a recent letter to this journal, Mungan noted that translational energy can be converted into gravitational potential energy when an object is projected vertically, but rotational energy is not usually converted in this manner. As an exception, he gave an example where "a ball initially rolling without slipping will travel higher up a…

  5. Real-time PCR protocols for the quantification of the begomovirus tomato yellow leaf curl Sardinia virus in tomato plants and in its insect vector.

    PubMed

    Noris, Emanuela; Miozzi, Laura

    2015-01-01

    Tomato yellow leaf curl Sardinia virus (TYLCSV) (Geminiviridae) is an important pathogen, transmitted by the whitefly Bemisia tabaci, that severely affects the tomato production in the Mediterranean basin. Here, we describe real-time PCR protocols suitable for relative and absolute quantification of TYLCSV in tomato plants and in whitefly extracts. Using primers and probe specifically designed for TYLCSV, the protocols for relative quantification allow to compare the amount of TYLCSV present in different plant or whitefly samples, normalized to the amount of DNA present in each sample using endogenous tomato or Bemisia genes as internal references. The absolute quantification protocol allows to calculate the number of genomic units of TYLCSV over the genomic units of the plant host (tomato), with a sensitivity of as few as ten viral genome copies per sample. The described protocols are potentially suitable for several applications, such as plant breeding for resistance, analysis of virus replication, and virus-vector interaction studies.

  6. Introgression of genes for cotton leaf curl virus resistance and increased fiber strength from Gossypium stocksii into upland cotton (G. hirsutum).

    PubMed

    Nazeer, W; Ahmad, S; Mahmood, K; Tipu, A L; Mahmood, A; Zhou, B

    2014-02-21

    Cotton leaf curl virus disease is a major hurdle for successful cotton production in Pakistan. There has been considerable economic loss due to this disease during the last decade. It would be desirable to have cotton varieties resistant to this disease. We explored the possibility of transferring virus resistant genes from the wild species Gossypium stocksii into MNH-786, a cultivar of G. hirsutum. Hybridization was done under field condition at the Cotton Research Station, Multan, during 2010-11. Boll shedding was controlled by application of exogenous hormones. F1 seeds were treated with 0.03% colchicine solution for 6 h and germinated. Cytological observations at peak squaring/flowering stage showed that these plants were hexaploid, having 2n = 6x = 78 chromosomes. The F1 plants showed intermediate expression for leaf size, leaf area, petiole length, bracteole number and size, bracteole area, bracteole dentation, flower size, pedicel size, and petal number and size. Moreover it possessed high fiber strength of 54.4 g/tex, which is 54% greater than that of the check variety, i.e. MNH-786 (G. hirsutum). The F1 population did not show any symptom of CLCuVD in the field, tested by grafting with CLCuVD susceptible rootstock (var. S12). We conclude that it is possible to transfer CLCuVD resistance and high fiber strength from G. stocksii to G. hirsutum.

  7. Expression of tomato yellow leaf curl virus coat protein using baculovirus expression system and evaluation of its utility as a viral antigen.

    PubMed

    Elgaied, Lamiaa; Salem, Reda; Elmenofy, Wael

    2017-08-01

    DNA encoding the coat protein (CP) of an Egyptian isolate of tomato yellow leaf curl virus (TYLCV) was inserted into the genome of Autographa californica nucleopolyhedrovirus (AcNPV) under the control of polyhedrin promoter. The generated recombinant baculovirus construct harboring the coat protein gene was characterized using PCR analysis. The recombinant coat protein expressed in infected insect cells was used as a coating antigen in an indirect Enzyme-linked immunosorbent assay (ELISA) and dot blot to test its utility for the detection of antibody generated against TYLCV virus particles. The results of ELISA and dot blot showed that the TYLCV-antibodies reacted positively with extracts of infected cells using the recombinant virus as a coating antigen with strong signals as well as the TYLCV infected tomato and beat plant extracts as positive samples. Scanning electron microscope examination showed that the expressed TYLCV coat protein was self-assembled into virus-like particles (VLPs) similar in size and morphology to TYLCV virus particles. These results concluded that, the expressed coat protein of TYLCV using baculovirus vector system is a reliable candidate for generation of anti-CP antibody for inexpensive detection of TYLCV-infected plants using indirect CP-ELISA or dot blot with high specificity.

  8. Clathrin-mediated endocytosis is involved in Tomato yellow leaf curl virus transport across the midgut barrier of its whitefly vector.

    PubMed

    Pan, Li-Long; Chen, Qun-Fang; Zhao, Juan-Juan; Guo, Tao; Wang, Xiao-Wei; Hariton-Shalev, Aliza; Czosnek, Henryk; Liu, Shu-Sheng

    2017-02-01

    Tomato yellow leaf curl virus (TYLCV) is a begomovirus transmitted by the whitefly Bemisia tabaci. The circulative translocation of the virus in the insect is known in its broad line. However, transit of TYLCV from the digestive tract into the haemolymph is poorly understood. We studied the involvement of clathrin in this process by disrupting the clathrin-mediated endocytosis and the endosome network using inhibitor feeding, antibody blocking and dsRNA silencing. We monitored the quantities of TYLCV in the whitefly and virus transmission efficiency. Following endocytosis and endosome network disruption, the quantity of virus was higher in the midgut relative to that of the whole insect body, and the quantity of virus in the haemolymph was reduced. The transmission efficiency of TYLCV by the treated insects was also reduced. These findings indicate that clathrin-mediated endocytosis and endosomes play an important role in the transport of TYLCV across the whitefly midgut. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  9. Molecular characterization and infectivity of a Tomato leaf curl New Delhi virus variant associated with newly emerging yellow mosaic disease of eggplant in India

    PubMed Central

    2011-01-01

    Background Begomoviruses have emerged as serious problem for vegetable and fiber crops in the recent past, frequently in tropical and subtropical region of the world. The association of begomovirus with eggplant yellow mosaic disease is hitherto unknown apart from one report from Thailand. A survey in Nagpur, Central India, in 2009-2010 showed severe incidence of eggplant yellow mosaic disease. Here, we have identified and characterized a begomovirus responsible for the newly emerging yellow mosaic disease of eggplant in India. Results The complete DNA-A and DNA-B genomic components of the causative virus were cloned and sequenced. Nucleotide sequence analysis of DNA-A showed that it shared highest 97.6% identity with Tomato leaf curl New Delhi virus-India[India:Udaipur:Okra:2007] and lowest 87.9% identity with Tomato leaf curl New Delhi virus-India[India:NewDelhi:Papaya:2005], while DNA-B showed highest 94.1% identity with ToLCNDV-IN[IN:UD:Ok:07] and lowest 76.2% identity with ToLCNDV-India[India:Lucknow]. Thus, it appears that this begomovirus is a variant of ubiquitous ToLCNDV and hence, we suggest the name ToLCNDV-India[India:Nagpur:Eggplant:2009] for this variant. The pathogenicity of ToLCNDV-IN[IN:Nag:Egg:09] isolate was confirmed by agroinfiltraion and dimeric clones of DNA-A and DNA-B induced characteristic yellow mosaic symptoms in eggplants and leaf curling in tomato plants. Conclusion This is the first report of a ToLCNDV variant moving to a new agriculturally important host, eggplant and causing yellow mosaic disease. This is also a first experimental demonstration of Koch's postulate for a begomovirus associated with eggplant yellow mosaic disease. PMID:21676270

  10. CONTROL FOR ROLLING MILL

    DOEpatents

    Shuck, A.B.; Shaw, W.C.

    1961-06-20

    A plutonium-rolling apparatus is patented that has two sets of feed rolls, shaping rolls between the feed rolls, and grippers beyond the feed rolls, which ready a workpiece for a new pass through the shaping rolls by angularly shifting the workpiece about its axis or transversely moving it on a line parallel to the axes of the shaping rolls. Actuation of each gripper for gripping or releasing the workpiece is produced by the relative positions assumed by the feed rolls adjacent to the gripper as the workpiece enters or leaves the feed rolls.

  11. Rolling Uphill

    NASA Astrophysics Data System (ADS)

    Cross, Rod

    2017-04-01

    In a recent letter to this journal, Mungan noted that translational energy can be converted into gravitational potential energy when an object is projected vertically, but rotational energy is not usually converted in this manner. As an exception, he gave an example where "a ball initially rolling without slipping will travel higher up a rough ramp than it will up a frictionless ramp." However, such a result is unlikely to be observed in practice. A better example would be a ball spinning rapidly forwards as it slides up the ramp, since the friction force on the ball then acts in a direction up the ramp.

  12. Reduction of leaf area and symptom severity as proxies of disease-induced plant mortality: the example of the Cauliflower mosaic virus infecting two Brassicaceae hosts.

    PubMed

    Doumayrou, Juliette; Leblaye, Sophie; Froissart, Rémy; Michalakis, Yannis

    2013-09-01

    Disease induced effects on host survival are important to understand the evolution of parasitic virulence and host resistance/tolerance. Unfortunately, experiments evaluating such effects are in most cases logistically demanding justifying the measurement of survival proxies. For plant hosts commonly used proxies are leaf area and the nature and severity of visual qualitative disease symptoms. In this study we tested whether these traits are indeed correlated to the host mortality rate induced by viral infection. We infected Brassica rapa and Arabidopsis thaliana plants with different natural isolates of Cauliflower mosaic virus (CaMV) and estimated over time the development of symptoms and the relative reduction of leaf area compared to healthy plants and followed plant mortality. We observed that the mortality of infected plants was correlated with the relative reduction of leaf area of both B. rapa and A. thaliana. Measures of mortality were also correlated with the severity of visual qualitative symptoms but the magnitude of the correlations and the time frame at which they were significant depended on the host plant: stronger and earlier correlations were observed on A. thaliana.

  13. First complete genomic characterization and phylogeny of a new recombinant of tomato yellow leaf curl virus (genus Begomovirus, family Geminiviridae) from Kuwait.

    PubMed

    Al-Ali, Ebtisam H M; Al-Hashash, Hanadi K; Ben-Hejji, Ahmed H; Al-Shayjji, Nabella; Al-Aqeel, Hamed A

    2015-07-01

    While whitefly-transmitted begomoviruses are economically important constraints to tomato production in Kuwait, little is known about genomic features of these viruses from Kuwait. A begomovirus isolated from severely diseased tomatoes, collected over a two-year period in the main tomato-growing areas of Kuwait, was characterized at the molecular level. The complete genomic sequence of the begomovirus was determined, and phlylogeographic studies were conducted to better understand genetic diversity of the virus in the region. Based on genome properties and phylogenetic analysis, the begomovirus was found to be a strain of tomato yellow leaf curl virus (TYLCV). The virus genome was monopartite, as neither DNA B nor satellite DNA molecules were detected. Two isolates characterized in this study shared 97% and 95% nucleotide sequence identity with a previously characterized Kuwaiti isolate, TYLCV-KISR. Among TYLCV isolates with known genome sequences, the Kuwaiti isolates shared highest sequence identity (95%) with TYLCV-Almeria (Spain). Genetic diversity and phylogenetic analysis showed that the three Kuwaiti isolates formed a distinct clade that was separate from those of known TYLCV sequences. One Kuwaiti isolate (KW 1-3) could be a novel variant of TYLCV. Two recombination events were detected in the genome sequence of KW 1-3, which appeared to be a recombinant derived from TYLCV parents from Oman and Kuwait.

  14. Water Balance, Hormone Homeostasis, and Sugar Signaling Are All Involved in Tomato Resistance to Tomato Yellow Leaf Curl Virus1[C][W][OPEN

    PubMed Central

    Sade, Dagan; Sade, Nir; Shriki, Oz; Lerner, Stephen; Gebremedhin, Alem; Karavani, Asaf; Brotman, Yariv; Osorio, Sonia; Fernie, Alisdair R.; Willmitzer, Lothar; Czosnek, Henryk; Moshelion, Menachem

    2014-01-01

    Vacuolar water movement is largely controlled by membrane channels called tonoplast-intrinsic aquaporins (TIP-AQPs). Some TIP-AQP genes, such as TIP2;2 and TIP1;1, are up-regulated upon exposure to biotic stress. Moreover, TIP1;1 transcript levels are higher in leaves of a tomato (Solanum lycopersicum) line resistant to Tomato yellow leaf curl virus (TYLCV) than in those of a susceptible line with a similar genetic background. Virus-induced silencing of TIP1;1 in the tomato resistant line and the use of an Arabidopsis (Arabidopsis thaliana) tip1;1 null mutant showed that resistance to TYLCV is severely compromised in the absence of TIP1:1. Constitutive expression of tomato TIP2;2 in transgenic TYLCV-susceptible tomato and Arabidopsis plants was correlated with increased TYLCV resistance, increased transpiration, decreased abscisic acid levels, and increased salicylic acid levels at the early stages of infection. We propose that TIP-AQPs affect the induction of leaf abscisic acid, which leads to increased levels of transpiration and gas exchange, as well as better salicylic acid signaling. PMID:24989233

  15. Analysis of the Mild strain of tomato yellow leaf curl virus, which overcomes Ty-2 gene-mediated resistance in tomato line H24.

    PubMed

    Ohnishi, Jun; Yamaguchi, Hirotaka; Saito, Atsushi

    2016-08-01

    In tomato line H24, an isolate of the Mild (Mld) strain of tomato yellow leaf curl virus (TYLCV-Mld [JR:Kis]) overcomes Ty-2 gene-mediated resistance and causes typical symptoms of tomato yellow leaf curl disease (TYLCD). No systemic infection with visible symptoms or accumulation of viral DNA in the upper leaves was observed in H24 challenged with another isolate, TYLCV-IL (TYLCV-IL [JR:Osaka]), confirming that H24 is resistant to the IL strain. To elucidate the genomic regions that cause the breakdown of the Ty-2 gene-mediated resistance, we constructed a series of chimeras by swapping genes between the two strains. A chimeric virus that had the overlapping C4/Rep region of the Mld strain in the context of the IL strain genome, caused severe TYLCD in H24 plants, suggesting that the overlapping C4/Rep region of the Mld strain is associated with the ability of this strain to overcome Ty-2 gene-mediated resistance.

  16. The complete genomic sequence of pepper yellow leaf curl virus (PYLCV) and its implications for our understanding of evolution dynamics in the genus polerovirus.

    PubMed

    Dombrovsky, Aviv; Glanz, Eyal; Lachman, Oded; Sela, Noa; Doron-Faigenboim, Adi; Antignus, Yehezkel

    2013-01-01

    We determined the complete sequence and organization of the genome of a putative member of the genus Polerovirus tentatively named Pepper yellow leaf curl virus (PYLCV). PYLCV has a wider host range than Tobacco vein-distorting virus (TVDV) and has a close serological relationship with Cucurbit aphid-borne yellows virus (CABYV) (both poleroviruses). The extracted viral RNA was subjected to SOLiD next-generation sequence analysis and used as a template for reverse transcription synthesis, which was followed by PCR amplification. The ssRNA genome of PYLCV includes 6,028 nucleotides encoding six open reading frames (ORFs), which is typical of the genus Polerovirus. Comparisons of the deduced amino acid sequences of the PYLCV ORFs 2-4 and ORF5, indicate that there are high levels of similarity between these sequences to ORFs 2-4 of TVDV (84-93%) and to ORF5 of CABYV (87%). Both PYLCV and Pepper vein yellowing virus (PeVYV) contain sequences that point to a common ancestral polerovirus. The recombination breakpoint which is located at CABYV ORF3, which encodes the viral coat protein (CP), may explain the CABYV-like sequences found in the genomes of the pepper infecting viruses PYLCV and PeVYV. Two additional regions unique to PYLCV (PY1 and PY2) were identified between nucleotides 4,962 and 5,061 (ORF 5) and between positions 5,866 and 6,028 in the 3' NCR. Sequence analysis of the pepper-infecting PeVYV revealed three unique regions (Pe1-Pe3) with no similarity to other members of the genus Polerovirus. Genomic analyses of PYLCV and PeVYV suggest that the speciation of these viruses occurred through putative recombination event(s) between poleroviruses co-infecting a common host(s), resulting in the emergence of PYLCV, a novel pathogen with a wider host range.

  17. Global Analysis of the Transcriptional Response of Whitefly to Tomato Yellow Leaf Curl China Virus Reveals the Relationship of Coevolved Adaptations▿ †

    PubMed Central

    Luan, Jun-Bo; Li, Jun-Min; Varela, Nélia; Wang, Yong-Liang; Li, Fang-Fang; Bao, Yan-Yuan; Zhang, Chuan-Xi; Liu, Shu-Sheng; Wang, Xiao-Wei

    2011-01-01

    The begomoviruses are the largest and most economically important group of plant viruses transmitted exclusively by the whitefly Bemisia tabaci in a circulative, persistent manner. The circulation of the viruses within the insect vectors involves complex interactions between virus and vector components; however, the molecular mechanisms of these interactions remain largely unknown. Here we investigated the transcriptional response of the invasive B. tabaci Middle East-Asia Minor 1 species to Tomato yellow leaf curl China virus (TYLCCNV) using Illumina sequencing technology. Results showed that 1,606 genes involved in 157 biochemical pathways were differentially expressed in the viruliferous whiteflies. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that TYLCCNV can perturb the cell cycle and primary metabolism in the whitefly, which explains the negative effect of this virus on the longevity and fecundity of B. tabaci. Our data also demonstrated that TYLCCNV can activate whitefly immune responses, such as autophagy and antimicrobial peptide production, which might lead to a gradual decrease of viral particles within the body of the viruliferous whitefly. Furthermore, PCR results showed that TYLCCNV can invade the ovary and fat body tissues of the whitefly, and Lysotracker and Western blot analyses revealed that the invasion of TYLCCNV induced autophagy in both the ovary and fat body tissues. Surprisingly, TYLCCNV also suppressed the whitefly immune responses by downregulating the expression of genes involved in Toll-like signaling and mitogen-activated protein kinase (MAPK) pathways. Taken together, these results reveal the relationship of coevolved adaptations between begomoviruses and whiteflies and will provide a road map for future investigations into the complex interactions between plant viruses and their insect vectors. PMID:21270146

  18. Global analysis of the transcriptional response of whitefly to tomato yellow leaf curl China virus reveals the relationship of coevolved adaptations.

    PubMed

    Luan, Jun-Bo; Li, Jun-Min; Varela, Nélia; Wang, Yong-Liang; Li, Fang-Fang; Bao, Yan-Yuan; Zhang, Chuan-Xi; Liu, Shu-Sheng; Wang, Xiao-Wei

    2011-04-01

    The begomoviruses are the largest and most economically important group of plant viruses transmitted exclusively by the whitefly Bemisia tabaci in a circulative, persistent manner. The circulation of the viruses within the insect vectors involves complex interactions between virus and vector components; however, the molecular mechanisms of these interactions remain largely unknown. Here we investigated the transcriptional response of the invasive B. tabaci Middle East-Asia Minor 1 species to Tomato yellow leaf curl China virus (TYLCCNV) using Illumina sequencing technology. Results showed that 1,606 genes involved in 157 biochemical pathways were differentially expressed in the viruliferous whiteflies. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that TYLCCNV can perturb the cell cycle and primary metabolism in the whitefly, which explains the negative effect of this virus on the longevity and fecundity of B. tabaci. Our data also demonstrated that TYLCCNV can activate whitefly immune responses, such as autophagy and antimicrobial peptide production, which might lead to a gradual decrease of viral particles within the body of the viruliferous whitefly. Furthermore, PCR results showed that TYLCCNV can invade the ovary and fat body tissues of the whitefly, and Lysotracker and Western blot analyses revealed that the invasion of TYLCCNV induced autophagy in both the ovary and fat body tissues. Surprisingly, TYLCCNV also suppressed the whitefly immune responses by downregulating the expression of genes involved in Toll-like signaling and mitogen-activated protein kinase (MAPK) pathways. Taken together, these results reveal the relationship of coevolved adaptations between begomoviruses and whiteflies and will provide a road map for future investigations into the complex interactions between plant viruses and their insect vectors.

  19. Dynamics of defense-related components in two contrasting genotypes of tomato upon infection with Tomato Leaf Curl New Delhi Virus.

    PubMed

    Sahu, Pranav Pankaj; Rai, Neeraj Kumar; Puranik, Swati; Roy, Anirban; Khan, Moinuddin; Prasad, Manoj

    2012-10-01

    Tomato leaf curl virus (ToLCV) disease is a serious threat for tomato cultivation in the tropics and subtropics. Despite serious efforts no immune commercial varieties or F(1) hybrids are available till date. In this study, the interaction between Solanum lycopersicum and ToLCV was characterized on molecular and biochemical basis. RNA silencing mediated by short interfering RNA (siRNA) and reactive oxygen species (ROS) has been proposed as central components of plant adaptation to several stresses. A comparative RNA interference study between two contrasting tomato genotypes, LA1777 (tolerant) and 15SBSB (susceptible) infected with Tomato Leaf Curl New Delhi Virus (ToLCNDV) revealed relatively higher accumulation of siRNA in the leaves of tolerant genotype. In LA1777, ToLCNDV produced chlorotic as well as necrotic areas at the inoculation sites 5-10 days post-inoculation. Caspase-9- and caspase-3-like activities were significantly increased in response to ToLCNDV infection in LA1777 at inoculated region. Activities of antioxidant enzymes involved in the detoxification of ROS were examined in both systemic and localized area of infection, and their expression level was further validated through quantitative real-time PCR of the corresponding transcripts. Expression patterns of three genes encoding pathogenesis-related proteins showed higher accumulation in tolerant genotype. Tolerance against the ToLCNDV in LA1777 can be attributed to the higher siRNA accumulation, localized cell death, altered levels of antioxidant enzymes and activation of pathogenesis-related genes at different durations of virus infection. Based on these direct and indirect evidences, we have proposed a putative mechanism for ToLCNDV tolerance in the tolerant genotype.

  20. Molecular characterization of Citrus tatter leaf virus historically associated with Meyer lemon trees: complete genome sequence and development of biologically active in vitro transcripts.

    PubMed

    Tatineni, Satyanarayana; Afunian, Mohammad R; Hilf, Mark E; Gowda, Siddarame; Dawson, William O; Garnsey, Stephen M

    2009-04-01

    Citrus tatter leaf virus isolated from Meyer lemon trees (CTLV-ML) from California and Florida induces bud union incompatibility of citrus trees grafted on the widely used trifoliate and trifoliate hybrid rootstocks. The complete genome sequence of CTLV-ML was determined to be 6,495 nucleotides (nts), with two overlapping open reading frames (ORFs) and a poly (A) tail at the 3' end. The genome organization is similar to other capilloviruses, with ORF1 (nts 37 to 6,354) encoding a putative 242-kDa polyprotein which contains replication-associated domains plus a coat protein (CP), and ORF2 (nts 4,788 to 5,750), which is located within ORF1 in a different reading frame and encodes a putative movement protein. Although the proteins encoded by CTLV-ML possesses 84 to 96% amino acid sequence identity with strains of Apple stem grooving virus (ASGV), we observed two strikingly different regions in ORF1: variable region I (amino acids 532 to 570) and variable region II (amino acids 1,583 to 1,868), with only 15 to 18 and 56 to 62% identities, respectively, with the corresponding regions of ASGV strains. Conditions for a herbaceous systemic assay host were optimized in which the wild-type virus induced systemic infection in Phaseolus vulgaris cv. Light Red Kidney (LRK) bean plants at 19 or 22 degrees C but not at higher temperatures. In vitro transcripts generated from full-length cDNA clones induced systemic symptoms on LRK bean plants similar to that of the wild-type virus. Replication of the recombinant virus was confirmed by hybridization of a 5' positive-stranded RNA-specific probe to a genome-sized RNA and by reverse-transcription polymerase chain reaction.

  1. Single-step multiplex RT-PCR for simultaneous and colourimetric detection of six RNA viruses in olive trees.

    PubMed

    Bertolini, E; Olmos, A; Martínez, M C; Gorris, M T; Cambra, M

    2001-07-01

    A single-step multiplex RT-PCR was developed for the simultaneous and colourimetric detection of six RNA viruses (Cucumber mosaic virus, Cherry leaf roll virus, strawberry latent ringspot virus, Arabis mosaic virus, Olive latent-1 virus and Olive latent-2 virus) which infect olive trees. Six compatible primer set for one-step RT-PCR amplification in a single closed-tube and 3' digoxigenin labelled probes were designed in optimal, specific and conserved regions. The method has been assessed with 195 Spanish field olive trees, suggesting that approximately 1.5% of the tested material was infected by Cucumber mosaic virus and 0.5% by Cherry leaf roll virus. This method saves time and reagent costs compared with monospecific RT-PCR which needs several reactions for the same number of tests. Using colourimetric detection, it is possible to analyse many samples, it increases sensitivity 10-fold, and whilst facilitating the interpretation of results, it avoids the use of gels and the toxic ethidium bromide. The method could be used routinely for sanitary and certification programmes.

  2. Mapping quantitative trait loci of resistance to Tomato spotted wilt virus and leaf spots in a recombinant inbred line population of peanut (Arachis hypogae L.) from SunOleic 97R and NC94022

    USDA-ARS?s Scientific Manuscript database

    Peanut is vulnerable to a range of diseases, such as Tomato spotted wilt virus (TSWV) and leaf spots. The most sustainable and economical solution for managing peanut diseases is development of resistance cultivars. The new breeding line NC94022, high resistance to TSWV and moderate resistance to le...

  3. Rolling-Element Bearings

    NASA Technical Reports Server (NTRS)

    Hamrock, B. J.; Anderson, W. J.

    1983-01-01

    Rolling element bearings are a precision, yet simple, machine element of great utility. A brief history of rolling element bearings is reviewed and the type of rolling element bearings, their geometry and kinematics, as well as the materials they are made from and the manufacturing processes they involve are described. Unloaded and unlubricated rolling element bearings, loaded but unlubricated rolling element bearings and loaded and lubricated rolling element bearings are considered. The recognition and understanding of elastohydrodynamic lubrication covered, represents one of the major development in rolling element bearings.

  4. MU06-857, a Green Leaf Lettuce Breeding Line with Resistance to Leafminer and Lettuce Mosaic Virus.

    USDA-ARS?s Scientific Manuscript database

    The Agricultural Research Service, United States Department of Agriculture announces the release of a breeding line of green leaf lettuce (Lactuca sativa L.) with resistance to leafminers (Liriomyza langei Frick) and lettuce mosaic. The line MU06-857 is similar to cultivar ‘Lolla Rossa’ (‘Lollo Ros...

  5. Silk gland gene expression during larval-pupal transitionin the cotton leaf roller Sylepta derogate (Lepidoptera: pyralidae)

    USDA-ARS?s Scientific Manuscript database

    The cotton leaf roller, Sylepta derogata, is a silk-producing insect pest. While young larvae feed on the underside of leaves, the older ones roll cotton leaves and feed on the leaf edges, which defoliates cotton plants. The larvae produce silk to stabilize the rolled leaf and to balloon from used t...

  6. The transmission efficiency of tomato yellow leaf curl virus by the whitefly Bemisia tabaci is correlated with the presence of a specific symbiotic bacterium species.

    PubMed

    Gottlieb, Yuval; Zchori-Fein, Einat; Mozes-Daube, Netta; Kontsedalov, Svetlana; Skaljac, Marisa; Brumin, Marina; Sobol, Iris; Czosnek, Henryk; Vavre, Fabrice; Fleury, Frédéric; Ghanim, Murad

    2010-09-01

    Tomato yellow leaf curl virus (TYLCV) (Geminiviridae: Begomovirus) is exclusively vectored by the whitefly Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). TYLCV transmission depends upon a 63-kDa GroEL protein produced by the vector's endosymbiotic bacteria. B. tabaci is a species complex comprising several genetically distinct biotypes that show different secondary-symbiont fauna. In Israel, the B biotype harbors Hamiltonella, and the Q biotype harbors Wolbachia and Arsenophonus. Both biotypes harbor Rickettsia and Portiera (the obligatory primary symbionts). The aim of this study was to determine which B. tabaci symbionts are involved in TYLCV transmission using B. tabaci populations collected in Israel. Virus transmission assays by B. tabaci showed that the B biotype efficiently transmits the virus, while the Q biotype scarcely transmits it. Yeast two-hybrid and protein pulldown assays showed that while the GroEL protein produced by Hamiltonella interacts with TYLCV coat protein, GroEL produced by Rickettsia and Portiera does not. To assess the role of Wolbachia and Arsenophonus GroEL proteins (GroELs), we used an immune capture PCR (IC-PCR) assay, employing in vivo- and in vitro-synthesized GroEL proteins from all symbionts and whitefly artificial feeding through membranes. Interaction between GroEL and TYLCV was found to occur in the B biotype, but not in the Q biotype. This assay further showed that release of virions protected by GroEL occurs adjacent to the primary salivary glands. Taken together, the GroEL protein produced by Hamiltonella (present in the B biotype, but absent in the Q biotype) facilitates TYLCV transmission. The other symbionts from both biotypes do not seem to be involved in transmission of this virus.

  7. 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. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  8. Apple chlorotic leaf spot virus 50 kDa protein is targeted to plasmodesmata and accumulates in sieve elements in transgenic plant leaves.

    PubMed

    Yoshikawa, N; Oogake, S; Terada, M; Miyabayashi, S; Ikeda, Y; Takahashi, T; Ogawa, K

    1999-01-01

    We investigated the in situ localization of the 50 kDa protein encoded by ORF2 of Apple chlorotic leaf spot virus (ACLSV) genome which is thought to be a movement protein. In immunogold electron microscopy of ACLSV-infected Chenopodium quinoa leaves, the 50 kDa protein was localized on plasmodesmata and nearby cytoplasm. Observation of transgenic Nicotiana occidentalis leaves expressing the 50 kDa protein fused to enhanced green fluorescent protein (EGFP) by fluorescence and confocal laser scanning microscopes revealed that green fluorescence was observed as spots on the cell wall or strands passing through the cell wall of several cell types, i.e., epidermal, palisade and spongy mesophyll and collenchyma cells. In transverse and longitudinal sections of leaf veins of transgenic plants showed that the 50K-EGFP fusion accumulated in sieve elements and formed an extensive interconnecting network of threadlike structure. These results indicated that ACLSV 50 kDa protein can target plasmodesmata and traffic into sieve elements.

  9. Amino Acids in the Capsid Protein of Tomato Yellow Leaf Curl Virus That Are Crucial for Systemic Infection, Particle Formation, and Insect Transmission

    PubMed Central

    Noris, E.; Vaira, A. M.; Caciagli, P.; Masenga, V.; Gronenborn, B.; Accotto, G. P.

    1998-01-01

    A functional capsid protein (CP) is essential for host plant infection and insect transmission in monopartite geminiviruses. We studied two defective genomic DNAs of tomato yellow leaf curl virus (TYLCV), Sic and SicRcv. Sic, cloned from a field-infected tomato, was not infectious, whereas SicRcv, which spontaneously originated from Sic, was infectious but not whitefly transmissible. A single amino acid change in the CP was found to be responsible for restoring infectivity. When the amino acid sequences of the CPs of Sic and SicRcv were compared with that of a closely related wild-type virus (TYLCV-Sar), differences were found in the following positions: 129 (P in Sic and SicRcv, Q in Sar), 134 (Q in Sic and Sar, H in SicRcv) and 152 (E in Sic and SicRcv, D in Sar). We constructed TYLCV-Sar variants containing the eight possible amino acid combinations in those three positions and tested them for infectivity and transmissibility. QQD, QQE, QHD, and QHE had a wild-type phenotype, whereas PHD and PHE were infectious but nontransmissible. PQD and PQE mutants were not infectious; however, they replicated and accumulated CP, but not virions, in Nicotiana benthamiana leaf discs. The Q129P replacement is a nonconservative change, which may drastically alter the secondary structure of the CP and affect its ability to form the capsid. The additional Q134H change, however, appeared to compensate for the structural modification. Sequence comparisons among whitefly-transmitted geminiviruses in terms of the CP region studied showed that combinations other than QQD are present in several cases, but never with a P129. PMID:9811744

  10. SlMAPK3 enhances tolerance to tomato yellow leaf curl virus (TYLCV) by regulating salicylic acid and jasmonic acid signaling in tomato (Solanum lycopersicum).

    PubMed

    Li, Yunzhou; Qin, Lei; Zhao, Jingjing; Muhammad, Tayeb; Cao, Hehe; Li, Hailiang; Zhang, Yan; Liang, Yan

    2017-01-01

    Several recent studies have reported on the role of mitogen-activated protein kinase (MAPK3) in plant immune responses. However, little is known about how MAPK3 functions in tomato (Solanum lycopersicum L.) infected with tomato yellow leaf curl virus (TYLCV). There is also uncertainty about the connection between plant MAPK3 and the salicylic acid (SA) and jasmonic acid (JA) defense-signaling pathways. The results of this study indicated that SlMAPK3 participates in the antiviral response against TYLCV. Tomato seedlings were inoculated with TYLCV to investigate the possible roles of SlMAPK1, SlMAPK2, and SlMAPK3 against this virus. Inoculation with TYLCV strongly induced the expression and the activity of all three genes. Silencing of SlMAPK1, SlMAPK2, and SlMAPK3 reduced tolerance to TYLCV, increased leaf H2O2 concentrations, and attenuated expression of defense-related genes after TYLCV infection, especially in SlMAPK3-silenced plants. Exogenous SA and methyl jasmonic acid (MeJA) both significantly induced SlMAPK3 expression in tomato leaves. Over-expression of SlMAPK3 increased the transcript levels of SA/JA-mediated defense-related genes (PR1, PR1b/SlLapA, SlPI-I, and SlPI-II) and enhanced tolerance to TYLCV. After TYLCV inoculation, the leaves of SlMAPK3 over-expressed plants compared with wild type plants showed less H2O2 accumulation and greater superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) activity. Overall, the results suggested that SlMAPK3 participates in the antiviral response of tomato to TYLCV, and that this process may be through either the SA or JA defense-signaling pathways.

  11. SlMAPK3 enhances tolerance to tomato yellow leaf curl virus (TYLCV) by regulating salicylic acid and jasmonic acid signaling in tomato (Solanum lycopersicum)

    PubMed Central

    Li, Yunzhou; Qin, Lei; Zhao, Jingjing; Muhammad, Tayeb; Cao, Hehe; Li, Hailiang; Zhang, Yan; Liang, Yan

    2017-01-01

    Several recent studies have reported on the role of mitogen-activated protein kinase (MAPK3) in plant immune responses. However, little is known about how MAPK3 functions in tomato (Solanum lycopersicum L.) infected with tomato yellow leaf curl virus (TYLCV). There is also uncertainty about the connection between plant MAPK3 and the salicylic acid (SA) and jasmonic acid (JA) defense-signaling pathways. The results of this study indicated that SlMAPK3 participates in the antiviral response against TYLCV. Tomato seedlings were inoculated with TYLCV to investigate the possible roles of SlMAPK1, SlMAPK2, and SlMAPK3 against this virus. Inoculation with TYLCV strongly induced the expression and the activity of all three genes. Silencing of SlMAPK1, SlMAPK2, and SlMAPK3 reduced tolerance to TYLCV, increased leaf H2O2 concentrations, and attenuated expression of defense-related genes after TYLCV infection, especially in SlMAPK3-silenced plants. Exogenous SA and methyl jasmonic acid (MeJA) both significantly induced SlMAPK3 expression in tomato leaves. Over-expression of SlMAPK3 increased the transcript levels of SA/JA-mediated defense-related genes (PR1, PR1b/SlLapA, SlPI-I, and SlPI-II) and enhanced tolerance to TYLCV. After TYLCV inoculation, the leaves of SlMAPK3 over-expressed plants compared with wild type plants showed less H2O2 accumulation and greater superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) activity. Overall, the results suggested that SlMAPK3 participates in the antiviral response of tomato to TYLCV, and that this process may be through either the SA or JA defense-signaling pathways. PMID:28222174

  12. Inhibitory activity of different medicinal extracts from Thuja leaves, ginger roots, Harmal seeds and turmeric rhizomes against Fig leaf mottle-associated virus 1 (FLMaV-1) infecting figs in Mecca region.

    PubMed

    Aldhebiani, Amal Y; Elbeshehy, Esam K F; Baeshen, Areej A; Elbeaino, Toufic

    2017-05-01

    Fig leaf mottle-associated virus-1 (FLMaV-1) is a closterovirus newly identified in fig trees, in the Mecca region, suffering from mosaic disease symptoms and apparently is compromising the fig plantation in the country. In the present study, we demonstrated the efficiency of two in vivo experiments including pre and post treatments using Thuja leaf, ginger roots, Harmal seeds and turmeric rhizome extracts on symptoms expression of rooted cuttings infected with FLMaV-1- and their impact on virus multiplication. Results showed that individual treatments with ginger roots and turmeric rhizomes in pre-grafting experiments and Thuja extract following Harmal seeds in post grafting experiments were efficient against symptom development. In addition, results showed that the total photosynthesis pigments; total soluble intracellular proteins and total phenol contents were higher in infected treated cuttings compared with healthy ones, thus it was taken as evidence on a mutual interaction between these extracts and virus multiplication.

  13. Genome-wide analysis of bHLH transcription factor and involvement in the infection by yellow leaf curl virus in tomato (Solanum lycopersicum).

    PubMed

    Wang, Jinyan; Hu, Zhongze; Zhao, Tongmin; Yang, Yuwen; Chen, Tianzi; Yang, Mali; Yu, Wengui; Zhang, Baolong

    2015-02-05

    The basic helix-loop-helix (bHLH) proteins are a superfamily of transcription factors that can bind to specific DNA target sites. They have been well characterized in model plants such as Arabidopsis and rice and have been shown to be important regulatory components in many different biological processes. However, no systemic analysis of the bHLH transcription factor family has yet been reported in tomatoes. Tomato yellow leaf curl virus (TYLCV) threatens tomato production worldwide by causing leaf yellowing, leaf curling, plant stunting and flower abscission. A total of 152 bHLH transcription factors were identified from the entire tomato genome. Phylogenetic analysis of bHLH domain sequences from Arabidopsis and tomato facilitated classification of these genes into 26 subfamilies. The evolutionary and possible functional relationships revealed during this analysis are supported by other criteria, including the chromosomal distribution of these genes, the conservation of motifs and exon/intron structural patterns, and the predicted DNA binding activities within subfamilies. Distribution mapping results showed bHLH genes were localized on the 12 tomato chromosomes. Among the 152 bHLH genes from the tomato genome, 96 bHLH genes were detected in the TYLCV-susceptible and resistant tomato breeding line before (0 dpi) and after TYLCV (357 dpi) infection. As anticipated, gene ontology (GO) analysis indicated that most bHLH genes are related to the regulation of macromolecule metabolic processes and gene expression. Only four bHLH genes were differentially expressed between 0 and 357 dpi. Virus-induced gene silencing (VIGS) of one bHLH genes SlybHLH131 in resistant lines can lead to the cell death. In the present study, 152 bHLH transcription factor genes were identified. One of which bHLH genes, SlybHLH131, was found to be involved in the TYLCV infection through qRT-PCR expression analysis and VIGS validation. The isolation and identification of these bHLH transcription

  14. The Tomato yellow leaf curl virus V2 protein forms aggregates depending on the cytoskeleton integrity and binds viral genomic DNA.

    PubMed

    Moshe, Adi; Belausov, Eduard; Niehl, Annette; Heinlein, Manfred; Czosnek, Henryk; Gorovits, Rena

    2015-05-05

    The spread of Tomato yellow leaf curl virus (TYLCV) was accompanied by the formation of coat protein (CP) aggregates of increasing size in the cytoplasm and nucleus of infected tomato (Solanum lycopersicum) cells. In order to better understand the TYLCV-host interaction, we investigated the properties and the subcellular accumulation pattern of the non-structural viral protein V2. CP and V2 are the only sense-oriented genes on the virus circular single-stranded DNA genome. Similar to CP, V2 localized to cytoplasmic aggregates of increasing size and as infection progressed was also found in nuclei, where it co-localized with CP. V2 was associated with viral genomic DNA molecules, suggesting that V2 functions as a DNA shuttling protein. The formation and the 26S proteasome-mediated degradation of V2 aggregates were dependent on the integrity of the actin and microtubule cytoskeleton. We propose that the cytoskeleton-dependent formation and growth of V2 aggregates play an important role during TYLCV infection, and that microtubules and actin filaments are important for the delivery of V2 to the 26S proteasome.

  15. The Tomato yellow leaf curl virus V2 protein forms aggregates depending on the cytoskeleton integrity and binds viral genomic DNA

    PubMed Central

    Moshe, Adi; Belausov, Eduard; Niehl, Annette; Heinlein, Manfred; Czosnek, Henryk; Gorovits, Rena

    2015-01-01

    The spread of Tomato yellow leaf curl virus (TYLCV) was accompanied by the formation of coat protein (CP) aggregates of increasing size in the cytoplasm and nucleus of infected tomato (Solanum lycopersicum) cells. In order to better understand the TYLCV-host interaction, we investigated the properties and the subcellular accumulation pattern of the non-structural viral protein V2. CP and V2 are the only sense-oriented genes on the virus circular single-stranded DNA genome. Similar to CP, V2 localized to cytoplasmic aggregates of increasing size and as infection progressed was also found in nuclei, where it co-localized with CP. V2 was associated with viral genomic DNA molecules, suggesting that V2 functions as a DNA shuttling protein. The formation and the 26S proteasome-mediated degradation of V2 aggregates were dependent on the integrity of the actin and microtubule cytoskeleton. We propose that the cytoskeleton-dependent formation and growth of V2 aggregates play an important role during TYLCV infection, and that microtubules and actin filaments are important for the delivery of V2 to the 26S proteasome. PMID:25940862

  16. Kinetics of interaction of Cotton Leaf Curl Kokhran Virus-Dabawali (CLCuKV-Dab) coat protein and its mutants with ssDNA

    SciTech Connect

    Priyadarshini, C.G. Poornima; Savithri, H.S.

    2009-04-10

    Gemini viral assembly and transport of viral DNA into nucleus for replication, essentially involve DNA-coat protein interactions. The kinetics of interaction of Cotton Leaf Curl Kokhran Virus-Dabawali recombinant coat protein (rCP) with DNA was studied by electrophoretic mobility shift assay (EMSA) and surface plasmon resonance (SPR). The rCP interacted with ssDNA with a K{sub A}, of 2.6 +- 0.29 x 10{sup 8} M{sup -1} in a sequence non-specific manner. The CP has a conserved C2H2 type zinc finger motif composed of residues C68, C72, H81 and H85. Mutation of these residues to alanine resulted in reduced binding to DNA probes. The H85A mutant rCP showed the least binding with approximately 756 fold loss in the association rate and a three order magnitude decrease in the binding affinity as compared to rCP. The CP-DNA interactions via the zinc finger motif could play a crucial role in virus assembly and in nuclear transport.

  17. The Whitefly Bemisia tabaci Knottin-1 Gene Is Implicated in Regulating the Quantity of Tomato Yellow Leaf Curl Virus Ingested and Transmitted by the Insect.

    PubMed

    Hariton Shalev, Aliza; Sobol, Iris; Ghanim, Murad; Liu, Shu-Sheng; Czosnek, Henryk

    2016-07-22

    The whitefly Bemisia tabaci is a major pest to agricultural crops. It transmits begomoviruses, such as Tomato yellow leaf curl virus (TYLCV), in a circular, persistent fashion. Transcriptome analyses revealed that B. tabaci knottin genes were responsive to various stresses. Upon ingestion of tomato begomoviruses, two of the four knottin genes were upregulated, knot-1 (with the highest expression) and knot-3. In this study, we examined the involvement of B. tabaci knottin genes in relation to TYLCV circulative transmission. Knottins were silenced by feeding whiteflies with knottin dsRNA via detached tomato leaves. Large amounts of knot-1 transcripts were present in the abdomen of whiteflies, an obligatory transit site of begomoviruses in their circulative transmission pathway; knot-1 silencing significantly depleted the abdomen from knot-1 transcripts. Knot-1 silencing led to an increase in the amounts of TYLCV ingested by the insects and transmitted to tomato test plants by several orders of magnitude. This effect was not observed following knot-3 silencing. Hence, knot-1 plays a role in restricting the quantity of virions an insect may acquire and transmit. We suggest that knot-1 protects B. tabaci against deleterious effects caused by TYLCV by limiting the amount of virus associated with the whitefly vector.

  18. The Whitefly Bemisia tabaci Knottin-1 Gene Is Implicated in Regulating the Quantity of Tomato Yellow Leaf Curl Virus Ingested and Transmitted by the Insect

    PubMed Central

    Hariton Shalev, Aliza; Sobol, Iris; Ghanim, Murad; Liu, Shu-Sheng; Czosnek, Henryk

    2016-01-01

    The whitefly Bemisia tabaci is a major pest to agricultural crops. It transmits begomoviruses, such as Tomato yellow leaf curl virus (TYLCV), in a circular, persistent fashion. Transcriptome analyses revealed that B. tabaci knottin genes were responsive to various stresses. Upon ingestion of tomato begomoviruses, two of the four knottin genes were upregulated, knot-1 (with the highest expression) and knot-3. In this study, we examined the involvement of B. tabaci knottin genes in relation to TYLCV circulative transmission. Knottins were silenced by feeding whiteflies with knottin dsRNA via detached tomato leaves. Large amounts of knot-1 transcripts were present in the abdomen of whiteflies, an obligatory transit site of begomoviruses in their circulative transmission pathway; knot-1 silencing significantly depleted the abdomen from knot-1 transcripts. Knot-1 silencing led to an increase in the amounts of TYLCV ingested by the insects and transmitted to tomato test plants by several orders of magnitude. This effect was not observed following knot-3 silencing. Hence, knot-1 plays a role in restricting the quantity of virions an insect may acquire and transmit. We suggest that knot-1 protects B. tabaci against deleterious effects caused by TYLCV by limiting the amount of virus associated with the whitefly vector. PMID:27455309

  19. The minimal sequence essential for replication and movement of Cotton leaf curl Multan betasatellite DNA by a helper virus in plant cells.

    PubMed

    Eini, Omid; Behjatnia, S A Akbar

    2016-10-01

    Betasatellites are single-stranded circular DNAs associated with a number of monopartite begomoviruses. Betasatellites rely on the helper begomoviruses for replication and movement in plant tissues and plant-to-plant transmission by vectors. Their genomes are approximately half the size of the helper viruses and consist of three main regions including the βC1 gene, an adenine-rich (A-rich) region, and the satellite conserved region (SCR). In this study, we investigated the minimal sequences required for Cotton leaf curl Multan betasatellite (CLCuMB) replication and movement. Mutational analysis of CLCuMB DNA genome indicated that βC1 gene and A-rich region were not required for trans-replication and movement of CLCuMB in host plants by a helper virus. Deletion of βC1 gene and a fragment (135 nt in length) upstream of this gene impaired CLCuMB replication. However, CLCuMB mutant with deletion of βC1 gene and a further 163 nucleotides replicated at a lower level as compared to the wild-type betasatellite. This suggests that there are essential elements in the fragment upstream of βC1 gene, which are required for the replication of CLCuMB rather than the size limitation of CLCuMB DNA.

  20. Effectiveness of gene silencing induced by viral vectors based on Citrus leaf blotch virus is different in Nicotiana benthamiana and citrus plants.

    PubMed

    Agüero, Jesus; Vives, María del Carmen; Velázquez, Karelia; Pina, José Antonio; Navarro, Luis; Moreno, Pedro; Guerri, Jose

    2014-07-01

    Virus induced gene silencing (VIGS) is an effective technology for gene function analysis in plants. We assessed the VIGS effectiveness in Nicotiana benthamiana and citrus plants of different Citrus leaf blotch virus (CLBV)-based vectors, using insets of the phytoene desaturase (pds) gene. While in N. benthamiana the silencing phenotype was induced only by the construct carrying a 58-nt pds hairpin, in citrus plants all the constructs induced the silencing phenotype. Differences in the generation of secondary small interfering RNAs in both species are believed to be responsible for differential host-species effects. The ability of CLBV-based vectors to silence different endogenous citrus genes was further confirmed. Since CLBV-based vectors are known to be stable and induce VIGS in successive flushes for several months, these vectors provide an important genomic tool and it is expected that they will be useful to analyze gene function by reverse genetics in the long-lived citrus plants. Copyright © 2014 Elsevier Inc. All rights reserved.

  1. Molecular cloning of Indian tomato leaf curl virus genome following a simple method of concentrating the supercoiled replicative form of viral DNA.

    PubMed

    Srivastava, K M; Hallan, V; Raizada, R K; Chandra, G; Singh, B P; Sane, P V

    1995-02-01

    DNA-A and DNA-B components of the genome of a whitefly transmitted virus causing yellowing and leaf curl in tomato (ITLCV) were cloned following a simple procedure for isolation of the double stranded replicative form of viral DNA from infected tomato plants. The method is based on extraction of total DNA from infected plants followed by concentration of the double stranded replicative form of viral DNA by an alkaline denaturation procedure identical to that used for isolation of plasmid DNA from Escherichia coli. The attempted cloning of DNA showed that 95% of the transformants contained plasmids with an insert of either DNA-A (2.75 kb) or DNA-B (2.55 kb). Cloned DNA-A and DNA-B when used as probes could detect DNA-A/DNA-B in total nucleic acid obtained from fresh diseased tissue. Both DNA-A and DNA-B are needed for infection and they have a common region of 166 bases with about 94% nucleotide sequence homology, a characteristic of all bipartite geminiviruses. Comparison of the amino acid sequence of the putative coat protein product of ITLCV with some other mono- and bipartite geminiviruses revealed a maximum of 86% homology with Indian cassava mosaic virus.

  2. Rapid Construction of Stable Infectious Full-Length cDNA Clone of Papaya Leaf Distortion Mosaic Virus Using In-Fusion Cloning.

    PubMed

    Tuo, Decai; Shen, Wentao; Yan, Pu; Li, Xiaoying; Zhou, Peng

    2015-12-01

    Papaya leaf distortion mosaic virus (PLDMV) is becoming a threat to papaya and transgenic papaya resistant to the related pathogen, papaya ringspot virus (PRSV). The generation of infectious viral clones is an essential step for reverse-genetics studies of viral gene function and cross-protection. In this study, a sequence- and ligation-independent cloning system, the In-Fusion(®) Cloning Kit (Clontech, Mountain View, CA, USA), was used to construct intron-less or intron-containing full-length cDNA clones of the isolate PLDMV-DF, with the simultaneous scarless assembly of multiple viral and intron fragments into a plasmid vector in a single reaction. The intron-containing full-length cDNA clone of PLDMV-DF was stably propagated in Escherichia coli. In vitro intron-containing transcripts were processed and spliced into biologically active intron-less transcripts following mechanical inoculation and then initiated systemic infections in Carica papaya L. seedlings, which developed similar symptoms to those caused by the wild-type virus. However, no infectivity was detected when the plants were inoculated with RNA transcripts from the intron-less construct because the instability of the viral cDNA clone in bacterial cells caused a non-sense or deletion mutation of the genomic sequence of PLDMV-DF. To our knowledge, this is the first report of the construction of an infectious full-length cDNA clone of PLDMV and the splicing of intron-containing transcripts following mechanical inoculation. In-Fusion cloning shortens the construction time from months to days. Therefore, it is a faster, more flexible, and more efficient method than the traditional multistep restriction enzyme-mediated subcloning procedure.

  3. Rapid Construction of Stable Infectious Full-Length cDNA Clone of Papaya Leaf Distortion Mosaic Virus Using In-Fusion Cloning

    PubMed Central

    Tuo, Decai; Shen, Wentao; Yan, Pu; Li, Xiaoying; Zhou, Peng

    2015-01-01

    Papaya leaf distortion mosaic virus (PLDMV) is becoming a threat to papaya and transgenic papaya resistant to the related pathogen, papaya ringspot virus (PRSV). The generation of infectious viral clones is an essential step for reverse-genetics studies of viral gene function and cross-protection. In this study, a sequence- and ligation-independent cloning system, the In-Fusion® Cloning Kit (Clontech, Mountain View, CA, USA), was used to construct intron-less or intron-containing full-length cDNA clones of the isolate PLDMV-DF, with the simultaneous scarless assembly of multiple viral and intron fragments into a plasmid vector in a single reaction. The intron-containing full-length cDNA clone of PLDMV-DF was stably propagated in Escherichia coli. In vitro intron-containing transcripts were processed and spliced into biologically active intron-less transcripts following mechanical inoculation and then initiated systemic infections in Carica papaya L. seedlings, which developed similar symptoms to those caused by the wild-type virus. However, no infectivity was detected when the plants were inoculated with RNA transcripts from the intron-less construct because the instability of the viral cDNA clone in bacterial cells caused a non-sense or deletion mutation of the genomic sequence of PLDMV-DF. To our knowledge, this is the first report of the construction of an infectious full-length cDNA clone of PLDMV and the splicing of intron-containing transcripts following mechanical inoculation. In-Fusion cloning shortens the construction time from months to days. Therefore, it is a faster, more flexible, and more efficient method than the traditional multistep restriction enzyme-mediated subcloning procedure. PMID:26633465

  4. The C2 protein of tomato leaf curl Taiwan virus is a pathogenicity determinant that interferes with expression of host genes encoding chromomethylases.

    PubMed

    Tu, Yu-Ching; Tsai, Wen-Shi; Wei, Jyuan-Yu; Chang, Kai-Ya; Tien, Chang-Ching; Hsiao, Hui-Yu; Fu, Shih-Feng

    2017-08-08

    Tomato (Solanum lycopersicum) is one of the most important crops worldwide and is severely affected by geminiviruses. Tomato leaf curl Taiwan virus (ToLCTWV), belonging to the geminiviruses, was isolated in Taiwan and causes tremendous crop loss. The geminivirus-encoded C2 proteins are crucial for a successful interaction between the virus and host plants. However, the exact functions of the viral C2 protein of ToLCTWV have not been investigated. We analyzed the molecular function(s) of the C2 protein by transient or stable expression in tomato cv. Micro-Tom and Nicotiana benthamiana. Severe stunting of tomato and N. benthamiana plants infected with ToLCTWV was observed. Expression of ToLCTWV C2-green fluorescent protein (GFP) fusion protein was predominately located in the nucleus and contributed to activation of coat protein promoter. Notably, the C2-GFP fluorescence was distributed in nuclear aggregates. Tomato and N. benthamiana plants inoculated with potato virus X (PVX)-C2 displayed chlorotic lesions and stunted growth. PVX-C2 elicited hypersensitive responses accompanied by production of reactive oxygen species in N. benthamiana plants, which suggests that the viral C2 was a potential recognition target to induce host-defense responses. In tomato and N. benthamiana, ToLCTWV C2 was found to interfere with expression of genes encoding chromomethylases. N. benthamiana plants with suppressed NbCMT3-2 expression were more susceptible to ToLCTWV infection. Transgenic N. benthamiana plants expressing the C2 protein showed decreased expression of the NbCMT3-2 gene and pNbCMT3-2::GUS (b-glucuronidase) promoter activity. C2 protein is an important pathogenicity determinant of ToLCTWV and interferes with host components involved in DNA methylation. This article is protected by copyright. All rights reserved.

  5. Prediction and characterization of Tomato leaf curl New Delhi virus (ToLCNDV) responsive novel microRNAs in Solanum lycopersicum.

    PubMed

    Pradhan, Bhubaneswar; Naqvi, Afsar Raza; Saraf, Shradha; Mukherjee, Sunil Kumar; Dey, Nrisingha

    2015-01-02

    Tomato leaf curl New Delhi virus (ToLCNDV) infects tomato (Solanum lycopersicum) plants and causes severe crop losses. As the microRNAs (miRNAs) are deregulated during stressful events, such as biotic stress, we wanted to study the effect of ToLCNDV infection on tomato miRNAs. We constructed two libraries, isolating small RNAs (sRNAs) from healthy (HT) and ToLCNDV infected (IT) tomato leaves, and sequenced the library-specific sRNAs using the next generation sequencing (NGS) approach. These data helped predict 112 mature miRNA sequences employing the miRDeep-P program. A substantial number (58) of the sequences were 24-mer in size, which was a bit surprising. Based on the calculation of precision values, 53 novel miRNAs were screened from the predicted sequences. Nineteen of these were chosen for expression analysis; a northern blot analysis showed 15 to be positive. Many of the predicted miRNAs were up-regulated following viral infection. The target genes of the miRNAs were also predicted and the expression analysis of selected transcripts showed a typical inverse relation between the accumulation of target transcripts and the abundance of corresponding miRNAs. Furthermore, the cleavage sites of the target transcripts for three novel miRNAs were mapped, confirming the correct annotation of the miRNA-targets. The sRNA deep sequencing clearly revealed that the virus modulated global miRNA expression in the host. The validated miRNAs (Tom_4; Tom_14; Tom_17; Tom_21; Tom_29; Tom_43) could be valuable tools for understanding the ToLCNDV-tomato interaction, ultimately leading to the development of a virus-resistant tomato plant. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Rolling-Contact Rheostat

    NASA Technical Reports Server (NTRS)

    Ruoff, C. F.

    1985-01-01

    Contact noise in rheostats and potentiometers reduced by rolling contact design. Smooth rolling action eliminates sporadic variations in resistance caused by bouncing and stick/slip motion of conventional sliding contacts.

  7. 4. ROLL LATHE OF THE MAIN ROLL SHOP. THE LATHE ...

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

    4. ROLL LATHE OF THE MAIN ROLL SHOP. THE LATHE WAS CLOSED WITH TWO ROLLS IN PLACE, AND THE LOWER ROLL WAS TURNED TO MATCH THE UPPER ROLL. - U.S. Steel Homestead Works, Main Roll Shop, Along Monongahela River, Homestead, Allegheny County, PA

  8. Internal roll compression system

    DOEpatents

    Anderson, Graydon E.

    1985-01-01

    This invention is a machine for squeezing water out of peat or other material of low tensile strength; the machine including an inner roll eccentrically positioned inside a tubular outer roll, so as to form a gradually increasing pinch area at one point therebetween, so that, as the rolls rotate, the material is placed between the rolls, and gets wrung out when passing through the pinch area.

  9. Effect of a single amino acid substitution in the NLS domain of Tomato yellow leaf curl virus-Israel (TYLCV-IL) capsid protein (CP) on its activity and on the virus life cycle.

    PubMed

    Yaakov, Noga; Levy, Yael; Belausov, Eduard; Gaba, Victor; Lapidot, Moshe; Gafni, Yedidya

    2011-06-01

    The capsid protein (CP) of Tomato yellow leaf curl virus-Israel (TYLCV-IL), encoded by the v1 gene, is the only known component of the viral capsid. Three point mutations introduced into the conserved NLS region of the CP were investigated. One mutant, in which the Arg at position 19 was converted to Leu, had the most significant effect on the CP-CP homotypic interaction as well as on CP's interaction with its nuclear receptor karyopherin α1 and with the protein GroEL. The latter has been suggested to protect the virions in the insect vector hemolymph. These effects were first observed by yeast two-hybrid assay and then confirmed in tobacco protoplasts by measuring fluorescence resonance energy transfer (FRET) between YFP- and CFP-tagged proteins. Most importantly, when the point mutation converting Arg 19 to Leu was introduced into the full-length TYLCV genome, it disrupted its ability to cause symptoms.

  10. Comparative transcriptome analysis in Bemisia tabaci in response to tomato yellow leaf curl virus and development of ribonucleic acid interference to manage whitefly-transmitted viruses

    USDA-ARS?s Scientific Manuscript database

    The whitefly, Bemisia tabaci transmits over 300 plant viruses, with the majority of them belonging to the Begomovirus genus. Begomoviruses are obligately transmitted to a wide range of agriculture crops, resulting in the loss of billions of dollars annually, while jeopardizing food security worldwid...

  11. Transcriptome analysis of Bemisia tabaci during tomato yellow leaf curl virus acquisition and ribonucleic acid interference to manage whitefly-transmitted viruses

    USDA-ARS?s Scientific Manuscript database

    Over 300 viruses are transmitted by the whitefly, Bemisia tabaci, with 90% of them belonging to the genus, Begomovirus. Begomoviruses are obligately transmitted by whiteflies to a wide range of agriculture crops, resulting in billions of dollars lost annually, while jeopardizing food security worldw...

  12. Molecular evolutionary history of Sugarcane yellow leaf virus based on sequence analysis of RNA-dependent RNA polymerase and putative aphid transmission factor-coding genes.

    PubMed

    ElSayed, Abdelaleim Ismail; Boulila, Moncef; Rott, Philippe

    2014-06-01

    RNA-dependent RNA polymerase (RdRp) encoded by ORF2 and putative aphid transmission factor (PATF) encoded by ORF5 of Sugarcane yellow leaf virus (SCYLV) were detected in six sugarcane cultivars affected by yellow leaf using RT-PCR and real-time RT-PCR assays. Expression of both genes varied among infected plants, but overall expression of RdRp was higher than expression of PATF. Cultivar H87-4094 from Hawaii yielded the highest transcript levels of RdRp, whereas cultivar C1051-73 from Cuba exhibited the lowest levels. Sequence comparisons among 25 SCYLV isolates from various geographical locations revealed an amino acid similarity of 72.1-99.4 and 84.7-99.8 % for the RdRp and PATF genes, respectively. The 25 SCYLV isolates were separated into three (RdRp) and two (PATF) phylogenetic groups using the MEGA6 program that does not account for genetic recombination. However, the SCYLV genome contained potential recombination signals in the RdRp and PATF coding genes based on the GARD genetic algorithm. Use of this later program resulted in the reconstruction of phylogenies on the left as well as on the right sides of the putative recombination breaking points, and the 25 SCYLV isolates were distributed into three distinct phylogenetic groups based on either RdRp or PATF sequences. As a result, recombination reshuffled the affiliation of the accessions to the different clusters. Analysis of selection pressures exerted on RdRp and PATF encoded proteins revealed that ORF 2 and ORF 5 underwent predominantly purifying selection. However, a few sites were also under positive selection as assessed by various models such as FEL, IFEL, REL, FUBAR, MEME, GA-Branch, and PRIME.

  13. Yellowing disease in zucchini squash produced by mixed infections of Cucurbit yellow stunting disorder virus and Cucumber vein yellowing virus.

    PubMed

    Gil-Salas, Francisco M; Peters, Jeff; Boonham, Neil; Cuadrado, Isabel M; Janssen, Dirk

    2011-11-01

    Zucchini squash is host to Cucurbit yellow stunting disorder virus (CYSDV), a member of the genus Crinivirus, and Cucumber vein yellowing virus (CVYV), a member of the genus Ipomovirus, both transmitted by the whitefly Bemisia tabaci. Field observations suggest the appearance of new symptoms observed on leaves of zucchini squash crops when both viruses were present. When infected during controlled experiments with CYSDV only, zucchini plants showed no obvious symptoms and the virus titer decreased between 15 and 45 days postinoculation (dpi), after which it was no longer detected. CVYV caused inconspicuous symptoms restricted to vein clearing on some of the apical leaves and the virus accumulated progressively between 15 and 60 dpi. Similar accumulations of virus followed single inoculations with the potyvirus Zucchini yellow mosaic virus (ZYMV) and plants showed severe stunting, leaf deformation, and mosaic yellowing. However, in mixed infections with CYSDV and CVYV, intermediate leaves showed chlorotic mottling which evolved later to rolling, brittleness, and complete yellowing of the leaf lamina, with exception of the veins. No consistent alteration of CVYV accumulation was detected but the amounts of CYSDV increased ≈100-fold and remained detectable at 60 dpi. Such synergistic effects on the titer of the crinivirus and symptom expression were not observed when co-infected with ZYMV.

  14. Intracellular distribution, cell-to-cell trafficking and tubule-inducing activity of the 50 kDa movement protein of Apple chlorotic leaf spot virus fused to green fluorescent protein.

    PubMed

    Satoh, H; Matsuda, H; Kawamura, T; Isogai, M; Yoshikawa, N; Takahashi, T

    2000-08-01

    The 50 kDa protein (50KP) encoded by ORF2 of Apple chlorotic leaf spot virus (ACLSV) fused to green fluorescent protein (GFP) was expressed transiently in cells of Nicotiana occidentalis and Chenopodium quinoa leaves. Its intracellular distribution, cell-to-cell trafficking in leaf epidermis and tubule formation on the surface of protoplasts were analysed. The 50KP-GFP fluorescence was distributed as small irregular spots or a fibrous network structure on the periphery of epidermal cells and protoplasts of both plant species. In leaf epidermis of N. occidentalis, the protein spread from the cells that produced it into neighbouring cells in both young and mature leaves and targetted plasmodesmata in these cells. In contrast, GFP was restricted to single cells in most cases in mature leaves. When 50KP and GFP were co-expressed in leaf epidermis of N. occidentalis, GFP spread more widely from the initial cells that produced it than when GFP was expressed alone, suggesting that 50KP facilitated the cell-to-cell trafficking of GFP. 50KP-GFP was able to complement local spread of 50KP-deficient virus when expressed transiently in leaf epidermis of C. quinoa. Expression of 50KP-GFP in protoplasts resulted in the production of tubular structures protruding from the surface. Mutational analyses showed that the C-terminal region (aa 287-457) was not essential for localization to plasmodesmata, cell-to-cell trafficking, complementation of movement of 50KP-deficient virus or tubule formation on protoplasts. In contrast, deletions in the N-terminal region resulted in the complete disruption of all these activities.

  15. Modelling pressure rolling of asymmetric rolling process

    NASA Astrophysics Data System (ADS)

    Alexa, V.; Ratiu, S. A.; Kiss, I.; Cioata, V. G.

    2017-05-01

    The paper presents a comparative analysis between experimental results and modelling in order to interpret the value of the contact pressure on the asymmetric longitudinal rolling. It is also intended action and the different behaviour of upper cylinder compared to the lower cylinder action in situations when both are driven, or only one operates. In the modelling will be presented on the basis of boundary conditions imposed rolling pressure variation in the degree of reduction and also re size arc length of contact. Determining a curve is also important to determine the locus of points which characterize symmetry conditions partial rolling process between unequal diameters cylinders.

  16. Geminivirus-Mediated Gene Silencing from Cotton Leaf Crumple Virus Is Enhanced by Low Temperature in Cotton1[C][OA

    PubMed Central

    Tuttle, John R.; Idris, A.M.; Brown, Judith K.; Haigler, Candace H.; Robertson, Dominique

    2008-01-01

    A silencing vector for cotton (Gossypium hirsutum) was developed from the geminivirus Cotton leaf crumple virus (CLCrV). The CLCrV coat protein gene was replaced by up to 500 bp of DNA homologous to one of two endogenous genes, the magnesium chelatase subunit I gene (ChlI) or the phytoene desaturase gene (PDS). Cotyledons of cotton cultivar ‘Deltapine 5415’ bombarded with the modified viral vectors manifested chlorosis due to silencing of either ChlI or PDS in approximately 70% of inoculated plants after 2 to 3 weeks. Use of the green fluorescence protein gene showed that replication of viral DNA was restricted to vascular tissue and that the viral vector could transmit to leaves, roots, and the ovule integument from which fibers originate. Temperature had profound effects on vector DNA accumulation and the spread of endogenous gene silencing. Consistent with reports that silencing against viruses increases at higher temperatures, plants grown at a 30°C/26°C day/night cycle had a greater than 10-fold reduction in viral DNA accumulation compared to plants grown at 22°C/18°C. However, endogenous gene silencing decreased at 30°C/26°C. There was an approximately 7 d delay in the onset of gene silencing at 22°C/18°C, but silencing was extensive and persisted throughout the life of the plant. The extent of silencing in new growth could be increased or decreased by changing temperature regimes at various times following the onset of silencing. Our experiments establish the use of the CLCrV silencing vector to study gene function in cotton and show that temperature can have a major impact on the extent of geminivirus-induced gene silencing. PMID:18621976

  17. Preference by a virus vector for infected plants is reversed after virus acquisition.

    PubMed

    Rajabaskar, Dheivasigamani; Bosque-Pérez, Nilsa A; Eigenbrode, Sanford D

    2014-06-24

    Pathogens and their vectors can interact either directly or indirectly via their shared hosts, with implications for the persistence and spread of the pathogen in host populations. For example, some plant viruses induce changes in host plants that cause the aphids that carry these viruses to settle preferentially on infected plants. Furthermore, relative preference by the vector for infected plants can change to a preference for noninfected plants after virus acquisition by the vector, as has recently been demonstrated in the wheat-Rhopalosiphum padi-Barley yellow dwarf virus pathosystem. Here we document a similar dynamic in the potato-Myzus persicae (Sulzer)-Potato leaf roll virus (PLRV) pathosystem. Specifically, in a dual choice bioassay, nonviruliferous apterous M. persicae settled preferentially on or near potato plants infected with PLRV relative to noninfected (sham-inoculated) control plants, whereas viruliferous M. persicae (carrying PLRV) preferentially settled on or near sham-inoculated potato plants relative to infected plants. The change in preference after virus acquisition also occurred in response to trapped headspace volatiles, and to synthetic mimics of headspace volatile blends from PLRV-infected and sham-inoculated potato plants. The change in preference we document should promote virus spread by increasing rates of virus acquisition and transmission by the vector. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. Strategies to facilitate the development of uncloned or cloned infectious full-length viral cDNAs: Apple chlorotic leaf spot virus as a case study

    PubMed Central

    2011-01-01

    Background Approaches to simplify and streamline the construction of full-length infectious cDNA clones (FL-cDNAs) are needed. Among desirable improvements are the ability to use total nucleic acids (TNA) extracts from infected hosts (to bypass viral purification limitations) for the direct one-step amplification of large FL-cDNAs, the possibility to inoculate plants with uncloned FL-cDNAs and the simplified cloning of these large molecules. Results Using the 7.55 kb genome of Apple chlorotic leaf spot trichovirus (ACLSV) approaches allowing the rapid generation from TNA extracts of FL-cDNAs under the control of the T7 promoter and the successful inoculation of plants using in vitro transcripts obtained from these uncloned amplification products have been developed. We also show that the yeast homologous recombination system permits efficient cloning of FL-cDNAs and the simultaneous one-step tailoring of a ternary Yeast-Escherichia coli-Agrobacterium tumefaciens shuttle vector allowing efficient inoculation of both herbaceous and woody host plants by agroinfiltration. Conclusions The fast and efficient strategies described here should have broad applications, in particular for the study of "difficult" plant viruses, such as those infecting woody hosts, and potentially for other, non plant-infecting viral agents. PMID:22040379

  19. Complete nucleotide sequences of the genomes of two isolates of apple chlorotic leaf spot virus from peach (Prunus persica) in China.

    PubMed

    Niu, Feiqing; Pan, Song; Wu, Zujian; Jiang, Dongmei; Li, Shifang

    2012-04-01

    The complete nucleotide sequences of two isolates of apple chlorotic leaf spot virus (Z1 and Z3) collected from peach in Henan Province, China, were determined. The genomes of both Z1 and Z3 were found to contain three open reading frames (ORFs). Sequence analysis showed that genomic sequences of Z1 and Z3 isolates shared 67.4%-82.9% and 67.2%-82.6% identity, respectively, with the other eight isolates of ACLSV that have been reported previously. Based on the putative amino acid sequences of the products of the three ORFs, Z1 and Z3 isolates showed the greatest identity to isolate PBM1 (GenBank accession number AJ243438) from plum and the least identity with isolate Ta Tao5 (GenBank Accession Number: EU223295) from peach. Considering the low level of sequence identity between Z1/Z3 isolate and Ta Tao5 isolate, two types of ACLSV may exist in peach.

  20. The Cucumber leaf spot virus p25 auxiliary replicase protein binds and modifies the endoplasmic reticulum via N-terminal transmembrane domains

    SciTech Connect

    Ghoshal, Kankana; Theilmann, Jane; Reade, Ron; Sanfacon, Helene; Rochon, D’Ann

    2014-11-15

    Cucumber leaf spot virus (CLSV) is a member of the Aureusvirus genus, family Tombusviridae. The auxiliary replicase of Tombusvirids has been found to localize to endoplasmic reticulum (ER), peroxisomes or mitochondria; however, localization of the auxiliary replicase of aureusviruses has not been determined. We have found that the auxiliary replicase of CLSV (p25) fused to GFP colocalizes with ER and that three predicted transmembrane domains (TMDs) at the N-terminus of p25 are sufficient for targeting, although the second and third TMDs play the most prominent roles. Confocal analysis of CLSV infected 16C plants shows that the ER becomes modified including the formation of punctae at connections between ER tubules and in association with the nucleus. Ultrastructural analysis shows that the cytoplasm contains numerous vesicles which are also found between the perinuclear ER and nuclear membrane. It is proposed that these vesicles correspond to modified ER used as sites for CLSV replication. - Highlights: • The CLSV p25 auxiliary replicase targets the endoplasmic reticulum (ER). • Targeting of CLSV p25 is associated with ER restructuring. • Restructuring of the ER occurs during CLSV infection. • CLSV p25 contains 3 predicted transmembrane domains 2 of which are required for ER targeting. • Vesicles derived from the ER may be sites of CLSV replication.

  1. Whitefly population dynamics and evaluation of whitefly-transmitted tomato yellow leaf curl virus (TYLCV)-resistant tomato genotypes as whitefly and TYLCV reservoirs.

    PubMed

    Srinivasan, Rajagopalbabu; Riley, David; Diffie, Stan; Sparks, Alton; Adkins, Scott

    2012-08-01

    Sweetpotato whitefly, Bemisia tabaci (Gennadius), and whitefly-transmitted tomato yellow leaf curl virus (TYLCV) are major threats to tomato production in the southeastern United States. TYLCV was introduced to Florida from the Caribbean islands and has spread to other southern states of the United States. In Georgia, in recent years, the incidence of TYLCV has been steadily increasing. Studies were conducted to monitor population dynamics of whiteflies in the vegetable production belt of Georgia, to evaluate TYLCV-resistant genotypes against whiteflies and TYLCV, and to assess the potential role of resistant genotypes in TYLCV epidemiology. Monitoring studies indicated that the peak incidence of whiteflies varied seasonally from year to year. In general, whitefly populations were not uniformly distributed. Tomato genotypes exhibited minor differences in their ability to support whitefly populations. TYLCV symptoms were visually undetectable in all but one resistant genotype. The infection rates (visually) in susceptible genotypes ranged from 40 to 87%. Greenhouse inoculations with viruliferous whiteflies followed by polymerase chain reaction (PCR) indicated that up to 100% of plants of resistant genotypes were infected, although predominantly symptomless. TYLCV acquisition by whiteflies from TYLCV-infected genotypes was tested by PCR; TYLCV acquisition rates from resistant genotypes were less than from susceptible genotypes. Nevertheless, this difference did not influence TYLCV transmission rates from resistant to susceptible genotypes. Results emphasize that resistant genotypes can serve as TYLCV and whitefly reservoirs and potentially influence TYLCV epidemics.

  2. Generation and characterization of a scFv against recombinant coat protein of the geminivirus tomato leaf curl New Delhi virus.

    PubMed

    Zakri, Adel M; Ziegler, Angelika; Torrance, Lesley; Fischer, Rainer; Commandeur, Ulrich

    2010-03-01

    We report the establishment of a hybridoma cell line secreting the monoclonal antibody (mAb) HAV, which recognizes the coat (AV1) protein of tomato leaf curl New Delhi virus (ToLCNDV), a begomovirus. The cell line was obtained following immunization of mice with purified recombinant AV1 fused to glutathione S-transferase (GST). A single-chain variable fragment (scFv-SAV) was assembled from hybridoma cDNA, but sequence analysis revealed a single nucleotide deletion causing a frame shift that resulted in a 21-residue N-terminal truncation. The missing nucleotide was restored by in vitro site-directed mutagenesis to create scFv-RWAV. The binding properties of mAb HAV and the corresponding scFvs were characterized by western blot, ELISA and surface plasmon resonance spectroscopy. MAb HAV bound to AV1 with nanomolar affinity but reacted neither with the N-terminal region of the protein nor with the GST fusion partner. This suggested that the antibody recognized a linear epitope in a region of the coat protein that is conserved among begomoviruses. Both scFvs retained the antigen specificity of mAb HAV, although the dissociation rate constant of scFv-RWAV was tenfold greater than that of scFv-SAV, showing the importance of restoring the 21 N-terminal amino acids.

  3. Transferring cucumber mosaic virus-white leaf strain coat protein gene into Cucumis melo L. and evaluating transgenic plants for protection against infections

    SciTech Connect

    Gonsalves, C.; Xue, B.; Yepes, M.; Fuchs, M.; Ling, K.; Namba, S. . Dept. of Plant Pathology)

    1994-03-01

    A single regeneration procedure using cotyledon examples effectively regenerated five commercially grown muskmelon cultivars. This regeneration scheme was used to facilitate gene transfers using either Agrobacterium tumefaciens or microprojectile bombardment methods. In both cases, the transferred genes were from the T-DNA region of the binary vector plasmid pGA482GG/cp cucumber mosaic virus-white leaf strain (CMV-WL), which contains genes that encode neomycin phosphotransferase II (NPT II), [beta]-glucuronidase (GUS), and the CMV-WL coat protein (CP). Explants treated with pGA482GG/cpCMV-WL regenerated shoots on Murashige and Skoog medium containing 4.4 [mu]m 6-benzylaminopurine (BA), kanamycin (Km) at 150 mg[center dot]liter[sup [minus]1] and carbenicillin (Cb) at 500 mg[center dot]liter[sup [minus]1]. The authors' comparison of A. tumefaciens- and microprojectile-mediated gene transfer procedures shows that both methods effectively produce nearly the same percentage of transgenic plants. R[sub 0] plants were first tested for GUS or NPT II expression, then the polymerase chain reaction (PCR) and other tests were used to verify the transfer of the NPT II, GUS, and CMV-WL CP genes.

  4. Cestrum yellow leaf curling virus (CmYLCV) promoter: a new strong constitutive promoter for heterologous gene expression in a wide variety of crops.

    PubMed

    Stavolone, Livia; Kononova, Maria; Pauli, Sandra; Ragozzino, Antonio; de Haan, Peter; Milligan, Steve; Lawton, Kay; Hohn, Thomas

    2003-11-01

    Appropriately regulated gene expression requires a suitable promoter. A number of promoters have been isolated and shown to be functional in plants, but only a few of them activate transcription of transgenes at high levels constitutively. We report here the cloning and characterization of a novel, constitutively expressed promoter isolated from Cestrum yellow leaf curling virus (CmYLCV), a double-stranded DNA plant pararetrovirus belonging to the Caulimoviridae family. The CmYLCV promoter is highly active in callus, meristems and vegetative and reproductive tissues in Arabidopsis thaliana, Nicotiana tabacum, Lycopersicon esculentum, Zea mays and Oryza sativa. Furthermore, the level of expression is comparable to, or higher than, that from the CaMV 35S, the 'super-promoter' or the maize ubiquitin 1 promoters, three frequently used promoters in agricultural biotechnology. The heritable, strong and constitutive activity in both monocotyledonous and dicotyledonous plants, combined with the extremely narrow CmYLCV host range, makes the CmYLCV promoter an attractive tool for regulating transgene expression in a wide variety of plant species.

  5. PCR-RFLP-based typing for differentiation of Tomato yellow leaf curl virus (TYLCV) genotypes from infected host plants in Korea.

    PubMed

    Oh, Sung; Kim, Seongdae; Vinod, Nagarajan; Koo, Jung Mo; Jang, Kyung Min; Choi, Chang Won; Kim, Seong Hwan; Kim, Young Shik

    2013-12-01

    A polymerase chain reaction (PCR) using two sets of primers designed from published Tomato yellow leaf curl virus (TYLCV) genomes was developed to distinguish from the TYLCV-IL groups. The specificity of the two sets of primers was proven by testing against control TYLCV genomes and the symptomatic leaves of 34 different tomato cultivars naturally infected with TYLCV in greenhouses. One set for TYLCV-IL strain-specific primers (TYLCV-UNI-F and TYLCV-UNI-R) amplified full-length genome fragments from all the 34 tomato cultivars. Another set for TYLCV-IL group-II strain-specific primers (TYLCV-GPII-F and TYLCV-GPII-R) amplified target DNA fragments from only 9 tomato cultivars. Digestion by BglII and EcoRV of the PCR amplicons produced restriction fragment length polymorphism pattern that distinguished the TYLCV-IL group-I with two fragments from the TYLCV-IL group-II with no digested fragment. PCR coupled with BglII and EcoRV digestion confirmed that the 9 tomato cultivars were infected with the TYLCV-IL group-II and the remained 25 tomato cultivars were infected with the TYLCV-IL group-I.

  6. Evaluation of management strategies for bean leaf beetles (Coleoptera: Chrysomelidae) and Bean pod mottle virus (Comoviridae) in soybean.

    PubMed

    Bradshaw, Jeffrey D; Rice, Marlin E; Hill, John H

    2008-08-01

    Cerotoma trifurcata Förster (Coleoptera: Chrysomelidae) and Bean pod mottle virus (Comoviridae) (BPMV) both can reduce yield and seed quality of soybean, Glycine max (L.) Merr. Field experiments were conducted to evaluate the effects of systemic, seed-applied, and foliar-applied insecticides for the management of this pest complex at three locations in central, northeastern, and northwestern Iowa during 2002-2004. Seed-applied insecticide was evaluated according to a currently recommended management program for Iowa (i.e., insecticide applications that target emerging overwintered beetles, F0, and the first seasonal generation, F1 ). The experimental treatments included seed-applied (thiamethoxam, 0.3-0.5 g [AI] kg(-1)] or clothianidin, 47.32 ml [AI] kg(-1)) and foliar-applied (A-cyhalothrin, 16.83-28.05 g [AI] ha(-1)) or esfenvalerate (43.74-54.69 g [AI] ha(-1)) insecticides. Applications of the foliar insecticides were timed to target F0, F1 or both F0 and F1 populations of C. trifurcata. Our results confirm that insecticides timed at F0 and F1 populations of C. trifurcata can reduce vector populations throughout the growing season, provide limited reduction in virus incidence, and improve both yield and seed coat color. Furthermore, seed-applied insecticides may be the more reliable option for an F0-targeted insecticide if used within this management strategy. An F0-targeted insecticide by itself only gave a yield improvement in one out of eight location-years. However, by adding an F1-targeted insecticide, there was a yield gain of 1.42-1.67 quintal ha(-1), based on contrast comparisons at three location-years.

  7. Origins of rolling friction

    NASA Astrophysics Data System (ADS)

    Cross, Rod

    2017-09-01

    When a hard object rolls on a soft surface, or vice versa, rolling friction arises from deformation of the soft object or the soft surface. The friction force can be described in terms of an offset in the normal reaction force or in terms of energy loss arising from the deformation. The origin of the friction force itself is not entirely clear. It is investigated qualitatively in this paper by rolling a steel ball on soft foam and by rolling a foam cylinder on a hard surface. The deformation of the foam was observed visually, providing simple insights into the origin of the friction force.

  8. Impact of aphid alarm pheromone release on virus transmission efficiency: When pest control strategy could induce higher virus dispersion.

    PubMed

    Lin, Fang-Jing; Bosquée, Emilie; Liu, Ying-Jie; Chen, Ju-Lian; Yong, Liu; Francis, Frédéric

    2016-09-01

    Aphids cause serious damages to crops not only by tacking sap but also by transmitting numerous viruses. To develop biological control, the aphid alarm pheromone, namely E-β-farnesene (EβF), has been demonstrated to be efficient to repel aphids and as attract beneficials, making it a potential tool to control aphid pests. Considering aphids also as virus vectors, changes of their behavior could also interfere with the virus acquisition and transmission process. Here, a combination of two aphid species and two potato virus models were selected to test the influence of EβF release on aphid and virus dispersion under laboratory conditions. EβF release was found to significantly decrease the population of Myzus persicae and Macrosiphum euphorbiae around the infochemical releaser but simultaneously also increasing the dispersal of Potato Virus Y (PVY). At the opposite, no significant difference for Potato Leaf Roll Virus (PLRV) transmission efficiency was observed with similar aphid alarm pheromone releases for none of the aphid species. These results provide some support to carefully consider infochemical releasers not only for push-pull strategy and pest control but also to include viral disease in a the plant protection to aphids as they are also efficient virus vectors. Impact of aphid kinds and transmission mechanisms will be discussed according to the large variation found between persistent and non persistent potato viruses and interactions with aphids and related infochemicals. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Effect of herbivore damage on broad leaf motion in wind

    NASA Astrophysics Data System (ADS)

    Burnett, Nicholas; Kothari, Adit

    2015-11-01

    Terrestrial plants regularly experience wind that imposes aerodynamic forces on the plants' leaves. Passive leaf motion (e.g. fluttering) and reconfiguration (e.g. rolling into a cone shape) in wind can affect the drag on the leaf. In the study of passive leaf motion in wind, little attention has been given to the effect of herbivory. Herbivores may alter leaf motion in wind by making holes in the leaf. Also, a small herbivore (e.g. snail) on a leaf can act as a point mass, thereby affecting the leaf's motion in wind. Conversely, accelerations imposed on an herbivore sitting on a leaf by the moving leaf may serve as a defense by dislodging the herbivore. In the present study, we investigated how point masses (>1 g) and holes in leaves of the tuliptree affected passive leaf motion in turbulent winds of 1 and 5 m s-1. Leaf motion was unaffected by holes in the leaf surface (about 10% of leaf area), but an herbivore's mass significantly damped the accelerations of fluttering leaves. These results suggest that an herbivore's mass, but not the damage it inflicts, can affect leaf motion in the wind. Furthermore, the damping of leaf fluttering from an herbivore's mass may prevent passive leaf motions from being an effective herbivore defense.

  10. Whitefly resistance traits derived from the wild tomato Solanum pimpinellifolium affect the preference and feeding behavior of Bemisia tabaci and reduce the spread of Tomato yellow leaf curl virus.

    PubMed

    Rodríguez-López, M J; Garzo, E; Bonani, J P; Fereres, A; Fernández-Muñoz, R; Moriones, E

    2011-10-01

    Breeding of tomato genotypes that limit whitefly (Bemisia tabaci) access and feeding might reduce the spread of Tomato yellow leaf curl virus (TYLCV), a begomovirus (genus Begomovirus, family Geminiviridae) that is the causal agent of tomato yellow leaf curl disease. TYLCV is restricted to the phloem and is transmitted in a persistent manner by B. tabaci. The tomato breeding line ABL 14-8 was developed by introgressing type IV leaf glandular trichomes and secretion of acylsucroses from the wild tomato Solanum pimpinellifolium accession TO-937 into the genetic background of the whitefly- and virus-susceptible tomato cultivar Moneymaker. Results of preference bioassays with ABL 14-8 versus Moneymaker indicated that presence of type IV glandular trichomes and the production of acylsucrose deterred the landing and settling of B. tabaci on ABL 14-8. Moreover, electrical penetration graph studies indicated that B. tabaci adults spent more time in nonprobing activities and showed a reduced ability to start probing. Such behavior resulted in a reduced ability to reach the phloem. The superficial type of resistance observed in ABL 14-8 against B. tabaci probing significantly reduced primary and secondary spread of TYLCV.

  11. Understanding Rolle's Theorem

    ERIC Educational Resources Information Center

    Parameswaran, Revathy

    2009-01-01

    This paper reports on an experiment studying twelfth grade students' understanding of Rolle's Theorem. In particular, we study the influence of different concept images that students employ when solving reasoning tasks related to Rolle's Theorem. We argue that students' "container schema" and "motion schema" allow for rich…

  12. Leaf Activities.

    ERIC Educational Resources Information Center

    Mingie, Walter

    Leaf activities can provide a means of using basic concepts of outdoor education to learn in elementary level subject areas. Equipment needed includes leaves, a clipboard with paper, and a pencil. A bag of leaves may be brought into the classroom if weather conditions or time do not permit going outdoors. Each student should pick a leaf, examine…

  13. MicroRNA profiling of the whitefly Bemisia tabaci Middle East-Aisa Minor I following the acquisition of Tomato yellow leaf curl China virus.

    PubMed

    Wang, Bi; Wang, Lanlan; Chen, Fangyuan; Yang, Xiuling; Ding, Ming; Zhang, Zhongkai; Liu, Shu-Sheng; Wang, Xiao-Wei; Zhou, Xueping

    2016-02-02

    The begomoviruses are the largest and most economically important group of plant viruses exclusively vectored by whitefly (Bemisia tabaci) in a circulative, persistent manner. During this process, begomoviruses and whitefly vectors have developed close relationships and complex interactions. However, the molecular mechanisms underlying these interactions remain largely unknown, and the microRNA profiles for viruliferous and nonviruliferous whiteflies have not been studied. Sequences of Argonaute 1(Ago1) and Dicer 1 (Dcr1) genes were cloned from B. tabaci MEAM1 cDNAs. Subsequently, deep sequencing of small RNA libraries from uninfected and Tomato yellow leaf curl China virus (TYLCCNV)-infected whiteflies was performed. The conserved and novel miRNAs were identified using the release of miRBase Version 19.0 and the prediction software miRDeep2, respectively. The sequencing results of selected deregulated and novel miRNAs were further confirmed using quantitative reverse transcription-PCR. Moreover, the previously published B. tabaci MEAM1 transcriptome database and the miRNA target prediction algorithm miRanda 3.1 were utilized to predict potential targets for miRNAs. Gene Ontology (GO) analysis was also used to classify the potential enriched functional groups of their putative targets. Ago1 and Dcr1orthologs with conserved domains were identified from B. tabaci MEAM1. BLASTn searches and sequence analysis identified 112 and 136 conserved miRNAs from nonviruliferous and viruliferous whitefly libraries respectively, and a comparison of the conserved miRNAs of viruliferous and nonviruliferous whiteflies revealed 15 up- and 9 down-regulated conserved miRNAs. 7 novel miRNA candidates with secondary pre-miRNA hairpin structures were also identified. Potential targets of conserved and novel miRNAs were predicted using GO analysis, for the targets of up- and down-regulated miRNAs, eight and nine GO terms were significantly enriched. We identified Ago1 and Dcr1 orthologs

  14. Development of a salicylic acid inducible minimal sub-genomic transcript promoter from Figwort mosaic virus with enhanced root- and leaf-activity using TGACG motif rearrangement.

    PubMed

    Kumar, Deepak; Patro, Sunita; Ghosh, Jayasish; Das, Abhimanyu; Maiti, Indu B; Dey, Nrisingha

    2012-07-15

    In Figwort mosaic virus sub-genomic transcript promoter (F-Sgt), function of the TGACG-regulatory motif, was investigated in the background of artificially designed promoter sequences. The 131bp (FS, -100 to +31) long F-Sgt promoter sequence containing one TGACG motif [FS-(TGACG)] was engineered to generate a set of three modified promoter constructs: [FS-(TGACG)(2), containing one additional TGACG motif at 7 nucleotides upstream of the original one], [FS-(TGACG)(3), containing two additional TGACG motifs at 7 nucleotides upstream and two nucleotides downstream of the original one] and [FS-(TGCTG)(mu), having a mutated TGACG motif]. EMSA and foot-printing analysis confirmed binding of tobacco nuclear factors with modified TGACG motif/s. The transcription-activation of the GUS gene by the TGACG motif/s in above promoter constructs was examined in transgenic tobacco and Arabidopsis plants and observed that the transcription activation was affected by the spacing/s and number/s of the TGACG motif/s. The FS-(TGACG)(2) promoter showed strongest root-activity compared to other modified and CaMV35S promoters. Also under salicylic acid (SA) stress, the leaf-activity of the said promoter was further enhanced. All above findings were confirmed by real-time and semi-qRT PCR analysis. Taken together, these results clearly demonstrated that the TGACG motif plays an important role in inducing the root-specific expression of the F-Sgt promoter. This study advocates the importance of genetic manipulation of functional cis-motif for amending the tissue specificity of a plant promoter. SA inducible FS-(TGACG)(2) promoter with enhanced activity could be a useful candidate promoter for developing plants with enhanced crop productivity.

  15. Evaluation of cotton leaf curl virus resistance in BC1, BC2, and BC3 progenies from an interspecific cross between Gossypium arboreum and Gossypium hirsutum.

    PubMed

    Nazeer, Wajad; Tipu, Abdul Latif; Ahmad, Saghir; Mahmood, Khalid; Mahmood, Abid; Zhou, Baoliang

    2014-01-01

    Cotton leaf curl virus disease (CLCuD) is an important constraint to cotton production. The resistance of G. arboreum to this devastating disease is well documented. In the present investigation, we explored the possibility of transferring genes for resistance to CLCuD from G. arboreum (2n = 26) cv 15-Mollisoni into G. hirsutum (2n = 52) cv CRSM-38 through conventional breeding. We investigated the cytology of the BC1 to BC3 progenies of direct and reciprocal crosses of G. arboreum and G. hirsutum and evaluated their resistance to CLCuD. The F1 progenies were completely resistant to this disease, while a decrease in resistance was observed in all backcross generations. As backcrossing progressed, the disease incidence increased in BC1 (1.7-2.0%), BC2 (1.8-4.0%), and BC3 (4.2-7.0%). However, the disease incidence was much lower than that of the check variety CIM-496, with a CLCuD incidence of 96%. Additionally, the disease incidence percentage was lower in the direct cross 2(G. arboreum)×G. hirsutum than in that of G. hirsutum×G. arboreum. Phenotypic resemblance of BC1 ∼BC3 progenies to G. arboreum confirmed the success of cross between the two species. Cytological studies of CLCuD-resistant plants revealed that the frequency of univalents and multivalents was high in BC1, with sterile or partially fertile plants, but low in BC2 (in both combinations), with shy bearing plants. In BC3, most of the plants exhibited normal bearing ability due to the high frequency of chromosome associations (bivalents). The assessment of CLCuD through grafting showed that the BC1 to BC3 progenies were highly resistant to this disease. Thus, this study successfully demonstrates the possibility of introgressing CLCuD resistance genes from G. arboreum to G. hirsutum.

  16. Evaluation of Cotton Leaf Curl Virus Resistance in BC1, BC2, and BC3 Progenies from an Interspecific Cross between Gossypium arboreum and Gossypium hirsutum

    PubMed Central

    Nazeer, Wajad; Tipu, Abdul Latif; Ahmad, Saghir; Mahmood, Khalid; Mahmood, Abid; Zhou, Baoliang

    2014-01-01

    Cotton leaf curl virus disease (CLCuD) is an important constraint to cotton production. The resistance of G. arboreum to this devastating disease is well documented. In the present investigation, we explored the possibility of transferring genes for resistance to CLCuD from G. arboreum (2n = 26) cv 15-Mollisoni into G. hirsutum (2n = 52) cv CRSM-38 through conventional breeding. We investigated the cytology of the BC1 to BC3 progenies of direct and reciprocal crosses of G. arboreum and G. hirsutum and evaluated their resistance to CLCuD. The F1 progenies were completely resistant to this disease, while a decrease in resistance was observed in all backcross generations. As backcrossing progressed, the disease incidence increased in BC1 (1.7–2.0%), BC2 (1.8–4.0%), and BC3 (4.2–7.0%). However, the disease incidence was much lower than that of the check variety CIM-496, with a CLCuD incidence of 96%. Additionally, the disease incidence percentage was lower in the direct cross 2(G. arboreum)×G. hirsutum than in that of G. hirsutum×G. arboreum. Phenotypic resemblance of BC1 ∼BC3 progenies to G. arboreum confirmed the success of cross between the two species. Cytological studies of CLCuD-resistant plants revealed that the frequency of univalents and multivalents was high in BC1, with sterile or partially fertile plants, but low in BC2 (in both combinations), with shy bearing plants. In BC3, most of the plants exhibited normal bearing ability due to the high frequency of chromosome associations (bivalents). The assessment of CLCuD through grafting showed that the BC1 to BC3 progenies were highly resistant to this disease. Thus, this study successfully demonstrates the possibility of introgressing CLCuD resistance genes from G. arboreum to G. hirsutum. PMID:25372141

  17. Effectiveness of Cyantraniliprole for Managing Bemisia tabaci (Hemiptera: Aleyrodidae) and Interfering with Transmission of Tomato Yellow Leaf Curl Virus on Tomato.

    PubMed

    Caballero, Rafael; Schuster, David J; Peres, Natalia A; Mangandi, Jozer; Hasing, Tomas; Trexler, Fred; Kalb, Steve; Portillo, Héctor E; Marçon, Paula C; Annan, I B

    2015-06-01

    Cyantraniliprole is the second xylem-systemic active ingredient in the new anthranilic diamide class. Greenhouse (2006), growth chamber (2007), and field studies (2009-2010) were conducted to determine the efficacy of cyantraniliprole for managing Bemisia tabaci (Gennadius) biotype B and in interfering with transmission of tomato yellow leaf curl virus (TYLCV) by this whitefly. Cyantraniliprole applied as soil treatments (200 SC) or foliar sprays (100 OD) provided excellent adult whitefly control, TYLCV suppression, and reduced oviposition and nymph survival, comparable to current standards. The positive results observed in these greenhouse experiments with a high level of insect pressure (10× the field threshold of one adult per plant) and disease pressure (five adults per plant, with a high level of confidence that TYLCV virulent adults were used), indicate a great potential for cyantraniliprole to be used in a whitefly management program. Field evaluations of soil drench treatments confirmed the suppression of TYLCV transmission demonstrated in the greenhouse studies. Field studies in 2009 and 2010 showed that cyantraniliprole (200 SC) provided TYLCV suppression for 2 wk after a drench application, when using a susceptible (2009) or imidacloprid-tolerant (2010) whitefly population. Cyantraniliprole was demonstrated to be a promising tool for management of TYLCV in tomato production, which is very difficult and expensive, and which has limited options. The integration of cyantraniliprole into a resistance management program will help to ensure the continued sustainability of this and current insecticides used for the management of insect vectors, including whiteflies and the TYLCV they spreads. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  18. Eugenol confers resistance to Tomato yellow leaf curl virus (TYLCV) by regulating the expression of SlPer1 in tomato plants.

    PubMed

    Sun, Wei-Jie; Lv, Wen-Jing; Li, Li-Na; Yin, Gan; Hang, Xiaofang; Xue, Yanfeng; Chen, Jian; Shi, Zhiqi

    2016-05-25

    Tomato yellow leaf curl virus (TYLCV) is one of the most devastating plant diseases, and poses a significant agricultural concern because of the lack of an efficient control method. Eugenol is a plant-derived natural compound that has been widely used as a food additive and in medicine. In the present study, we demonstrated the potential of eugenol to enhance the resistance of tomato plants to TYLCV. The anti-TYLCV efficiency of eugenol was significantly higher than that of moroxydine hydrochloride (MH), a widely used commercial antiviral agent. Eugenol application stimulated the production of endogenous nitric oxide (NO) and salicylic acid (SA) in tomato plants. The full-length cDNA of SlPer1, which has been suggested to be a host R gene specific to TYLCV, was isolated from tomato plants. A sequence analysis suggested that SlPer1 might be a nucleobase-ascorbate transporter (NAT) belonging to the permease family. The transcript levels of SlPer1 increased markedly in response to treatment with eugenol or TYLCV inoculation. The results of this study also showed that SlPer1 expression was strongly induced by SA, MeJA (jasmonic acid methyl ester), and NO. Thus, we propose that the increased transcription of SlPer1 contributed to the high anti-TYLCV efficiency of eugenol, which might involve in the generation of endogenous SA and NO. Such findings provide the basis for the development of eugenol as an environmental-friendly agricultural antiviral agent. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Host-induced gene silencing of wheat leaf rust fungus Puccinia triticina pathogenicity genes mediated by the Barley stripe mosaic virus.

    PubMed

    Panwar, Vinay; McCallum, Brent; Bakkeren, Guus

    2013-04-01

    Rust fungi are devastating plant pathogens and several Puccinia species have a large economic impact on wheat production worldwide. Disease protection, mostly offered by introgressed host-resistance genes, is often race-specific and rapidly overcome by newly-emerging virulent strains. Extensive new genomic resources have identified vital pathogenicity genes but their study is hampered because of the biotrophic life styles of rust fungi. In cereals, Barley stripe mosaic virus (BSMV)-induced RNAi has emerged as a useful tool to study loss-of-function phenotypes of candidate genes. Expression of pathogen-derived gene fragments in this system can be used to obtain in planta-generated silencing of corresponding genes inside biotrophic pathogens, a technique termed host-induced gene silencing (HIGS). Here we test the effectiveness of BSMV-mediated HIGS in the wheat leaf rust fungus Puccinia triticina (Pt) by targeting three predicted pathogenicity genes, a MAPK, a cyclophilin, and a calcineurin regulatory subunit. Inoculation of BSMV RNAi constructs generated fungal gene-specific siRNA molecules in systemic leaves of wheat plant. Subsequent Pt inoculation resulted in a suppressed disease phenotype and a reduction in endogenous transcript levels of the targeted fungal genes indicating translocation of siRNA molecules from host to fungal cells. Efficiency of this host-generated trans-specific RNAi was enhanced by using BSMV silencing vectors defective in coat protein coupled with introducing fungal gene sequences simultaneously in sense and antisense orientation. The disease suppression indicated the likely involvement of these fungal genes in pathogenicity. This study demonstrates that BSMV-mediated in planta-generated RNAi is an effective strategy for functional genomics in rust fungi.

  20. Rolling circle amplification, a powerful tool for genetic and functional studies of complete hepatitis B virus genomes from low-level infections and for directly probing covalently closed circular DNA.

    PubMed

    Margeridon, Séverine; Carrouée-Durantel, Sandra; Chemin, Isabelle; Barraud, Luc; Zoulim, Fabien; Trépo, Christian; Kay, Alan

    2008-09-01

    Complete characterization of the biological properties of hepatitis B virus (HBV) variants requires the generation of full-length genomes. The aim of this study was to develop new tools for the efficient full-length genome amplification of virus from samples with low viral loads. Rolling circle amplification (RCA) was used to amplify full-length HBV genomes from both sera and liver biopsy samples from chronic HBV carriers. Serum-derived relaxed circular HBV DNA could be amplified only after completion and ligation of plus-strand DNA. Covalently closed circular DNA (cccDNA) from liver biopsies could be amplified directly from as few as 13 copies, using RCA, followed by a full-length HBV PCR. Three serial liver biopsy samples were obtained from a lamivudine-resistant patient who cleared detectable serum HBV after adefovir dipivoxil was added to the lamivudine therapy and then seroconverted to anti-HBs. Only the genomes from the last biopsy specimen obtained after the emergence of lamivudine resistance contained the lamivudine resistance-associated mutations rtL180M and rtM204V ("rt" indicates reverse transcriptase domain). Defective genomes were also found in this biopsy sample. Genomes cloned from the liver biopsy specimens were transfected into HuH7 cells to study their replication competence and their susceptibility to lamivudine. RCA is a powerful tool for amplifying full-length HBV genomes and will be especially useful for the study of occult or inactive HBV infections and patients undergoing antiviral treatment. It can also be used to probe HBV cccDNA, the crucial intermediate in viral persistence and the archive of resistance mutations.

  1. A recombinant Tobacco curly shoot virus causes leaf curl disease in tomato in a north-eastern state of India and has potentiality to trans-replicate a non-cognate betasatellite.

    PubMed

    Shilpi, S; Kumar, Alok; Biswas, S; Roy, Anirban; Mandal, Bikash

    2015-02-01

    Leaf curl disease is a serious constraint in tomato production throughout India. Several begomoviruses were reported from different parts of the country; however, identity of begomovirus associated with leaf curl disease in tomato in north-eastern states of India was obscured. In the present study, the complete genome of an isolate (To-Ag-1) of begomovirus was generated from a leaf curl sample collected from Tripura state. However, no DNA-B and betasatellite were detected in the field samples. The genome of To-Ag-1 isolate contained 2,755 nucleotides that shared 94.7 % sequence identity with Tobacco curly shoot virus (TbCSV) and 71.3-90.1 % sequence identity with the other tomato-infecting begomoviruses occurring in the Indian subcontinent. Several inter-specific recombination events among different tomato-infecting begomoviruses from India and intra-specific recombination among different isolates of TbCSV reported from China were observed in the genome of To-Ag-1 isolate. Agroinoculation of the virus alone produced leaf curl symptoms in tomato and Nicotiana benthamiana. However, co-inoculation with a non-cognate betasatellite, Croton yellow vein mosaic betasatellite (CroYVMB) with the TbCSV resulted in increased severity of the symptoms both in tomato and N. benthamiana. Systemic distribution of the TbCSV and CroYVMB was detected in the newly developed leaves of tobacco and tomato, which showed ability of TbCSV to trans-replicate CroYVMB. The present study for the first time provides evidence of occurrence of TbCSV in tomato in north-eastern region of India and showed increased virulence of TbCSV with a non-cognate betasatellite.

  2. OsLBD3-7 Overexpression Induced Adaxially Rolled Leaves in Rice

    PubMed Central

    Zhang, Chunyu; Shao, Qinghao; Liu, Jun; Liu, Bin; Li, Hongyu

    2016-01-01

    Appropriate leaf rolling enhances erect-leaf habits and photosynthetic efficiency, which consequently improves grain yield. Here, we reported the novel lateral organ boundaries domain (LBD) gene OsLBD3-7, which is involved in the regulation of leaf rolling. OsLBD3-7 works as a transcription activator and its protein is located on the plasma membrane and in the nucleus. Overexpression of OsLBD3-7 leads to narrow and adaxially rolled leaves. Microscopy of flag leaf cross-sections indicated that overexpression of OsLBD3-7 led to a decrease in both bulliform cell size and number. Transcriptional analysis showed that key genes that had been reported to be negative regulators of bulliform cell development were up-regulated in transgenic plants. These results indicated that OsLBD3-7 might acts as an upstream regulatory gene of bulliform cell development to regulate leaf rolling, which will give more insights on the leaf rolling regulation mechanism. PMID:27258066

  3. Effect of Roll Material on Surface Quality of Rolled Aluminum

    NASA Astrophysics Data System (ADS)

    Zhao, Qi

    The surface defects of aluminum alloys that have undergone hot rolling were studied. The effects of different roll materials, of the number of rolling passes and of lubrication on surface defects of hot rolled aluminum alloys were investigated by laboratory hot rolling. Two different aluminum alloys, Al-Mn and Al-Mg, were each rolled against three different steel alloy rolls, AISI 52100, AISI 440C and AISI D2. The results showed that different roll materials do affect the morphology of the mating aluminum alloy surface with apparent surface defects, which included magnesium and oxygen rich dark regions on both alloys. The carbide protrusions in 440C and D2 steel rolls are confirmed to be responsible for the dark, rich magnesium and oxygen regions on both the rolled Al-Mn and Al-Mg alloy surfaces. As the number of passes increases, Mg and O deposit in the form of patches and grain boundaries near the surface area.

  4. Project LEAF

    EPA Pesticide Factsheets

    Project LEAF has a goal of educating farmworkers about how to reduce pesticide exposure to their families from pesticide residues they may be inadvertently taking home on their clothing, etc. Find outreach materials.

  5. Biomechanics of leukocyte rolling

    PubMed Central

    Sundd, Prithu; Pospieszalska, Maria K.; Cheung, Luthur Siu-Lun; Konstantopoulos, Konstantinos; Ley, Klaus

    2011-01-01

    Leukocyte rolling on endothelial cells and other P-selectin substrates is mediated by P-selectin binding to P-selectin glycoprotein ligand-1 expressed on the tips of leukocyte microvilli. Leukocyte rolling is a result of rapid, yet balanced formation and dissociation of selectin-ligand bonds in the presence of hydrodynamic shear forces. The hydrodynamic forces acting on the bonds may either increase (catch bonds) or decrease (slip-bonds) their lifetimes. The force-dependent ‘catch-slip’ bond kinetics are explained using the ‘two pathway model’ for bond dissociation. Both the ‘sliding-rebinding’ and the ‘allosteric’ mechanisms attribute ‘catch-slip’ bond behavior to the force-induced conformational changes in the lectin-EGF domain hinge of selectins. Below a threshold shear stress, selectins cannot mediate rolling. This ‘shear-threshold’ phenomenon is a consequence of shear-enhanced tethering and catch-bond enhanced rolling. Quantitative dynamic footprinting microscopy has revealed that leukocytes rolling at venular shear stresses (> 0.6 Pa) undergo cellular deformation (large footprint) and form long tethers. The hydrodynamic shear force and torque acting on the rolling cell are thought to be synergistically balanced by the forces acting on tethers and stressed microvilli, however, their relative contribution remains to be determined. Thus, improvement beyond the current understanding requires in silico models that can predict both cellular and microvillus deformation and experiments that allow measurement of forces acting on individual microvilli and tethers. PMID:21515934

  6. Leaf Development

    PubMed Central

    Tsukaya, Hirokazu

    2002-01-01

    The shoot system is the basic unit of development of seed plants and is composed of a leaf, a stem, and a lateral bud that differentiates into a lateral shoot. The most specialized organ in angiosperms, the flower, can be considered to be part of the same shoot system since floral organs, such as the sepal, petal, stamen, and carpel, are all modified leaves. Scales, bracts, and certain kinds of needle are also derived from leaves. Thus, an understanding of leaf development is critical to an understanding of shoot development. Moreover, leaves play important roles in photosynthesis, respiration and photoperception. Thus, a full understanding of leaves is directly related to a full understanding of seed plants. The details of leaf development remain unclear. The difficulties encountered in studies of leaf development, in particular in dicotyledonous plants such as Arabidopsis thaliana (L.) Henyn., are derived from the complex process of leaf development, during which the division and elongation of cells occur at the same time and in the same region of the leaf primordium (Maksymowych, 1963; Poethig and Sussex, 1985). Thus, we cannot divide the entire process into unit processes in accordance with the tenets of classical anatomy. Genetic approaches in Arabidopsis, a model plant (Meyerowitz and Pruitt, 1985), have provided a powerful tool for studies of mechanisms of leaf development in dicotyledonous plants, and various aspects of the mechanisms that control leaf development have been revealed in recent developmental and molecular genetic studies of Arabidopsis (for reviews, see Tsukaya, 1995 and 1998; Van Lijsebettens and Clarke, 1998; Sinha, 1999; Van Volkenburgh, 1999; Tsukaya, 2000; Byrne et al., 2001; Dengler and Kang, 2001; Dengler and Tsukaya, 2001; Tsukaya, 2001). In this review, we shall examine the information that is currently available about various mechanisms of leaf development in Arabidopsis. Vascular patterning is also an important factor in the

  7. Scavenging of reactive oxygen species in apoplastic and symplastic areas of rolled leaves in Ctenanthe setosa under drought stress.

    PubMed

    Saruhan, Neslihan; Terzi, Rabiye; Sağlam, Aykut; Kadioğlu, Asim

    2010-09-01

    The correspondence among apoplastic and symplastic antioxidant status, stomatal conductance and water potential was investigated during leaf rolling in Ctenanthe setosa (Rosc.) Eichler (Marantaceae) under drought stress. Apoplastic and symplastic extractions of leaf and petiole were performed at different visual leaf rolling scores from 1 to 4 (1 is unrolled, 4 is tightly rolled and the others are intermediate form). In the leaf symplast, the highest changes were found in catalase (CAT) and guaiacol peroxidase (GPX) activities when compared to score 1 during leaf rolling. No significant change was observed in superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities in the symplast of leaf during the rolling. The same phenomenon was also present in the symplast of petiole except APX activity. In the leaf apoplast, the highest increase occurred in APX and GPX activities, whilst a slight increase in CAT and SOD activities. In the apoplast of petiole, the highest increment was found only in GPX activity, while there were small increases in SOD, APX and CAT activities. Hydrogen peroxide content increased up to score 3 in the apoplast and symplast of leaf and petiole but then slightly decreased. Also, superoxide production increased in the leaf and petiole apoplast but its quantity in the apoplast was much more than that of the symplast. On the other hand, NAD(P)H oxidase activity increased in the leaf but no change was observed in the petiole. In conclusion, as a result of water deficit during leaf rolling antioxidant enzymes are induced to scavenging of ROS produced in symplast and apoplast.

  8. Other Viruses and Viruslike Agents

    USDA-ARS?s Scientific Manuscript database

    The diseases reported under 'Virus and Virus-like Agents' in the first volume of this compendium, with the exception of Cherry rasp leaf virus and Rubus chinese seed-borne virus, should be considered oddities since there are no known type isolates available for these reported viruses. Without a po...

  9. Storage effects on genomic DNA in rolled and mature coca leaves.

    PubMed

    Johnson, Emanuel L; Kim, Soo-Hyung; Emche, Stephen D

    2003-08-01

    Rolled and mature leaf tissue was harvested from Erythroxylum coca var. coca Lam. (coca) to determine a method for storage that would maintain DNA with high quality and content up to 50 days. Harvesting coca leaf tissue under Andean field conditions often requires storage from 3 to 10 days before extraction where tissue integrity is lost. All samples of rolled and mature coca leaf tissue were harvested and separately stored fresh in RNAlater for 50 days at 4 degrees, -20 degrees, and 23 degrees C, while similar samples were air-dried for 72 h at 23 degrees C or oven-dried for 72 h at 40 degrees C after storage, before extraction. Triplicate samples of each tissue type were extracted for DNA at 10-day intervals and showed that DNA integrity and content were preserved in leaf tissue stored at 4 degrees and -20 degrees C for 50 days. Rolled and mature leaf tissue stored at 4 degrees, -20 degrees, and 23 degrees C showed insignificant degradation of DNA after 10 days, and by day 50, only leaf tissue stored at 4 degrees and -20 degrees C had not significantly degraded. All air- and oven-dried leaf tissue extracts showed degradation upon drying (day 0) and continuous degradation up to day 50, despite storage conditions. Amplified fragment length polymorphism analysis of DNA from rolled and mature leaf tissue of coca stored at 4 degrees and -20 degrees C for 0, 10, and 50 days showed that DNA integrity and content were preserved. We recommend that freshly harvested rolled or mature coca leaf tissue be stored at 4 degrees, -20 degrees, and 23 degrees C for 10 days after harvest, and if a longer storage is required, then store at 4 degrees or -20 degrees C.

  10. Making a friend from a foe: expressing a GroEL gene from the whitefly Bemisia tabaci in the phloem of tomato plants confers resistance to tomato yellow leaf curl virus.

    PubMed

    Akad, F; Eybishtz, A; Edelbaum, D; Gorovits, R; Dar-Issa, O; Iraki, N; Czosnek, H

    2007-01-01

    Some (perhaps all) plant viruses transmitted in a circulative manner by their insect vectors avoid destruction in the haemolymph by interacting with GroEL homologues, ensuring transmission. We have previously shown that the phloem-limited begomovirus tomato yellow leaf curl virus (TYLCV) interacts in vivo and in vitro with GroEL produced by the whitefly vector Bemisia tabaci. In this study, we have exploited this phenomenon to generate transgenic tomato plants expressing the whitefly GroEL in their phloem. We postulated that following inoculation, TYLCV particles will be trapped by GroEL in the plant phloem, thereby inhibiting virus replication and movement, thereby rendering the plants resistant. A whitefly GroEL gene was cloned in an Agrobacterium vector under the control of an Arabidopsis phloem-specific promoter, which was used to transform two tomato genotypes. During three consecutive generations, plants expressing GroEL exhibited mild or no disease symptoms upon whitefly-mediated inoculation of TYLCV. In vitro assays indicated that the sap of resistant plants contained GroEL-TYLCV complexes. Infected resistant plants served as virus source for whitefly-mediated transmission as effectively as infected non-transgenic tomato. Non-transgenic susceptible tomato plants grafted on resistant GroEL-transgenic scions remained susceptible, although GroEL translocated into the grafted plant and GroEL-TYLCV complexes were detected in the grafted tissues.

  11. Spray Rolling Aluminum Strip

    SciTech Connect

    Lavernia, E.J.; Delplanque, J-P; McHugh, K.M.

    2006-05-10

    Spray forming is a competitive low-cost alternative to ingot metallurgy for manufacturing ferrous and non-ferrous alloy shapes. It produces materials with a reduced number of processing steps, while maintaining materials properties, with the possibility of near-net-shape manufacturing. However, there are several hurdles to large-scale commercial adoption of spray forming: 1) ensuring strip is consistently flat, 2) eliminating porosity, particularly at the deposit/substrate interface, and 3) improving material yield. Through this program, a new strip/sheet casting process, termed spray rolling, has been developed, which is an innovative manufacturing technique to produce aluminum net-shape products. Spray rolling combines the benefits of twin-roll casting and conventional spray forming, showing a promising potential to overcome the above hurdles associated with spray forming. Spray rolling requires less energy and generates less scrap than conventional processes and, consequently, enables the development of materials with lower environmental impacts in both processing and final products. Spray Rolling was developed as a collaborative project between the University of California-Davis, the Colorado School of Mines, the Idaho National Engineering and Environmental Laboratory, and an industry team. The following objectives of this project were achieved: (1) Demonstration of the feasibility of the spray rolling process at the bench-scale level and evaluation of the materials properties of spray rolled aluminum strip alloys; and (2) Demonstration of 2X scalability of the process and documentation of technical hurdles to further scale up and initiate technology transfer to industry for eventual commercialization of the process.

  12. Leaf Development

    PubMed Central

    2013-01-01

    Leaves are the most important organs for plants. Without leaves, plants cannot capture light energy or synthesize organic compounds via photosynthesis. Without leaves, plants would be unable perceive diverse environmental conditions, particularly those relating to light quality/quantity. Without leaves, plants would not be able to flower because all floral organs are modified leaves. Arabidopsis thaliana is a good model system for analyzing mechanisms of eudicotyledonous, simple-leaf development. The first section of this review provides a brief history of studies on development in Arabidopsis leaves. This history largely coincides with a general history of advancement in understanding of the genetic mechanisms operating during simple-leaf development in angiosperms. In the second section, I outline events in Arabidopsis leaf development, with emphasis on genetic controls. Current knowledge of six important components in these developmental events is summarized in detail, followed by concluding remarks and perspectives. PMID:23864837

  13. Roll cooling and its relationship to roll life

    NASA Astrophysics Data System (ADS)

    Tseng, A. A.; Lin, F. H.; Gunderia, A. S.; Ni, D. S.

    1989-11-01

    Combined experimental and numerical research has been conducted to investigate the roll cooling system used in steel rolling mills and its relationship to roll life. Roll cooling has been identified as a critical factor in the problems of excessive roll wear or spalling, which cause shortened roll life. A special laboratory apparatus resembling the cooling systems used in the steel mills has been developed to evaluate the corresponding heat transfer coefficients. These coefficients have then been utilized for numerical simulation of the rolling processes. In simulation, the thermal behavior of both the roll and the strip have been considered with emphasis on roll temperature and the induced cyclic thermal stresses. An understanding of the cyclic stress can be utilized to evaluate roll wear, and leads to reduction of the roll spalling, or to extension of the roll life by minimizing the cyclic stress or the resultant fatigue stress. As indicated by the present study, in order to minimize the cyclic or fatigue stresses, the roll should be subjected to uniform cooling, as the sharper the heat transfer coefficient distribution, the higher the thermal stress induced.

  14. Wear of hot rolling mill rolls: An overview

    NASA Astrophysics Data System (ADS)

    Spuzic, S.; Strafford, K. N.; Subramanian, C.; Savage, G.

    1994-08-01

    Rolling is today one of the most important industrial processes because a greater volume of material is worked by rolling than by any other technique. Roll wear is a multiplex process where mechanical and thermal fatigue combines with impact, abrasion, adhesion and corrosion, which all depend on system interactions rather than material characteristics only. The situation is more complicated in section rolling because of the intricacy of roll geometry. Wear variables and modes are reviewed along with published methods and models used in the study and testing of roll wear. This paper reviews key aspects of roll wear control - roll material properties, roll pass design, and system factors such as temperature, loads and sliding velocity. An overview of roll materials is given including adamites, high Cr materials, high speed tool steels and compound rolls. Non-uniform wear, recognized as the most detrimental phenomenon in section rolling, can be controlled by roll pass design. This can be achieved by computer-aided graphical and statistical analyses of various pass series. Preliminary results obtained from pilot tests conducted using a two-disc hot wear rig and a scratch tester are discussed.

  15. Stochastic disks that roll.

    PubMed

    Holmes-Cerfon, Miranda

    2016-11-01

    We study a model of rolling particles subject to stochastic fluctuations, which may be relevant in systems of nano- or microscale particles where rolling is an approximation for strong static friction. We consider the simplest possible nontrivial system: a linear polymer of three disks constrained to remain in contact and immersed in an equilibrium heat bath so the internal angle of the polymer changes due to stochastic fluctuations. We compare two cases: one where the disks can slide relative to each other and the other where they are constrained to roll, like gears. Starting from the Langevin equations with arbitrary linear velocity constraints, we use formal homogenization theory to derive the overdamped equations that describe the process in configuration space only. The resulting dynamics have the formal structure of a Brownian motion on a Riemannian or sub-Riemannian manifold, depending on if the velocity constraints are holonomic or nonholonomic. We use this to compute the trimer's equilibrium distribution with and without the rolling constraints. Surprisingly, the two distributions are different. We suggest two possible interpretations of this result: either (i) dry friction (or other dissipative, nonequilibrium forces) changes basic thermodynamic quantities like the free energy of a system, a statement that could be tested experimentally, or (ii) as a lesson in modeling rolling or friction more generally as a velocity constraint when stochastic fluctuations are present. In the latter case, we speculate there could be a "roughness" entropy whose inclusion as an effective force could compensate the constraint and preserve classical Boltzmann statistics. Regardless of the interpretation, our calculation shows the word "rolling" must be used with care when stochastic fluctuations are present.

  16. Stochastic disks that roll

    NASA Astrophysics Data System (ADS)

    Holmes-Cerfon, Miranda

    2016-11-01

    We study a model of rolling particles subject to stochastic fluctuations, which may be relevant in systems of nano- or microscale particles where rolling is an approximation for strong static friction. We consider the simplest possible nontrivial system: a linear polymer of three disks constrained to remain in contact and immersed in an equilibrium heat bath so the internal angle of the polymer changes due to stochastic fluctuations. We compare two cases: one where the disks can slide relative to each other and the other where they are constrained to roll, like gears. Starting from the Langevin equations with arbitrary linear velocity constraints, we use formal homogenization theory to derive the overdamped equations that describe the process in configuration space only. The resulting dynamics have the formal structure of a Brownian motion on a Riemannian or sub-Riemannian manifold, depending on if the velocity constraints are holonomic or nonholonomic. We use this to compute the trimer's equilibrium distribution with and without the rolling constraints. Surprisingly, the two distributions are different. We suggest two possible interpretations of this result: either (i) dry friction (or other dissipative, nonequilibrium forces) changes basic thermodynamic quantities like the free energy of a system, a statement that could be tested experimentally, or (ii) as a lesson in modeling rolling or friction more generally as a velocity constraint when stochastic fluctuations are present. In the latter case, we speculate there could be a "roughness" entropy whose inclusion as an effective force could compensate the constraint and preserve classical Boltzmann statistics. Regardless of the interpretation, our calculation shows the word "rolling" must be used with care when stochastic fluctuations are present.

  17. Extended slow-roll conditions and rapid-roll conditions

    SciTech Connect

    Chiba, Takeshi; Yamaguchi, Masahide E-mail: gucci@phys.aoyama.ac.jp

    2008-10-15

    We derive slow-roll conditions for a scalar field which is non-minimally coupled with gravity in a consistent manner and express spectral indices of scalar/tensor perturbations in terms of the slow-roll parameters. The conformal invariance of the curvature perturbation is proved without linear approximations. Rapid-roll conditions are also derived, and the relation with the slow-roll conditions is discussed.

  18. Rolling through a vacuum

    NASA Astrophysics Data System (ADS)

    van der Schaar, Jan Pieter; Yang, I.-Sheng

    2013-12-01

    We clarify under what conditions slow-roll inflation can continue almost undisturbed, while briefly evolving through a (semi-classically) metastable false vacuum. Furthermore, we look at potential signatures in the primordial power spectrum that could point towards the existence of traversed metastable false vacua. Interestingly, the theoretical constraints for the existence of traversable metastable vacua imply that Planck should be able to detect the resulting features in the primordial power spectrum. In other words, if Planck does not see features this immediately implies the non-existence of metastable false vacua rolled through during the inflationary epoch.

  19. METHOD OF ROLLING URANIUM

    DOEpatents

    Smith, C.S.

    1959-08-01

    A method is described for rolling uranium metal at relatively low temperatures and under non-oxidizing conditions. The method involves the steps of heating the uranium to 200 deg C in an oil bath, withdrawing the uranium and permitting the oil to drain so that only a thin protective coating remains and rolling the oil coated uranium at a temperature of 200 deg C to give about a 15% reduction in thickness at each pass. The operation may be repeated to accomplish about a 90% reduction without edge cracking, checking or any appreciable increase in brittleness.

  20. Virion Stability Is Important for the Circulative Transmission of Tomato Yellow Leaf Curl Sardinia Virus by Bemisia tabaci, but Virion Access to Salivary Glands Does Not Guarantee Transmissibility▿ †

    PubMed Central

    Caciagli, Piero; Medina Piles, Vicente; Marian, Daniele; Vecchiati, Manuela; Masenga, Vera; Mason, Giovanna; Falcioni, Tania; Noris, Emanuela

    2009-01-01

    The capsid protein (CP) of the monopartite begomovirus Tomato yellow leaf curl Sardinia virus (TYLCSV), family Geminiviridae, is indispensable for plant infection and vector transmission. A region between amino acids 129 and 152 is critical for virion assembly and insect transmissibility. Two previously described mutants, one with a double Q129P Q134H mutation (PNHD) and another with a further D152E change (PNHE), were found nontransmissible (NT). Another NT mutant with a single N130D change (QDQD) was retrieved from a new mutational analysis. In this study, these three NT mutants and the wild-type (wt) virus were compared in their relationships with the whitefly vector Bemisia tabaci and the nonvector Trialeurodes vaporariorum. Retention kinetics of NT mutants were analyzed by quantitative dot blot hybridization in whiteflies fed on infected plants. The QDQD mutant, whose virions appeared nongeminate following purification, was hardly detectable in either whitefly species at any sampling time. The PNHD mutant was acquired and circulated in both whitefly species for up to 10 days, like the wt virus, while PNHE circulated in B. tabaci only. Using immunogold labeling, both PNHD and PNHE CPs were detected in B. tabaci salivary glands (SGs) like the wt virus, while no labeling was found in any whitefly tissue with the QDQD mutant. Significant inhibition of transmission of the wt virus was observed after prior feeding of the insects on plants infected with the PNHE mutant, but not on plants infected with the other mutants. Virion stability and ability to cross the SG barrier are necessary for TYLCSV transmission, but interactions with molecular components inside the SGs are also critical for transmissibility. PMID:19321611

  1. Rolling Spot Welder

    NASA Technical Reports Server (NTRS)

    Wagner, Garret E.; Fonteyne, Steve L.

    1990-01-01

    Wheeled tool speeds tack-welding operations. Spotwelds foil to parts in preparation for brazing. Includes electrode wheel rolling across foil. Welding current in electrode pulsed as electrode moves along, making series of uniformly-spaced low-current spot welds.

  2. Rolling Spot Welder

    NASA Technical Reports Server (NTRS)

    Wagner, Garret E.; Fonteyne, Steve L.

    1990-01-01

    Wheeled tool speeds tack-welding operations. Spotwelds foil to parts in preparation for brazing. Includes electrode wheel rolling across foil. Welding current in electrode pulsed as electrode moves along, making series of uniformly-spaced low-current spot welds.

  3. Viruses

    USDA-ARS?s Scientific Manuscript database

    Lytic bacteriophages, viruses which infect and lyse bacterial cells, can provide a natural method to reduce bacterial pathogens on produce commodities. The use of multi-phage cocktails is most likely to be effective against bacterial pathogens on produce commodities, and minimize the development of...

  4. Aircraft roll steering command system

    NASA Technical Reports Server (NTRS)

    Lambregts, Antonius A. (Inventor)

    1985-01-01

    Aircraft roll command signals are generated as a function of the Microwave Landing System based azimuth, groundtrack, groundspeed and azimuth rate or range distance input parameters. On initial approach, roll command signals are inhibited until a minimum roll command requirement is met. As the aircraft approaches the centerline of the runway, the system reverts to a linear track control.

  5. Identification of a New Cotton Disease Caused by an Atypical Cotton Leafroll Dwarf Virus in Argentina.

    PubMed

    Agrofoglio, Yamila C; Delfosse, Verónica C; Casse, María F; Hopp, Horacio E; Kresic, Iván Bonacic; Distéfano, Ana J

    2017-03-01

    An outbreak of a new disease occurred in cotton (Gossypium hirsutum) fields in northwest Argentina starting in the 2009-10 growing season and is still spreading steadily. The characteristic symptoms of the disease included slight leaf rolling and a bushy phenotype in the upper part of the plant. In this study, we determined the complete nucleotide sequences of two independent virus genomes isolated from cotton blue disease (CBD)-resistant and -susceptible cotton varieties. This virus genome comprised 5,866 nucleotides with an organization similar to that of the genus Polerovirus and was closely related to cotton leafroll dwarf virus, with protein identity ranging from 88 to 98%. The virus was subsequently transmitted to a CBD-resistant cotton variety using Aphis gossypii and symptoms were successfully reproduced. To study the persistence of the virus, we analyzed symptomatic plants from CBD-resistant varieties from different cotton-growing fields between 2013 and 2015 and showed the presence of the same virus strain. In addition, a constructed full-length infectious cDNA clone from the virus caused disease symptoms in systemic leaves of CBD-resistant cotton plants. Altogether, the new leafroll disease in CBD-resistant cotton plants is caused by an atypical cotton leafroll dwarf virus.

  6. Roll Eccentricity Control Using Identified Eccentricity of Top/Bottom Rolls by Roll Force

    NASA Astrophysics Data System (ADS)

    Imanari, Hiroyuki; Koshinuma, Kazuyoshi

    Roll eccentricity is a periodic disturbance caused by a structure of back up rolls in rolling mills, and it affects product thickness accuracy. It cannot be measured directly by sensors, so it should be identified by measured thickness or measured roll force. When there is a large difference of diameters between top and bottom back up roll, the performance of roll eccentricity control using feedback signals of roll force or thickness has not been so good. Also it has been difficult for the control to be applied from the most head end because it is necessary to identify the roll eccentricity during rolling. A new roll eccentricity control has been developed to improve these disadvantages and to get better performance. The method identifies top and bottom roll eccentricity respectively from one signal of roll force and it can start the control from head end. In this paper the new control method is introduced and actual application results to a hot strip mill are shown.

  7. A new virus discovered by immunocapture of double-stranded RNA, a rapid method for virus enrichment in metagenomic studies.

    PubMed

    Blouin, Arnaud G; Ross, Howard A; Hobson-Peters, Jody; O'Brien, Caitlin A; Warren, Ben; MacDiarmid, Robin

    2016-09-01

    Next-generation sequencing technologies enable the rapid identification of viral infection of diseased organisms. However, despite a consistent decrease in sequencing costs, it is difficult to justify their use in large-scale surveys without a virus sequence enrichment technique. As the majority of plant viruses have an RNA genome, a common approach is to extract the double-stranded RNA (dsRNA) replicative form, to enrich the replicating virus genetic material over the host background. The traditional dsRNA extraction is time-consuming and labour-intensive. We present an alternative method to enrich dsRNA from plant extracts using anti-dsRNA monoclonal antibodies in a pull-down assay. The extracted dsRNA can be amplified by reverse transcriptase-polymerase chain reaction and sequenced by next-generation sequencing. In our study, we have selected three distinct plant hosts: Māori potato (Solanum tuberosum), rengarenga (Arthropodium cirratum) and broadleaved dock (Rumex obtusifolius) representing a cultivated crop, a New Zealand-native ornamental plant and a weed, respectively. Of the sequence data obtained, 31-74% of the reads were of viral origin, and we identified five viruses including Potato virus Y and Potato virus S in potato; Turnip mosaic virus in rengarenga (a new host record); and in the dock sample Cherry leaf roll virus and a novel virus belonging to the genus Macluravirus. We believe that this new assay represents a significant opportunity to upscale virus ecology studies from environmental, primary industry and/or medical samples.

  8. GRCop-84 Rolling Parameter Study

    NASA Technical Reports Server (NTRS)

    Loewenthal, William S.; Ellis, David L.

    2008-01-01

    This report is a section of the final report on the GRCop-84 task of the Constellation Program and incorporates the results obtained between October 2000 and September 2005, when the program ended. NASA Glenn Research Center (GRC) has developed a new copper alloy, GRCop-84 (Cu-8 at.% Cr-4 at.% Nb), for rocket engine main combustion chamber components that will improve rocket engine life and performance. This work examines the sensitivity of GRCop-84 mechanical properties to rolling parameters as a means to better define rolling parameters for commercial warm rolling. Experiment variables studied were total reduction, rolling temperature, rolling speed, and post rolling annealing heat treatment. The responses were tensile properties measured at 23 and 500 C, hardness, and creep at three stress-temperature combinations. Understanding these relationships will better define boundaries for a robust commercial warm rolling process. The four processing parameters were varied within limits consistent with typical commercial production processes. Testing revealed that the rolling-related variables selected have a minimal influence on tensile, hardness, and creep properties over the range of values tested. Annealing had the expected result of lowering room temperature hardness and strength while increasing room temperature elongations with 600 C (1112 F) having the most effect. These results indicate that the process conditions to warm roll plate and sheet for these variables can range over wide levels without negatively impacting mechanical properties. Incorporating broader process ranges in future rolling campaigns should lower commercial rolling costs through increased productivity.

  9. Mapping Quantitative Trait Loci of Resistance to Tomato Spotted Wilt Virus and Leaf Spots in a Recombinant Inbred Line Population of Peanut (Arachis hypogaea L.) from SunOleic 97R and NC94022

    PubMed Central

    Feng, Suping; Qiao, Lixian; Culbreath, Albert K.; Kale, Sandip; Wang, Jianping; Holbrook, C. Corley; Zhuang, Weijian; Varshney, Rajeev K.; Guo, Baozhu

    2016-01-01

    Peanut is vulnerable to a range of diseases, such as Tomato spotted wilt virus (TSWV) and leaf spots which will cause significant yield loss. The most sustainable, economical and eco-friendly solution for managing peanut diseases is development of improved cultivars with high level of resistance. We developed a recombinant inbred line population from the cross between SunOleic 97R and NC94022, named as the S-population. An improved genetic linkage map was developed for the S-population with 248 marker loci and a marker density of 5.7 cM/loci. This genetic map was also compared with the physical map of diploid progenitors of tetraploid peanut, resulting in an overall co-linearity of about 60% with the average co-linearity of 68% for the A sub-genome and 47% for the B sub-genome. The analysis using the improved genetic map and multi-season (2010–2013) phenotypic data resulted in the identification of 48 quantitative trait loci (QTLs) with phenotypic variance explained (PVE) from 3.88 to 29.14%. Of the 48 QTLs, six QTLs were identified for resistance to TSWV, 22 QTLs for early leaf spot (ELS) and 20 QTLs for late leaf spot (LLS), which included four, six, and six major QTLs (PVE larger than 10%) for each disease, respectively. A total of six major genomic regions (MGR) were found to have QTLs controlling more than one disease resistance. The identified QTLs and resistance gene-rich MGRs will facilitate further discovery of resistance genes and development of molecular markers for these important diseases. PMID:27427980

  10. Mapping Quantitative Trait Loci of Resistance to Tomato Spotted Wilt Virus and Leaf Spots in a Recombinant Inbred Line Population of Peanut (Arachis hypogaea L.) from SunOleic 97R and NC94022.

    PubMed

    Khera, Pawan; Pandey, Manish K; Wang, Hui; Feng, Suping; Qiao, Lixian; Culbreath, Albert K; Kale, Sandip; Wang, Jianping; Holbrook, C Corley; Zhuang, Weijian; Varshney, Rajeev K; Guo, Baozhu

    2016-01-01

    Peanut is vulnerable to a range of diseases, such as Tomato spotted wilt virus (TSWV) and leaf spots which will cause significant yield loss. The most sustainable, economical and eco-friendly solution for managing peanut diseases is development of improved cultivars with high level of resistance. We developed a recombinant inbred line population from the cross between SunOleic 97R and NC94022, named as the S-population. An improved genetic linkage map was developed for the S-population with 248 marker loci and a marker density of 5.7 cM/loci. This genetic map was also compared with the physical map of diploid progenitors of tetraploid peanut, resulting in an overall co-linearity of about 60% with the average co-linearity of 68% for the A sub-genome and 47% for the B sub-genome. The analysis using the improved genetic map and multi-season (2010-2013) phenotypic data resulted in the identification of 48 quantitative trait loci (QTLs) with phenotypic variance explained (PVE) from 3.88 to 29.14%. Of the 48 QTLs, six QTLs were identified for resistance to TSWV, 22 QTLs for early leaf spot (ELS) and 20 QTLs for late leaf spot (LLS), which included four, six, and six major QTLs (PVE larger than 10%) for each disease, respectively. A total of six major genomic regions (MGR) were found to have QTLs controlling more than one disease resistance. The identified QTLs and resistance gene-rich MGRs will facilitate further discovery of resistance genes and development of molecular markers for these important diseases.

  11. A geminiviral amplicon (VA) derived from Tomato leaf curl virus (ToLCV) can replicate in a wide variety of plant species and also acts as a VIGS vector

    PubMed Central

    Pandey, Prerna; Choudhury, Nirupam R; Mukherjee, Sunil K

    2009-01-01

    Background The Tomato leaf curl virus (ToLCV) belongs to the genus begomoviridae of the family Geminiviridae. The 2.7 kb DNA genome of the virus encodes all the information required for viral DNA replication, transcription and transmission across the plant cells. However, all of the genome sequences are not required for viral DNA replication. We attempted to reveal the minimal essential region required for DNA replication and stable maintenance. The phenomenon of Virus Induced Gene Silencing (VIGS) has recently been observed with several geminiviruses. We investigated whether the minimal replicating region was also capable of producing siRNAs in planta and a VIGS vector could be constructed using the same minimal sequences. Results We have constructed vectors containing various truncated portions of the Tomato leaf curl virus (ToLCV) genome and established that a segment spanning from common region (CR) to AC3 (ORF coding for a replication enhancer) was the minimal portion which could efficiently replicate in a variety of both monocot and dicot plants. A viral amplicon (VA) vector was constructed using this region that produced siRNAs from various sites of the vector, in a temporal manner in plants, and hence can be used as a VIGS vector. The tomato endogene PCNA was silenced using this vector. Introduction of a mutation in the ORF AC2 (a silencing suppressor) increased the silencing efficiency of the newly constructed vector several folds. Conclusion Our study reveals that the vector is capable of replicating in diverse plant species and is highly efficient in silencing endogenes like PCNA of the host plant, thus acting as a VIGS vector. We observed that the geminiviral ORF AC2 functioned as a silencing suppressor and a null mutation in this ORF increased the efficiency of silencing several fold. This is the first report of construction of improved VIGS vector by mutation of the resident silencing suppressor gene. The present study opens up the possibility of

  12. The C2 protein of tomato yellow leaf curl Sardinia virus acts as a pathogenicity determinant and a 16-amino acid domain is responsible for inducing a hypersensitive response in plants.

    PubMed

    Matić, Slavica; Pegoraro, Mattia; Noris, Emanuela

    2016-04-02

    The role of the C2 protein in the pathogenicity of tomato yellow leaf curl Sardinia virus (TYLCSV) was investigated. Here we report that Agrobacterium-mediated transient expression of TYLCSV C2 resulted in a strong hypersensitive response (HR) in Nicotiana benthamiana, N. tabacum, and Arabidopsis thaliana, with induction of plant cell death and production of H2O2. Since HR is not evident in plants infected by TYLCSV, it is expected that TYLCSV encodes a gene (or genes) that counters this response. HR was partially counteracted by co-agroinfiltration of TYLCSV V2 and Rep, leading to chlorotic reaction, with no HR development. Considering that the corresponding C2 protein of the closely related tomato yellow leaf curl virus (TYLCV) did not induce HR, alignment of the C2 proteins of TYLCSV and TYLCV were carried out and a hypervariable region of 16 amino acids was identified. Its role in the induction of HR was demonstrated using TYLCSV-TYLCV C2 chimeric genes, encoding two TYLCSV C2 variants with a complete (16 aa) or a partial (10 aa only) swap of the corresponding sequence of TYLCV C2. Furthermore, using NahG transgenic N. benthamiana lines compromised in the accumulation of salicylic acid (SA), a key regulator of HR, only a chlorotic response occurred in TYLCSV C2-infiltrated tissue, indicating that SA participates in such plant defense process. These findings demonstrate that TYLCSV C2 acts as a pathogenicity determinant and induces host defense responses controlled by the SA pathway.

  13. Virus-induced gene silencing of the two squalene synthase isoforms of apple tree (Malus × domestica L.) negatively impacts phytosterol biosynthesis, plastid pigmentation and leaf growth.

    PubMed

    Navarro Gallón, Sandra M; Elejalde-Palmett, Carolina; Daudu, Dimitri; Liesecke, Franziska; Jullien, Frédéric; Papon, Nicolas; Dugé de Bernonville, Thomas; Courdavault, Vincent; Lanoue, Arnaud; Oudin, Audrey; Glévarec, Gaëlle; Pichon, Olivier; Clastre, Marc; St-Pierre, Benoit; Atehortùa, Lucia; Yoshikawa, Nobuyuki; Giglioli-Guivarc'h, Nathalie; Besseau, Sébastien

    2017-07-01

    The use of a VIGS approach to silence the newly characterized apple tree SQS isoforms points out the biological function of phytosterols in plastid pigmentation and leaf development. Triterpenoids are beneficial health compounds highly accumulated in apple; however, their metabolic regulation is poorly understood. Squalene synthase (SQS) is a key branch point enzyme involved in both phytosterol and triterpene biosynthesis. In this study, two SQS isoforms were identified in apple tree genome. Both isoforms are located at the endoplasmic reticulum surface and were demonstrated to be functional SQS enzymes using an in vitro activity assay. MdSQS1 and MdSQS2 display specificities in their expression profiles with respect to plant organs and environmental constraints. This indicates a possible preferential involvement of each isoform in phytosterol and/or triterpene metabolic pathways as further argued using RNAseq meta-transcriptomic analyses. Finally, a virus-induced gene silencing (VIGS) approach was used to silence MdSQS1 and MdSQS2. The concomitant down-regulation of both MdSQS isoforms strongly affected phytosterol synthesis without alteration in triterpene accumulation, since triterpene-specific oxidosqualene synthases were found to be up-regulated to compensate metabolic flux reduction. Phytosterol deficiencies in silenced plants clearly disturbed chloroplast pigmentation and led to abnormal development impacting leaf division rather than elongation or differentiation. In conclusion, beyond the characterization of two SQS isoforms in apple tree, this work brings clues for a specific involvement of each isoform in phytosterol and triterpene pathways and emphasizes the biological function of phytosterols in development and chloroplast integrity. Our report also opens the door to metabolism studies in Malus domestica using the apple latent spherical virus-based VIGS method.

  14. Walk and roll robot

    NASA Technical Reports Server (NTRS)

    Wilson, Andrew (Inventor); Punnoose, Andrew (Inventor); Strausser, Katherine (Inventor); Parikh, Neil (Inventor)

    2011-01-01

    A mobile robotic unit features a main body, a plurality of legs for supporting the main body on and moving the main body in forward and reverse directions about a base surface, and a drive assembly. According to an exemplary embodiment each leg includes a respective pivotal hip joint, a pivotal knee joint, and a wheeled foot adapted to roll along the base surface. Also according to an exemplary embodiments the drive assembly includes a motor operatively associated with the hip and knee joints and the wheeled foot for independently driving pivotal movement of the hip joint and the knee joint and rolling motion of the wheeled foot. The hip joint may include a ball-and-socket-type joint interconnecting top portion of the leg to the main body, such that the hip joint is adapted to pivot said leg in a direction transverse to a forward-and-reverse direction.

  15. Shear Roll Mill Reactivation

    DTIC Science & Technology

    2012-09-13

    pneumatically operated paste dumper and belt conveyor system, the loss in weight feeder system, the hydraulically operated shear roll mill, the pellet...out feed belt conveyor , and the pack out system comprised of the metal detector, scale, and pack out empty and full drum roller conveyors . Page | 4...feed hopper and conveyor supplying the loss in weight feeder were turned on, and it was verified that these items functioned as designed. The

  16. Investigation of thermomechanical behavior of a work roll and of roll life in hot strip rolling

    NASA Astrophysics Data System (ADS)

    Sun, C. G.; Hwang, S. M.; Yun, C. S.; Chung, J. S.

    1998-09-01

    An integrated finite element-based model is presented for the prediction of the steady-state thermomechanical behavior of the roll-strip system and of roll life in hot strip rolling. The model is comprised of basic finite-element models, which are incorporated into an iterative-solution procedure to deal with the interdependence between the thermomechanical behavior of the strip and that of the work roll, which arises from roll-strip contact, as well as with the interdependence between the thermal and mechanical behavior. Comparison is made between the predictions and the measurements to assess solution accuracy. Then, the effect of various process parameters on the detailed aspects of thermomechanical behavior of the work roll and on roll life is investigated via a series of process simulations.

  17. Diversity of beet curly top Iran virus isolated from different hosts in Iran.

    PubMed

    Gharouni Kardani, Sara; Heydarnejad, Jahangir; Zakiaghl, Mohammad; Mehrvar, Mohsen; Kraberger, Simona; Varsani, Arvind

    2013-06-01

    Beet curly top Iran virus (BCTIV) is a major pathogen of sugar beet in Iran. In order to study diversity of BCTIV, we sampled 68 plants in Iran during the summer of 2010 with curly top disease symptoms on beans (Phaseolus vulgaris), cowpeas (Vigna unguiculata), tomatoes (Solanum lycopersicum L.), sea beets (Beta vulgaris subsp. maritima), and sugar beets (Beta vulgaris). Plant samples showing leaf curling, yellowing, and/or swelling of veins on the lower leaf surfaces were collected from various fields in Khorasan Razavi, Northern Khorasan (north-eastern Iran), East Azarbayejan, West Azarbayejan (north-western Iran), and Fars (southern Iran) provinces. Using rolling circle amplification coupled with restriction digests, cloning, and Sanger sequencing, we determined the genomes of nine new BCTIV isolates from bean, cowpea, tomato, sea beet, and sugar beet in Iran. Our analysis reveals ~11 % diversity amongst BCTIV isolates and we detect evidence of recombination within these genomes.

  18. Roll-to-Roll production of carbon nanotubes based supercapacitors

    NASA Astrophysics Data System (ADS)

    Zhu, Jingyi; Childress, Anthony; Karakaya, Mehmet; Roberts, Mark; Arcilla-Velez, Margarita; Podila, Ramakrishna; Rao, Apparao

    2014-03-01

    Carbon nanomaterials provide an excellent platform for electrochemical double layer capacitors (EDLCs). However, current industrial methods for producing carbon nanotubes are expensive and thereby increase the costs of energy storage to more than 10 Wh/kg. In this regard, we developed a facile roll-to-roll production technology for scalable manufacturing of multi-walled carbon nanotubes (MWNTs) with variable density on run-of-the-mill kitchen Al foils. Our method produces MWNTs with diameter (heights) between 50-100 nm (10-100 μm), and a specific capacitance as high as ~ 100 F/g in non-aqueous electrolytes. In this talk, the fundamental challenges involved in EDLC-suitable MWNT growth, roll-to-roll production, and device manufacturing will be discussed along with electrochemical characteristics of roll-to-roll MWNTs. Research supported by NSF CMMI Grant1246800.

  19. Mechanical inoculation of plant viruses.

    PubMed

    Hull, Roger

    2009-05-01

    This technique is for the mechanical inoculation of viruses to plants. It is used to diagnose a virus by its reactions in a variety of plant species, to test the infectivity of virus samples using local lesion hosts, and to propagate viruses. The virus preparation is rubbed onto the surface of the leaf in such a way as to break the surface cells without causing too much mechanical damage. The preparation of the virus sample, its application to the leaf, and the care of the plants before and after inoculation are described.

  20. Explicit Nonlinear Finite Element Geometric Analysis of Parabolic Leaf Springs under Various Loads

    PubMed Central

    Kong, Y. S.; Omar, M. Z.; Chua, L. B.; Abdullah, S.

    2013-01-01

    This study describes the effects of bounce, brake, and roll behavior of a bus toward its leaf spring suspension systems. Parabolic leaf springs are designed based on vertical deflection and stress; however, loads are practically derived from various modes especially under harsh road drives or emergency braking. Parabolic leaf springs must sustain these loads without failing to ensure bus and passenger safety. In this study, the explicit nonlinear dynamic finite element (FE) method is implemented because of the complexity of experimental testing A series of load cases; namely, vertical push, wind-up, and suspension roll are introduced for the simulations. The vertical stiffness of the parabolic leaf springs is related to the vehicle load-carrying capability, whereas the wind-up stiffness is associated with vehicle braking. The roll stiffness of the parabolic leaf springs is correlated with the vehicle roll stability. To obtain a better bus performance, two new parabolic leaf spring designs are proposed and simulated. The stress level during the loadings is observed and compared with its design limit. Results indicate that the newly designed high vertical stiffness parabolic spring provides the bus a greater roll stability and a lower stress value compared with the original design. Bus safety and stability is promoted, as well as the load carrying capability. PMID:24298209

  1. Virus movement within grafted watermelon plants

    USDA-ARS?s Scientific Manuscript database

    Watermelon production in Florida is impacted by several viruses including whitefly-transmitted Squash vein yellowing virus (SqVYV), Cucurbit yellow stunting disorder virus and Cucurbit leaf crumple virus, and aphid-transmitted Papaya ringspot virus type W (PRSV-W). While germplasm resistant to some...

  2. Ecosystem engineers on plants: indirect facilitation of arthropod communities by leaf-rollers at different scales.

    PubMed

    Vieira, Camila; Romero, Gustavo Q

    2013-07-01

    Ecosystem engineering is a process by which organisms change the distribution of resources and create new habitats for other species via non-trophic interactions. Leaf-rolling caterpillars can act as ecosystem engineers because they provide shelter to secondary users. In this study, we report the influence of leaf-rolling caterpillars on speciose tropical arthropod communities along both spatial scales (leaf-level and plant-level effects) and temporal scales (dry and rainy seasons). We predict that rolled leaves can amplify arthropod diversity at both the leaf and plant levels and that this effect is stronger in dry seasons, when arthropods are prone to desiccation. Our results show that the abundance, richness, and biomass of arthropods within several guilds increased up to 22-fold in naturally and artificially created leaf shelters relative to unaltered leaves. These effects were observed at similar magnitudes at both the leaf and plant scales. Variation in the shelter architecture (funnel, cylinders) did not influence arthropod parameters, as diversity, abundance, orbiomass, but rolled leaves had distinct species composition if compared with unaltered leaves. As expected, these arthropod parameters on the plants with rolled leaves were on average approximately twofold higher in the dry season. Empty leaf rolls and whole plants were rapidly recolonized by arthropods over time, implying a fast replacement of individuals; within 15-day intervals the rolls and plants reached a species saturation. This study is the first to examine the extended effects of engineering caterpillars as diversity amplifiers at different temporal and spatial scales. Because shelter-building caterpillars are ubiquitous organisms in tropical and temperate forests, they can be considered key structuring elements for arthropod communities on plants.

  3. The 35-amino acid C2 protein of Cotton leaf curl Kokhran virus, Burewala, implicated in resistance breaking in cotton, retains some activities of the full-length protein.

    PubMed

    Akbar, Fazal; Iqbal, Zafar; Briddon, Rob W; Vazquez, Franck; Saeed, Muhammad

    2016-10-01

    With one exception, all the begomoviruses characterized so far encode an ~134-amino acid (aa) (A)C2 protein. The exception is the "Burewala" strain of Cotton leaf curl Kokhran virus (CLCuKoV-Bu), associated with resistance breaking in cotton across Pakistan and northwestern India, that encodes a truncated 35-aa C2. The C2 protein encoded by begomoviruses performs multiple functions including suppression of post-transcriptional gene silencing (PTGS), modulating microRNA (miRNA) expression and may be a pathogenicity determinant. The study described here was designed to investigate whether the CLCuKoV-Bu 35-aa C2 retains the activities of the full-length C2 protein. The results showed the 35-aa C2 of CLCuKoV-Bu acts as a pathogenicity determinant, suppresses PTGS and upregulates miRNA expression when expressed from a Potato virus X vector in Nicotiana benthamiana. The symptoms induced by expression of full-length C2 were more severe than those induced by the 35-aa C2. The accumulation of most developmental miRNAs decreases with the full-length C2 protein and increases with the 35-aa peptide of CLCuKoV-Bu. The study also revealed that 35-aa peptide of CLCuKoV-Bu maintains suppressor of silencing activity at a level equal to that of full-length C2. The significance of the results with respect to virus fitness and resistance breaking is discussed.

  4. Tomato yellow leaf curl Sardinia virus-resistant tomato plants expressing the multifunctional N-terminal domain of the replication-associated protein show transcriptional changes resembling stress-related responses.

    PubMed

    Lucioli, Alessandra; Berardi, Alessandra; Gatti, Francesca; Tavazza, Raffaela; Pizzichini, Daniele; Tavazza, Mario

    2014-01-01

    The N-terminal domain (amino acids 1-130) of the replication-associated protein (Rep130 ) of Tomato yellow leaf curl Sardinia virus (TYLCSV) retains the ability of full-length Rep to localize to the nucleus and to down-regulate C1 transcription when ectopically expressed in plants, both functions being required to inhibit homologous viral replication. In this study, we analysed the effect of Rep130 expression on virus resistance and the plant transcriptome in the natural and agronomically important host species of TYLCSV, Solanum lycopersicum. Tomato plants accumulating high levels of Rep130 were generated and proved to be resistant to TYLCSV. Using an in vitro assay, we showed that plant-expressed Rep130 also retains the catalytic activity of Rep, thus supporting the notion that this protein domain is fully functional. Interestingly, Rep130 -expressing tomatoes were characterized by an altered transcriptional profile resembling stress-related responses. Notably, the serine-type protease inhibitor (Ser-PI) category was over-represented among the 20 up-regulated genes. The involvement of Rep130 in the alteration of host mRNA steady-state levels was confirmed using a distinct set of virus-resistant transgenic tomato plants expressing the same TYLCSV Rep130 , but from a different, synthetic, gene. Eight genes were found to be up-regulated in both types of transgenic tomato and two encoded Ser-PIs. Four of these eight genes were also up-regulated in TYLCSV-infected wild-type tomato plants. Implications with regard to the ability of this Rep domain to interfere with viral infections and to alter the host transcriptome are discussed. © 2013 BSPP AND JOHN WILEY & SONS LTD.

  5. Rolling cuff flexible bellows

    DOEpatents

    Lambert, Donald R.

    1985-01-01

    A flexible connector apparatus used to join two stiff non-deformable members, such as piping. The apparatus is provided with one or more flexible sections or assemblies each utilizing a bellows of a rolling cuff type connected between two ridge members, with the bellows being supported by a back-up ring, such that only the curved end sections of the bellows are unsupported. Thus, the bellows can be considered as being of a tube-shaped configuration and thus have high pressure resistance. The components of the flexible apparatus are sealed or welded one to another such that it is fluid tight.

  6. VIEW OF HANDOPERATED ROLLING MILLS ROLLING STANDS FROM LEFT TO ...

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

    VIEW OF HAND-OPERATED ROLLING MILLS ROLLING STANDS FROM LEFT TO RIGHT: THREE HIGH; THREE HIGH; THREE HIGH; THREE HIGH (OPERATED AS A TWO-HIGH); TWO HIGH TWO HIGH MANUFACTURED BY BLAW-KNOX THREE HIGH MANUFACTURED BY LEWIS FOUNDRY AND MACHINE CO. - Cambria Iron Company, Gautier Works, 12" Mill, Clinton Street & Little Conemaugh River, Johnstown, Cambria County, PA

  7. Expression of a mutant maize gene in the ventral leaf epidermis is sufficient to signal a switch of the leaf's dorsoventral axis.

    PubMed

    Nelson, Jennifer M; Lane, Barbara; Freeling, Michael

    2002-10-01

    Maize leaves are initiated from the shoot apex with an inherent leaf dorsoventral polarity; the leaf surface closest to the meristem is the adaxial (upper, dorsal) surface whereas the opposite leaf surface is the abaxial (lower, ventral) surface. The Rolled leaf1 (Rld1) semi-dominant maize mutations affect dorsoventral patterning by causing adaxialization of abaxial leaf regions. This adaxialization is sometimes associated with abaxialization of the adaxial leaf regions, which constitutes a "switch". Dosage analysis indicates Rld1 mutants are antimorphs. We mapped Rld1's action to a single cell layer using a mosaic analysis and show Rld1 acts non cell-autonomously along the dorsoventral axis. The presence of Rld1 mutant product in the abaxial epidermis is necessary and sufficient to induce the Rolled leaf1 phenotype within the lower epidermis as well as in other leaf layers along the dorsoventral axis. These results support a model for the involvement of wild-type RLD1 in the maintenance of dorsoventral features of the leaf. In addition, they demonstrate the abaxial epidermis sends/receives a cell fate determining signal to/from the adaxial epidermis and controls the dorsoventral patterning of the maize leaf.

  8. Ocimum sanctum leaf extract induces drought stress tolerance in rice

    PubMed Central

    Pandey, Veena; Ansari, M.W.; Tula, Suresh; Sahoo, R.K.; Bains, Gurdeep; Kumar, J.; Tuteja, Narendra; Shukla, Alok

    2016-01-01

    ABSTRACT Ocimum leaves are highly enriched in antioxidant components. Thus, its leaf extract, if applied in plants, is believed to efficiently scavenge ROS, thereby preventing oxidative damage under drought stress. Thus, the present study was performed in kharif 2013 and rabi 2014 season to evaluate the effect of aqueous leaf extract of Ocimum sanctum against drought stress in 2 rice genotype under glass house conditions. Here we show that various morpho- physiological (chlorophyll fluorescence, leaf rolling score, leaf tip burn, number of senesced leaves and total dry matter) and biochemical parameters (proline, malondialdehyde and superoxide dismutase content) were amended by Ocimum treatment in both the seasons. Application of Ocimum extract increased expression of dehydrin genes, while reducing expression of aquaporin genes in drought stressed rice plant. Thus, application of Ocimum leaf extract under drought stress can be suggested as a promising strategy to mitigate drought stress in economical, accessible and ecofriendly manner. PMID:26890603

  9. Molecular Characterization of Watermelon Chlorotic Stunt Virus (WmCSV) from Palestine

    PubMed Central

    Ali-Shtayeh, Mohammed S.; Jamous, Rana M.; Mallah, Omar B.; Abu-Zeitoun, Salam Y.

    2014-01-01

    The incidence of watermelon chlorotic stunt disease and molecular characterization of the Palestinian isolate of Watermelon chlorotic stunt virus (WmCSV-[PAL]) are described in this study. Symptomatic leaf samples obtained from watermelon Citrullus lanatus (Thunb.), and cucumber (Cucumis sativus L.) plants were tested for WmCSV-[PAL] infection by polymerase chain reaction (PCR) and Rolling Circle Amplification (RCA). Disease incidence ranged between 25%–98% in watermelon fields in the studied area, 77% of leaf samples collected from Jenin were found to be mixed infected with WmCSV-[PAL] and SLCV. The full-length DNA-A and DNA-B genomes of WmCSV-[PAL] were amplified and sequenced, and the sequences were deposited in the GenBank. Sequence analysis of virus genomes showed that DNA-A and DNA-B had 97.6%–99.42% and 93.16%–98.26% nucleotide identity with other virus isolates in the region, respectively. Sequence analysis also revealed that the Palestinian isolate of WmCSV shared the highest nucleotide identity with an isolate from Israel suggesting that the virus was introduced to Palestine from Israel. PMID:24956181

  10. Video Analysis of Rolling Cylinders

    ERIC Educational Resources Information Center

    Phommarach, S.; Wattanakasiwich, P.; Johnston, I.

    2012-01-01

    In this work, we studied the rolling motion of solid and hollow cylinders down an inclined plane at different angles. The motions were captured on video at 300 frames s[superscript -1], and the videos were analyzed frame by frame using video analysis software. Data from the real motion were compared with the theory of rolling down an inclined…

  11. Video Analysis of Rolling Cylinders

    ERIC Educational Resources Information Center

    Phommarach, S.; Wattanakasiwich, P.; Johnston, I.

    2012-01-01

    In this work, we studied the rolling motion of solid and hollow cylinders down an inclined plane at different angles. The motions were captured on video at 300 frames s[superscript -1], and the videos were analyzed frame by frame using video analysis software. Data from the real motion were compared with the theory of rolling down an inclined…

  12. Lubrication of rolling element bearings

    NASA Technical Reports Server (NTRS)

    Parker, R. J.

    1980-01-01

    This paper is a broad survey of the lubrication of rolling-element bearings. Emphasis is on the critical design aspects related to speed, temperature, and ambient pressure environment. Types of lubrication including grease, jets, mist, wick, and through-the-race are discussed. The paper covers the historical development, present state of technology, and the future problems of rolling-element bearing lubrication.

  13. The C2 Protein from the Geminivirus Tomato Yellow Leaf Curl Sardinia Virus Decreases Sensitivity to Jasmonates and Suppresses Jasmonate-Mediated Defences.

    PubMed

    Rosas-Díaz, Tábata; Macho, Alberto P; Beuzón, Carmen R; Lozano-Durán, Rosa; Bejarano, Eduardo R

    2016-01-15

    An increasing body of evidence points at a role of the plant hormones jasmonates (JAs) in determining the outcome of plant-virus interactions. Geminiviruses, small DNA viruses infecting a wide range of plant species worldwide, encode a multifunctional protein, C2, which is essential for full pathogenicity. The C2 protein has been shown to suppress the JA response, although the current view on the extent of this effect and the underlying molecular mechanisms is incomplete. In this work, we use a combination of exogenous hormone treatments, microarray analysis, and pathogen infections to analyze, in detail, the suppression of the JA response exerted by C2. Our results indicate that C2 specifically affects certain JA-induced responses, namely defence and secondary metabolism, and show that plants expressing C2 are more susceptible to pathogen attack. We propose a model in which C2 might interfere with the JA response at several levels.

  14. The C2 Protein from the Geminivirus Tomato Yellow Leaf Curl Sardinia Virus Decreases Sensitivity to Jasmonates and Suppresses Jasmonate-Mediated Defences

    PubMed Central

    Rosas-Díaz, Tábata; Macho, Alberto P.; Beuzón, Carmen R.; Lozano-Durán, Rosa; Bejarano, Eduardo R.

    2016-01-01

    An increasing body of evidence points at a role of the plant hormones jasmonates (JAs) in determining the outcome of plant-virus interactions. Geminiviruses, small DNA viruses infecting a wide range of plant species worldwide, encode a multifunctional protein, C2, which is essential for full pathogenicity. The C2 protein has been shown to suppress the JA response, although the current view on the extent of this effect and the underlying molecular mechanisms is incomplete. In this work, we use a combination of exogenous hormone treatments, microarray analysis, and pathogen infections to analyze, in detail, the suppression of the JA response exerted by C2. Our results indicate that C2 specifically affects certain JA-induced responses, namely defence and secondary metabolism, and show that plants expressing C2 are more susceptible to pathogen attack. We propose a model in which C2 might interfere with the JA response at several levels. PMID:27135228

  15. 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.

  16. Critical rolling angle of microparticles

    NASA Astrophysics Data System (ADS)

    Farzi, Bahman; Vallabh, Chaitanya K. P.; Stephens, James D.; Cetinkaya, Cetin

    2016-03-01

    At the micrometer-scale and below, particle adhesion becomes particularly relevant as van der Waals force often dominates volume and surface proportional forces. The rolling resistance of microparticles and their critical rolling angles prior to the initiation of free-rolling and/or complete detachment are critical in numerous industrial processes and natural phenomenon involving particle adhesion and granular dynamics. The current work describes a non-contact measurement approach for determining the critical rolling angle of a single microparticle under the influence of a contact-point base-excitation generated by a transient displacement field of a prescribed surface acoustic wave pulse and reports the critical rolling angle data for a set of polystyrene latex microparticles.

  17. Roll mill for milling coal

    SciTech Connect

    Brundiek, H.; Werner, L.

    1984-02-21

    A roll mill, more specially for coal, has a turning pan and a number of rolls running thereon for producing a milling effect. Each roll is supported on a rocker arm, able to be turned about a horizontal axis which is roughly tangential to the pan. The rocker arm and the roll on it are forced down against the pan by a hydraulic cylinder joined with a fork which, in turn, is joined with the rocker arm for turning it about the turnpin. The fork may be unjoined from the rocker arm for upkeep work on the roll. Each rocker arm has a gas-tight cover structure which is part of the casing of the mill.

  18. Deep sequencing analysis of apple infecting viruses in Korea

    USDA-ARS?s Scientific Manuscript database

    Deep sequencing of viruses isolated from eight symptomatic apple trees in Korea has generated 52 contigs derived from five viruses: Apple chlorotic leaf spot virus (ACLSV), Apple stem grooving virus (ASGV), Apple stem pitting virus (ASPV), Apple green crinkle associated virus (AGCaV) and Apricot lat...

  19. Interaction of Arabidopsis TGA3 and WRKY53 transcription factors on Cestrum yellow leaf curling virus (CmYLCV) promoter mediates salicylic acid-dependent gene expression in planta.

    PubMed

    Sarkar, Shayan; Das, Abhimanyu; Khandagale, Prashant; Maiti, Indu B; Chattopadhyay, Sudip; Dey, Nrisingha

    2017-09-14

    This paper highlighted a salicylic acid-inducible Caulimoviral promoter fragment from Cestrum yellow leaf curling virus (CmYLCV). Interaction of Arabidopsis transcription factors TGA3 and WRKY53 on CmYLCV promoter resulted in the enhancement of the promoter activity via NPR1-dependent salicylic acid signaling. Several transcriptional promoters isolated from plant-infecting Caulimoviruses are being presently used worldwide as efficient tools for plant gene expression. The CmYLCV promoter has been isolated from the Cestrum yellow leaf curling virus (Caulimoviruses) and characterized more than 12 years ago; also we have earlier reported a near-constitutive, pathogen-inducible CmYLCV promoter fragment (-329 to +137 from transcription start site; TSS) that enhances stronger (3×) expression than the previously reported fragments; all these fragments are highly efficient in monocot and dicot plants (Sahoo et al. Planta 240: 855-875, 2014). Here, we have shown that the full-length CmYLCV promoter fragment (-729 to +137 from TSS) is salicylic acid (SA) inducible. In this context, we have performed an in-depth study to elucidate the factors responsible for SA-inducibility of the CmYLCV promoter. We found that the as-1 1 and W-box1 elements (located at -649 and -640 from the TSS) of the CmYLCV promoter are required for SA-induced activation by recruiting Arabidopsis TGA3 and WRKY53 transcription factors. Consequently, as a nascent observation, we established the physical interaction between TGA3 and WYKY53; also demonstrated that the N-terminal domain of TGA3 is sufficient for the interaction with the full-length WRKY53. Such interaction synergistically activates the CmYLCV promoter activity in planta. Further, we found that activation of the CmYLCV promoter by SA through TGA3 and WRKY53 interaction depends on NPR1. Finally, the findings presented here provide strong support for the direct regulatory roles of TGA3 and WRKY53 in the SA and NPR1-dependent activation of a

  20. Analysis of sequences from field samples reveals the presence of the recently described pepper vein yellows virus (genus Polerovirus) in six additional countries.

    PubMed

    Knierim, Dennis; Tsai, Wen-Shi; Kenyon, Lawrence

    2013-06-01

    Polerovirus infection was detected by reverse transcription polymerase chain reaction (RT-PCR) in 29 pepper plants (Capsicum spp.) and one black nightshade plant (Solanum nigrum) sample collected from fields in India, Indonesia, Mali, Philippines, Thailand and Taiwan. At least two representative samples for each country were selected to generate a general polerovirus RT-PCR product of 1.4 kb length for sequencing. Sequence analysis of the partial genome sequences revealed the presence of pepper vein yellows virus (PeVYV) in all 13 samples. A 1990 Australian herbarium sample of pepper described by serological means as infected with capsicum yellows virus (CYV) was identified by sequence analysis of a partial CP sequence as probably infected with a potato leaf roll virus (PLRV) isolate.

  1. Avoiding the parametric roll

    NASA Astrophysics Data System (ADS)

    Acomi, Nicoleta; Ancuţa, Cristian; Andrei, Cristian; Boştinǎ, Alina; Boştinǎ, Aurel

    2016-12-01

    Ships are mainly built to sail and transport cargo at sea. Environmental conditions and state of the sea are communicated to vessels through periodic weather forecasts. Despite officers being aware of the sea state, their sea time experience is a decisive factor when the vessel encounters severe environmental conditions. Another important factor is the loading condition of the vessel, which triggers different behaviour in similar marine environmental conditions. This paper aims to analyse the behaviour of a port container vessel in severe environmental conditions and to estimate the potential conditions of parametric roll resonance. Octopus software simulation is employed to simulate vessel motions under certain conditions of the sea, with possibility to analyse the behaviour of ships and the impact of high waves on ships due to specific wave encounter situations. The study should be regarded as a supporting tool during the decision making process.

  2. First report of association of potato virus X and potato virus Y and 'Candidatus Phytoplasma trifolii' in brinjal in India.

    PubMed

    Kumar, Manish; Katiyar, Akshay; Madhupriya; Rao, G P

    2016-06-01

    Symptoms of little leaf, leaf chlorosis and leaf malformations with mosaic mottling symptoms were observed in two brinjal varieties (Pusa Shyamla and Pusa Purple Cluster) in fields of IARI, New Delhi, India during 2014-2015. Electron microscopy, PCR and sequence analysis first time provided evidence of association of Candidatus Phytoplasma trifolii with potato virus X and potato virus Y in brinjal in India.

  3. Transcriptomics of the interaction between the monopartite phloem-limited geminivirus tomato yellow leaf curl Sardinia virus and Solanum lycopersicum highlights a role for plant hormones, autophagy and plant immune system fine tuning during infection.

    PubMed

    Miozzi, Laura; Napoli, Chiara; Sardo, Luca; Accotto, Gian Paolo

    2014-01-01

    Tomato yellow leaf curl Sardinia virus (TYLCSV), a DNA virus belonging to the genus Begomovirus, causes severe losses in tomato crops. It infects only a limited number of cells in the vascular tissues, making difficult to detect changes in host gene expression linked to its presence. Here we present the first microarray study of transcriptional changes induced by the phloem-limited geminivirus TYLCSV infecting tomato, its natural host. The analysis was performed on the midrib of mature leaves, a material naturally enriched in vascular tissues. A total of 2206 genes were up-regulated and 1398 were down-regulated in infected plants, with an overrepresentation of genes involved in hormone metabolism and responses, nucleic acid metabolism, regulation of transcription, ubiquitin-proteasome pathway and autophagy among those up-regulated, and in primary and secondary metabolism, phosphorylation, transcription and methylation-dependent chromatin silencing among those down-regulated. Our analysis showed a series of responses, such as the induction of GA- and ABA-responsive genes, the activation of the autophagic process and the fine tuning of the plant immune system, observed only in TYLCSV-tomato compatible interaction so far. On the other hand, comparisons with transcriptional changes observed in other geminivirus-plant interactions highlighted common host responses consisting in the deregulation of biotic stress responsive genes, key enzymes in the ethylene biosynthesis and methylation cycle, components of the ubiquitin proteasome system and DNA polymerases II. The involvement of conserved miRNAs and of solanaceous- and tomato-specific miRNAs in geminivirus infection, investigated by integrating differential gene expression data with miRNA targeting data, is discussed.

  4. Blueberry virus A

    USDA-ARS?s Scientific Manuscript database

    Leaf yellowing on highbush blueberry ‘Spartan’ prompted Isogai et al. to investigate whether a virus was the causal agent of the disorder. After double-stranded RNA extraction from symptomatic material they identified a single band of 17 Kb, indicative of virus infection. Shotgun cloning and sequenc...

  5. Molecular diagnosis of Papaya meleira virus (PMeV) from leaf samples of Carica papaya L. using conventional and real-time RT-PCR.

    PubMed

    Abreu, Paolla M V; Piccin, João G; Rodrigues, Silas P; Buss, David S; Ventura, José A; Fernandes, Patricia M B

    2012-03-01

    Papaya meleira virus (PMeV) is the causal agent of papaya sticky disease. This study describes two methods for molecular diagnosis of PMeV using conventional and real-time PCR. These methods were shown to be more efficient than current methods of viral detection using extraction of PMeV dsRNA and observation of symptoms in the field. The methods described here were used to evaluate the effect of inoculation of papaya plants with purified PMeV dsRNA on the progress of PMeV infection. A single inoculation with PMeV dsRNA was observed to delay the progress of the virus infection by several weeks. The possibility of vertical transmission of PMeV was also investigated. No evidence was found for PMeV transmission through seeds collected from diseased fruit. The implications of these results for the epidemiology of PMeV and the management of papaya sticky disease are discussed.

  6. Label-free Proteomic Reveals that Cowpea Severe Mosaic Virus Transiently Suppresses the Host Leaf Protein Accumulation During the Compatible Interaction with Cowpea (Vigna unguiculata [L.] Walp.).

    PubMed

    Paiva, Ana L S; Oliveira, Jose T A; de Souza, Gustavo A; Vasconcelos, Ilka M

    2016-12-02

    Viruses are important plant pathogens that threaten diverse crops worldwide. Diseases caused by Cowpea severe mosaic virus (CPSMV) have drawn attention because of the serious damages they cause to economically important crops including cowpea. This work was undertaken to quantify and identify the responsive proteins of a susceptible cowpea genotype infected with CPSMV, in comparison with mock-inoculated controls, using label-free quantitative proteomics and databanks, aiming at providing insights on the molecular basis of this compatible interaction. Cowpea leaves were mock- or CPSMV-inoculated and 2 and 6 days later proteins were extracted and analyzed. More than 3000 proteins were identified (data available via ProteomeXchange, identifier PXD005025) and 75 and 55 of them differentially accumulated in response to CPSMV, at 2 and 6 DAI, respectively. At 2 DAI, 76% of the proteins decreased in amount and 24% increased. However, at 6 DAI, 100% of the identified proteins increased. Thus, CPSMV transiently suppresses the synthesis of proteins involved particularly in the redox homeostasis, protein synthesis, defense, stress, RNA/DNA metabolism, signaling, and other functions, allowing viral invasion and spread in cowpea tissues.

  7. Combinations of two amino acids (Ala40 and Phe75 or Ser40 and Tyr75) in the coat protein of apple chlorotic leaf spot virus are crucial for infectivity.

    PubMed

    Yaegashi, Hajime; Isogai, Masamichi; Tajima, Hiroko; Sano, Teruo; Yoshikawa, Nobuyuki

    2007-09-01

    Amino acid sequences of apple chlorotic leaf spot virus (ACLSV) coat protein (CP) were compared between 12 isolates from apple, plum and cherry, and 109 cDNA clones that were amplified directly from infected apple tissues. Phylogenetic analysis based on the amino acid sequences of CP showed that the isolates and cDNA clones were separated into two major clusters in which the combinations of the five amino acids at positions 40, 59, 75, 130 and 184 (Ala(40)-Val(59)-Phe(75)-Ser(130)-Met(184) or Ser(40)-Leu(59)-Tyr(75)-Thr(130)-Leu(184)) were highly conserved within each cluster. Site-directed mutagenesis using an infectious cDNA clone of ACLSV indicated that the combinations of two amino acids (Ala(40) and Phe(75) or Ser(40) and Tyr(75)) are necessary for infectivity to Chenopodium quinoa plants by mechanical inoculation. Moreover, an agroinoculation assay indicated that the substitution of a single amino acid (Ala(40) to Ser(40) or Phe(75) to Tyr(75)) resulted in extreme reduction in the accumulation of viral genomic RNA, double-stranded RNAs and viral proteins (movement protein and CP) in infiltrated tissues, suggesting that the combinations of the two amino acids at positions 40 and 75 are important for effective replication in host plant cells.

  8. Mechanism of regulation of tomato TRN1 gene expression in late infection with tomato leaf curl New Delhi virus (ToLCNDV).

    PubMed

    Mandal, Arunava; Sarkar, Deepti; Kundu, Surekha; Kundu, Pallob

    2015-12-01

    Tomato leaf curl disease caused by geminiviruses is manifested by curling and puckering of leaves and thickening of veins, resembling developmental defects. This is probably due to the long-term altered regulation of expression of development related gene(s). Our results show that in the infected leaves the transcript level of TORNADO1 (SlTRN1), a gene important for cell expansion and vein formation, increased significantly. SlTRN1 is transcribed from two start sites. The preferential usage of one start site governs its expression in viral-stressed plants. To investigate the role of specific promoter elements in mediating differential expression of SlTRN1, we performed SlTRN1 promoter analysis. The promoter-regulatory sequences harbor multiple W-boxes. The SlWRKY16 transcription factor actively interacts with one of the W-boxes. WRKY proteins are commonly induced by salicylic acid (SA), and consequently SA treatment increased transcript level of SlWRKY16 and SlTRN1. Further mutational analyses confirmed the role of W-boxes in mediating SlTRN1 induction during ToLCNDV infection or SA treatment. We postulate that the activation of SA pathway during stress-response in tomato induces WRKY16, which in turn modulates transcription of SlTRN1 gene. This study unravels the mechanism of regulation of a developmental gene during stress-response, which may affect the severity of symptoms.

  9. Genetic variability of Cotton leaf curl betasatellite in Northern India

    PubMed Central

    Sohrab, Sayed Sartaj; Azhar, Esam I.; Kamal, Mohammad A.; Bhattacharya, P.S.; Rana, D.

    2014-01-01

    Cotton is an important crop and its production is affected by various disease pathogens. Monopartite begomovirus associated betasatellites cause Cotton leaf curl disease (CLCuD) in Northern India. In order to access the occurrence and genetic variability of Cotton leaf curl betasatellites, an extensive field survey was conducted in states of Rajasthan, Punjab and Haryana. We selected the betasatellite sequence for analysis as they are reported as important for disease severity and sequence variability. Based on the field observations, the disease incidence ranged from 30% to 80% during the survey. Full genome and DNA β were amplified from various samples while no amplicon was obtained in some samples. The nucleotide sequence homology ranged from 90.0% to 98.7% with Cotton leaf curl virus (CLCuV), 55.2–55.5% with Bhendi yellow vein mosaic virus, 55.8% with Okra leaf curl virus and 51.70% with Tomato leaf curl virus isolates. The lowest similarity (47.8%) was found in CLCuV-Sudan isolate. Phylogenetic analysis showed that analyzed isolates formed a close cluster with various CLCuV isolates reported earlier. The analysis results show sequence variation in Cotton leaf curl betasatellite which could be the result of recombination. The results obtained by genome amplification and sequence variability indicate that some new variants are circulating and causing leaf curl disease in Rajasthan, Punjab and Haryana. PMID:25473373

  10. Genetic variability of Cotton leaf curl betasatellite in Northern India.

    PubMed

    Sohrab, Sayed Sartaj; Azhar, Esam I; Kamal, Mohammad A; Bhattacharya, P S; Rana, D

    2014-12-01

    Cotton is an important crop and its production is affected by various disease pathogens. Monopartite begomovirus associated betasatellites cause Cotton leaf curl disease (CLCuD) in Northern India. In order to access the occurrence and genetic variability of Cotton leaf curl betasatellites, an extensive field survey was conducted in states of Rajasthan, Punjab and Haryana. We selected the betasatellite sequence for analysis as they are reported as important for disease severity and sequence variability. Based on the field observations, the disease incidence ranged from 30% to 80% during the survey. Full genome and DNA β were amplified from various samples while no amplicon was obtained in some samples. The nucleotide sequence homology ranged from 90.0% to 98.7% with Cotton leaf curl virus (CLCuV), 55.2-55.5% with Bhendi yellow vein mosaic virus, 55.8% with Okra leaf curl virus and 51.70% with Tomato leaf curl virus isolates. The lowest similarity (47.8%) was found in CLCuV-Sudan isolate. Phylogenetic analysis showed that analyzed isolates formed a close cluster with various CLCuV isolates reported earlier. The analysis results show sequence variation in Cotton leaf curl betasatellite which could be the result of recombination. The results obtained by genome amplification and sequence variability indicate that some new variants are circulating and causing leaf curl disease in Rajasthan, Punjab and Haryana.

  11. Leaf curl diseases of two solanaceous species in Southwest Arabia are caused by a monopartite begomovirus evolutionarily most closely related to a species from the Nile Basin and unique suite of betasatellites.

    PubMed

    Idris, Ali M; Abdullah, N M; Brown, J K

    2012-10-01

    The complete genome of 2780 bases was amplified using rolling circle amplification, and cloned, and sequenced for two distinct strains of the monopartite begomovirus Tomato leaf curl Sudan virus (ToLCSDV). The two strains shared 86-91% identity with the previously described ToLCSDV from the Nile Basin, and 90-91% identity with one another. One strain was cloned from symptomatic tomato plants from Tihamah (ToLCSDV-YE[YE:Tih:05]) while the other was cloned from symptomatic tobacco plants collected from Wadi Hadramaut (ToLCSDV-YE[YE:Had:89]). A distinct full-length betasatellite molecule (1352 bases) was cloned from the respective field-infected tomato and tobacco plants. Agro-inoculation of tomato and Nicotiana benthamiana plants with cloned partial tandem repeats of ToLCSDV-YE[YE:Tih11:05]) and the associated betasatellite, Tomato leaf curl Yemen betasatellite (ToLCYEB-[Tih:tom:137:05]), resulted in the reproduction of leaf curl disease symptoms in test plants like those observed in the field-infected plants. The betasatellite contributed to symptom severity in N. benthamiana test plants when it was co-inoculated with ToLCSDV-YE, compared to the milder symptoms that were observed in tobacco plants infected with the helper virus alone.

  12. Manufacturing Demonstration Facility: Roll-to-Roll Processing

    SciTech Connect

    Datskos, Panos G; Joshi, Pooran C; List III, Frederick Alyious; Duty, Chad E; Armstrong, Beth L; Ivanov, Ilia N; Jacobs, Christopher B; Graham, David E; Moon, Ji Won

    2015-08-01

    This Manufacturing Demonstration Facility (MDF)e roll-to-roll processing effort described in this report provided an excellent opportunity to investigate a number of advanced manufacturing approaches to achieve a path for low cost devices and sensors. Critical to this effort is the ability to deposit thin films at low temperatures using nanomaterials derived from nanofermentation. The overarching goal of this project was to develop roll-to-roll manufacturing processes of thin film deposition on low-cost flexible substrates for electronics and sensor applications. This project utilized ORNL s unique Pulse Thermal Processing (PTP) technologies coupled with non-vacuum low temperature deposition techniques, ORNL s clean room facility, slot dye coating, drop casting, spin coating, screen printing and several other equipment including a Dimatix ink jet printer and a large-scale Kyocera ink jet printer. The roll-to-roll processing project had three main tasks: 1) develop and demonstrate zinc-Zn based opto-electronic sensors using low cost nanoparticulate structures manufactured in a related MDF Project using nanofermentation techniques, 2) evaluate the use of silver based conductive inks developed by project partner NovaCentrix for electronic device fabrication, and 3) demonstrate a suite of low cost printed sensors developed using non-vacuum deposition techniques which involved the integration of metal and semiconductor layers to establish a diverse sensor platform technology.

  13. Rolling Process Modeling Report: Finite-Element Prediction of Roll Separating Force and Rolling Defects

    SciTech Connect

    Soulami, Ayoub; Lavender, Curt A.; Paxton, Dean M.; Burkes, Douglas

    2014-04-23

    Pacific Northwest National Laboratory (PNNL) has been investigating manufacturing processes for the uranium-10% molybdenum (U-10Mo) alloy plate-type fuel for the U.S. high-performance research reactors. This work supports the Convert Program of the U.S. Department of Energy’s National Nuclear Security Administration (DOE/NNSA) Global Threat Reduction Initiative. This report documents modeling results of PNNL’s efforts to perform finite-element simulations to predict roll separating forces and rolling defects. Simulations were performed using a finite-element model developed using the commercial code LS-Dyna. Simulations of the hot rolling of U-10Mo coupons encapsulated in low-carbon steel have been conducted following two different schedules. Model predictions of the roll-separation force and roll-pack thicknesses at different stages of the rolling process were compared with experimental measurements. This report discusses various attributes of the rolled coupons revealed by the model (e.g., dog-boning and thickness non-uniformity).

  14. Body roll in swimming: a review.

    PubMed

    Psycharakis, Stelios G; Sanders, Ross H

    2010-02-01

    In this article, we present a critical review of the swimming literature on body roll, for the purposes of summarizing and highlighting existing knowledge, identifying the gaps and limitations, and stimulating further research. The main research findings can be summarized as follows: swimmers roll their shoulders significantly more than their hips; swimmers increase hip roll but maintain shoulder roll when fatigued; faster swimmers roll their shoulders less than slower swimmers during a 200-m swim; roll asymmetries, temporal differences in shoulder roll and hip roll, and shoulder roll side dominance exist in front crawl swimming, but there is no evidence to suggest that they affect swimming performance; and buoyancy contributes strongly to generating body roll in front crawl swimming. Based on and stimulated by current knowledge, future research should focus on the following areas: calculation of body roll for female swimmers and for backstroke swimming; differences in body roll between breathing and non-breathing cycles; causes of body roll asymmetries and their relation to motor laterality; body roll analysis across a wide range of velocities and swimming distances; exploration of the association between body roll and the magnitude and direction of propulsive/resistive forces developed during the stroke cycle; and the influence of kicking actions on the generation of body roll.

  15. A review of the mechanisms and components that determine the transmission efficiency of Tomato yellow leaf curl virus (Geminiviridae; Begomovirus) by its whitefly vector.

    PubMed

    Ghanim, Murad

    2014-06-24

    Begomoviruses are a group of icosahedral single stranded DNA viruses exclusively transmitted by the sweet potato whitefly Bemisia tabaci in a persistent, circulative manner. In this mode of transmission, begomoviruses are acquired by their insect vector as intact virions from the plant phloem, move along the food canal, foregut and esophagus and reach the midgut where they are absorbed into the hemolymph via the filter chamber. The filter chamber is the site where most of the ingested food is filtered, and the first site where the majority of begomoviruses appear to be translocated into the hemolymph via unknown proteins or receptors. Transport from the filter chamber to the hemolymph is aided by a Heat Shock Protein 70. Virus particles not translocated across the filter chamber circulate in the midgut loop but it is not known whether absorption into the hemolymph occurs along this loop. Localization studies have confirmed that begomoviruses are not associated with the hindgut and absorption of virions in this organ is unlikely. In the hemolymph, virions have been shown to interact with a GroEL chaperone produced by the whitefly's endosymbiontic bacteria for ensuring their safe journey to the salivary glands. Virions penetrate the primary salivary glands via unknown proteins or receptors and are transported and secreted outside the whitefly to the plant with salivary secretions. Several recent studies have demonstrated the implications of insect and endosymbiont proteins such as the heat shock protein 70 and the bacterial GroEL protein, in the transmission of begomoviruses by B. tabaci. Additional studies attempting to identify other proteins that aid or interact with begomoviruses along their circulation pathway in the whitefly are reviewed in this paper. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Frontal Impact of Rolling Spheres.

    ERIC Educational Resources Information Center

    Domenech, A.; Casasus, E.

    1991-01-01

    A model of the inelastic collision between two spheres rolling along a horizontal track is presented, taking into account the effects of frictional forces at impact. This experiment makes possible direct estimates of the coefficients of restitution and friction. (Author)

  17. Rolling Stitch Welder For Foil

    NASA Technical Reports Server (NTRS)

    Gilbert, Jeffery L.; Morgan, Gene E.

    1992-01-01

    Hand-operated rolling spotwelder stitch-welds foil faster and more consistently than single-spotwelding gun without damaging it. Internal spring reacts against roller frame, exerting force on welding wheel when rollers contact workpiece.

  18. Chemoprevention of skin cancer: effect of Lawsonia inermis L. (Henna) leaf powder and its pigment artifact, lawsone in the Epstein- Barr virus early antigen activation assay and in two-stage mouse skin carcinogenesis models.

    PubMed

    Kapadia, Govind J; Rao, G Subba; Sridhar, Rajagopalan; Ichiishi, Eiichiro; Takasaki, Midori; Suzuki, Nobutaka; Konoshima, Takao; Iida, Akira; Tokuda, Harukuni

    2013-12-01

    In continuation of our studies with chemoprevention potential of plant-derived naphthoquinone derivatives, leaf powder of the medicinal plant Lawsonia inermis L, commonly known as 'henna', was evaluated by its inhibition of the Epstein-Barr virus early antigen (EBV-EA) activation induced by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) in Raji cells. Lawsone (2-hydroxy- 1,4-naphthoquinone), the reddish orange pigment artifact formed during the extraction or preparation of the dye from henna leaves and believed to be the active component, was also assessed in this in vitro assay. Both showed a profound inhibition (>88%) of EBV-EA activation. In the in vivo two-stage mouse skin carcinogenesis study using UV-B radiation for initiation and TPA for tumor promotion, oral feeding of henna (0.0025%) in drinking water ad libitum decreased tumor incidence by 66% and multiplicity by 40% when compared to the positive control at 10 weeks of treatment. Similarly, in the above mouse model, orally fed lawsone (0.0025%) decreased tumor incidence by 72% and multiplicity by 50%. The tumor inhibitory trend continued throughout the 20-week test period. Similar antitumor activities were observed when henna (0.5 mg/ml) was applied topically on the back skin in the UV-B initiated, TPA promoted and peroxynitrite initiated, TPA promoted mouse skin carcinogenesis models. Topically applied lawsone (0.015 mg/ml) also exhibited similar protection against tumor formation in the 7,12-dimtehylbenz(a)anthracene induced and TPA promoted skin cancer in mice. Also, there was a delay of 1 to 2 weeks in tumor appearance in both henna and lawsone treated groups compared to control in all three test models. This study ascertains the skin cancer chemopreventive activity of henna leaf powder and lawsone when administered by either oral (through drinking water) or topical (by application on the back skin) routes. Further, it emphasizes the need for the evaluation of these henna-derived green

  19. Development of roll-to-roll hot embossing system with induction heater for micro fabrication.

    PubMed

    Yun, Dongwon; Son, Youngsu; Kyung, Jinho; Park, Heechang; Park, Chanhun; Lee, Sunghee; Kim, Byungin

    2012-01-01

    In this paper, a hot embossing heating roll with induction heater inside the roll is proposed. The induction heating coil is installed inside a roll that is used as a heating roll of a roll-to-roll (R2R) hot embossing apparatus. Using an inside installed heating coil gives the roll-to-roll hot embossing system a more even temperature distribution on the surface of the heating roll compared to that of previous systems, which used an electric wire for heating. This internal induction heating roll can keep the working environment much cleaner because there is no oil leakage compared to the oiled heating roll. This paper describes the principles and provides an analysis of this proposed system; some evaluation has also been performed for the system. A real R2R hot embossing heating roll system was fabricated and some experiments on micro-pattering have been performed. After that, evaluation has been performed on the results.

  20. Fundamental phenomena governing heat transfer during rolling

    NASA Astrophysics Data System (ADS)

    Chen, W. C.; Samarasekera, I. V.; Hawbolt, E. B.

    1993-06-01

    To quantify the effect of roll chilling on the thermal history of a slab during hot rolling, tests were conducted at the Canada Center for Mineral and Energy Technology (CANMET) and at the University of British Columbia (UBC). In these tests, the surface and the interior temperatures of specimens were recorded during rolling using a data acquisition system. The corresponding heat-transfer coefficients in the roll bite were back-calculated by a trial-and-error method using a heat-transfer model. The heat-transfer coefficient was found to increase along the arc of contact and reach a maximum, followed by a decrease, until the exit of the roll bite. Its value was influenced by rolling parameters, such as percent reduction, rolling speed, rolling temperature, material type, etc. It was shown that the heat-transfer coefficient in the roll gap was strongly dependent on the roll pressure, and the effect of different variables on the interfacial heat-transfer coefficient can be related to their influence on pressure. At low mean roll pressure, such as in the case of rolling plain carbon steels at elevated temperature, the maximum heat-transfer coefficient in the roll bite was in the 25 to 35 kW/m2 °C range. As the roll pressure increased with lower rolling temperature and higher deformation resistance of stainless steel and microalloyed grades, the maximum heat-transfer coefficient reached a value of 620 kW/m2 °C. Obviously, the high pressure improved the contact between the roll and the slab surface, thereby reducing the resistance to heat flow. The mean roll-gap heat-transfer coefficient at the interface was shown to be linearly related to mean roll pressure. This finding is important because it permitted a determination of heat-transfer coefficients applicable to industrial rolling from pilot mill data. Thus, the thermal history of a slab during rough rolling was computed using a model in which the mean heat-transfer coefficient between the roll and the slab was

  1. 75 FR 64254 - Certain Hot-Rolled Flat-Rolled Carbon Quality Steel Products From Brazil; Final Results of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-19

    ... International Trade Administration Certain Hot-Rolled Flat-Rolled Carbon Quality Steel Products From Brazil... order on certain hot-rolled, flat-rolled carbon quality steel products (hot-rolled steel) from Brazil. See Certain Hot- Rolled Flat-Rolled Carbon Quality Steel Products From Brazil: Preliminary Results of...

  2. The 2b silencing suppressor of a mild strain of Cucumber mosaic virus alone is sufficient for synergistic interaction with Tobacco mosaic virus and induction of severe leaf malformation in 2b-transgenic tobacco plants.

    PubMed

    Siddiqui, Shahid A; Valkonen, Jari P T; Rajamäki, Minna-Liisa; Lehto, Kirsi

    2011-06-01

    Tobacco plants infected simultaneously by Tobacco mosaic virus (TMV) and Cucumber mosaic virus (CMV) are known to produce a specific synergistic disease in which the emerging leaves are filiformic. Similar developmental malformations are also caused to a lesser extent by the severe strains (e.g., Fny) of CMV alone, but mild strains (e.g., Kin) cause them only in mixed infection with TMV. We show here that transgenic tobacco plants expressing 2b protein of CMV-Kin produce filiformic symptoms when infected with TMV, indicating that only 2b protein is needed from CMV-Kin for this synergistic relationship. On the other hand, transgenic plants that express either the wild-type TMV genome or a modified TMV genome with its coat protein deleted or movement protein (MP) inactivated also develop filiformic or at least distinctly narrow leaves, while plants expressing the MP alone do not develop any malformations when infected with CMV-Kin. These results show that either TMV helicase/replicase protein or active TMV replication are required for this synergistic effect. The effect appears to be related to an efficient depletion of silencing machinery, caused jointly by both viral silencing suppressors, i.e., CMV 2b protein and the TMV 126-kDa replicase subunit.

  3. Hot rolling of thick uranium molybdenum alloys

    DOEpatents

    DeMint, Amy L.; Gooch, Jack G.

    2015-11-17

    Disclosed herein are processes for hot rolling billets of uranium that have been alloyed with about ten weight percent molybdenum to produce cold-rollable sheets that are about one hundred mils thick. In certain embodiments, the billets have a thickness of about 7/8 inch or greater. Disclosed processes typically involve a rolling schedule that includes a light rolling pass and at least one medium rolling pass. Processes may also include reheating the rolling stock and using one or more heavy rolling passes, and may include an annealing step.

  4. First report of Persimmon cryptic virus and Persimmon virus A in Korea

    USDA-ARS?s Scientific Manuscript database

    In 2014, a total of 77 persimmon (Diospyros kaki Thunb.) trees from Korean commercial persimmon orchards were surveyed for Persimmon cryptic virus (PeCV) and Persimmon virus A (PeVA). Leaf samples were collected from symptomatic trees with necrosis (two), or mosaic and leaf malformations (one) and 7...

  5. Improvement of rolling 6 mm thin plates in plate rolling mill PT. Krakatau Posco

    NASA Astrophysics Data System (ADS)

    Pujiyanto, Hamdani

    2017-01-01

    A 6-mm thin plate is difficult to produce especially if the product requires wide size and high strength. Flatness is the main quality issue in rolling 6-mm plate using a 4-high reversing mill which use ±1100-mm work roll. Thus some methods are applied to overcome such issue in order to comply to customer quality requirement. Pre-rolling, rolling, and post-rolling conditions have to be considered comprehensively. Roll unit management will be the key factor before rolling condition. The roll unit itself has a significant impact on work roll crown wearness in relation with work roll intial crown and thermal crown. Work roll crown along with the modification of hydraulic gap control (HGC) could directly alter the flatness of the plate.

  6. Begomovirus diversity in tomato crops and weeds in Ecuador and the detection of a recombinant isolate of rhynchosia golden mosaic Yucatan virus infecting tomato.

    PubMed

    Paz-Carrasco, Lenin C; Castillo-Urquiza, Gloria P; Lima, Alison T M; Xavier, Cesar A D; Vivas-Vivas, Leticia M; Mizubuti, Eduardo S G; Zerbini, F Murilo

    2014-08-01

    Viral diseases caused by begomoviruses are of economic importance due to their adverse effects on the production of tropical and subtropical crops. In Ecuador, despite reports of significant infestations of Bemisia tabaci in the late 1990s, only very recently has a begomovirus, tomato leaf deformation virus (ToLDeV, also present in Peru), been reported in tomato. ToLDeV is the first monopartite begomovirus discovered that originated in the Americas, and its presence in Ecuador highlights the need for a wider survey of tomato-infecting begomoviruses in this country. Tomato and weed samples were collected in 2010 and 2011 in six provinces of Ecuador, and begomovirus genomes were cloned and sequenced using a rolling-circle-amplification-based approach. Most tomato samples from the provinces of Guayas, Loja, Manabi and Santa Elena were infected with tomato leaf deformation virus (ToLDeV). One sample from Manabi had a triple infection with ToLDeV, rhynchosia golden mosaic Yucatan virus (RhGMYuV) and an isolate that was a recombinant between the two. A new begomovirus was detected in another tomato sample from Manabi. Samples of Rhynchosia sp. from the provinces of Guayas and Manabi were infected by RhGMYuV. These results indicate not only the prevalence of ToLDeV in tomato in Ecuador but also the presence of other viruses, albeit at a much lower frequency.

  7. Roll-to-Roll Nanoimprint Lithography Simulations for Flexible Substrates

    NASA Astrophysics Data System (ADS)

    Spann, Andrew; Jain, Akhilesh; Bonnecaze, Roger

    2015-11-01

    UV roll-to-roll nanoimprint lithography enables the patterning of features onto a flexible substrate for bendable electronics in a continuous process. One of the most important design goals in this process is to make the residual layer thickness of the photoresist in unpatterned regions as thin and uniform as possible. Another important goal is to minimize the imprint time to maximize throughput. We develop a multi-scale model to simulate the spreading of photoresist drops as the template is pressed against the substrate. We include the effect of capillary pressure on the bending of the substrate and show how this distorts uniformity in the residual thickness layer. Our simulation code is parallelized and can simulate the flow and merging of thousands of drops. We investigate the effect of substrate tension and the initial arrangement of drops on the residual layer thickness and imprint time. We find that for a given volume of photoresist, distributing that volume to more drops initially decreases the imprint time. We conclude with recommendations for scale-up and optimal operations of roll-to-roll nanoimprint lithography systems. The authors acknowledge the Texas Advanced Computing Center at The University of Texas at Austin for providing high performance computing resources.

  8. Climate Ready Estuaries Rolling Easements Primer

    EPA Pesticide Factsheets

    Rolling easements enable wetlands and beaches to migrate inland and allow society to avoid the costs and hazards of protecting low lands from rising sea levels. This document provides a primer on more than a dozen rolling easement approaches.

  9. High performance rolling element bearing

    NASA Technical Reports Server (NTRS)

    Bursey, Jr., Roger W. (Inventor); Olinger, Jr., John B. (Inventor); Owen, Samuel S. (Inventor); Poole, William E. (Inventor); Haluck, David A. (Inventor)

    1993-01-01

    A high performance rolling element bearing (5) which is particularly suitable for use in a cryogenically cooled environment, comprises a composite cage (45) formed from glass fibers disposed in a solid lubricant matrix of a fluorocarbon polymer. The cage includes inserts (50) formed from a mixture of a soft metal and a solid lubricant such as a fluorocarbon polymer.

  10. Rolling Tachyon in Nonlocal Cosmology

    SciTech Connect

    Joukovskaya, L.

    2007-11-20

    Nonlocal cosmological models derived from String Field Theory are considered. A new method for constructing rolling tachyon solutions in the FRW metric in two field configuration is proposed and solutions of the Friedman equations with nonlocal operator are presented. The cosmological properties of these solutions are discussed.

  11. One-zone rolling of composite materials

    NASA Astrophysics Data System (ADS)

    Kokhan, L. S.; Morozov, Yu. A.; Slavgorodskaya, Yu. B.

    2016-12-01

    The energy-force parameters of free rolling of a strip without its tension and rolling with one backward or forward creep zone in the deformation zone are compared. The limiting backward or forward tensions are determined, and the change in the linear sizes of a composite billet during deformation in a rolling mill is considered.

  12. Prediction of Rolling Force Using AN Adaptive Neural Network Model during Cold Rolling of Thin Strip

    NASA Astrophysics Data System (ADS)

    Xie, H. B.; Jiang, Z. Y.; Tieu, A. K.; Liu, X. H.; Wang, G. D.

    Customers for cold rolled strip products expect the good flatness and surface finish, consistent metallurgical properties and accurate strip thickness. These requirements demand accurate prediction model for rolling parameters. This paper presents a set-up optimization system developed to predict the rolling force during cold strip rolling. As the rolling force has the very nonlinear and time-varying characteristics, conventional methods with simple mathematical models and a coarse learning scheme are not sufficient to achieve a good prediction for rolling force. In this work, all the factors that influence the rolling force are analyzed. A hybrid mathematical roll force model and an adaptive neural network have been improved by adjusting the adaptive learning algorithm. A good agreement between the calculated results and measured values verifies that the approach is applicable in the prediction of rolling force during cold rolling of thin strips, and the developed model is efficient and stable.

  13. Identification of extrachromosomal circular DNA in hop via rolling circle amplification

    USDA-ARS?s Scientific Manuscript database

    During a survey looking for viruses affecting hop plants in the Willamette Valley of Oregon, a circular DNA molecule was identified via rolling circle amplification (RCA) and later characterized. The 5.7 kb long molecule only matched in a minor cover to a microsatellite region in the Humulus lupulus...

  14. The rolling-circle melting-pot model for porcine circovirus DNA replication

    USDA-ARS?s Scientific Manuscript database

    A stem-loop structure, formed by a pair of inverted repeats during DNA replication, is a conserved feature at the origin of DNA replication (Ori) among plant and animal viruses, bacteriophages and plasmids that replicate their genomes via the rolling-circle replication (RCR) mechanism. Porcine circo...

  15. The Discovery of Rolling Circle Amplification and Rolling Circle Transcription.

    PubMed

    Mohsen, Michael G; Kool, Eric T

    2016-11-15

    Nucleic acid amplification is a hugely important technology for biology and medicine. While the polymerase chain reaction (PCR) has been highly useful and effective, its reliance on heating and cooling cycles places some constraints on its utility. For example, the heating step of PCR can destroy biological molecules under investigation and heat/cool cycles are not applicable in living systems. Thus, isothermal approaches to DNA and RNA amplification are under widespread study. Perhaps the simplest of these are the rolling circle approaches, including rolling circle amplification (RCA) and rolling circle transcription (RCT). In this strategy, a very small circular oligonucleotide (e.g., 25-100 nucleotides in length) acts as a template for a DNA or an RNA polymerase, producing long repeating product strands that serve as amplified copies of the circle sequence. Here we describe the early developments and studies involving circular oligonucleotides that ultimately led to the burgeoning rolling circle technologies currently under development. This Account starts with our studies on the design of circular oligonucleotides as novel DNA- and RNA-binding motifs. We describe how we developed chemical and biochemical strategies for synthesis of well-defined circular oligonucleotides having defined sequence and open (unpaired) structure, and we outline the unusual ways in which circular DNAs can interact with other nucleic acids. We proceed next to the discovery of DNA and RNA polymerase activity on these very small cyclic DNAs. DNA polymerase "rolling circle" activities were discovered concurrently in our laboratory and that of Andrew Fire. We describe the surprising efficiency of this process even on shockingly small circular DNAs, producing repeating DNAs thousands of nucleotides in length. RNA polymerase activity on circular oligonucleotides was first documented in our group in 1995; especially surprising in this case was the finding that the process occurs efficiently

  16. A review of roll-to-roll nanoimprint lithography.

    PubMed

    Kooy, Nazrin; Mohamed, Khairudin; Pin, Lee Tze; Guan, Ooi Su

    2014-01-01

    Since its introduction in 1995, nanoimprint lithography has been demonstrated in many researches as a simple, low-cost, and high-throughput process for replicating micro- and nanoscale patterns. Due to its advantages, the nanoimprint lithography method has been rapidly developed over the years as a promising alternative to conventional nanolithography processes to fulfill the demands generated from the recent developments in the semiconductor and flexible electronics industries, which results in variations of the process. Roll-to-roll (R2R) nanoimprint lithography (NIL) is the most demanded technique due to its high-throughput fulfilling industrial-scale application. In the present work, a general literature review on the various types of nanoimprint lithography processes especially R2R NIL and the methods commonly adapted to fabricate imprint molds are presented to provide a clear view and understanding on the nanoimprint lithography technique as well as its recent developments. 81.16.Nd.

  17. A review of roll-to-roll nanoimprint lithography

    PubMed Central

    2014-01-01

    Since its introduction in 1995, nanoimprint lithography has been demonstrated in many researches as a simple, low-cost, and high-throughput process for replicating micro- and nanoscale patterns. Due to its advantages, the nanoimprint lithography method has been rapidly developed over the years as a promising alternative to conventional nanolithography processes to fulfill the demands generated from the recent developments in the semiconductor and flexible electronics industries, which results in variations of the process. Roll-to-roll (R2R) nanoimprint lithography (NIL) is the most demanded technique due to its high-throughput fulfilling industrial-scale application. In the present work, a general literature review on the various types of nanoimprint lithography processes especially R2R NIL and the methods commonly adapted to fabricate imprint molds are presented to provide a clear view and understanding on the nanoimprint lithography technique as well as its recent developments. PACS 81.16.Nd PMID:25024682

  18. A rolling 3-UPU parallel mechanism

    NASA Astrophysics Data System (ADS)

    Miao, Zhihuai; Yao, Yan'an; Kong, Xianwen

    2013-12-01

    A novel rolling mechanism is proposed based on a 3-UPU parallel mechanism in this paper. The rolling mechanism is composed of two platforms connected by three UPU (universal-prismatic-universal) serial-chain type limbs. The degree-of-freedom of the mechanism is analyzed using screw theory. Gait analysis and stability analysis are presented in detail. Four rolling modes of the mechanism are discussed and simulated. The feasibility of the rolling mechanism is verified by means of a physical prototype. Finally, its terrain adaptability is enhanced through planning the rolling gaits.

  19. Reconstruction of constant slow-roll inflation

    NASA Astrophysics Data System (ADS)

    Gao, Qing

    2017-09-01

    Using the relations between the slow-roll parameters and the power spectra for the single field slow-roll inflation, we derive the scalar spectral tilt n s and the tensor to scalar ratio r for the constant slow-roll inflation, and obtain the constraint on the slow-roll parameter η from the Planck 2015 results. The inflationary potential for the constant slow-roll inflation is then reconstructed in the framework of both general relativity and the scalar-tensor theory of gravity, and compared with the recently reconstructed E model potential. In the strong coupling limit, we show that the η attractor is reached.

  20. 75 FR 65453 - Certain Hot-Rolled Flat-Rolled Carbon Quality Steel Flat Products From Brazil: Notice of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-25

    ... International Trade Administration Certain Hot-Rolled Flat-Rolled Carbon Quality Steel Flat Products From Brazil... duty order on certain hot-rolled flat-rolled carbon quality steel flat products (hot-rolled steel) from Brazil. The review covers four producers/exporters of hot-rolled steel from Brazil, all mandatory...

  1. 76 FR 22868 - Certain Hot-Rolled Flat-Rolled Carbon-Quality Steel Products From Brazil: Final Results of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-25

    ... International Trade Administration Certain Hot-Rolled Flat-Rolled Carbon-Quality Steel Products From Brazil... certain hot-rolled flat-rolled carbon- quality steel products (HRS) from Brazil for the period January 1...: Background Since the issuance of Certain Hot-Rolled Flat-Rolled Carbon-Quality Steel Products From Brazil...

  2. Towards roll-to-roll manufacturing of polymer photonic devices

    NASA Astrophysics Data System (ADS)

    Subbaraman, Harish; Lin, Xiaohui; Ling, Tao; Guo, L. Jay; Chen, Ray T.

    2014-03-01

    Traditionally, polymer photonic devices are fabricated using clean-room processes such as photolithography, e-beam lithography, reactive ion etching (RIE) and lift-off methods etc, which leads to long fabrication time, low throughput and high cost. We have utilized a novel process for fabricating polymer photonic devices using a combination of imprinting and ink jet printing methods, which provides high throughput on a variety of rigid and flexible substrates with low cost. We discuss the manufacturing challenges that need to be overcome in order to realize true implementation of roll-to-roll manufacturing of flexible polymer photonic systems. Several metrology and instrumentation challenges involved such as availability of particulate-free high quality substrate, development and implementation of high-speed in-line and off-line inspection and diagnostic tools with adaptive control for patterned and unpatterned material films, development of reliable hardware, etc need to be addressed and overcome in order to realize a successful manufacturing process. Due to extreme resolution requirements compared to print media, the burden of software and hardware tools on the throughput also needs to be carefully determined. Moreover, the effect of web wander and variations in web speed need to accurately be determined in the design of the system hardware and software. In this paper, we show the realization of solutions for few challenges, and utilizing these solutions for developing a high-rate R2R dual stage ink-jet printer that can provide alignment accuracy of <10μm at a web speed of 5m/min. The development of a roll-to-roll manufacturing system for polymer photonic systems opens limitless possibilities for the deployment of high performance components in a variety of applications including communication, sensing, medicine, agriculture, energy, lighting etc.

  3. Rubber rolling over a sphere

    NASA Astrophysics Data System (ADS)

    Koiller, J.; Ehlers, K.

    2007-04-01

    “Rubber” coated bodies rolling over a surface satisfy a no-twist condition in addition to the no slip condition satisfied by “marble” coated bodies [1]. Rubber rolling has an interesting differential geometric appeal because the geodesic curvatures of the curves on the surfaces at corresponding points are equal. The associated distribution in the 5 dimensional configuration space has 2 3 5 growth (these distributions were first studied by Cartan; he showed that the maximal symmetries occurs for rubber rolling of spheres with 3:1 diameters ratio and materialize the exceptional group G 2). The 2 3 5 nonholonomic geometries are classified in a companion paper [2] via Cartan’s equivalence method [3]. Rubber rolling of a convex body over a sphere defines a generalized Chaplygin system [4 8] with SO(3) symmetry group, total space Q = SO(3) × S 2 and base S 2, that can be reduced to an almost Hamiltonian system in T* S 2 with a non-closed 2-form ωNH. In this paper we present some basic results on the sphere-sphere problem: a dynamically asymmetric but balanced sphere of radius b (unequal moments of inertia I j but with center of gravity at the geometric center), rubber rolling over another sphere of radius a. In this example ωNH is conformally symplectic [9]: the reduced system becomes Hamiltonian after a coordinate dependent change of time. In particular there is an invariant measure, whose density is the determinant of the reduced Legendre transform, to the power p = 1/2( b/a - 1). Using sphero-conical coordinates we verify the result by Borisov and Mamaev [10] that the system is integrable for p = -1/2 (ball over a plane). They have found another integrable case [11] corresponding to p = -3/2 (rolling ball with twice the radius of a fixed internal ball). Strikingly, a different set of sphero-conical coordinates separates the Hamiltonian in this case. No other integrable cases with different I j are known.

  4. Roll Casting of Al-25%Si

    SciTech Connect

    Haga, Toshio; Harada, Hideto; Watari, Hisaki

    2011-05-04

    Strip casting of Al-25%Si strip was tried using an unequal diameter twin roll caster. The diameter of the lower roll (large roll) was 1000 mm and the diameter of the upper roll (small roll) was 250 mm. Roll material was mild steel. The sound strip could be cast at the speeds ranging from 8 m/min to 12 m/min. The strip did not stick to the roll without the parting material. The primary Si, which existed at centre area of the thickness direction, was larger than that which existed at other area. The size of the primary Si was smaller than 0.2 mm. Eutectic Si was smaller 5 {mu}m. The as-cast strip was ranging from 2 mm to 3 mm thick and its width was 100 mm. The as-cast strip could be hot rolled down to 1 mm. The hot rolled strip was cold rolled. The primary Si became smaller and the pore occurred around the primary Si after the rolling.

  5. Leaf Size in Swietenia

    Treesearch

    Charles B. Briscoe; F. Bruce. Lamb

    1962-01-01

    A study was made of the putative hybrid of bigleaf and small-leaf mahoganies. Initial measurements indicated that bigleaf mahogany can be distinguished from small-leaf mahogany by gross measurements of leaflets. Isolated mother trees yield typical progeny. Typical mother trees in mixed stands yield like progeny plus, usually, mediumleaf progeny. Mediumleaf mother trees...

  6. Tribological Testing of Anti-Adhesive coatings for Cold Rolling Mill Rolls--Application to TiN-Coated Rolls

    SciTech Connect

    Ould, Choumad; Montmitonnet, Pierre; Gachon, Yves; Badiche, Xavier

    2011-05-04

    Roll life is a major issue in cold strip rolling. Roll wear may result either in too low roll roughness, bringing friction below the minimum requested for strip entrainment; or it may degrade strip surface quality. On the contrary, adhesive wear and transfer (''roll coating'', ''pick up'') may form a thick metallic deposits on the roll which increases friction excessively and degrades strip surface again [1]. The roll surface, with the help of a materials-adapted lubricant, must therefore possess anti-wear and anti-adhesive properties. Thus, High Speed Steeel (HSS) rolls show superior properties compared with standard Cr-steel rolls due to their high carbide surface coverage. Another way to improve wear and adhesion properties of surfaces is to apply hard metallic (hard-Cr) or ceramic coatings. Chromium is renowned for its excellent anti-wear and anti-adhesive properties and may serve as a reference. Here, as a first step towards alternative, optimised coatings, a PVD TiN coating has been deposited on tool steels, as previous attempts have proved TiN to be rather successful in cold rolling experiments [2,3]. Different tribological tests are reported here, giving insight in both anti-adhesive properties and fatigue life improvement.

  7. Slow-roll thawing quintessence

    SciTech Connect

    Chiba, Takeshi

    2009-04-15

    We derive slow-roll conditions for thawing quintessence. We solve the equation of motion of {phi} for a Taylor expanded potential (up to the quadratic order) in the limit where the equation of state w is close to -1 to derive the equation of state as a function of the scale factor. We find that the evolution of {phi} and hence w are described by only two parameters. The expression for w(a), which can be applied to general thawing models, coincides precisely with that derived recently by Dutta and Scherrer for hilltop quintessence. The consistency conditions of |w+1|<<1 are derived. The slow-roll conditions for freezing quintessence are also derived.

  8. Rolling process and its influence analysis on hot continuous rolling mill vibration

    NASA Astrophysics Data System (ADS)

    Fan, Xiaobin; Zang, Yong; Jin, Ke

    2016-12-01

    Rolling mill vibration is a worldwide problem, although it has been found that the mill structure defects are the main cause of vibration, but the inhibition measures are difficult to implement. So we discussed the rolling force parameters influence on rolling mill vibration and suppression measures from rolling technology in this article. The results show that the rolling force is increased as the reduction ratio and speed increases, and decreases with the increase in temperature; reduction ratio has more obvious influence than rolling temperature and speed, so we should give priority to the reduction allocation in production. Rolled strip is thinner; the self-excited relations in mill system are stronger, namely the stability of the rolling mill is poor and mill vibration would more likely produce. The comprehensive field tests and analysis were carried out, and it shows that roll horizontal vibration and torsional vibration have less relationship and the mechanism between horizontal and vertical vibration is same.

  9. Roll force, torque, lever arm coefficient, and strain distribution in edge rolling

    NASA Astrophysics Data System (ADS)

    Lundberg, S.-E.; Gustafsson, T.

    1993-12-01

    Due to the growing importance of width control in strip and plate mills, edge rolling is currently an im-portant process in hot rolling mills. Research in edge rolling has been carried out, and in the present ar-ticle, models for roll force, torque, and lever arm coefficient are derived using the upper bound method. A simple, kinematically admissible deformation zone and velocity field, independent of friction in the roll gap, is proposed, and the energy dissipation rate is derived. The formula for energy dissipation rate has, in practice, no limitation because all edge rolling geometries are safely in the area where the formula is valid. Roll force and torque are derived by means of two independent integrals. Thus, the lever arm coef-ficient is evaluated from the expressions for roll force and torque using conventional rolling theory. Roll-ing trials report good agreement with theory. Measured roll forces are similar to calculated forces. Furthermore, the shape of the dogbone that arises during edge rolling is in fairly good agreement with the calculated dogbone shape. Deviations are due to the deviation from ideally plastic material in the ex-periments. Also, the strain distribution over the dogbone is similar to the proposed deformation zone. Thus, a new formula has been developed to a stage that it can be implemented in width control systems for edge rolling stands in hot strip and plate mills.

  10. Slow-roll extended quintessence

    SciTech Connect

    Chiba, Takeshi; Siino, Masaru; Yamaguchi, Masahide

    2010-04-15

    We derive the slow-roll conditions for a nonminimally coupled scalar field (extended quintessence) during the radiation/matter dominated era extending our previous results for thawing quintessence. We find that the ratio {phi}e/3H{phi} becomes constant but negative, in sharp contrast to the ratio for the minimally coupled scalar field. We also find that the functional form of the equation of state of the scalar field asymptotically approaches that of the minimally coupled thawing quintessence.

  11. Rolling Mill Hill, Nashville, TN

    EPA Pesticide Factsheets

    Rolling Mill Hill was the home to Nashville General Hospital from 1890 to the 1990s and encompassed several buildings and structures. These existing buildings of historical significance were re-used in the form of apartments. The original Trolley Barns on the site are now artists’ lofts and are home to several companies and non-profit offices. Nance Place, which entails additional buildings built on-site, is a Tax Credit Workforce Housing Development and is Platinum LEED certified.

  12. Rolling-Friction Robotic Gripper

    NASA Technical Reports Server (NTRS)

    Vranish, John M.

    1992-01-01

    Robotic gripper using rolling-friction fingers closes in on object with interface designed to mate with rollers somewhat misaligned initially, aligns object with respect to itself, then holds object securely in uniquely determined position and orientation. Operation of gripper causes minimal wear and burring of gripper and object. Exerts minimal friction forces on object when grasping and releasing. Releases object easily and reliably even when side forces and torques are between itself and object.

  13. Roll formed pan solar module

    SciTech Connect

    Jester, T.L.; Bottenberg, W.R.; Gay, C.F.; Yerkes, J.W.

    1984-02-21

    A solar module comprising a solar cell string laminated between layers of pottant material and a transparent superstrate and a steel substrate. The steel substrate is roll formed to provide stiffening flanges on its edges while simultaneously forming a pan-shaped structure to hold other portions of the laminate in position during the laminating process. An improved terminal provides high voltage protection and improved mechanical strength. A conduit element provides protected raceways for external wires connected to module terminals.

  14. Nanogrid rolling circle DNA sequencing

    DOEpatents

    Church, George M.; Porreca, Gregory J.; Shendure, Jay; Rosenbaum, Abraham Meir

    2017-04-18

    The present invention relates to methods for sequencing a polynucleotide immobilized on an array having a plurality of specific regions each having a defined diameter size, including synthesizing a concatemer of a polynucleotide by rolling circle amplification, wherein the concatemer has a cross-sectional diameter greater than the diameter of a specific region, immobilizing the concatemer to the specific region to make an immobilized concatemer, and sequencing the immobilized concatemer.

  15. Mechanisms of rolling contact spalling

    NASA Technical Reports Server (NTRS)

    Kumar, A. M.; Kulkarni, S. M.; Bhargava, V.; Hahn, G. T.; Rubin, C. A.

    1987-01-01

    The results of a study aimed at analyzing the mechanical material interactions responsible for rolling contact spalling of the 440 C steel, high pressure oxygen turbopump bearings are presented. A coupled temperature displacement finite element analysis of the effects of friction heating under the contact is presented. The contact is modelled as a stationary, heat generating, 2 dimensional indent in an elastic perfectly plastic half-space with heat fluxes up to 8.6 x 10000 KW/m sq comparable to those generated in the bearing. Local temperatures in excess of 1000 C are treated. The calculations reveal high levels of residual tension after the contact is unloaded and cools. Efforts to promote Mode 2/Mode 3 fatigue crack growth under cyclic torsion in hardened 440 C steel are described. Spalls produced on 440 C steel by a 3 ball/rod rolling contact testing machine were studied with scanning microscopy. The shapes of the cyclic, stress strain hysteresis loops displayed by hardened 440 C steel in cyclic torsion at room temperature are defined for the plastic strain amplitudes encountered in rolling/sliding contact. Results of these analyses are discussed in detail.

  16. Rolling Contact Fatigue of Ceramics

    SciTech Connect

    Wereszczak, Andrew A; Wang, W.; Wang, Y.; Hadfield, M.; Kanematsu, W.; Kirkland, Timothy Philip; Jadaan, Osama M.

    2006-09-01

    High hardness, low coefficient of thermal expansion and high temperature capability are properties also suited to rolling element materials. Silicon nitride (Si{sub 3}N{sub 4}) has been found to have a good combination of properties suitable for these applications. However, much is still not known about rolling contact fatigue (RCF) behavior, which is fundamental information to assess the lifetime of the material. Additionally, there are several test techniques that are employed internationally whose measured RCF performances are often irreconcilable. Due to the lack of such information, some concern for the reliability of ceramic bearings still remains. This report surveys a variety of topics pertaining to RCF. Surface defects (cracks) in Si{sub 3}N{sub 4} and their propagation during RCF are discussed. Five methods to measure RCF are then briefly overviewed. Spalling, delamination, and rolling contact wear are discussed. Lastly, methods to destructively (e.g., C-sphere flexure strength testing) and non-destructively identify potential RCF-limiting flaws in Si{sub 3}N{sub 4} balls are described.

  17. Virus Strains Causing Mosaic in Louisiana and Florida Sugarcane

    USDA-ARS?s Scientific Manuscript database

    Mosaic caused by Sugarcane mosaic virus (SCMV) and Sorghum mosaic virus (SrMV), respectively, affects sugarcane in Louisiana and Florida. Between 2004 and 2007, surveys were conducted in both states to determine which virus and virus strains were causing mosaic of sugarcane. In Louisiana, leaf sam...

  18. A Semianalytic Model of Leukocyte Rolling

    PubMed Central

    Krasik, Ellen F.; Hammer, Daniel A.

    2004-01-01

    Rolling allows leukocytes to maintain adhesion to vascular endothelium and to molecularly coated surfaces in flow chambers. Using insights from adhesive dynamics, a computational method for simulating leukocyte rolling and firm adhesion, we have developed a semianalytic model for the steady-state rolling of a leukocyte. After formation in a force-free region of the contact zone, receptor-ligand bonds are transported into the trailing edge of the contact zone. Rolling velocity results from a balance of the convective flux of bonds and the rate of dissociation at the back edge of the contact zone. We compare the model's results to that of adhesive dynamics and to experimental data on the rolling of leukocytes, with good agreement. We calculate the dependence of rolling velocity on shear rate, intrinsic forward and reverse reaction rates, bond stiffness, and reactive compliance, and use the model to calculate a state diagram relating molecular parameters and the dynamic state of adhesion. A dimensionless form of the analytic model permits exploration of the parameters that control rolling. The chemical affinity of a receptor-ligand pair does not uniquely determine rolling velocity. We elucidate a fundamental relationship between off-rate, ligand density, and reactive compliance at the transition between firm and rolling adhesion. The model provides a rapid method for screening system parameters for the potential to mediate rolling. PMID:15315955

  19. QTL analysis of disease resistance to leaf spots and TSWV in peanut (Arachis hypogaea)

    USDA-ARS?s Scientific Manuscript database

    Early leaf spot (ELS), caused by Cercospora arachidicola, late leaf spot (LLS), caused by Cercosporidium personatum, and Tomato spotted wilt virus (TSWV) result in great losses in yield in peanut (Arachis hypogaea L.). In order to identify quantitative trait loci (QTL) for resistance to these dise...

  20. Wheat curl mite resistance: interactions of mite feeding with wheat streak mosaic virus infection.

    PubMed

    Murugan, M; Sotelo Cardona, P; Duraimurugan, P; Whitfield, A E; Schneweis, D; Starkey, S; Smith, C M

    2011-08-01

    The majority of plant viruses are dependent on arthropod vectors for spread between plants. Wheat streak mosaic virus (family Potyviridae, genus Tritimovirus, WSMV) is transmitted by the wheat curl mite, Aceria tosichella Keifer, and this virus and vector cause extensive yield losses in most major wheat (Triticum aestivum L.)-growing regions of the world. Many cultivars in use are susceptible to this vector-virus complex, and yield losses of 10-99% have been documented. wheat curl mite resistance genes have been identified in goat grass, Aegilops tauschii (Coss) Schmal., and transferred to hexaploid wheat, but very few varieties contain effectively wheat curl mite resistance, due to virulent wheat curl mite populations. However, wheat curl mite resistance remains an effective strategy to reduce losses due to WSMV. The goal of our project was to identify the most effective, reproducible, and rapid method for assessing wheat curl mite resistance. We also wanted to determine whether mite resistance is affected by WSMV infection, because the pathogen and pest commonly occur together. Single and group wheat curl mite infestations produced similar amounts of leaf rolling and folding on wheat curl mite-susceptible wheat varieties that were independent of initial wheat curl mite infestation. This finding will allow accurate, efficient, large-scale screening of wheat germplasm for wheat curl mite resistance by infesting plants with sections of wheat leaf tissue containing mixed stages of wheat curl mite. The wheat curl mite-resistant breeding line 'OK05312' displayed antibiosis (reduced wheat curl mite population development). The effect of WSMV infection on wheat curl mite reproduction was genotype-dependent. Mite populations increased on infected wheat curl mite- and WSMV-susceptible plants compared with uninfected plants, but WSMV infection had no significant effect on wheat curl mite populations on resistant plants. OK05312 is a strong source of wheat curl mite

  1. Leaf growth is conformal

    NASA Astrophysics Data System (ADS)

    Alim, Karen; Armon, Shahaf; Shraiman, Boris I.; Boudaoud, Arezki

    2016-10-01

    Growth pattern dynamics lie at the heart of morphogenesis. Here, we investigate the growth of plant leaves. We compute the conformal transformation that maps the contour of a leaf at a given stage onto the contour of the same leaf at a later stage. Based on the mapping we predict the local displacement field in the leaf blade and find it to agree with the experimentally measured displacement field to 92%. This approach is applicable to any two-dimensional system with locally isotropic growth, enabling the deduction of the whole growth field just from observation of the tissue contour.

  2. Diversity, Mutation and Recombination Analysis of Cotton Leaf Curl Geminiviruses

    PubMed Central

    Saleem, Huma; Nahid, Nazia; Shakir, Sara; Ijaz, Sehrish; Murtaza, Ghulam; Khan, Asif Ali; Mubin, Muhammad; Nawaz-ul-Rehman, Muhammad Shah

    2016-01-01

    The spread of cotton leaf curl disease in China, India and Pakistan is a recent phenomenon. Analysis of available sequence data determined that there is a substantial diversity of cotton-infecting geminiviruses in Pakistan. Phylogenetic analyses indicated that recombination between two major groups of viruses, cotton leaf curl Multan virus (CLCuMuV) and cotton leaf curl Kokhran virus (CLCuKoV), led to the emergence of several new viruses. Recombination detection programs and phylogenetic analyses showed that CLCuMuV and CLCuKoV are highly recombinant viruses. Indeed, CLCuKoV appeared to be a major donor virus for the coat protein (CP) gene, while CLCuMuV donated the Rep gene in the majority of recombination events. Using recombination free nucleotide datasets the substitution rates for CP and Rep genes were determined. We inferred similar nucleotide substitution rates for the CLCuMuV-Rep gene (4.96X10-4) and CLCuKoV-CP gene (2.706X10-4), whereas relatively higher substitution rates were observed for CLCuMuV-CP and CLCuKoV-Rep genes. The combination of sequences with equal and relatively low substitution rates, seemed to result in the emergence of viral isolates that caused epidemics in Pakistan and India. Our findings also suggest that CLCuMuV is spreading at an alarming rate, which can potentially be a threat to cotton production in the Indian subcontinent. PMID:26963635

  3. Diversity, Mutation and Recombination Analysis of Cotton Leaf Curl Geminiviruses.

    PubMed

    Saleem, Huma; Nahid, Nazia; Shakir, Sara; Ijaz, Sehrish; Murtaza, Ghulam; Khan, Asif Ali; Mubin, Muhammad; Nawaz-Ul-Rehman, Muhammad Shah

    2016-01-01

    The spread of cotton leaf curl disease in China, India and Pakistan is a recent phenomenon. Analysis of available sequence data determined that there is a substantial diversity of cotton-infecting geminiviruses in Pakistan. Phylogenetic analyses indicated that recombination between two major groups of viruses, cotton leaf curl Multan virus (CLCuMuV) and cotton leaf curl Kokhran virus (CLCuKoV), led to the emergence of several new viruses. Recombination detection programs and phylogenetic analyses showed that CLCuMuV and CLCuKoV are highly recombinant viruses. Indeed, CLCuKoV appeared to be a major donor virus for the coat protein (CP) gene, while CLCuMuV donated the Rep gene in the majority of recombination events. Using recombination free nucleotide datasets the substitution rates for CP and Rep genes were determined. We inferred similar nucleotide substitution rates for the CLCuMuV-Rep gene (4.96X10-4) and CLCuKoV-CP gene (2.706X10-4), whereas relatively higher substitution rates were observed for CLCuMuV-CP and CLCuKoV-Rep genes. The combination of sequences with equal and relatively low substitution rates, seemed to result in the emergence of viral isolates that caused epidemics in Pakistan and India. Our findings also suggest that CLCuMuV is spreading at an alarming rate, which can potentially be a threat to cotton production in the Indian subcontinent.

  4. Elimination of five sugarcane viruses from sugarcane using in vitro culture of axillary bud and apical meristem

    USDA-ARS?s Scientific Manuscript database

    Procedures were developed for the in vitro elimination of Sugarcane mosaic virus (SCMV), Sorghum mosaic virus (SrMV), Sugarcane streak mosaic virus (SCSMV), Sugarcane yellow leaf virus (SCYLV) and Fiji disease virus (FDV) from infected sugarcane. In vitro shoot regeneration, elongation and virus el...

  5. Project LEAF Documents

    EPA Pesticide Factsheets

    Project LEAF has a goal of educating farmworkers about how to reduce pesticide exposure to their families from pesticide residues they may be inadvertently taking home on their clothing, etc. Find outreach materials.

  6. How rolling forecasting facilitates dynamic, agile planning.

    PubMed

    Miller, Debra; Allen, Michael; Schnittger, Stephanie; Hackman, Theresa

    2013-11-01

    Rolling forecasting may be used to replace or supplement the annual budget process. The rolling forecast typically builds on the organization's strategic financial plan, focusing on the first three years of plan projections and comparing the strategic financial plan assumptions with the organization's expected trajectory. Leaders can then identify and respond to gaps between the rolling forecast and the strategic financial plan on an ongoing basis.

  7. Numerical investigation of the temperature distribution in a roll system

    SciTech Connect

    Eriksson, D.: Sunden, B.; Postoaca, I.

    1996-12-31

    An analysis of heat transfer in a fast rotating roll system is carried out. A hot film is transported between a steel roll and a roll of a soft material. The soft roll is supported by another steel roll at the opposite side. The inner surfaces of the rolls are water-cooled while the outer surfaces are cooled by combined convection and thermal radiation. The soft roll is also cooled by an impinging jet. The surface temperature of the soft roll is very important and the influence of various cooling parameters is assessed by numerical solutions to the problem. The importance of modelling the whole system of rolls has been proved. Rolling processes are widely used in forming of metals and polymeric materials. The adequate cooling of the rolls and the rolled products is a major concern at the design stage and during operation. Non-adequate cooling may shorten the roll life and severely affect the processed material.

  8. Fluid management in roll-to-roll nanoimprint lithography

    NASA Astrophysics Data System (ADS)

    Jain, A.; Bonnecaze, R. T.

    2013-06-01

    The key process parameters of UV roll-to-roll nanoimprint lithography are identified from an analysis of the fluid, curing, and peeling dynamics. The process includes merging of droplets of imprint material, curing of the imprint material from a viscous liquid to elastic solid resist, and pattern replication and detachment of the resist from template. The time and distances on the web or rigid substrate over which these processes occur are determined as function of the physical properties of the uncured liquid, the cured solid, and the roller configuration. The upper convected Maxwell equation is used to model the viscoelastic liquid and to calculate the force on the substrate and the torque on the roller. The available exposure time is found to be the rate limiting parameter and it is O(√Rho /uo), where R is the radius of the roller, ho is minimum gap between the roller and web, and uo is the velocity of the web. The residual layer thickness of the resist should be larger than the gap between the roller and the substrate to ensure complete feature filling and optimal pattern replication. For lower residual layer thickness, the droplets may not merge to form a continuous film for pattern transfer.

  9. METHOD OF HOT ROLLING URANIUM METAL

    DOEpatents

    Kaufmann, A.R.

    1959-03-10

    A method is given for quickly and efficiently hot rolling uranium metal in the upper part of the alpha phase temperature region to obtain sound bars and sheets possessing a good surface finish. The uranium metal billet is heated to a temperature in the range of 1000 deg F to 1220 deg F by immersion iii a molten lead bath. The heated billet is then passed through the rolls. The temperature is restored to the desired range between successive passes through the rolls, and the rolls are turned down approximately 0.050 inch between successive passes.

  10. Spray Rolling Aluminum Strip for Transportation Applications

    SciTech Connect

    Kevin M. McHugh; Y. Lin; Y. Zhou; E. J. Lavernia; J.-P. Delplanque; S. B. Johnson

    2005-02-01

    Spray rolling is a novel strip casting technology in which molten aluminum alloy is atomized and deposited into the roll gap of mill rolls to produce aluminum strip. A combined experimental/modeling approach has been followed in developing this technology with active participation from industry. The feasibility of this technology has been demonstrated at the laboratory scale and it is currently being scaled-up. This paper provides an overview of the process and compares the microstructure and properties of spray-rolled 2124 aluminum alloy with commercial ingot-processed material

  11. Interactions between stably rolling leukocytes in vivo

    NASA Astrophysics Data System (ADS)

    King, Michael R.; Ruscio, Aimee D.; Kim, Michael B.; Sarelius, Ingrid H.

    2005-03-01

    We have characterized the two-dimensional spatial dependence of the hydrodynamic interactions between two adhesively rolling leukocytes in a live venule in the mouse cremaster muscle. Two rolling leukocytes were observed to slow each other down when rolling together in close proximity due to mutual sheltering from the external blood flow in the vessel lumen. A previous study of leukocyte rolling interactions using carbohydrate-coated beads in a parallel-plate flow chamber and a detailed computer model of adhesion in a multicellular environment is in qualitative agreement with the current in vivo results.

  12. Complete genomic characterization of milk vetch dwarf virus isolates from cowpea and broad bean in Anhui province, China.

    PubMed

    Zhang, Chenhua; Zheng, Hongying; Yan, Dankan; Han, Kelei; Song, Xijiao; Liu, Yong; Zhang, Dongfang; Chen, Jianping; Yan, Fei

    2017-08-01

    Cowpea and broad bean plants showing severe stunting and leaf rolling symptoms were observed in Hefei city, Anhui province, China, in 2014. Symptomatic plants from both species were shown to be infected with milk vetch dwarf virus (MDV) by PCR. The complete genomes of MDV isolates from cowpea and broad bean were sequenced. Each of them had eight genomic DNAs that differed between the two isolates by 10.7% in their overall nucleotide sequences. In addition, the MDV genomes from cowpea and broad bean were associated with two and three alphasatellite DNAs, respectively. This is the first report of MDV on cowpea in China and the first complete genome sequences of Chinese MDV isolates.

  13. Rolling-cuff flexible bellows

    DOEpatents

    Lambert, D.R.

    1982-09-27

    A flexible connector apparatus used to join two stiff non-deformable members, such as piping, is described. The apparatus is provided with one or more flexible sections or assemblies each utilizing a bellows of a rolling cuff type connected between two ridge members, with the bellows being supported by a back-up ring, such that only the curved end sections of the bellows are unsupported. Thus, the bellows can be considered as being of a tube-shaped configuration and thus have high pressure resistance. The components of the flexible apparatus are sealed or welded one to another such that it is fluid tight.

  14. Effect of temper rolling on final shape defects in a V-section roll forming process

    NASA Astrophysics Data System (ADS)

    Abvabi, Akbar; Rolfe, Bernard; Hodgson, Peter D.; Weiss, Matthias

    2013-12-01

    Roll forming is a continuous process in which a flat strip is shaped to the desired profile by sequential bending in a series of roll stands. Because of the large variety of applications of roll forming in the industry, Finite Element Analysis (FEA) is increasingly utilized for roll forming process design. Bending is the dominant deformation mode in roll forming. Sheet materials used in this process are generally temper rolled, roller- or tension- leveled. These processes introduce residual stresses into the material, and recent studies have shown that those affect the material behavior in bending. In this study a numerical model of the temper rolling (skin passing) process was used to determine a residual stress distribution in a dual phase, DP780, steel strip. A 5-stand roll forming process for the forming of a V-section was modeled, and the effect of various thickness reduction levels in the temper rolling process on the final shape defects was analyzed. The results show that a small thickness reduction in the temper rolling process decreases the maximum bow height but the final springback angle increases. It is also shown that reasonable model accuracy can be achieved by including the residual stress information due to temper rolling as initial condition in the numerical modeling of a roll forming process.

  15. Deer predation on leaf miners via leaf abscission

    NASA Astrophysics Data System (ADS)

    Yamazaki, Kazuo; Sugiura, Shinji

    2008-03-01

    The evergreen oak Quercus gilva Blume sheds leaves containing mines of the leaf miner Stigmella sp. (Lepidoptera: Nepticulidae) earlier than leaves with no mines in early spring in Nara, central Japan. The eclosion rates of the leaf miner in abscised and retained leaves were compared in the laboratory to clarify the effects of leaf abscission on leaf miner survival in the absence of deer. The leaf miner eclosed successfully from both fallen leaves and leaves retained on trees. However, sika deer ( Cervus nippon centralis Kishida) feed on the fallen mined leaves. Field observations showed that deer consume many fallen leaves under Q. gilva trees, suggesting considerable mortality of leaf miners due to deer predation via leaf abscission. This is a previously unreported relationship between a leaf miner and a mammalian herbivore via leaf abscission.

  16. Deer predation on leaf miners via leaf abscission.

    PubMed

    Yamazaki, Kazuo; Sugiura, Shinji

    2008-03-01

    The evergreen oak Quercus gilva Blume sheds leaves containing mines of the leaf miner Stigmella sp. (Lepidoptera: Nepticulidae) earlier than leaves with no mines in early spring in Nara, central Japan. The eclosion rates of the leaf miner in abscised and retained leaves were compared in the laboratory to clarify the effects of leaf abscission on leaf miner survival in the absence of deer. The leaf miner eclosed successfully from both fallen leaves and leaves retained on trees. However, sika deer (Cervus nippon centralis Kishida) feed on the fallen mined leaves. Field observations showed that deer consume many fallen leaves under Q. gilva trees, suggesting considerable mortality of leaf miners due to deer predation via leaf abscission. This is a previously unreported relationship between a leaf miner and a mammalian herbivore via leaf abscission.

  17. Tobacco rattle virus (TRV) based silencing of cotton enoyl-CoA reductase (ECR) gene and the role of very long chain fatty acids in normal leaf development and resistance to wilt disease

    USDA-ARS?s Scientific Manuscript database

    A Tobacco rattle virus (TRV) based virus-induced gene silencing (VIGS) assay was employed as a reverse genetic approach to study gene function in cotton (Gossypium hirsutum). This approach was used to investigate the function of Enoyl-CoA reductase (GhECR) in pathogen defense. Amino acid sequence al...

  18. 49 CFR 393.134 - What are the rules for securing roll-on/roll-off or hook lift containers?

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... or hook lift containers? 393.134 Section 393.134 Transportation Other Regulations Relating to... for securing roll-on/roll-off or hook lift containers? (a) Applicability. The rules in this section apply to the transportation of roll-on/roll-off or hook lift containers. (b) Securement of a roll-on...

  19. 75 FR 47263 - Certain Hot-Rolled Flat-Rolled Carbon-Quality Steel Products From the Russian Federation; Final...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-05

    ... International Trade Administration Certain Hot-Rolled Flat-Rolled Carbon-Quality Steel Products From the Russian... carbon- quality steel products from the Russian Federation; final results. SUMMARY: On April 1, 2010, the... antidumping duty investigation of certain hot-rolled flat- rolled carbon-quality steel products (``hot-rolled...

  20. Ebola Virus and Marburg Virus

    MedlinePlus

    Diseases and Conditions Ebola virus and Marburg virus By Mayo Clinic Staff Ebola virus and Marburg virus are related viruses that cause hemorrhagic ... Africa, where sporadic outbreaks have occurred for decades. Ebola virus and Marburg virus live in animal hosts, ...

  1. 76 FR 62894 - Following Procedures When Going Between Rolling Equipment

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-11

    ... Federal Railroad Administration Following Procedures When Going Between Rolling Equipment AGENCY: Federal... of following procedures when going ] between rolling equipment. This safety advisory contains various... who, in the course of their work, place themselves between rolling equipment. The railroad...

  2. Model development of work roll wear in hot strip mill

    NASA Astrophysics Data System (ADS)

    Liu, Ziying; Guan, Yingping; Wang, Fengqin

    2017-06-01

    This paper, based on the analysis of the main factors(specific roll force, mean roll surface temperature, irregular edge wear and contact arc length) affecting roll wear, designed a new work roll wear model, the test data shows that the model can more accurately reflect the work roll wear, can be on-line prediction of work roll wear. The roll wear curve, including constant wear and irregular edge wear, presents a box shape, and the reasons also are showed in this paper. The top roll wear and bottom roll wear in the same mill are inconsistent, and the reasons are also analysed in this paper. Results show that the construction of the work roll mathematical model accords with the general law of work roll wear and tear; it can more accurately forecast roll wear online.

  3. Characterization of a new begomovirus and betasatellite associated with chilli leaf curl disease in India.

    PubMed

    Khan, Zainul A; Khan, Jawaid A

    2017-02-01

    Chilli leaf curl disease (ChiLCD) is a serious problem and a major limitation to chilli (Capsicum spp.) cultivation in India. Leaves of a chilli plant showing leaf curl symptoms were collected from the Gonda district in Uttar Pradesh, India, in April, 2013. Full-length genomes of a begomovirus and an associated betasatellite were amplified, cloned and sequenced. The size of the begomovirus genome and the betasatellite were 2760 bp and 1374 bp, respectively. The nucleotide sequence of the begomovirus genome shared maximum identity (89 %) with pepper leaf curl Bangladesh virus-India isolate Chhapra (PepLCBV, JN663853), below the threshold for species demarcation. Sequence analysis showed that the begomovirus is a potential recombinant between viruses related to PepLCBV and chilli leaf curl virus (ChiLCV). The name chilli leaf curl Gonda virus (ChiLCGV) is being proposed. The betasatellite associated with ChiLCGV was identified as tomato leaf curl Bangladesh betasatellite (ToLCBDB). Agroinoculation of the viral genome along with betasatellite induced severe leaf curl symptoms in Nicotiana benthamiana. ChiLCGV and ToLCBDB characterized in this study represent a new begomovirus-betasatellite complex infecting Capsicum.

  4. Lubrication of rolling-element bearings

    NASA Technical Reports Server (NTRS)

    Parker, R. J.

    1980-01-01

    The lubrication of rolling element bearings is surveyed. Emphasis is on the critical design aspects related to speed, temperature, and ambient pressure environment. Types of lubrication including grease, jets, mist, wick, and through the race are discussed. The historical development, present state of technology, and the future problems of rolling element bearing lubrication are discussed.

  5. School Roll Forecasting Methods: A Review.

    ERIC Educational Resources Information Center

    Simpson, Stephen

    1987-01-01

    A review of the literature concerning local school roll forecasting describes the theoretical model common to most local education agency (LEA) forecasts, identifies a variety of issues relevant to this area of LEA planning, and suggests some opportunities for improvement in LEA school roll forecasting. (Author/CB)

  6. Roll splitting for field processing of biomass

    Treesearch

    Dennis T. Curtin; Donald L. Sirois; John A. Sturos

    1987-01-01

    The concept of roll splitting wood originated in 1967 when the Tennessee Valley Authority (TVA) forest products specialists developed a wood fibrator. The objective of that work was to produce raw materials for reconstituted board products. More recently, TVA focused on roll splitting as a field process to accelerate drying of small trees (3-15 cm diameter), much...

  7. Roll-forming tubes to header plates

    NASA Technical Reports Server (NTRS)

    Kramer, K.

    1976-01-01

    Technique has been developed for attaching and sealing tubes to header plates using a unique roll-forming tool. Technique is useful for attaching small tubes which are difficult to roll into conventional grooves in header plate tube holes, and for attaching when welding, brazing, or soldering is not desirable.

  8. School Roll Forecasting Methods: A Review.

    ERIC Educational Resources Information Center

    Simpson, Stephen

    1987-01-01

    A review of the literature concerning local school roll forecasting describes the theoretical model common to most local education agency (LEA) forecasts, identifies a variety of issues relevant to this area of LEA planning, and suggests some opportunities for improvement in LEA school roll forecasting. (Author/CB)

  9. Numerical analysis of Swiss roll metamaterials.

    PubMed

    Demetriadou, A; Pendry, J B

    2009-08-12

    A Swiss roll metamaterial is a resonant magnetic medium, with a negative magnetic permeability for a range of frequencies, due to its self-inductance and self-capacitance components. In this paper, we discuss the band structure, S-parameters and effective electromagnetic parameters of Swiss roll metamaterials, with both analytical and numerical results, which show an exceptional convergence.

  10. 33 CFR 159.107 - Rolling test.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 33 Navigation and Navigable Waters 2 2013-07-01 2013-07-01 false Rolling test. 159.107 Section 159.107 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) POLLUTION MARINE SANITATION DEVICES Design, Construction, and Testing § 159.107 Rolling test. (a) The device,...

  11. Rolling maneuver load alleviation using active controls

    NASA Technical Reports Server (NTRS)

    Woods-Vedeler, Jessica A.; Pototzky, Anthony S.

    1992-01-01

    Rolling Maneuver Load Alleviation (RMLA) was demonstrated on the Active Flexible Wing (AFW) wind tunnel model in the LaRC Transonic Dynamics Tunnel. The design objective was to develop a systematic approach for developing active control laws to alleviate wing incremental loads during roll maneuvers. Using linear load models for the AFW wind-tunnel model which were based on experimental measurements, two RMLA control laws were developed based on a single-degree-of-freedom roll model. The RMLA control laws utilized actuation of outboard control surface pairs to counteract incremental loads generated during rolling maneuvers and roll performance. To evaluate the RMLA control laws, roll maneuvers were performed in the wind tunnel at dynamic pressures of 150, 200, and 250 psf and Mach numbers of .33, .38, and .44, respectively. Loads obtained during these maneuvers were compared to baseline maneuver loads. For both RMLA controllers, the incremental torsion moments were reduced by up to 60 percent at all dynamic pressures and performance times. Results for bending moment load reductions during roll maneuvers varied. In addition, in a multiple function test, RMLA and flutter suppression system control laws were operated simultaneously during roll maneuvers at dynamic pressures 11 percent above the open-loop flutter dynamic pressure.

  12. Power roll 2007: A five year summary

    USDA-ARS?s Scientific Manuscript database

    The powered roll gin stand (PRT – Powered Roll Technology) was first tested in a field application on seed cotton during the 2002 ginning season at Servico, Inc., Courtland Alabama. During the 2003 season, Servico installed and operated PRT stands in all three lines. In subsequent years, mechanical ...

  13. Damped leaf flexure hinge

    NASA Astrophysics Data System (ADS)

    Chen, Zhong; Chen, Guisheng; Zhang, Xianmin

    2015-05-01

    Flexure-based mechanism like compliant actuation system embeds complex dynamics that will reduce the control bandwidth and limits their dynamic positioning precision. This paper presents a theoretical model of a leaf flexure hinge with damping layers using strain energy method and Kelvin damping model. The modified loss factor of the damped leaf flexure hinge is derived, and the equivalent viscous damping coefficient of the damped leaf hinge is obtained, which could be used to improve the pseudo-rigid-model. The free vibration signals of the hinge in three different damping configurations are measured. The experimental modal analysis also is performed on the three kinds of damped leaf flexure hinges in order to evaluate their 1st order bending natural frequency and vibration-suppressing effects. The evaluation of modified loss factor model also is performed. The experimental results indicate that the constrained layer damping can enhance the structure damping of the hinge even if only single damping layer each side, the modified loss factor model can get good predicts of a damped leaf flexure hinge in the frequency range below 1st order natural frequency, and it is necessary that the dimensional parameters of the damping layers and basic layer of the hinge should be optimized for simplification at the mechanism's design stage.

  14. Damped leaf flexure hinge.

    PubMed

    Chen, Zhong; Chen, Guisheng; Zhang, Xianmin

    2015-05-01

    Flexure-based mechanism like compliant actuation system embeds complex dynamics that will reduce the control bandwidth and limits their dynamic positioning precision. This paper presents a theoretical model of a leaf flexure hinge with damping layers using strain energy method and Kelvin damping model. The modified loss factor of the damped leaf flexure hinge is derived, and the equivalent viscous damping coefficient of the damped leaf hinge is obtained, which could be used to improve the pseudo-rigid-model. The free vibration signals of the hinge in three different damping configurations are measured. The experimental modal analysis also is performed on the three kinds of damped leaf flexure hinges in order to evaluate their 1st order bending natural frequency and vibration-suppressing effects. The evaluation of modified loss factor model also is performed. The experimental results indicate that the constrained layer damping can enhance the structure damping of the hinge even if only single damping layer each side, the modified loss factor model can get good predicts of a damped leaf flexure hinge in the frequency range below 1st order natural frequency, and it is necessary that the dimensional parameters of the damping layers and basic layer of the hinge should be optimized for simplification at the mechanism's design stage.

  15. Roll-to-Roll Atomic Layer Deposition for Ultrabarriers

    NASA Astrophysics Data System (ADS)

    Yersak, Alexander

    Atomic layer deposition (ALD) is a bottom-up, gas phase, thin film deposition technique based on sequential, self-limiting binary surface reactions. The precise sub-nanometer film thickness control and conformal nature of this process have led to various commercial applications of ALD. However, ALD films are most commonly deposited in batch processes at low pressures, which raises throughput and/or cost concerns for many otherwise promising applications. This problem can be solved by spatial ALD (S-ALD) which is a version of the ALD technique where the precursors are separated in space rather than time. We have demonstrated the first atmospheric pressure roll-to-roll (R2R) ALD web coating system. A thickness uniformity of +/-2% was achieved across the web. ALD cycle times as low as 76 ms were demonstrated with a web speed of 1 m/s and a vertical gap height of 0.5 mm. Extrinsic defects in the ALD films were investigated, and a predictive cluster model was proposed, and was demonstrated with a residual (i.e. difference between the actual defect counts and those predicted by the cluster model) of <10%. A R2R ALD web coating tool with molecular layer deposition (MLD) capabilities was investigated and achieved a defect density <10 /cm2. A hyperbaric corrosion chamber with in situ monitoring of film thickness was demonstrated with the ability to characterize R2R ALD films using water dissolution as a metric. ALD SiO2 films were determined to be dissolution-predictable with a predicted dissolution rate of 3.7 nm/year at physiological temperatures. ALD TiO2 films were observed with no measurable dissolution in 150 °C water over the measurement period of 12 days.

  16. Advances in roll-to-roll imprint lithography for display applications

    NASA Astrophysics Data System (ADS)

    Jeans, Albert; Almanza-Workman, Marcia; Cobene, Robert; Elder, Richard; Garcia, Robert; Gomez-Pancorbo, Fernando; Jackson, Warren; Jam, Mehrban; Kim, Han-Jun; Kwon, Ohseung; Luo, Hao; Maltabes, John; Mei, Ping; Perlov, Craig; Smith, Mark; Taussig, Carl; Jeffrey, Frank; Braymen, Steve; Hauschildt, Jason; Junge, Kelly; Larson, Don; Stieler, Dan

    2010-03-01

    A solution to the problems of roll-to-roll lithography on flexible substrates is presented. We have developed a roll-toroll imprint lithography technique to fabricate active matrix transistor backplanes on flexible webs of polyimide that have a blanket material stack of metals, dielectrics, and semiconductors. Imprint lithography produces a multi-level 3- dimensional mask that is then successively etched to pattern the underlying layers into the desired structures. This process, Self-Aligned Imprint Lithography (SAIL), solves the layer-to-layer alignment problem because all masking levels are created with one imprint step. The processes and equipment required for complete roll-to-roll SAIL fabrication will be described. Emphasis will be placed on the advances in the roll-to-roll imprint process which have enabled us to produce working transistor arrays.

  17. Computational Analysis of Ares I Roll Control System Jet Interaction Effects on Rolling Moment

    NASA Technical Reports Server (NTRS)

    Deere, Karen A.; Pao, S. Paul; Abdol-Hamid, Khaled S.

    2011-01-01

    The computational flow solver USM3D was used to investigate the jet interaction effects from the roll control system on the rolling moment of the Ares I full protuberance configuration at wind tunnel Reynolds numbers. Solutions were computed at freestream Mach numbers from M = 0.5 to M = 5 at the angle of attack 0deg, at the angle of attack 3.5deg for a roll angle of 120deg, and at the angle of attack 7deg for roll angles of 120deg and 210deg. Results indicate that the RoCS housing provided a beneficial jet interaction effect on vehicle rolling moment for M > or = 0.9. Most of the components downstream of the roll control system housing contributed to jet interaction penalties on vehicle rolling moment.

  18. Automated Procedure for Roll Pass Design

    NASA Astrophysics Data System (ADS)

    Lambiase, F.; Langella, A.

    2009-04-01

    The aim of this work has been to develop an automatic roll pass design method, capable of minimizing the number of roll passes. The adoption of artificial intelligence technologies, particularly expert systems, and a hybrid model for the surface profile evaluation of rolled bars, has allowed us to model the search for the minimal sequence with a tree path search. This approach permitted a geometrical optimization of roll passes while allowing automation of the roll pass design process. Moreover, the heuristic nature of the inferential engine contributes a great deal toward reducing search time, thus allowing such a system to be employed for industrial purposes. Finally, this new approach was compared with other recently developed automatic systems to validate and measure possible improvements among them.

  19. Aerodynamics of a rolling airframe missile

    NASA Astrophysics Data System (ADS)

    Tisserand, L. E.

    1981-05-01

    For guidance-related reasons, there is considerable interest in rolling missiles having single-plane steering capability. To aid the aerodynamic design of these airframes, a unique investigation into the aerodynamics of a rolling, steering missile has been carried out. It represents the first known attempt to measure in a wind tunnel the aerodynamic forces and moments that act on a spinning body-canard-tail configuration that exercises canard steering in phase with body roll position. Measurements were made with the model spinning at steady-state roll rates ranging from 15 to 40 Hz over an angle-of-attack range up to about 16 deg. This short, exploratory investigation has demonstrated that a better understanding and a more complete definition of the aerodynamics of rolling, steering vehicles can be developed by way of simulative wind-tunnel testing.

  20. Roll Casting of Aluminum Alloy Clad Strip

    SciTech Connect

    Nakamura, R.; Tsuge, H.; Haga, T.; Watari, H.; Kumai, S.

    2011-01-17

    Casting of aluminum alloy three layers of clad strip was tried using the two sets of twin roll casters, and effects of the casting parameters on the cladding conditions were investigated. One twin roll caster was mounted on the other twin roll caster. Base strip was 8079 aluminum alloy and overlay strips were 6022 aluminum alloy. Effects of roll-load of upper and lower casters and melt temperature of the lower caster were investigated. When the roll-load of the upper and lower caster was large enough, the overlay strip could be solidified and be connected. The overlay strip could be connected when the melt of the overlay strip cast by the lower caster was low enough. Sound three layers of clad strip could be cast by proper conditions.