利用荧光定量PCR验证与比较豇豆耐旱表达谱  

Validation and comparison of drought-responsive microarray with real-time fluorescent quantitative PCR in asparagus bean

作　　者：王莎[1,2] 徐沛[2] 汪宝根[2] 吴晓花[2] 黄芸萍[3] 鲁忠富[2] 刘永华[2] 李国景[2] 

机构地区：[1]浙江师范大学化学与生命科学学院,浙江金华321004 [2]浙江省农业科学院蔬菜研究所,浙江杭州310021 [3]宁波市农业科学研究院蔬菜研究所,浙江宁波330200

出　　处：《浙江农业学报》2013年第1期43-47,共5页Acta Agriculturae Zhejiangensis

基　　金：国家自然科学基金面上项目(31272183);浙江省蔬菜农业新品种选育重大科技专项(2012C12903);省科技厅项目(201122010);宁波市科技计划项目(201001C1011011)

摘　　要：豇豆[Vigna unguiculata(L.)Walp]起源于非洲,是我国重要的蔬菜作物。干旱是制约长豇豆生产的重要因素,研究和挖掘长豇豆种质中蕴含的耐旱基因是改良长豇豆耐旱性的必要基础。荧光定量PCR是定量检测基因表达的重要手段之一。本实验室前期开发了首张豇豆cDNA表达谱芯片,并利用该芯片研究了长豇豆耐旱表达谱。在此基础上,利用荧光定量PCR对其中11个基因的表达模式进行了验证和比较。试验结果表明,在干旱处理的长豇豆2个品种的根和叶中,供试基因的表达谱芯片分析和qPCR结果一致的共计34组,其中上调27组,下调7组,不一致的共计10组,一致率为0.773。本试验证明了高通量cDNA芯片与荧光定量PCR在检测豇豆干旱耐旱表达谱方面的一致性,为进一步细致分析和挖掘长豇豆耐旱相关基因奠定了基础。Originating from Mrica and domesticated in Asia, asparagus bean [ Vigna unguiculata ssp. sesquipedialis （L.） Verdc] is an important vegetable crop in China. Drought is an important environmental factor restricting the safety of asparagus bean production ; therefore it is fundamental to identify drought tolerance genes from the asparagus bean germplasms. Fluorescence quantitative PCR is a useful tool for quantitative assay of gene expression. In this study, we validated and compared expression pattern of a subset of 11 genes from an in-house cowpea cDNA microar- ray with fluorescence quantitative PCR. It turned out that in roots and leaves of two different asparagus bean cuhivars subject to drought stress, qPCR and microarray generally got similar results. The gene/tissue combinations showed congruent expression regulations between the two methods included 27 up-regulated and7 down-regulated genes, relative to 10 inconsistently observed combinations, giving a rate of consistence of 0. 773. This experiment verified the confidence of high-throughput eDNA microarray in monitoring drought-related gene expressions, which lays a founda- tion for mining and utilizing drought-tolerant genes in asparagus bean. The causes of differential gene expression pat- terns between the two methods were discussed.

关 键 词：长豇豆 耐旱 qPCR 表达谱 cDNA基因芯片 

分 类 号：S643.4[农业科学—蔬菜学]