【摘要】 为了获得抗马铃薯Y病毒(PVY)转基因烟草,分别扩增PVY HC-Pro基因3′端正、反向片段,构建反向重复植物表达载体。利用农杆菌介导法转化普通烟草(Nicotiana tabacum)云烟85,经抗性筛选及抗病性鉴定,获得T0代转基因抗病烟草株系。繁殖T0代抗病株系,抗病性鉴定发现,部分抗病株系的T1代烟株发生了一定比例的抗感分离。Real-time PCR检测发生抗感分离的T1代烟株,发现抗病烟株中PVY HC-Pro基因RNA积累水平显著低于感病烟株,说明抗病烟株中HC-Pro基因发生了RNA沉默。利用dsRNA技术沉默HC-Pro基因,获得了烟草品种云烟85的T1代转基因抗PVY株系。更多还原

【Abstract】 In order to obtain transgenic tobacco plants resistant to Potato virus Y(PVY),sense and antisense fragments of 3’ end of gene HC-Pro of PVY were amplified respectively to construct an inverted repeat plant expression vector.Tobacco(Nicotiana tabacum)cultivar Yunyan 85 was transformed via Agrobacterium tumefaciens mediation,and T0 generation transgenic tobacco lines were acquired by resistance screening and resistance identification.T0 resistance transgenic tobacco lines were propagated.It was found that T1 progeny of partial resistant lines generated a certain proportion of segregation of resistance to susceptibility by resistance identification.T1 generation tobacco plants generating segregation of resistance to susceptibility were detected by real-time PCR.It was found that RNA accumulation level of gene HC-Pro in transgenic resistant tobacco plants was significantly lower than that in susceptible transgenic tobacco plants,and it was illustrated that gene HC-Pro generated RNA silencing in transgenic resistant tobacco plants.In our research work,gene HC-Pro was silenced by using dsRNA technique and T1 generation transgenic plants of tobacco cultivar Yunyan 85 resistant to PVY was obtained.更多还原