进境苹果重要病原真菌高通量测序排查及检测技术研究

摘要：我国是苹果生产及贸易进口大国。进境苹果可能携带十余种检疫性有害生物及几十种危险性病原真菌，通过贸易及旅客携带传入我国的风险极高，开展苹果病原菌的检测技术研究对保护我国苹果等水果生产和贸易具有重要的检疫意义。本实验使用分子生物学和生物信息学相关技术，就227种苹果病原真菌构建ITS信息数据库，再使用SSU、LSU以及EF-1α三种辅助基因构建PCR检测数据库，最后构建苹果病原菌高通量测序文库，通过测序及数据分析来加以验证。此种结合新一代高通量测序技术的分子技术体系，可实现快速、高效、多病原并行检测，该技术的发展将对我国口岸植物病原菌的快速、准确、高效地检出具有重要意义。32149
毕业论文关键词：苹果病原真菌；信息数据库；高通量测序技术
The high-throughput sequencing of Import apple’s important pathogenic fungi and the research of pathogen detection techniques
Abstract：Our country is apple production and trade importer. Import apple may carry more than 10 kinds of quarantine pests and dozens of pathogenic fungi, which have a high risk of being introduced into China through trade and passengers carrying. It is of great quarantine significance to research on apple pathogen detection technology for protecting China's apple and other fruit production and trade. With the use of molecular biology and bioinformatics technology, the ITS information database of 227 kinds of apple pathogenic fungi can be set up, and then the PCR test database using three auxiliary genes which include SSU, LSU and EF-1α can also be built .Finally we build high throughput sequencing Library of apple pathogenic fungi. Through the analysis of sequencing and data, it can be verified. Combined with a new generation of high-throughput sequencing technology system, integrated molecular detection technology can realize rapid, efficient and parallel pathogen detection. The development of which will be of great significance to our country’s rapid, accurate, efficient detection of port plant pathogenic bacteria.
Key words: apple pathogenic fungi；information database；high-throughput sequencing technology
目录
摘要   2
关键词   2
Abstract   2
Key words   2
引言   2
第一节 227种苹果病原菌ITS信息数据库的构建   3
1材料与方法   3
1.1供试材料与菌株   3
1.2菌株DNA提取   3
1.3 PCR扩增、测序   3
1.4序列差异分析   4
1.5 ITS高通量数据库构建   4
2结果与分析   4
2.1 ITS基因PCR及测序结果   4
2.2 ITS高通量数据库   11
第二节 SSU、LSU以及EF-1α三种辅助基因PCR检测数据库构建   11
1材料与方法   11
1.1供试材料与菌株   11
1.2菌株DNA提取   11
1.3 PCR扩增、测序   12
1.4序列差异分析   12
1.5辅助基因数据库构建   12
2 结果与分析   13
2.1辅助基因PCR及测序结果   13
2.2 SSU辅助基因数据库   18
第三节苹果病原菌高通量测序文库构建、测序及数据分析   18
1 材料与方法   18
1.1供试材料与菌株   18
1.2菌株DNA提取   18
1.3菌株混合   18
1. 4 ITSHiSeq测序   20
2 结果与分析   21
2.1测序原始数据   21
2.2数据拼接结果   21 进境苹果重要病原真菌高通量测序排查及检测技术研究:http://www.751com.cn/shengwu/lunwen_28593.html