灰霉菌转录因子Zn（2）-Cys（6）基因敲除载体的构建.docxVIP

PAGE \* MERGEFORMAT PAGE \* MERGEFORMAT II 中文摘要 灰霉菌是一种全球性植物致病真菌，在引起宿??主植物的灰霉病。病原体可以侵染500多种植物，每年会造成巨大的经济损失。我们从具有超过50,000个T-DNA标记的转化子的灰霉菌突变体文库筛选致病性减弱的突变体，使用热不对称交错PCR（Tail-PCR）和序列分析方法，我们发现T-DNA标签插入到灰霉菌Zn（2）-Cys（6）转录因子的基因启动子区域，导致在病原体毒力的减弱。含有Zn(2)-Cys(6)锌指结构域的转录因子是灰霉菌中基因数量较多的转录因子家族。目前仅在真菌中发现了该转录因子，其对真菌生长发育的转录调控有积极作用。为了进一步阐明Zn（2）-Cys（6）转录因子基因在灰霉菌生长发育过程中的调节作用，我们构建了灰霉菌Zn（2）-Cys（6）转录因子的敲除载体，并希望获得Zn（2）-Cys（6）转录因子基因的缺失突变体进行进一步研究。 关键词：灰霉菌；Tail-PCR；转录因子；Zn(2)-Cys(6)；敲除载体 Abstract Botrytis cinerea is a global plant pathogenic fungus that causes gray mold on host plants. The pathogen can infect more than 500 plant species and causes huge economic losses each year. A pathogenicity-attenuated mutant was screened from Botrytis cinerea mutant library that contains over 50,000 T-DNA tagged transformants. Using the methods of thermal asymmetric interlaced PCR (Tail-PCR) and the sequence analyses, we found that the T-DNA tag inserted into the gene promoter region of B. cinerea Zn (2) -Cys (6) transcription factor, which resulted in the attenuation of the pathogen virulence. The transcription factor containing the Zn (2) -Cys (6) zinc finger domain is a family of transcription factors with many gene members in the Botrytis cinerea. The transcription factors, currently only found in fungi, have a positive effect on the transcriptional regulation of fungal growth and development. In order to further elucidate the regulation of Zn (2) -Cys (6) transcription factor gene in the growth and pathogenesis of B. cinerea, we generated the knockout vector of B. cinerea Zn (2) -Cys (6) transcription factor, and hoped to obtain a gene deletion mutant of the Zn (2) -Cys (6) transcription factor for our further study. Key words: Botrytis cinerea; thermal asymmetric interlaced PCR (Tail-PCR)； transcription factor；Zn (2) -Cys (6)；knockout vector 目录 TOC \o 1-3 \h \z \u HYPERLINK \l _Toc483051445 前言 1 HYPERLINK \l _Toc483051446 第一节 灰葡萄孢菌与灰霉病 1 HYPERLINK \l _Toc483051447 第二节 Zn(2)-Cys(6)转录因子研究进展 3 H