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Generation of transgenic wheat resistant to wheat yellow mosaic virus and identification of gene silence induced by virus infection
Authors:Jin Dong  Zhentian He  Chenggui Han  Xiulan Chen  Lingdi Zhang  Weihua Liu  Yuepeng Han  Jinrong Wang  Yafeng Zhai  Jialin Yu  Yi Liu  Yueyan Xiao
Institution:(1) State Key Laboratory for Agrobiotechnology, China Agricultural University, 100094 Beijing, China;(2) Agricultural Institute of Lixiahe District, 225207 Yangzhou, China
Abstract:The plasmid containing the promoter Act1, the coat protein (cp) gene of wheat yellow mosaic virus (WYMV) and the selectable bar gene, was delivered via particle bombardment, directly into immature embryos of a wheat cultivars. PCR and PCR-RFLP were employed to screen the existence of the cp gene in T0 and T1 generations. Seeds from the positive T1 plants were sowed in fields heavily contaminated with WYMV to detect their resistance. In field trial of virus infection, one of the transgenic wheat lines, P8-T2, exhibited highly disease-resistance. Western blot and RT-PCR analysis showed that the expression level of cp gene in the resistant transgenic line was reduced greatly compared to those susceptible to WYMV infection. This provided evidence to presume that the resistance obtained by the transgenic wheat line was stimulated by the mechanism of the virus induced gene silencing.
Keywords:wheat yellow mosaic virus  coat protein gene  transgenic wheat  resistance  gene silencing
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