紫外线胁迫的花生幼果cDNA文库构建与RS cDNA的分离

Construction of cDNA library with RNA isolated from young fruit of peanut challenged with UV,and screening of cDNA library for RS cDNA

紫外线胁迫可导致花生产生白藜芦醇的诱导合成防御反应,本研究在利用UV处理花生幼果、激发花生幼果防御反应的基础上提取总RNA,应用SMART-cDNA文库构建试剂盒构建了花生幼果的cDNA文库.该文库含有1.0×106独立克隆,插入片段平均1 kb左右.利用所构建的cDNA文库,采用96孔板PCR筛选方法,分离获得2个白藜芦醇合成酶cDNA.

Defence reaction, such as synthesis of resveratrol, can be induced by UV in grape and peanut, and application of the inducible defense reaction mechanism is a promising approach of genetic improvement of crop stress resistance. To isolate the full length cDNA of resveratrol synthase for its application in crop biosynthetic pathway engineering, the total RNA was extracted from immature pods of peanut challenged with UV, and the cDNA library with more than 1.0 × 10^6 individual clones was constructed with SMARTTM cDNA construction kit. By a method of PCR-based 96-hole screening, 2 positive candidates of resveratrol synthase cDNAs were isolated, which turned out to be two members of resveratrol synthase family by sequence analysis. The results of the study laid important base for the pathway engineering of resveratrol in crops and the isolation of genes expressed in young pods of peanut challenged with UV.