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、βactin、αtubulin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，αtubulin2和βactin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，βactin和αtubulin2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αtubulin和βactin作为内参基因。
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,*
. 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
.
DOI: 10.3969/j.issn.10017216.2012.05.015
A total of five traditional reference genes of Ustilaginoidea virens were systematically compared by using realtime quantitative PCR, including 18S rRNA, GAPDH, ubiquitin, βactin and αtubulin. 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 αtubulin2 and βactin were stably expressed when the fungus was at different days of age. For the case of 0.4 mol/L NaCl stress, βactin and αtubulin2 were also expressed stably. As a conclusion, βactin and αtubulin could be used as the reference genes for normalization of realtime quantitative PCR data.
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