hrpZ_(Psg12)转基因烟草及其抗性评价

Transgenic tobacco with hrpZ_(Psg12) gene and evaluation of its resistance

【目的】将来源于大豆细菌性斑点病菌(Pseudomonas syringae pv.glycinea)Psg12菌株的hrpZPsg12基因导入烟草"云烟87",对转化植株进行烟草普通花叶病毒(TMV)、烟草马铃薯Y病毒(PVY)和烟草野火病菌(Pseudomonas syringae pv.tabaci)的抗病性鉴定,为培育多抗烟草新品系奠定基础。【方法】采用农杆菌介导法,将hrpZPsg12基因转入烟草"云烟87"中,对T0和T1代转基因植株进行PCR检测、Southern杂交,并对T1代转基因植株的抗病性进行评价。【结果】hrpZPsg12基因成功转入"云烟87"中,PCR检测表明共获得7株T0代阳性植株、35株T1代阳性植株。对T1代PCR阳性植株进行Southern检测,表明外源目的基因hrpZPsg12已经整合进烟草基因组中,并可在转基因后代植株中稳定遗传。抗病性测定表明,T1代转基因烟草对TMV和PVY表现高抗,对烟草野火病表现中抗。【结论】获得了高抗TMV和PVY及中抗烟草野火病菌的转hrpZPsg12基因植株20和15株。

【Objective】hrpZPsg12gene fromPseudomonas syringae pv.glycineaPsg12 strain was imported to tobacco variety＂Yunyan 87＂and the disease resistance to TMV,PVY and Pseudomonas syringae pv.tabaci was identified to provide reference for cultivating resistant tobacco varieties.【Method】hrpZPsg12gene was transferred into ＂Yunyan 87＂by agrobacterium-mediated transformation method.Transgenic plants of T0 generation and T1 generation were detected by PCR,Southern blot and resistance assay.The disease resistance of T1 generation plants was also evaluated.【Result】PCR detection showed that this study obtained 7positive transgenic plant lines in T0 generation and 35 plant lines in T1 generation,indicating that hrpZPsg12 gene could inherit in transgenic plants.Southern blot for T1 generation confirmed that hrpZPsg12 gene was integrated into tobacco.Resistance assay confirmed that T1 generation had high resistance against TMV and PVY,and moderate resistance against Pseudomonas syringae pv.tabaci.【Conclusion】A total of 20 transgenic plants lines with high resistance against TMV and PVY and 15 transgenic plants lines with moderate resistance against Pseudomonas syringae pv.tabaci were obtained.