中国水稻科学 ›› 2012, Vol. 26 ›› Issue (5): 615-618.DOI: 10.3969/j.issn.10017216.2012.05.015

• 实验技术 • 上一篇    下一篇

实时荧光定量PCR筛选稻曲病菌内参基因

顾志敏1,2,丁正中1,陈析丰1,郭龙彪2 ,曾大力2 ,钱前2,* ,马伯军1,*   

  1. 1 浙江师范大学 化学与生命科学学院, 浙江 金华 321004;2 中国水稻研究所 水稻生物学国家重点实验室, 浙江 杭州310006;
  • 收稿日期:2012-01-16 修回日期:2012-03-12 出版日期:2012-09-10 发布日期:2012-09-10
  • 通讯作者: 钱前2,* ,马伯军1,*
  • 基金资助:

    国家自然科学基金资助项目(31171519, 31101130); 浙江省自然科学基金资助项目(LY12CD6001)。

Reference Gene Selection of Ustilaginoidea virens by Realtime  PCR

GU  Zhimin 1,2, DING Zhengzhong 1, CHEN Xifeng 1, GUO Longbiao 2, ZENG Dali 2 , QIAN Qian 2,* , MA Bojun 1,*   

  1. 1 College of Chemistry and Life Science, Zhejiang Normal University, Jinhua  321004, China; 2 State Key Laboratory of Rice Biology, China National Rice Research Institute, Hangzhou 310006, China;
  • Received:2012-01-16 Revised:2012-03-12 Online:2012-09-10 Published:2012-09-10
  • Contact: QIAN Qian2,*, MA Bojun1,*

摘要: 通过实时定量PCR分析了稻曲病菌中5个传统内参基因18S rRNA、GAPDH、ubiquitin、βactin、αtubulin的mRNA差异表达情况,并利用geNorm软件分析了它们在4个发育时期和NaCl胁迫处理中的表达稳定性。结果表明,在菌龄为7 d、8 d、9 d、10 d的稻曲病菌中,αtubulin2和βactin表达稳定;在0.4 mol/L  NaCl溶液处理0 h、4 h、8 h、12 h、16 h后,βactin和αtubulin2表达稳定。因此,当利用荧光定量PCR分析比较稻曲病菌基因表达差异时,可选择αtubulin和βactin作为内参基因。

关键词: 稻曲病菌, 实时定量PCR, 内参基因, geNorm软件

Abstract: A total of five traditional reference genes of Ustilaginoidea virens were systematically compared by using realtime quantitative PCR, including 18S rRNA, GAPDH, ubiquitin, βactin and αtubulin. The expression stability of the 5 candidate reference genes was evaluated with geNorm program when the fungus was 7, 8, 9 and 10 days old or exposed to 0.4 mol/L NaCl for 0 h,4 h, 8 h, 12 h, 16 h. The results showed that αtubulin2 and βactin were stably expressed when the fungus was at different days of age. For the case of 0.4 mol/L NaCl stress, βactin and αtubulin2 were also expressed stably. As a conclusion, βactin and αtubulin could be used as the reference genes for normalization of realtime quantitative PCR data.

Key words: Ustilaginoidea virens, realtime quantitative PCR, reference gene, geNorm program

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