Virus isolate Y1 was obtained from tobacco showing curly shoot symptoms in Baoshan, Yunnan Province. Whitefly transmission test and virion morphology observa-tion showed that it is a begomovirus. In reactions with 14 ...Virus isolate Y1 was obtained from tobacco showing curly shoot symptoms in Baoshan, Yunnan Province. Whitefly transmission test and virion morphology observa-tion showed that it is a begomovirus. In reactions with 14 monoclonal antibodies raised against begomoviruses, Y1 was readily differentiated from begomoviruses reported in China, Pakistan and India. The complete nucleotide sequence of DNA-A was determined, it contains 2746 nucleotides, with two ORFs in virion-sense DNA and four ORFs in comple-mentary-sense DNA. Comparisons with total DNA-A, inter-genie region and deduced amino acid sequences of individual ORFs showed that Y1 is a distinct Begomovirus species, for which the name Tobacco curly shoot virus (TCSV) is pro-posed. The total DNA-A of TCSV is most closely related to that of Tomato leaf curl virus from India (85% sequence identity). In contrast, the deduced coat protein of TCSV is most like that of Cotton leaf curl virus 72b isolate from Paki-stan (98% amino acid sequence identity).展开更多
Tomato yellow leaf curl China virus Y10 isolate (TYLCCNV-Y10) alone could systemically infect host plants such as Nicotiana benthamiana without symptoms. In con- trast, Tobacco curly shoot virus Y35 isolate (TbCSV-Y35...Tomato yellow leaf curl China virus Y10 isolate (TYLCCNV-Y10) alone could systemically infect host plants such as Nicotiana benthamiana without symptoms. In con- trast, Tobacco curly shoot virus Y35 isolate (TbCSV-Y35) alone induces leaf curl symptoms in N. benthamiana. When inoculated into transgenic N. benthamiana plants expressing GFP gene (line 16c), TYLCCNV-Y10 neither reverses the established GFP silencing nor blocks the onset of GFP si- lencing. In contrast, TbCSV-Y35 can partially reverse the established GFP silencing and block the onset of GFP silenc- ing in new leaves. In the patch co-infiltration assays, the AC2 and AC4 proteins of TYLCCNV-Y10 and TbCSV-Y35 could suppress local GFP silencing and delay systemic GFP silenc- ing, suggesting that they are suppressors of RNA silencing. Comparison of the accumulation levels of GFP mRNA in the co-infiltration patches showed that Y10 AC2 and Y35 AC2 proteins had similar efficiency for suppression of RNA si- lencing. However, Y35 AC4 protein functioned as a stronger suppressor of RNA silencing than Y10 AC4 protein. There- fore, the pathogenicity difference between TbCSV-Y35 and TYLCCNV-Y10 may be related to the functional difference in their AC4 proteins.展开更多
The molecular variation of satellite DNA associated with Tobacco curly shoot virus (TbCSV) was compared by DNA sequencing of progenies of TbCSV DNA in Nicotiana benthamiana and N. glutinosa plants agroinoculated with ...The molecular variation of satellite DNA associated with Tobacco curly shoot virus (TbCSV) was compared by DNA sequencing of progenies of TbCSV DNA in Nicotiana benthamiana and N. glutinosa plants agroinoculated with infectious clones of TbCSV and its associated DNA. Comparisons indicated that TbCSV DNA populations had the higher levels of mutation frequency in N. glutinosa than that in N. benthamiana, and mutations generated in N. benthamiana were found upstream or downstream of C1 ORF whereas those in N. glutinosa were mostly located in A-rich region. No mutation was found in C1 ORF and satellite conserved region of DNA. The biological significance of the mutation accumulation and its role in disease epidemic were discussed.展开更多
基金This work was partially supported by the National Science Fund for Distinguished Young Scholars (Grant No. 30125032) the Teaching and Research Award Program for Outstanding Young Teachers in Higher Education Institutions of MOE The work was also sup
文摘Virus isolate Y1 was obtained from tobacco showing curly shoot symptoms in Baoshan, Yunnan Province. Whitefly transmission test and virion morphology observa-tion showed that it is a begomovirus. In reactions with 14 monoclonal antibodies raised against begomoviruses, Y1 was readily differentiated from begomoviruses reported in China, Pakistan and India. The complete nucleotide sequence of DNA-A was determined, it contains 2746 nucleotides, with two ORFs in virion-sense DNA and four ORFs in comple-mentary-sense DNA. Comparisons with total DNA-A, inter-genie region and deduced amino acid sequences of individual ORFs showed that Y1 is a distinct Begomovirus species, for which the name Tobacco curly shoot virus (TCSV) is pro-posed. The total DNA-A of TCSV is most closely related to that of Tomato leaf curl virus from India (85% sequence identity). In contrast, the deduced coat protein of TCSV is most like that of Cotton leaf curl virus 72b isolate from Paki-stan (98% amino acid sequence identity).
文摘Tomato yellow leaf curl China virus Y10 isolate (TYLCCNV-Y10) alone could systemically infect host plants such as Nicotiana benthamiana without symptoms. In con- trast, Tobacco curly shoot virus Y35 isolate (TbCSV-Y35) alone induces leaf curl symptoms in N. benthamiana. When inoculated into transgenic N. benthamiana plants expressing GFP gene (line 16c), TYLCCNV-Y10 neither reverses the established GFP silencing nor blocks the onset of GFP si- lencing. In contrast, TbCSV-Y35 can partially reverse the established GFP silencing and block the onset of GFP silenc- ing in new leaves. In the patch co-infiltration assays, the AC2 and AC4 proteins of TYLCCNV-Y10 and TbCSV-Y35 could suppress local GFP silencing and delay systemic GFP silenc- ing, suggesting that they are suppressors of RNA silencing. Comparison of the accumulation levels of GFP mRNA in the co-infiltration patches showed that Y10 AC2 and Y35 AC2 proteins had similar efficiency for suppression of RNA si- lencing. However, Y35 AC4 protein functioned as a stronger suppressor of RNA silencing than Y10 AC4 protein. There- fore, the pathogenicity difference between TbCSV-Y35 and TYLCCNV-Y10 may be related to the functional difference in their AC4 proteins.
文摘The molecular variation of satellite DNA associated with Tobacco curly shoot virus (TbCSV) was compared by DNA sequencing of progenies of TbCSV DNA in Nicotiana benthamiana and N. glutinosa plants agroinoculated with infectious clones of TbCSV and its associated DNA. Comparisons indicated that TbCSV DNA populations had the higher levels of mutation frequency in N. glutinosa than that in N. benthamiana, and mutations generated in N. benthamiana were found upstream or downstream of C1 ORF whereas those in N. glutinosa were mostly located in A-rich region. No mutation was found in C1 ORF and satellite conserved region of DNA. The biological significance of the mutation accumulation and its role in disease epidemic were discussed.
