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Conserved Sequences of Replicase Gene-Mediated Resistance to Potyvirus through RNA Silencing
Li Xu,YunZhi Song,JunHua Zhu,XingQi Guo,ChangXiang Zhu,FuJiang Wen 한국식물학회 2009 Journal of Plant Biology Vol.52 No.6
Nuclear inclusion protein b (NIb) genes of three Potato virus Y isolates PVY-SD1 (O strain), PVY-SD4 (N strain), PVY-SD5 (NTN strain), and Tobacco etch virus isolate TEV-SD1 in Shandong Province were cloned and sequenced. Sequence analysis showed that the sequence homology of the entire NIb gene among the four viruses ranged from 65% to 95%. Hairpin RNA (hpRNA) constructs were designed based on five conserved regions derived from PVY-SD1 and introduced into tobacco plants. After asexual propagation, the transgenic plants were analyzed for resistance to PVY-SD1, PVY-SD4, PVY-SD5, and TEV-SD1. We obtained resistance ratios of 26.2%, 22.7%, 36.4%, 20.3%, and 21.7% to PVY-SD1. When inoculated with the PVY-SD5 virus, the transgenic plants had resistance ratios ranging from 2.4% to 15.9%, but no resistance at all to the other viruses, PVY-SD4 and TEV-SD1. No correlation was found between resistance of transgenic plants and the transgene copy numbers. Northern blot and small interfering RNA (siRNA) analysis demonstrated that the resistance was attributable to RNA silencing. Genetic analysis demonstrated that virus resistance was stably inherited in progeny T1 and T2. These results indicate that the siRNA molecules against conserved regions can confer virus resistance but are restricted to viruses with more than 90% sequence homology.
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Chengguo Pei,Xu Liu,Wenying Wang,Hanfeng Ding,Mingsong Jiang,Guangxian Li,ChangXiang Zhu,FuJiang Wen,Fangyin Yao 한국식물학회 2011 Journal of Plant Biology Vol.54 No.3
Heading date is one of the importance agronomic traits. A library consisting of 1,123 single segment substitution lines (SSSLs) in the same genetic background of an elite rice variety Huajingxian 74 (HJX74) was evaluated for heading date (HD). From this library, the SSSL W06-26-35-1-5-2 with the substituted interval of PSM152–PSM154–PSM155–RM25–RM547–RM72–RM404 was found having a gene, which performed stable and late heading in the different environments of Shandong and Hainan provinces. To map the gene governing heading date, the SSSL W06-26-35-1-5-2 was crossed with the recipient HJX74 to develop an F_2 segregating population. The distribution of late and early heading plants in this population fitted a segregation ratio of 3:1, indicating the late heading was controlled by a dominant gene. The gene locus for heading date was tentatively designated as qHD8-1. Using a random sample of 460 individuals from the F_2population, the qHD8-1 was narrowed down to a region flanking by two SSR markers PSM155 and RM547. For fine mapping of qHD8-1, a large F_2:3 segregating population of 3,000 individuals were developed from F_2 plants heterozygous in the PSM155–RM547 region. Recombinants analysis further mapped qHD8-1 to an interval of region 26 kb with markers RM22492 and P23 bounded on the left and right sides, respectively. Sequence analysis of this 26-kb fragment revealed that it contains five putative open reading frames,which were regarded as candidates of qHD8-1. These results will be useful in cloning of the qHD8-1 gene
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