马铃薯晚疫病菌快速分子检测与群体遗传多样性分析植物病理学专业论文.docxVIP

优秀毕业论文 精品参考文献资料 Rapid Rapid molecular detection and population genetic diversity infestans Master candidate：Gao Xue Major：Plant pathology Supervisor：Professor Yang Zhi—hui Professor Zhu Jie．hua Potato late blight，which is caused by Phytophthora infestans，is the most devastating disease of potato in the most of potato product areas in China．This disease could kill all potato crops when it happened in growing seasons．At the same time，the potato late blight could lead potatoes decomposed in the basement．It was the huge obstacle of potato industry in the world．We developed two molecular detection methods．These methods not only could be offer the technical support for the detection of transmission ratio of potato seeds but also could be offer the early diagnosis of P infestans．In the meanwhile，the study of the change of population genetic structure directly affected the disease occurrence and popular．T11is study could provide a basis of P infstans group evolution and disease epidemic，and then effectively promote the healthy development of potato industry in China． 1．The real．time fluorescent loop．mediated isothermal amplification detection method was set up in this study．Primers were designed according the Yptl gene．A pair of primers was designed for detection P infestans，the primers were two outer primers F3 and B3，two inner primers FIP and BIP and two loop primers LB and LF． We established and optimized the reaction system．The system were 1．2 pM inner primers，0．32 gM outer primers，0．68 mM loop primers，1．8 mM dNTPs，1．5 mM MgS04，3．2 gL 8U／gL Bst DNA polymerase，2．5 gL 1 0×ThermoPol Reaction Buffer， 2 uL DNA，0．3 pL 50×SYBR Green I and added ddH20 up to 25 gL．In our study， the detection limit was 37 1 pg／gL，which was 1 0 times more sensitive than conventional PCR method． 2．In this study．we set up the real．time fluorescent quantitative PCR detection method．Primers were designed according the Yptl gene．The reaction system was 0．5 gL primers，1 0 gL Bestar SybrGreen Qpcr ma