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Makoto Hattori - One of the best experts on this subject based on the ideXlab platform.

  • characterization of multicopper oxidase genes in the green rice leafhopper Nephotettix Cincticeps hemiptera cicadellidae with focus on salivary gland specific genes
    Archives of Insect Biochemistry and Physiology, 2019
    Co-Authors: Yukiko Matsumoto, Makoto Hattori
    Abstract:

    Multicopper oxidase (MCO) enzymes are present ubiquitously and act on diverse substrates. Recently, the presence of multiple MCO genes has been described in many insects. Based on sialotranscriptome data, we identified and comprehensively characterized six MCO genes: NcLac1S, 1G, and 2-5 in the green rice leafhopper, Nephotettix Cincticeps (Hemiptera: Cicadellidae). NcLac1S and NcLac1G belong to the MCO1 ortholog of other insects. NcLac2 forms a clade with MCO2s involved in the sclerotization and pigmentation of the cuticle. NcLac3 and NcLac4 form a clade with NlMCO3 -5 of the hemipteran Nilaparvata luges. NcLac5 forms a clade with MCORPs (MCO-related proteins) that lack amino acid residues normally highly conserved in copper-coordinated MCOs. NcLac1S and NcLac3 were specifically expressed in the salivary glands; whereas NcLac5 was primarily expressed in the salivary glands. Only NcLac3 protein is considered to have laccase activity in the salivary glands and salivary sheaths ejected by the insect. NcLac1G expression was relatively high in the testis. NcLac2 and NcLac4 were specifically expressed in the integument and in Malpighian tubules, respectively. Knockdown by RNA interference (RNAi) of either NcLac2 and NcLac5 in nymphs caused high mortality. All NcLac2-knockdown nymphs showed depigmentation and soft cuticle, and eventually died, as did other MCO2-knockdown insects. DsNcLac5-injected nymphs (third, fourth, and fifth-instar) showed high mortality, but injection into adults had no effect on survival or number of eggs deposited, suggesting that NcLac5 is not essential for survival after molting (eclosion). NcLac5 could be a promising target gene for control of N. Cincticeps.

  • the green rice leafhopper Nephotettix Cincticeps hemiptera cicadellidae salivary protein ncsp75 is a key effector for successful phloem ingestion
    PLOS ONE, 2018
    Co-Authors: Yukiko Matsumoto, Makoto Hattori
    Abstract:

    Nephotettix Cincticeps, a prevalent rice pest, injects gelling and watery saliva into plant tissues during the sucking process. Certain components within the saliva are believed to interact with plant cellular constituents and play important roles in overcoming host plant defense responses. Based on our previous analysis of the salivary gland transcriptome and secreted saliva proteome of N. Cincticeps, in this study, we analyzed the biological functions of salivary protein, NcSP75 (N. Cincticeps salivary protein 75 kD). NcSP75, a salivary glands-specific gene, showed low similarities to any previously reported sequences. Knockdown of NcSP75 by RNA interference (RNAi) reduced the longevity of treated nymphs to approximately half of the longevity of controls and caused severe developmental retardation. Furthermore, the knockdown of NcSP75 decreased the survival rate of adults, and reduced the number of deposited eggs and hatched nymphs. Thus, the adverse effects caused by the knockdown of NcSP75 were observed throughout the lifetime of N. Cincticeps, when feeding on rice plants. In contrast, no reduction was observed in the survival rate of the knockdown of NcSP75 adults fed on an artificial diet. Electrical penetration graph measurements taken from adult females feeding on rice plants showed a significantly shorter duration of phloem ingestion associated with the knockdown of NcSP75 than the knockdown of the enhanced green fluorescent protein (EGFP). Furthermore, the total sugar content of the honeydew was lower when NcSP75 was knocked down. These results suggest that the NcSP75 protein contribute to successful and sustainable ingestion from the sieve elements of rice plants. The NcSP75 protein of N. Cincticeps can, accordingly, be considered as a key effector for establishing compatible interaction with rice plants and could be a potential target for controlling this species.

  • RESEARCH ARTICLE Proteome Analysis of Watery Saliva Secreted
    2016
    Co-Authors: Makoto Hattori
    Abstract:

    The green rice leafhopper, Nephotettix Cincticeps, is a vascular bundle feeder that dis-charges watery and gelling saliva during the feeding process. To understand the potential functions of saliva for successful and safe feeding on host plants, we analyzed the complex-ity of proteinaceous components in the watery saliva of N. Cincticeps. Salivary proteins were collected from a sucrose diet that adult leafhoppers had fed on through a membrane of stretched parafilm. Protein concentrates were separated using SDS-PAGE under reducing and non-reducing conditions. Six proteins were identified by a gas-phase protein sequencer and two proteins were identified using LC-MS/MS analysis with reference to expressed se-quence tag (EST) databases of this species. Full-length cDNAs encoding these major proteins were obtained by rapid amplification of cDNA ends-PCR (RACE-PCR) and degen-erate PCR. Furthermore, gel-free proteome analysis that was performed to cover the broad range of salivary proteins with reference to the latest RNA-sequencing data from the sali-vary gland of N. Cincticeps, yielded 63 additional protein species. Out of 71 novel proteins identified from the watery saliva, about 60 % of those were enzymes or other functional proteins, including GH5 cellulase, transferrin, carbonic anhydrases, aminopeptidase, regu-calcin, and apolipoprotein. The remaining proteins appeared to be unique and species-specific. This is the first study to identify and characterize the proteins in watery saliva of Auchenorrhyncha species, especially sheath-producing, vascular bundle-feeders

  • Proteome Analysis of Watery Saliva Secreted by Green Rice Leafhopper, Nephotettix Cincticeps.
    PloS one, 2015
    Co-Authors: Makoto Hattori, Hiroaki Noda, Setsuko Komatsu, Yukiko Matsumoto
    Abstract:

    The green rice leafhopper, Nephotettix Cincticeps, is a vascular bundle feeder that discharges watery and gelling saliva during the feeding process. To understand the potential functions of saliva for successful and safe feeding on host plants, we analyzed the complexity of proteinaceous components in the watery saliva of N. Cincticeps. Salivary proteins were collected from a sucrose diet that adult leafhoppers had fed on through a membrane of stretched parafilm. Protein concentrates were separated using SDS-PAGE under reducing and non-reducing conditions. Six proteins were identified by a gas-phase protein sequencer and two proteins were identified using LC-MS/MS analysis with reference to expressed sequence tag (EST) databases of this species. Full -length cDNAs encoding these major proteins were obtained by rapid amplification of cDNA ends-PCR (RACE-PCR) and degenerate PCR. Furthermore, gel-free proteome analysis that was performed to cover the broad range of salivary proteins with reference to the latest RNA-sequencing data from the salivary gland of N. Cincticeps, yielded 63 additional protein species. Out of 71 novel proteins identified from the watery saliva, about 60 % of those were enzymes or other functional proteins, including GH5 cellulase, transferrin, carbonic anhydrases, aminopeptidase, regucalcin, and apolipoprotein. The remaining proteins appeared to be unique and species- specific. This is the first study to identify and characterize the proteins in watery saliva of Auchenorrhyncha species, especially sheath-producing, vascular bundle-feeders.

  • Multiple alignment of GH5 family protein sequences identified from Nephotettix Cincticeps with GH5 proteins of five coleopteran insects and two bacterial GH5 proteins.
    2015
    Co-Authors: Makoto Hattori, Setsuko Komatsu, Yukiko Matsumoto
    Abstract:

    Multiple alignments was performed using deposited data from Hemiptera: Nephotettix Cincticeps (NcSP38:LC009514, and TsukubaH.comp12770), Coleoptera: Hypothenemus hampei (ACU52526), Apriona japonica (AHI15747), Anoplophora chinensis (AFN89566), Psacothea hilaris (BAB86867), Oncideres albomarginata chamela (ADI24131), Bacteria: Paenibacillus mucilaginosus (WP_013917961), Streptomyce rimosus (WP_030596221). Asterisk (*) and number sign (#) on the amino acid residues represent the catalytic nucleophile and catalytic proton donor, respectively (based on Eyun et [58]).

Hideshi Yasui - One of the best experts on this subject based on the ideXlab platform.

  • four resistance alleles derived from oryza longistaminata a chev roehrich against green rice leafhopper Nephotettix Cincticeps uhler identified using novel introgression lines
    Breeding Science, 2019
    Co-Authors: Hnin Wah Thein, Yoshiyuki Yamagata, Tan Van Mai, Hideshi Yasui
    Abstract:

    The green rice leafhopper (GRH, Nephotettix Cincticeps Uhler) is a serious insect pest of rice (Oryza sativa L.) in temperate regions of Asia. Wild Oryza species are the main source of resistance to insects. The W1413 accession of African wild rice (O. longistaminata A. Chev. & Roehrich) is resistant to GRH. To analyze its resistance, we developed 28 BC3F3 introgression lines carrying W1413 segments in the genetic background of Nipponbare, a susceptible rice cultivar, and evaluated their GRH resistance. Five BC3F3 populations were used for quantitative trait locus (QTL) analysis and seven BC3F4 populations for QTL validation. Four significant QTLs on the long arm of chromosome 2 (qGRH2), short arm of chromosome 4 (qGRH4), short arm of chromosome 5 (qGRH5), and long arm of chromosome 11 (qGRH11) were identified. The contribution of the W1413 allele at qGRH11 was the largest among the four QTLs; the other QTLs also contributed to GRH resistance. Chromosomal locations suggested that qGRH11 corresponds to the previously reported GRH resistance gene Grh2, qGRH4 to Grh6, and qGRH5 to Grh1. qGRH2 is a novel QTL for resistance to GRH. Thus, resistance of O. longistaminata to GRH can be explained by at least four QTLs.

  • high resolution mapping of grh6 a gene from oryza nivara sharma et shastry conferring resistance to green rice leafhopper Nephotettix Cincticeps uhler
    Breeding Science, 2019
    Co-Authors: Cong Nguyen Phi, Daisuke Fujita, Atsushi Yoshimura, Yoshiyuki Yamagata, Hideshi Yasui
    Abstract:

    The green rice leafhopper (GRH), Nephotettix Cincticeps Uhler, is a major insect pest of cultivated rice, Oryza sativa L., throughout the temperate regions of East Asia. GRH resistance had been reported in the wild species Oryza nivara but genetic basis of GRH resistance in wild rice accession has not been clarified. Here, we found a major QTL, qGRH4.2, on chromosome 4 conferred GRH resistance with 14.1 of the logarithm of odds (LOD) score explaining 67.6% of phenotypic variance in the BC1F1 population derived from a cross between the susceptible japonica cultivar ‘Taichung 65’ (T65) and O. nivara accession IRGC105715. qGRH4.2 has been identified as GRH6 between the markers RM5414 and {"type":"entrez-nucleotide","attrs":{"text":"C60248","term_id":"2418953","term_text":"C60248"}}C60248 in a BC3F2 population derived from two BC3F1 plants resistant to GRH. In a high-resolution mapping, the GRH6 region was delimited between the markers G6-c60k and 7L16f, and corresponded to an 31.2-kbp region of the ‘Nipponbare’ genome. Understanding the genetic basis of GRH resistance will facilitate the use of GRH resistance genes in marker-assisted breeding in rice.

  • characterization of resistance to the green rice leafhopper Nephotettix Cincticeps uhler in a core collection of landraces in rice oryza sativa l
    American Journal of Plant Sciences, 2017
    Co-Authors: Tan Van Mai, Atsushi Yoshimura, Hideshi Yasui
    Abstract:

    The green rice leafhopper (GRH; Nephotettix Cincticeps Uhler) is one of the most devastating insect pests of cultivated rice (Oryza sativa L.) in temperate regions in Asia. Using the rice germplasms with biotic stress resistance is the most effective and environmentally-friendly way to control the insect pests in the paddy. Sixty accessions from a core set of worldwide collection of rice were characterized for resistance to the GRH by antibiosis test both at the seedling and at the booting stages. The positive correlations of average nymph mortality (ANM) were observed between at the seedling stage and at the booting stage on 3 days after infestation (DAI) (r = 0.684**), 5DAI (r = 0.680**), and 7DAI (r = 0.652**), respectively. This result will give us the opportunity to screen resistance to the GRH with the cost-efficient way using rice seedlings in a growth chamber. To classify the 60 accessions evaluated, the ANM of the GRH of each accession was compared to the respective ANM of resistant and susceptible controls with the least significant difference (LSD) value. Based on the statistical difference or similarity of the ANMs to the resistant and the susceptible controls, we proposed the four groups of resistance to the GRH, (I) high level of resistance, (II) considerable level of resistance, (III) moderate level of resistance, and (IV) susceptibility. At the seedling stage, a total of 26 accessions were highly resistant in addition to other 6 for considerable level of resistance and other 10 for moderate level of resistance. At the booting stage, on the other hand, a total of 18 accessions were highly resistant in addition to other 3 for considerable level of resistance and other 5 for moderate level of resistance. A total of 42 accessions with high to moderate level of resistance were distributed across 16 countries in Asia in addition to each one for Madagascar and USA. The classification of landraces based on the present protocol for screening resistance to the insect provided fundamental information for genetics and breeding on resistance to the GRH in rice.

  • a major qtl for resistance to green rice leafhopper Nephotettix Cincticeps uhler derived from african rice oryza glaberrima steud
    Breeding Science, 2010
    Co-Authors: Daisuke Fujita, Atsushi Yoshimura, Kazuyuki Doi, Hideshi Yasui
    Abstract:

    The green rice leafhopper (GRH), Nephotettix Cincticeps Uhler, is a serious insect pest of cultivated rice (Oryza sativa L.) in temperate East Asia. An African rice cultivar, Oryza glaberrima Steud. (IRGC104038), was shown to be highly GRH-resistant at the booting stage. To reveal the genetic basis of the GRH resistance in O. glaberrima, a BC1F1 population derived from a cross between a susceptible japonica variety, Taichung 65 (T65), and O. glaberrima was analyzed by quantitative trait locus (QTL) analysis. A single major QTL for GRH resistance, designated qGRH9, was detected on rice chromosome 9, and three minor QTLs were detected on rice chromosomes 3, 7, and 10. A series of O. glaberrima introgression lines (GILs) containing IRGC104038 chromosome segments in the T65 genetic background were evaluated to confirm the genetic effects of the QTLs, and three GILs carrying qGRH9 showed resistance to GRH. Substitution mapping using the GILs revealed that qGRH9 was located between simple sequence repeat (SSR) markers RM215 and RM2482 in a 1.39-Mbp region on the distal region of the long arm of chromosome 9, and tightly linked to RM7306. These SSR markers maybe useful for marker-assisted selection of qGRH9 for improvement of GRH resistance in rice.

  • development of near isogenic lines and pyramided lines carrying resistance genes to green rice leafhopper Nephotettix Cincticeps uhler with the taichung 65 genetic background in rice oryza sativa l
    Breeding Science, 2010
    Co-Authors: Daisuke Fujita, Atsushi Yoshimura, Hideshi Yasui
    Abstract:

    The green rice leafhopper (GRH), Nephotettix Cincticeps Uhler, is a serious insect pest of cultivated rice (Oryza sativa L.) in temperate East Asia. Six GRH-resistance genes (Grh1, Grh2, Grh3, Grh4, Grh5, and Grh6) and one quantitative trait locus (QTL; qGRH4) have been identified. We selected near-isogenic lines (NILs) carrying Grh1, Grh2, Grh4, Grh5, Grh6, and qGRH4 with the japonica genetic background (Taichung 65 cultivar) by means of marker-assisted selection using new simple sequence repeat markers flanking the GRH-resistance genes and QTL. We also developed three pyramided lines (PYLs; Grh2/Grh6-PYL, Grh4/Grh6-PYL, and Grh5/qGRH4-PYL) using each NIL that carried a GRH-resistance gene or QTL. The NILs, PYLs, and donor parents were evaluated by using an antibiosis test. The resistance of Grh1-NIL and Grh5-NIL did not differ significantly from those of the donor parents, whereas the resistances of Grh2-NIL and Grh6-NIL were significantly lower than those of the donor parents. Grh4-NIL and qGRH4-NIL were highly susceptible. The resistance levels of the pyramided lines for Grh2 and Grh6, Grh4 and Grh6, and Grh5 and qGRH4 demonstrated a gene pyramiding effect that significantly increased their resistance. The developed NILs and PYLs should be useful genetic resources for rice improvement and deployment of the resistance genes.

Xiao-feng Zhang - One of the best experts on this subject based on the ideXlab platform.

  • Rice Yellow Stunt Nucleorhabdovirus Matrix Protein Mediates Viral Axonal Transport in the Central Nervous System of Its Insect Vector.
    Frontiers in Microbiology, 2019
    Co-Authors: Haitao Wang, Juan Wang, Qian Zhang, Tianbao Zeng, Yuemin Zheng, Hongyan Chen, Xiao-feng Zhang
    Abstract:

    Abstract: Persistently transmitted plant viruses encounter multiple membrane and tissue barriers in the process of completing their infection routes within their insect vectors. Some of these viruses have been reported to overcome the elaborate barriers of the central nervous system (CNS) to travel through the nervous tissues, but the specific mechanisms of this process remain unknown. Here, we report the axonal transport mechanism of rice yellow stunt virus (RYSV), a nucleorhabdovirus, in the CNS of the green rice leafhopper (Nephotettix Cincticeps). Infection route of RYSV in the internal organs of its insect vector after ingestion of virus was investigated by immunofluorescence microscopy. RYSV was first detected in the epithelial cells of intestinal regions, from where it proceeded to the nervous system, and finally into the salivary glands. We then utilized immunofluorescence and electron microscopy to investigate the distribution of RYSV particles within the leafhopper CNS, demonstrating that non-enveloped viral particles distributed along the microtubule-based neurofilaments in the axon cytoplasm following the direct interaction of leafhopper α-tubulin with the RYSV M protein. Tubulin inhibitors inhibited the dissemination of RYSV to the CNS then into the salivary glands in leafhoppers. We therefore describe a mechanism of plant virus transport through CNS axons as a alternative means of rapid viral dissemination in an insect vector.

  • Data_Sheet_1_Rice Yellow Stunt Nucleorhabdovirus Matrix Protein Mediates Viral Axonal Transport in the Central Nervous System of Its Insect Vector.PDF
    2019
    Co-Authors: Haitao Wang, Juan Wang, Xiao-feng Zhang, Qian Zhang, Tianbao Zeng, Yuemin Zheng, Hongyan Chen, Taiyun Wei
    Abstract:

    Persistently transmitted plant viruses encounter multiple membrane and tissue barriers in the process of completing their infection routes within their insect vectors. Some of these viruses have been reported to overcome the elaborate barriers of the central nervous system (CNS) to travel through the nervous tissues, but the specific mechanisms of this process remain unknown. Here, we report the axonal transport mechanism of rice yellow stunt virus (RYSV), a nucleorhabdovirus, in the CNS of the green rice leafhopper (Nephotettix Cincticeps). The infection route of RYSV in the internal organs of its insect vector after ingestion of the virus was investigated by immunofluorescence microscopy. RYSV was first detected in the epithelial cells of midgut regions, from where it proceeded to the nervous system, and finally into the salivary glands. We then utilized immunofluorescence and electron microscopy to investigate the distribution of RYSV particles within the leafhopper CNS, demonstrating that non-enveloped viral particles distributed along the microtubule-based neurofilaments in the axon cytoplasm following the direct interaction of leafhopper α-tubulin with the RYSV M protein. Tubulin inhibitors inhibited the dissemination of RYSV to the CNS, then into the salivary glands in leafhoppers. We therefore describe a mechanism of plant virus transport through CNS axons as an alternative means of rapid viral dissemination in an insect vector.

  • Exploration of an Actin Promoter-Based Transient Expression Vector to Trace the Cellular Localization of Nucleorhabdovirus Proteins in Leafhopper Cultured Cells
    Frontiers in Microbiology, 2018
    Co-Authors: Xiao-feng Zhang, Haitao Wang, Juan Wang, Tianbao Zeng, Hongyan Chen, Yunjie Xie, Yibing Zhao, Taiyun Wei
    Abstract:

    Abstract: Continuously cultured cell lines derived from planthopper and leafhopper have greatly facilitated the investigation of rice viruses transmitted by these insects. However, the lack of a suitable transient expression vector has limited their utility. Here, by cloning and analyzing the promoter sequence of the gene encoding cytoplasmic actin from the leafhopper Nephotettix Cincticeps, we successfully developed the first efficient transient expression vector for cultured leafhopper cells, which can also be used to express exogenous proteins in other insect culture cell lines, including those derived from Recilia dorsalis leafhopper and Spodoptera frugiperda (Sf9). Furthermore, insertion of the Hr5 viral enhancer element and knockdown of the endogenous Dicer2 gene notably improved the vector’s expression efficiency in leafhopper cells. Using the optimized vector, we have for the first time traced the cellular localization of the proteins encoded by rice yellow stunt virus (RYSV) in cells of its insect vector and demonstrated that P6 protein is a component of the viroplasm.

  • development of leafhopper cell culture to trace the early infection process of a nucleorhabdovirus rice yellow stunt virus in insect vector cells
    Virology Journal, 2018
    Co-Authors: Haitao Wang, Juan Wang, Taiyun Wei, Yunjie Xie, Xiao-feng Zhang
    Abstract:

    In China, the rice pathogen Rice yellow stunt virus (RYSV), a member of the genus Nucleorhabdovirus in the family Rhabdoviridae, was a severe threat to rice production during the1960s and1970s. Fundamental aspects of the biology of this virus such as protein localization and formation of the RYSV viroplasm during infection of insect vector cells are largely unexplored. The specific role(s) of the structural proteins nucleoprotein (N) and phosphoprotein (P) in the assembly of the viroplasm during RYSV infection in insect vector is also unclear. In present study, we used continuous leafhopper cell culture, immunocytochemical techniques, and transmission electron microscopy to investigate the subcellular distributions of N and P during RYSV infection. Both GST pull-down assay and yeast two-hybrid assay were used to assess the in vitro interaction of N and P. The dsRNA interference assay was performed to study the functional roles of N and P in the assembly of RYSV viroplasm. Here we demonstrated that N and P colocalized in the nucleus of RYSV-infected Nephotettix Cincticeps cell and formed viroplasm-like structures (VpLSs). The transiently expressed N and P are sufficient to form VpLSs in the Sf9 cells. In addition, the interactions of N/P, N/N and P/P were confirmed in vitro. More interestingly, the accumulation of RYSV was significantly reduced when the transcription of N gene or P gene was knocked down by dsRNA treatment. In summary, our results suggest that N and P are the main viral factors responsible for the formation of viroplasm in RYSV-infected insect cells. Early during RYSV infection in the insect vector, N and P interacted with each other in the nucleus to form viroplasm-like structures, which are essential for the infection of RYSV.

  • Development of leafhopper cell culture to trace the early infection process of a nucleorhabdovirus, rice yellow stunt virus, in insect vector cells
    BMC, 2018
    Co-Authors: Haitao Wang, Juan Wang, Taiyun Wei, Yunjie Xie, Xiao-feng Zhang
    Abstract:

    Abstract Background In China, the rice pathogen Rice yellow stunt virus (RYSV), a member of the genus Nucleorhabdovirus in the family Rhabdoviridae, was a severe threat to rice production during the1960s and1970s. Fundamental aspects of the biology of this virus such as protein localization and formation of the RYSV viroplasm during infection of insect vector cells are largely unexplored. The specific role(s) of the structural proteins nucleoprotein (N) and phosphoprotein (P) in the assembly of the viroplasm during RYSV infection in insect vector is also unclear. Methods In present study, we used continuous leafhopper cell culture, immunocytochemical techniques, and transmission electron microscopy to investigate the subcellular distributions of N and P during RYSV infection. Both GST pull-down assay and yeast two-hybrid assay were used to assess the in vitro interaction of N and P. The dsRNA interference assay was performed to study the functional roles of N and P in the assembly of RYSV viroplasm. Results Here we demonstrated that N and P colocalized in the nucleus of RYSV-infected Nephotettix Cincticeps cell and formed viroplasm-like structures (VpLSs). The transiently expressed N and P are sufficient to form VpLSs in the Sf9 cells. In addition, the interactions of N/P, N/N and P/P were confirmed in vitro. More interestingly, the accumulation of RYSV was significantly reduced when the transcription of N gene or P gene was knocked down by dsRNA treatment. Conclusions In summary, our results suggest that N and P are the main viral factors responsible for the formation of viroplasm in RYSV-infected insect cells. Early during RYSV infection in the insect vector, N and P interacted with each other in the nucleus to form viroplasm-like structures, which are essential for the infection of RYSV

Daisuke Fujita - One of the best experts on this subject based on the ideXlab platform.

  • high resolution mapping of grh6 a gene from oryza nivara sharma et shastry conferring resistance to green rice leafhopper Nephotettix Cincticeps uhler
    Breeding Science, 2019
    Co-Authors: Cong Nguyen Phi, Daisuke Fujita, Atsushi Yoshimura, Yoshiyuki Yamagata, Hideshi Yasui
    Abstract:

    The green rice leafhopper (GRH), Nephotettix Cincticeps Uhler, is a major insect pest of cultivated rice, Oryza sativa L., throughout the temperate regions of East Asia. GRH resistance had been reported in the wild species Oryza nivara but genetic basis of GRH resistance in wild rice accession has not been clarified. Here, we found a major QTL, qGRH4.2, on chromosome 4 conferred GRH resistance with 14.1 of the logarithm of odds (LOD) score explaining 67.6% of phenotypic variance in the BC1F1 population derived from a cross between the susceptible japonica cultivar ‘Taichung 65’ (T65) and O. nivara accession IRGC105715. qGRH4.2 has been identified as GRH6 between the markers RM5414 and {"type":"entrez-nucleotide","attrs":{"text":"C60248","term_id":"2418953","term_text":"C60248"}}C60248 in a BC3F2 population derived from two BC3F1 plants resistant to GRH. In a high-resolution mapping, the GRH6 region was delimited between the markers G6-c60k and 7L16f, and corresponded to an 31.2-kbp region of the ‘Nipponbare’ genome. Understanding the genetic basis of GRH resistance will facilitate the use of GRH resistance genes in marker-assisted breeding in rice.

  • a major qtl for resistance to green rice leafhopper Nephotettix Cincticeps uhler derived from african rice oryza glaberrima steud
    Breeding Science, 2010
    Co-Authors: Daisuke Fujita, Atsushi Yoshimura, Kazuyuki Doi, Hideshi Yasui
    Abstract:

    The green rice leafhopper (GRH), Nephotettix Cincticeps Uhler, is a serious insect pest of cultivated rice (Oryza sativa L.) in temperate East Asia. An African rice cultivar, Oryza glaberrima Steud. (IRGC104038), was shown to be highly GRH-resistant at the booting stage. To reveal the genetic basis of the GRH resistance in O. glaberrima, a BC1F1 population derived from a cross between a susceptible japonica variety, Taichung 65 (T65), and O. glaberrima was analyzed by quantitative trait locus (QTL) analysis. A single major QTL for GRH resistance, designated qGRH9, was detected on rice chromosome 9, and three minor QTLs were detected on rice chromosomes 3, 7, and 10. A series of O. glaberrima introgression lines (GILs) containing IRGC104038 chromosome segments in the T65 genetic background were evaluated to confirm the genetic effects of the QTLs, and three GILs carrying qGRH9 showed resistance to GRH. Substitution mapping using the GILs revealed that qGRH9 was located between simple sequence repeat (SSR) markers RM215 and RM2482 in a 1.39-Mbp region on the distal region of the long arm of chromosome 9, and tightly linked to RM7306. These SSR markers maybe useful for marker-assisted selection of qGRH9 for improvement of GRH resistance in rice.

  • development of near isogenic lines and pyramided lines carrying resistance genes to green rice leafhopper Nephotettix Cincticeps uhler with the taichung 65 genetic background in rice oryza sativa l
    Breeding Science, 2010
    Co-Authors: Daisuke Fujita, Atsushi Yoshimura, Hideshi Yasui
    Abstract:

    The green rice leafhopper (GRH), Nephotettix Cincticeps Uhler, is a serious insect pest of cultivated rice (Oryza sativa L.) in temperate East Asia. Six GRH-resistance genes (Grh1, Grh2, Grh3, Grh4, Grh5, and Grh6) and one quantitative trait locus (QTL; qGRH4) have been identified. We selected near-isogenic lines (NILs) carrying Grh1, Grh2, Grh4, Grh5, Grh6, and qGRH4 with the japonica genetic background (Taichung 65 cultivar) by means of marker-assisted selection using new simple sequence repeat markers flanking the GRH-resistance genes and QTL. We also developed three pyramided lines (PYLs; Grh2/Grh6-PYL, Grh4/Grh6-PYL, and Grh5/qGRH4-PYL) using each NIL that carried a GRH-resistance gene or QTL. The NILs, PYLs, and donor parents were evaluated by using an antibiosis test. The resistance of Grh1-NIL and Grh5-NIL did not differ significantly from those of the donor parents, whereas the resistances of Grh2-NIL and Grh6-NIL were significantly lower than those of the donor parents. Grh4-NIL and qGRH4-NIL were highly susceptible. The resistance levels of the pyramided lines for Grh2 and Grh6, Grh4 and Grh6, and Grh5 and qGRH4 demonstrated a gene pyramiding effect that significantly increased their resistance. The developed NILs and PYLs should be useful genetic resources for rice improvement and deployment of the resistance genes.

  • Molecular mapping of a novel gene, Grh5, conferring resistance to green rice leafhopper (Nephotettix Cincticeps Uhler) in rice, Oryza sativa L.
    Theoretical and Applied Genetics, 2006
    Co-Authors: Daisuke Fujita, Atsushi Yoshimura, Kazuyuki Doi, Hideshi Yasui
    Abstract:

    The green rice leafhopper (GRH), Nephotettix Cincticeps Uhler, is one of the most serious insect pests affecting cultivated rice ( Oryza sativa L.) in temperate regions of East Asia. An accession of the wild rice species, Oryza rufipogon Griff. (W1962), was found to be highly resistant to GRH by an antibiosis test. To understand the genetic basis of the GRH resistance, a BC_1F_1 population derived from a cross between a susceptible Japonica variety, Taichung 65 (T65), and a highly resistant accession W1962 was analyzed by quantitative trait loci (QTL) mapping. A single major QTL for GRH resistance was detected on rice chromosome 8. A nearly isogenic population containing segments of the targeted QTL region derived from W1962 was then developed through advanced backcrossing with marker-assisted selection. Further molecular mapping using a BC_4F_2 population revealed that a new resistance gene, designated as Green rice leafhopper resistance 5 ( Grh5 ), was located on the distal region of the long arm of chromosome 8 and tightly linked to the simple sequence repeat markers RM3754 and RM3761. A nearly isogenic line (NIL) carrying Grh5 was subsequently developed in the progeny of the mapping population. The resistance level of Grh5- NIL was compared with those of developed NILs for GRH resistance and was found to have the highest resistance. The DNA markers found to be closely linked to Grh5 would be useful for marker-assisted selection for the improvement of resistance to GRH in rice.

Taiyun Wei - One of the best experts on this subject based on the ideXlab platform.

  • Data_Sheet_1_Rice Yellow Stunt Nucleorhabdovirus Matrix Protein Mediates Viral Axonal Transport in the Central Nervous System of Its Insect Vector.PDF
    2019
    Co-Authors: Haitao Wang, Juan Wang, Xiao-feng Zhang, Qian Zhang, Tianbao Zeng, Yuemin Zheng, Hongyan Chen, Taiyun Wei
    Abstract:

    Persistently transmitted plant viruses encounter multiple membrane and tissue barriers in the process of completing their infection routes within their insect vectors. Some of these viruses have been reported to overcome the elaborate barriers of the central nervous system (CNS) to travel through the nervous tissues, but the specific mechanisms of this process remain unknown. Here, we report the axonal transport mechanism of rice yellow stunt virus (RYSV), a nucleorhabdovirus, in the CNS of the green rice leafhopper (Nephotettix Cincticeps). The infection route of RYSV in the internal organs of its insect vector after ingestion of the virus was investigated by immunofluorescence microscopy. RYSV was first detected in the epithelial cells of midgut regions, from where it proceeded to the nervous system, and finally into the salivary glands. We then utilized immunofluorescence and electron microscopy to investigate the distribution of RYSV particles within the leafhopper CNS, demonstrating that non-enveloped viral particles distributed along the microtubule-based neurofilaments in the axon cytoplasm following the direct interaction of leafhopper α-tubulin with the RYSV M protein. Tubulin inhibitors inhibited the dissemination of RYSV to the CNS, then into the salivary glands in leafhoppers. We therefore describe a mechanism of plant virus transport through CNS axons as an alternative means of rapid viral dissemination in an insect vector.

  • Exploration of an Actin Promoter-Based Transient Expression Vector to Trace the Cellular Localization of Nucleorhabdovirus Proteins in Leafhopper Cultured Cells
    Frontiers in Microbiology, 2018
    Co-Authors: Xiao-feng Zhang, Haitao Wang, Juan Wang, Tianbao Zeng, Hongyan Chen, Yunjie Xie, Yibing Zhao, Taiyun Wei
    Abstract:

    Abstract: Continuously cultured cell lines derived from planthopper and leafhopper have greatly facilitated the investigation of rice viruses transmitted by these insects. However, the lack of a suitable transient expression vector has limited their utility. Here, by cloning and analyzing the promoter sequence of the gene encoding cytoplasmic actin from the leafhopper Nephotettix Cincticeps, we successfully developed the first efficient transient expression vector for cultured leafhopper cells, which can also be used to express exogenous proteins in other insect culture cell lines, including those derived from Recilia dorsalis leafhopper and Spodoptera frugiperda (Sf9). Furthermore, insertion of the Hr5 viral enhancer element and knockdown of the endogenous Dicer2 gene notably improved the vector’s expression efficiency in leafhopper cells. Using the optimized vector, we have for the first time traced the cellular localization of the proteins encoded by rice yellow stunt virus (RYSV) in cells of its insect vector and demonstrated that P6 protein is a component of the viroplasm.

  • development of leafhopper cell culture to trace the early infection process of a nucleorhabdovirus rice yellow stunt virus in insect vector cells
    Virology Journal, 2018
    Co-Authors: Haitao Wang, Juan Wang, Taiyun Wei, Yunjie Xie, Xiao-feng Zhang
    Abstract:

    In China, the rice pathogen Rice yellow stunt virus (RYSV), a member of the genus Nucleorhabdovirus in the family Rhabdoviridae, was a severe threat to rice production during the1960s and1970s. Fundamental aspects of the biology of this virus such as protein localization and formation of the RYSV viroplasm during infection of insect vector cells are largely unexplored. The specific role(s) of the structural proteins nucleoprotein (N) and phosphoprotein (P) in the assembly of the viroplasm during RYSV infection in insect vector is also unclear. In present study, we used continuous leafhopper cell culture, immunocytochemical techniques, and transmission electron microscopy to investigate the subcellular distributions of N and P during RYSV infection. Both GST pull-down assay and yeast two-hybrid assay were used to assess the in vitro interaction of N and P. The dsRNA interference assay was performed to study the functional roles of N and P in the assembly of RYSV viroplasm. Here we demonstrated that N and P colocalized in the nucleus of RYSV-infected Nephotettix Cincticeps cell and formed viroplasm-like structures (VpLSs). The transiently expressed N and P are sufficient to form VpLSs in the Sf9 cells. In addition, the interactions of N/P, N/N and P/P were confirmed in vitro. More interestingly, the accumulation of RYSV was significantly reduced when the transcription of N gene or P gene was knocked down by dsRNA treatment. In summary, our results suggest that N and P are the main viral factors responsible for the formation of viroplasm in RYSV-infected insect cells. Early during RYSV infection in the insect vector, N and P interacted with each other in the nucleus to form viroplasm-like structures, which are essential for the infection of RYSV.

  • Development of leafhopper cell culture to trace the early infection process of a nucleorhabdovirus, rice yellow stunt virus, in insect vector cells
    BMC, 2018
    Co-Authors: Haitao Wang, Juan Wang, Taiyun Wei, Yunjie Xie, Xiao-feng Zhang
    Abstract:

    Abstract Background In China, the rice pathogen Rice yellow stunt virus (RYSV), a member of the genus Nucleorhabdovirus in the family Rhabdoviridae, was a severe threat to rice production during the1960s and1970s. Fundamental aspects of the biology of this virus such as protein localization and formation of the RYSV viroplasm during infection of insect vector cells are largely unexplored. The specific role(s) of the structural proteins nucleoprotein (N) and phosphoprotein (P) in the assembly of the viroplasm during RYSV infection in insect vector is also unclear. Methods In present study, we used continuous leafhopper cell culture, immunocytochemical techniques, and transmission electron microscopy to investigate the subcellular distributions of N and P during RYSV infection. Both GST pull-down assay and yeast two-hybrid assay were used to assess the in vitro interaction of N and P. The dsRNA interference assay was performed to study the functional roles of N and P in the assembly of RYSV viroplasm. Results Here we demonstrated that N and P colocalized in the nucleus of RYSV-infected Nephotettix Cincticeps cell and formed viroplasm-like structures (VpLSs). The transiently expressed N and P are sufficient to form VpLSs in the Sf9 cells. In addition, the interactions of N/P, N/N and P/P were confirmed in vitro. More interestingly, the accumulation of RYSV was significantly reduced when the transcription of N gene or P gene was knocked down by dsRNA treatment. Conclusions In summary, our results suggest that N and P are the main viral factors responsible for the formation of viroplasm in RYSV-infected insect cells. Early during RYSV infection in the insect vector, N and P interacted with each other in the nucleus to form viroplasm-like structures, which are essential for the infection of RYSV

  • Additional file 1: of Development of leafhopper cell culture to trace the early infection process of a nucleorhabdovirus, rice yellow stunt virus, in insect vector cells
    2018
    Co-Authors: Haitao Wang, Juan Wang, Taiyun Wei, Yunjie Xie, Xiao-feng Zhang
    Abstract:

    Sequences of the forward primer and reverse primer for RYSV N, P, M and Nephotettix Cincticeps actin genes. (PDF 10 kb