The wheat_ Thinopyrum intermedium addition lines Z1,Z2 contain a pair of Th. intermedium chromosomes 2Ai_2 carrying the gene with resistance to barley yellow dwarf virus (BYDV). Genomic in situ hybridizat...The wheat_ Thinopyrum intermedium addition lines Z1,Z2 contain a pair of Th. intermedium chromosomes 2Ai_2 carrying the gene with resistance to barley yellow dwarf virus (BYDV). Genomic in situ hybridization (GISH) was used to analyze the chromosome constitution of Z1,Z2 by using genomic DNA probes from Th. intermedium and Pseudoroegneria strigosa . The results showed that the chromosome constitution of either Z1 or Z2 composes of 42 wheat chromosomes and two Th. intermedium chromosomes (2Ai_2). The 2Ai_2 chromosome is St_E intercalary translocation, in which the E genomic chromosome segment translocated into the middle region of the long arm of chromosome belonging to St genome. With the genomic DNA probe of Ps. strigosa , the GISH pattern specific to the 2Ai_2 chromosome may be used as a molecular cytogenetic marker. A detailed RFLP analysis on Z1, Z2 and their parents was carried out by using 12 probes on the wheat group 2 chromosomes. Twenty RFLP markers specific to the 2Ai_2 chromosome were identified. Two RAPD markers of OPR16 -350 and OPH09 -1580 , specific to the 2Ai_2 chromosome, were identified from 280 RAPD primers. These molecular markers could be used to assisted_select translocation lines with small segment of the 2Ai_2 chromosome and provide tools to localize the BYDV resistance.展开更多
Barley yellow dwarf virus(BYDV)can infect wheat(Triticum aestivum L.),leading to yield loss.Among four BYDV strains(GAV,GPV,PAV,and RMV)identified in China,BYDV-GAV is the prevailing isolate.YW642,a wheat–Thinopyrum ...Barley yellow dwarf virus(BYDV)can infect wheat(Triticum aestivum L.),leading to yield loss.Among four BYDV strains(GAV,GPV,PAV,and RMV)identified in China,BYDV-GAV is the prevailing isolate.YW642,a wheat–Thinopyrum intermedium translocation line,is resistant to BYDV isolates at both seedling and adult stages.Zhong 8601 is the wheat recurrent parent of YW642 and is susceptible to BYDV.In this study,we investigated the adult-stage resistance mechanism of YW642,measured BYDV titer and hydrogen peroxide(H_2O_2) in adult-stage leaves of YW642 and Zhong 8601 inoculated with BYDV-GAV,and identified transcriptional differences between YW642 and Zhong 8601 using microarray-based comparative transcriptomics.Enzyme-linked immunosorbent assay and H_2O_2assay showed that both BYDV titer and H_2O_2 content were markedly lower in YW642 than in Zhong 8601 at 21,28,35,and 40 days post-inoculation(dpi).The transcriptomic comparison revealed that many types of genes were significantly up-regulated at 35 dpi in adult-stage leaves of YW642 compared to Zhong 8601.The important up-regulated genes associated with the adult-stage resistance encoded 15 resistance-like proteins,pathogenesis-related proteins(such as defensin and lipid transfer proteins),protein kinase homologs,transcription factors,reactive oxygen species scavenging-related proteins,and jasmonic acid and gibberellic acid biosynthesis enzymes.These results suggest that precise expression regulation of these proteins plays a crucial role in adult-stage resistance of YW642 against BYDV infection.展开更多
[Objective]The aim of this study was to identify transient expression of movement protein （MP） gene in Nicotinana benthaminana rapidly and further investigate the function of this exogenous gene. [Method]The movemen...[Objective]The aim of this study was to identify transient expression of movement protein （MP） gene in Nicotinana benthaminana rapidly and further investigate the function of this exogenous gene. [Method]The movement protein gene of barley yellow dwarf virus （BYDV） was cloned into potato virus X （PVX） viral vector of pGR107,and PVX-recombinant vector was obtained. After electroporation of Agrobacterium tumefaciens,PVX was inoculated into the lower leaves of tobacco by Agrobacterium infiltration assay to observe the infection of virus on tobacco. [Result]After infection for 7 days,upper non-inoculated leaves of tobacco infected by the PVX-recombinant vector showed the virus infection symptoms,while the control group had no viral infection phenomenon. Daily follow-up observations for two groups revealed that tobacco infected by PVX-recombinant vector had severe symptoms of virus infection and curling leaves,or even led to necrosis both in infiltrated and systemic leaves in late period. However,tobacco infected by PVX vector had only slight symptoms of virus infection and could recover from infection. RT-PCR of the infected tobacco indicated that exogenous gene BYDV-MP had a normal transcription and expression in tobacco. [Conclusion]As a determinant factor for viral disease,BYDV-MP promotes the systemic infection rate of PVX and its symptom. In addition,it is feasible to express exogenous MP gene in Nicotiana benthaminan via PVX expression vector.展开更多
Barley yellow dwarf virus (BYDV) is one of the most serious wheat diseases in China. So far no resistance has been described in common wheat. A certain level of BYDV resistance was found in thirteen Triticeae species....Barley yellow dwarf virus (BYDV) is one of the most serious wheat diseases in China. So far no resistance has been described in common wheat. A certain level of BYDV resistance was found in thirteen Triticeae species. Thinopyrum intermedium, two octoploids derived from TH. intermedium/wheat, Zhong 4 awnless and TAF46, and one disomic addition line, L1 derived from TAF46, showed good resistance to BYDV by enzyme linked immunosorbent assay (ELISA). Two wheat/TA. intermedium translocation lines, CPI 119880 and CPI 119899, showing good BYDV resistance were developed from L1 by using both CSph mutant and tissue culture. It is found that their BYDV resistance was controlled by a single dominant gene. Two cDNA probes pEleAcc3 and pPJN8 (E1-T1) were screened for detecting Th. intermedium DNA in wheat background. A specific band for the DNA of Th. intermedium and its derivatives was found in Southern hybridization. It is also possible to determine the size of the alien segment by comparing the relative density of the specific band. Therefore, this can be used as a marker to identify the BYDV resistance in wheat breeding program.展开更多
Life tables of brown and green color morphs of the English grain aphid, Sitobion avenae （Fabricius） reared on barley under laboratory conditions at 20 ＋ 1℃,65% ± 5% relative humidity and a photoperiod of 16 ...Life tables of brown and green color morphs of the English grain aphid, Sitobion avenae （Fabricius） reared on barley under laboratory conditions at 20 ＋ 1℃,65% ± 5% relative humidity and a photoperiod of 16 ： 8 h （L ： D） were compared. The plants were either： （i） infected with the Barley yellow dwarf virus （BYDV）; （ii） not infectedwith virus but previously infested with aphids; or （iii） healthy barley plants, which were not previously infested with aphids. Generally, both color morphs of S. avenae performedsignificantly better when fed on BYDV-infected plants than on plants that were virus free but had either not been or had been previously infested with aphids. Furthermore,when fed on BYDV-infected plants, green S. avenae developed significantly faster and had a significantly shorter reproductive period than the brown color morph. There wereno significant differences in this respect between the two color morphs ofS. avenae when they were reared on virus-free plants that either had been or not been previously infestedwith aphids. These results indicate that barley infected with BYDV is a more favorable host plant than uninfected barley for both the color morphs ofS. avenae tested, particularly the green color morph.展开更多
The complete nucleotide sequence of genomic RNA of BYDV-GAV was determined. It comprised 5685 nucleotides and contained six open reading frames and four un-translated regions. The size and organization of BYDV-GAV gen...The complete nucleotide sequence of genomic RNA of BYDV-GAV was determined. It comprised 5685 nucleotides and contained six open reading frames and four un-translated regions. The size and organization of BYDV-GAV genome were similar to those of BYDV PAV-aus. The nucleotide and deduced amino acid sequences of the six ORFs were aligned and compared with those of other luteoviruses. The results showed that there was a high degree of identity between BYDV-GAV and MAV-PS1 in all ORFs except ORF5 and ORF6, which had only 87.4% and 70.2% identities respectively. The reported genomic nucleotide sequence of MAV was shorter than that of BYDV-GAV, but the comparison of the genomic nucleotide sequences for MAV-PS1 and GAV showed 90.4% sequence identity for the same region of the genome. Ac-cording to the level of sequence similarities, BYDV-GAV should be closely related to BYDV-MAV.展开更多
Using 2-D electrophoresis and virus overlay assay, a 50-kDa protein (P50) exhibiting specific binding to purified virus particles of BYDV-GAV was found in the protein extracts from Schizaphis graminum and Sitobion ave...Using 2-D electrophoresis and virus overlay assay, a 50-kDa protein (P50) exhibiting specific binding to purified virus particles of BYDV-GAV was found in the protein extracts from Schizaphis graminum and Sitobion avenae, two aphid species transmitting BYDV-GAV. P50 in the extracts of S. graminum was isolated by preparation electrophoresis and electro-eluted proteins from the gel slices for antiserum preparation. After feeding the antiserum through membrane, the transmission efficiencies of S. graminum and S. avenae for BYDV-GAV decreased significantly. It was suggested that P50 should be related with transmission pro- cess. Location of P50 was found at the plasma membrane surrounding the accessory salivary gland (ASG) in the head tissues of S. graminum by immunogold-labelling experiment. The ascertainment of the protein associated with virus transmission has a significance influence on further understanding the transmission mechanism and genetic engineering for resistant to vector transmission.展开更多
文摘The wheat_ Thinopyrum intermedium addition lines Z1,Z2 contain a pair of Th. intermedium chromosomes 2Ai_2 carrying the gene with resistance to barley yellow dwarf virus (BYDV). Genomic in situ hybridization (GISH) was used to analyze the chromosome constitution of Z1,Z2 by using genomic DNA probes from Th. intermedium and Pseudoroegneria strigosa . The results showed that the chromosome constitution of either Z1 or Z2 composes of 42 wheat chromosomes and two Th. intermedium chromosomes (2Ai_2). The 2Ai_2 chromosome is St_E intercalary translocation, in which the E genomic chromosome segment translocated into the middle region of the long arm of chromosome belonging to St genome. With the genomic DNA probe of Ps. strigosa , the GISH pattern specific to the 2Ai_2 chromosome may be used as a molecular cytogenetic marker. A detailed RFLP analysis on Z1, Z2 and their parents was carried out by using 12 probes on the wheat group 2 chromosomes. Twenty RFLP markers specific to the 2Ai_2 chromosome were identified. Two RAPD markers of OPR16 -350 and OPH09 -1580 , specific to the 2Ai_2 chromosome, were identified from 280 RAPD primers. These molecular markers could be used to assisted_select translocation lines with small segment of the 2Ai_2 chromosome and provide tools to localize the BYDV resistance.
基金supported by the National Key Research and Development Program of China(2016YFD0101802)
文摘Barley yellow dwarf virus(BYDV)can infect wheat(Triticum aestivum L.),leading to yield loss.Among four BYDV strains(GAV,GPV,PAV,and RMV)identified in China,BYDV-GAV is the prevailing isolate.YW642,a wheat–Thinopyrum intermedium translocation line,is resistant to BYDV isolates at both seedling and adult stages.Zhong 8601 is the wheat recurrent parent of YW642 and is susceptible to BYDV.In this study,we investigated the adult-stage resistance mechanism of YW642,measured BYDV titer and hydrogen peroxide(H_2O_2) in adult-stage leaves of YW642 and Zhong 8601 inoculated with BYDV-GAV,and identified transcriptional differences between YW642 and Zhong 8601 using microarray-based comparative transcriptomics.Enzyme-linked immunosorbent assay and H_2O_2assay showed that both BYDV titer and H_2O_2 content were markedly lower in YW642 than in Zhong 8601 at 21,28,35,and 40 days post-inoculation(dpi).The transcriptomic comparison revealed that many types of genes were significantly up-regulated at 35 dpi in adult-stage leaves of YW642 compared to Zhong 8601.The important up-regulated genes associated with the adult-stage resistance encoded 15 resistance-like proteins,pathogenesis-related proteins(such as defensin and lipid transfer proteins),protein kinase homologs,transcription factors,reactive oxygen species scavenging-related proteins,and jasmonic acid and gibberellic acid biosynthesis enzymes.These results suggest that precise expression regulation of these proteins plays a crucial role in adult-stage resistance of YW642 against BYDV infection.
基金Supported by National Natural Science Foundation of China(30870109)~~
文摘[Objective]The aim of this study was to identify transient expression of movement protein （MP） gene in Nicotinana benthaminana rapidly and further investigate the function of this exogenous gene. [Method]The movement protein gene of barley yellow dwarf virus （BYDV） was cloned into potato virus X （PVX） viral vector of pGR107,and PVX-recombinant vector was obtained. After electroporation of Agrobacterium tumefaciens,PVX was inoculated into the lower leaves of tobacco by Agrobacterium infiltration assay to observe the infection of virus on tobacco. [Result]After infection for 7 days,upper non-inoculated leaves of tobacco infected by the PVX-recombinant vector showed the virus infection symptoms,while the control group had no viral infection phenomenon. Daily follow-up observations for two groups revealed that tobacco infected by PVX-recombinant vector had severe symptoms of virus infection and curling leaves,or even led to necrosis both in infiltrated and systemic leaves in late period. However,tobacco infected by PVX vector had only slight symptoms of virus infection and could recover from infection. RT-PCR of the infected tobacco indicated that exogenous gene BYDV-MP had a normal transcription and expression in tobacco. [Conclusion]As a determinant factor for viral disease,BYDV-MP promotes the systemic infection rate of PVX and its symptom. In addition,it is feasible to express exogenous MP gene in Nicotiana benthaminan via PVX expression vector.
基金This research is supported by the Australian Centre for International Agricultural Research(Projects 8379 and 8813)by the National Science and Technology Committee of China on China's side.
文摘Barley yellow dwarf virus (BYDV) is one of the most serious wheat diseases in China. So far no resistance has been described in common wheat. A certain level of BYDV resistance was found in thirteen Triticeae species. Thinopyrum intermedium, two octoploids derived from TH. intermedium/wheat, Zhong 4 awnless and TAF46, and one disomic addition line, L1 derived from TAF46, showed good resistance to BYDV by enzyme linked immunosorbent assay (ELISA). Two wheat/TA. intermedium translocation lines, CPI 119880 and CPI 119899, showing good BYDV resistance were developed from L1 by using both CSph mutant and tissue culture. It is found that their BYDV resistance was controlled by a single dominant gene. Two cDNA probes pEleAcc3 and pPJN8 (E1-T1) were screened for detecting Th. intermedium DNA in wheat background. A specific band for the DNA of Th. intermedium and its derivatives was found in Southern hybridization. It is also possible to determine the size of the alien segment by comparing the relative density of the specific band. Therefore, this can be used as a marker to identify the BYDV resistance in wheat breeding program.
文摘Life tables of brown and green color morphs of the English grain aphid, Sitobion avenae （Fabricius） reared on barley under laboratory conditions at 20 ＋ 1℃,65% ± 5% relative humidity and a photoperiod of 16 ： 8 h （L ： D） were compared. The plants were either： （i） infected with the Barley yellow dwarf virus （BYDV）; （ii） not infectedwith virus but previously infested with aphids; or （iii） healthy barley plants, which were not previously infested with aphids. Generally, both color morphs of S. avenae performedsignificantly better when fed on BYDV-infected plants than on plants that were virus free but had either not been or had been previously infested with aphids. Furthermore,when fed on BYDV-infected plants, green S. avenae developed significantly faster and had a significantly shorter reproductive period than the brown color morph. There wereno significant differences in this respect between the two color morphs ofS. avenae when they were reared on virus-free plants that either had been or not been previously infestedwith aphids. These results indicate that barley infected with BYDV is a more favorable host plant than uninfected barley for both the color morphs ofS. avenae tested, particularly the green color morph.
基金This work was supported by the National Key Basic Research of China(973 contract TG2000016201)the National Natural Science Foundation of China(Grant No.39970034).
文摘The complete nucleotide sequence of genomic RNA of BYDV-GAV was determined. It comprised 5685 nucleotides and contained six open reading frames and four un-translated regions. The size and organization of BYDV-GAV genome were similar to those of BYDV PAV-aus. The nucleotide and deduced amino acid sequences of the six ORFs were aligned and compared with those of other luteoviruses. The results showed that there was a high degree of identity between BYDV-GAV and MAV-PS1 in all ORFs except ORF5 and ORF6, which had only 87.4% and 70.2% identities respectively. The reported genomic nucleotide sequence of MAV was shorter than that of BYDV-GAV, but the comparison of the genomic nucleotide sequences for MAV-PS1 and GAV showed 90.4% sequence identity for the same region of the genome. Ac-cording to the level of sequence similarities, BYDV-GAV should be closely related to BYDV-MAV.
基金This work was supported by the National Key Basic Research of China(Grant No.TG2000016201)the National Natural Science Foundation of China(Grant No.30070498).
文摘Using 2-D electrophoresis and virus overlay assay, a 50-kDa protein (P50) exhibiting specific binding to purified virus particles of BYDV-GAV was found in the protein extracts from Schizaphis graminum and Sitobion avenae, two aphid species transmitting BYDV-GAV. P50 in the extracts of S. graminum was isolated by preparation electrophoresis and electro-eluted proteins from the gel slices for antiserum preparation. After feeding the antiserum through membrane, the transmission efficiencies of S. graminum and S. avenae for BYDV-GAV decreased significantly. It was suggested that P50 should be related with transmission pro- cess. Location of P50 was found at the plasma membrane surrounding the accessory salivary gland (ASG) in the head tissues of S. graminum by immunogold-labelling experiment. The ascertainment of the protein associated with virus transmission has a significance influence on further understanding the transmission mechanism and genetic engineering for resistant to vector transmission.
