›› 2013, Vol. 25 ›› Issue (1): 0-47.

• 论文 •    

利用荧光定量PCR验证与比较豇豆耐旱表达谱

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,*   

  1. 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
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2013-01-25 发布日期:2013-01-25

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

WANG Sha;XYU Pei;WANG Baogen;WU Xiaohua;HUANG Yunping;LU Zhongfu;LIU Yonghua;LI Guojing;*   

  1. 1College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004,China; 2Institute of Vegetables, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021,China; 3Institute of Vegetables, Ningbo Academy of Agricultural Sciences, Ningbo 330200,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2013-01-25 Published:2013-01-25

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。

关键词: 长豇豆, 耐旱, qPCR, 表达谱, cDNA基因芯片

Abstract: Originating from Africa 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 microarray with fluorescence quantitative PCR. It turned out that in roots and leaves of two different asparagus bean cultivars 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 and 7 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 cDNA microarray in monitoring drought-related gene expressions, which lays a foundation for mining and utilizing drought-tolerant genes in asparagus bean. The causes of differential gene expression patterns between the two methods were discussed.

Key words: asparagus bean, drought tolerance, qPCR, expression patterns, cDNA