稻瘟病菌发育cDNA文库构建与表达序列标签分析  

Magnaporthe grisesa Development cDNA Library Construction and Expressed Sequence Tags Analysis

作　　者：金庆超[1] 董海涛[1] 彭友良[2] 陈保善[3] 邓晔[1] 戴承恩[1] 方永启[1] 邵菁[1] 娄沂春[1] 李有志[3] 李德葆[1] 

机构地区：[1]浙江大学农业与生物技术学院生物技术研究所,杭州310029 [2]中国农业大学农业部分子植物病理学重点实验室,北京100094 [3]广西大学亚热带生物资源保护和利用实验室,南宁530004

出　　处：《农业生物技术学报》2006年第6期963-969,共7页Journal of Agricultural Biotechnology

基　　金：国家高技术研究与发展计划(863)项目(No.2002BA711A15)资助。

摘　　要：利用稻瘟病菌(Magnaporthegriesa)连续6个发育时期的材料构建了一个混合cDNA文库。文库滴度,重组率和插入片段长度等质量分析表明,构建的文库包含完整的稻瘟病菌基因,可用于病菌基因表达分析。利用该文库获得了7456条5′端表达序列标签(ESTs)(GenBank收录号:(CK909944￣CK913666和CK928583￣CK932582),生物信息分析表明:EST序列拼接出2975个假定独立转录本(TUTs),冗余度为60.1%;从cDNA文库中筛选出大量的低丰度表达基因,约占TUT总数的79.8%,说明在文库中基因组成类型的复杂性较高;在所有TUTs中,功能未知基因约占85.5%,编码ECM33蛋白和疏水蛋白等病菌致病相关的注释基因高丰度表达,进一步表明该cDNA文库反映了病菌侵染和发育过程中基因表达的状况。The infection model of mechanical penetration of plant surfaces by Magnaporthe grisea has become a focus of molecular mechanism of the fungal pathogenesis. In order to overall analyze gene expression during infection and development, a mixed cDNA library was constructed with materials from contiguous six development stages of M. grisea. Some quality analysis, such as the titer, the recombinant rate and insert cDNA length of the cDNA library, indicated that the library contained intact genes and could be used for gene expression analysis of M.grisea Total 7 456 expressed sequence tags （ESTs）（GenBank（CK909944-CK913666 and CK928583-CK932582） of 5＇ ends were obtained from the cDNA library. Results of bioinformatics analysis for all ESTs data showed that EST sequences assembled out 2 975 tentative unique transcripts （TUTs） and endued 60.1% redundancy; most genes expressed with low abundance generated from the cDNA library and occupied 79.8% of all TUTs, indicating the library had a good complexity of gene composition; about 85.5% TUTs could not be assigned functional description and infection related genes, such as ECM33 protein and hydrophobin expressed at high abundance level among the remained annotated genes, furtherly indicated that the cDNA library reflected correctly gene expression during M. grisea development. So the mixed cDNA library provides an effective resource for functional study of the fungus and is suitable for further study for molecular mechanism of infection and development of M.grisea.

关 键 词：稻瘟病菌 CDNA文库 表达序列标签 

分 类 号：S188[农业科学—农业基础科学]