华北农学报
華北農學報
화북농학보
ACTA AGRICULTURAE BOREALI-SINICA
2013年
3期
227-233
,共7页
常佳迎%寇宏达%乔丹娜%张艳杰%赤国彤%李亚宁%李兴红%刘大群
常佳迎%寇宏達%喬丹娜%張豔傑%赤國彤%李亞寧%李興紅%劉大群
상가영%구굉체%교단나%장염걸%적국동%리아저%리흥홍%류대군
灰葡萄孢菌%Flipper 转座子%PCR 扩增体系%单因素筛选
灰葡萄孢菌%Flipper 轉座子%PCR 擴增體繫%單因素篩選
회포도포균%Flipper 전좌자%PCR 확증체계%단인소사선
Botrytis cinerea%Flipper transposable elements%PCR amplification system%Single factor secreening
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1159 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。
通過對模闆 DNA、Primer、dNTPs、10×Taq 10 Buffer、Taq 酶的不同濃度進行單因素篩選，併對退火溫度和循環次數進行摸索，建立和優化瞭灰葡萄孢菌 Flipper 轉座子 PCR 擴增體繫。採用該體繫及 Flipper 轉座子引物擴增得到瞭1159 bp 的片段，通過比對髮現，該片段與 Botryotinia fuckeliana 的 Flipper 轉座因子轉座酶基因相似性達99％，錶明擴增得到的是預期目的片段。 Flipper 轉座子擴增體繫的建立，為深入研究我國葡萄灰黴病菌菌株的轉座子類型及其與緻病力、抗藥性等因素之間的關繫奠定瞭重要的基礎。
통과대모판 DNA、Primer、dNTPs、10×Taq 10 Buffer、Taq 매적불동농도진행단인소사선，병대퇴화온도화순배차수진행모색，건립화우화료회포도포균 Flipper 전좌자 PCR 확증체계。채용해체계급 Flipper 전좌자인물확증득도료1159 bp 적편단，통과비대발현，해편단여 Botryotinia fuckeliana 적 Flipper 전좌인자전좌매기인상사성체99％，표명확증득도적시예기목적편단。 Flipper 전좌자확증체계적건립，위심입연구아국포도회매병균균주적전좌자류형급기여치병력、항약성등인소지간적관계전정료중요적기출。
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 sys-tem of grape in China,including template DNA,primer,dNTPs,10 ×Taq 10 Buffer,Taq DNA polymerase,annea-ling temperature,cycle indexthe 1 159 bp fragment was obtained ,which has 99% similarity with Botryotinia fuckeli-ana Flipper transposable element transposase gene through NCBI blastn .So the amplified fragment is verified as ex-pected 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 trans -posable elements,pathogenicity and resistant.
