野生烟草（Nicotiana alata）中抗番茄斑萎病毒基因的克隆和鉴定

番茄斑萎病毒属（Tospovirus）病毒是寄主范围最广、发生最为严重的一类植物病毒，其代表种番茄斑萎病毒（Tomato Spotted Wilt Virus, TSWV）已对云南烟区的烟叶生产构成了巨大的威胁，并有扩大和加重的趋势。发掘抗病基因培育抗病烟草品种是防治该病害最有效的防治手段。最近研究发现，野生烟草Nicotiana alata 对TSWV及Tospovirus属其他病毒具有很好的抗性，通过遗传学和分子标记筛选发现该抗性由显性单基因控制, 并已初步确定其对应的无毒基因（avr）。本项目将通过同源基因克隆、RNA-seq转录组测序以及瞬时沉默候选抗病基因技术从N. alata中分离抗TSWV的关键候选基因，并通过转基因过表达和RNA干扰进行候选抗性基因功能鉴定。项目结果对于明确新抗性基因抗TSWV的机理具有重要科学意义，还为培育抗TSWV的新品种提供基因资源具有重要实践意义。

Tomato spotted wilt virus (TSWV), is the type member of the genus Tospovirus, which causes severe economic losses in many crops worldwide. Now TSWV is a serious threat to tobacco production in the Yunnan province, China. Use of TSWV resistance cultivar offers an effective control strategy and significant potential in reducing fungicide. Recent studies have found that Nicotiana alata has broad-spectrum resistance to TSWV and other Tospoviruses. In addition, N. alata resistance to TSWV infection is controlled by a single dominant gene which corresponds to the NSm avirulence gene in the TSWV. This project will attempt to clone the TSWV resistance gene from N. alata by using the resistance gene analogous sequences, RNA-seq transcriptome sequencing and/or candidate gene transient silencing strategy. This project will also identify the resistance gene by using transgenic plants to overexpress and silence the candidate genes. This research will contribute to an understanding of the mechanism of TSWV disease resistance and the development of improved tobacco varieties with disease resistance traits.