Reference Gene Selection of Ustilaginoidea virens by Realtime  PCR

doi:10.3969/j.issn.10017216.2012.05.015

Chinese Journal of Rice Science ›› 2012, Vol. 26 ›› Issue (5): 615-618.DOI: 10.3969/j.issn.10017216.2012.05.015

• Experimental Techniques • Previous Articles Next Articles

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 College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China; ²State 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,*

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 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；

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）。

Abstract

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

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

CLC Number:

GU Zhimin1,2, DING Zhengzhong1， CHEN Xifeng1， GUO Longbiao2， ZENG Dali2， QIAN Qian2,*， MA Bojun1,*. Reference Gene Selection of Ustilaginoidea virens by Realtime PCR[J]. Chinese Journal of Rice Science, 2012, 26(5): 615-618.

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,* . 实时荧光定量PCR筛选稻曲病菌内参基因[J]. 中国水稻科学, 2012, 26(5): 615-618.

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Reference Gene Selection of Ustilaginoidea virens by Realtime PCR

实时荧光定量PCR筛选稻曲病菌内参基因

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