摘要： 通过对模板DNA、Primer、dNTPs、10×Taq 10 Buffer、Taq酶的不同浓度进行单因素筛选,并对退火温度和循环次数进行摸索,建立和优化了灰葡萄孢菌Flipper转座子PCR扩增体系。采用该体系及Flipper转座子引物扩增得到了1 159 bp的片段,通过比对发现,该片段与Botryotinia fuckeliana的Flipper转座因子转座酶基因相似性达99%,表明扩增得到的是预期目的片段。Flipper转座子扩增体系的建立,为深入研究我国葡萄灰霉病菌菌株的转座子类型及其与致病力、抗药性等因素之间的关系奠定了重要的基础。
关键词:
灰葡萄孢菌,
Flipper转座子,
PCR扩增体系,
单因素筛选
Abstract: Grape gray mold caused by Botrytic cinerea Pers． is one of the most important diseases on grapes worldwide． The genetic variation of Botrytis cinerea is widely and adaptable，which is easy to produce drug-resistant strains． It is reported that Boty and Flipper transposon of Botrytis cinerea are related to the presence or absence of resistant strains． By setting different gradient and optimize Botrytis cinerea Flipper transposon PCR amplification system of grape in China， including template DNA，primer，dNTPs， 10 × Taq 10 Buffer，Taq DNA polymerase，annealing temperature，cycle indexthe 1 159 bp fragment was obtained ，which has 99% similarity with Botryotinia fuckeliana Flipper transposable element transposase gene through NCBI blastn． So the amplified fragment is verified as expected target fragment． The establishment and optimization of Flipper transposon PCR amplification system of the grape Botrytis cinerea in China provide an important foundation to further study of the relationship between the transposable elements，pathogenicity and resistant
Key words:
Botrytis cinerea,
Flipper transposable elements,
PCR amplification system,
Single factor secreening
中图分类号:
常佳迎, 寇宏达, 乔丹娜, 张艳杰, 赤国彤, 李亚宁, 李兴红, 刘大群. 葡萄灰霉病菌Flipper转座子扩增体系的建立和优化[J]. 华北农学报, 2013, 28(3): 227-233. doi: 10.3969/j.issn.1000-7091.2013.03.041.
Chang Jiaying, Kou Hongda, Qiao Danna, Zhang Yanjie, Chi Guotong, Li Yaning, Li Xinghong, Liu Daqun. Establishment and Optimization of Flipper Transposon PCR Amplification System of the Grape Botrytis cinerea[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2013, 28(3): 227-233. doi: 10.3969/j.issn.1000-7091.2013.03.041.