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草地学报  2014 

鸭茅根组织实时定量PCR分析中候选内参基因的筛选

DOI: 10.11733/j.issn.1007-0435.2014.04.026, PP. 847-853

Keywords: 鸭茅,实时荧光定量PCR,非生物胁迫,候选内参基因,根组织

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Abstract:

实时荧光定量PCR(qRT-PCR)技术已成为研究基因表达的重要技术之一。以鸭茅(Dactylisglomerata)根组织为材料,利用qRT-PCR技术检测ACTIN(肌动蛋白酶)、TCTP(翻译控制肿瘤蛋白酶)、UBC(泛素结合酶)、ZTL(ZTL蛋白酶)、GAPDH(甘油醛-3-磷酸-脱氢酶)和SAMDC(腺苷甲硫氨酸脱羧酶)6个候选内参基因在不同非生物胁迫环境下的表达情况,并运用Genorm3.5,Delta-Ctmethod,NormFinder0.953,Bestkeeper1.0和RefFinder软件分析各候选内参基因在鸭茅根组织中的表达稳定性。结果表明6个候选内参基因综合表达稳定性从高到低排序为ACTIN,TCTP,GAPDH,UBC,ZTL,SAMDC;在干旱胁迫、盐胁迫、涝胁迫、热胁迫与ABA处理条件下,表达稳定性最高的基因是ACTIN;在涝胁迫条件下,表达稳定性最高的基因是TCTP;在不同的胁迫条件下,表达稳定性最低的基因是SAMDC。综合分析可知,基因ACTIN和TCTP最适合作为鸭茅非生物胁迫研究的候选内参基因,这可为鸭茅非生物胁迫下各种基因表达研究奠定基础。

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