中国农业科学2012，Vol.45Issue(8)：1505-1512,8.DOI:10.3864/j.issn.0578-1752.2012.08.006
野生茄子托鲁巴姆StLTPa7的克隆与分析
Cloning and Analysis of StLTPa7 from Solanum torvum
摘要
Abstract
[Objective] The objective of this study is to clone non-specific lipid transfer protein gene StLTPal from Solarium torvum and to analyze the function of the gene. [Method] Primers were designed according to homologous cloning method. In order to clone StLTPal, the gene cDNA was amplified by RT-PCR. Over-expression transgenic tobacco plants were generated via Agrobacterium-medhted transformation. Analysis on the inhibitory activity of proteins extracted from transgenic tobacco plants was done using mycelia growth rate method.[Result]StZ,TPa7 cDNA contained an ORF of 345 bp long and encoded a putative protein of 114 amino acids with a molecular weight of 11.42 kD and a theoretical pi of 9.01. StLTPal was induced to express by both salicylic acid (SA) and Verticillium dahlias and the level of transcript was the highest 24 to 48 h after treatment. To analyze the resistance of StLTPal to V. dahliae, the plants over-expressing StLTPa7 were generated by Agrobacterium-mediat&i transformation and 4 transgenic lines were identified by PCR. Quantitative RT-PCR indicated that the gene over-expressed in transgenic lines L5 and L7. Anti-fungal assay revealed that the inhibitory rate of the proteins extracted from the transgenic line L5 to V. dahliae was 2.5 times compared to the control.[Conclusion\SiLTPal exhibits associated to inhibitory effect of V. dahliae growth, suggesting that it may be involved in plant defense against V. dahliae.关键词
野生茄子/StLTPa7/基因克隆/遗传转化/功能分析Key words
wild eggplant/StLTPal/gene cloning/genetic transformation/function analysis引用本文复制引用
谢超,杨清,史策,决登伟,朱艳平,刘水平..野生茄子托鲁巴姆StLTPa7的克隆与分析[J].中国农业科学,2012,45(8):1505-1512,8.基金项目
国家“863”项目(2010AA10A108)、江苏省农业科技自主创新资金项目(cx(11)1020)、国家产业技术体系分子育种岗位项目(CARS-29) （2010AA10A108）
