桉树焦枯病菌内参基因的筛选

Screening and evaluation of reference genes of Calonectria pseudoreteaudii

为筛选出桉树焦枯病菌在盐胁迫及侵染桉树叶片两种条件下表达稳定的内参基因,采用实时荧光定量聚合酶链式扩增反应(RT-q PCR)技术分析了桉树焦枯病菌中7个常用内参基因(GAPDH、α-tubulin-1、α-tubulin-2、18S rRNA、ubiquitin、β-actin、β-tubulin)的mRNA差异表达情况,并采用ge Norm软件分析了它们在两种条件下的表达稳定性。结果表明,ubiquitin和α-tubulin-2为桉树焦枯病菌在NaCl胁迫下表达最稳定的基因,且不需要引入第3个基因;18S rRNA和α-tubulin-1为侵染桉树叶片下表达最稳定的基因,但需引入第3个基因GAPDH。因此,ubiquitin和α-tubulin-2为桉树焦枯病菌在NaCl胁迫下较适合的内参基因,而18S rRNA、α-tubulin-1和GAPDH为侵染桉树叶片较适合的内参基因。

In order to screen out the best reference genes for（ real-time quantitative polymerase chain reaction,RT-q PCR）normalization in Calonectria pseudoreteaudii under salt stress and infection. The mRNA differential expression of 7 traditional reference genes,including GAPDH,α-tubulin-1,α-tubulin-2,18 S rRNA,ubiquitin,β-actin and β-tubulin of C. pseudoreteaudii,were systematically compared by RT-q PCR. The expression stability of the 7 candidate reference genes were evaluated with ge Norm program under different salt stress and injection. The results showed that ubiquitin and α-tubulin-2 were the most stable genes of C. pseudoreteaudii under salt stress while the third gene was not need to introduce; 18 S rRNA and α-tubulin-1 were the most stable genes under infection when GAPDH was introduced. Therefore,ubiquitin and α-tubulin-2 were the appropriate reference gene as the stability expression under salt stress while 18 S rRNA,α-tubulin-1 and GAPDH were the suitable reference gene under infection.