【摘要】 大豆花叶病毒病是危害大豆产量和品质的主要病害之一 ,通过分子手段研究大豆花叶病毒病基因的抗病相关基因表达可以辅助抗病育种工作。利用SSH技术 ,对抗大豆花叶病毒病的大豆品种 814 3进行分析 ,构建大豆花叶病毒胁迫诱导表达的cDNA文库 ,并对文库进行初步分析。更多还原

【Abstract】 SMV is one of main diseases of soybean, which could affec t yields and quality of soybean seriously. It was effective to soybean breeding by studying the expression of resistant gene to SMV with molecular technology. I n this study, a soybean resistance line, DongNong 8143, was used to construct a subtractive cDNA library by SSH from soybean leaves inoculated by SMV No.1 at pr imary stage. cDNA dominantly or specifically expressed in infected leaves was pu rified using PCR Purification Kit and cloned into pGEM-T easy vector. Colonies w ere grown on LB-agar plates containing ampicillin, X-gal and IPTG. A subtractive plasmid library was constructed by SSH. Then the library was transformed to hos t bacteria E. coli DH5α, and the titer of the library was measured as 2×10 3 . 64 clones were picked up randomly and sequenced. Of them there is 50 clones which result of sequenced were good. The length of EST fragment varied from 136b p to 691bp, and the average length is 456bp. Among them, 41 sequences has poly(A ). Through ESTs were compared with sequences in unigene database of GeneBank wit h BLASTn and BLASTx algorithm, 38 ESTs of them had comparatively clear results a nd the percent of them in acquired ESTs is 74%. The EST expression profile showe d that the resistance-related genes include cell protection, signal transduction , restrict pathogen growth, system acquired resistance, and house-keeping gene. There are 12 ESTs, which have not comparatively clear results, that maybe new ge nes.更多还原