喀西茄内参基因实时荧光定量PCR表达稳定性评价

园艺学报 ›› 2014, Vol. 41 ›› Issue (8): 1731-1738.

喀西茄内参基因实时荧光定量PCR表达稳定性评价

周晓慧，刘军，庄勇*

（江苏省农业科学院蔬菜研究所，江苏省高效园艺作物遗传改良重点实验室，南京 210014）

收稿日期:2014-04-14 出版日期:2014-08-25 发布日期:2014-08-25

Selection of Appropriate Reference Genes in Solanum aculeatissimum for Quantitative Gene Expression Studies Under Different Experimental Conditions

ZHOU Xiao-hui，LIU Jun，and ZHUANG Yong*

（Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement，Institute of Vegetable Crops，Jiangsu Academy of Agricultural Sciences，Nanjing 210014，China）

Received:2014-04-14 Online:2014-08-25 Published:2014-08-25

摘要/Abstract

摘要： 为了筛选在喀西茄（Solanum aculeatissimum）中表达稳定的内参基因，选取7个常用内参基因GAPDH、ACTIN、18sRNA、UBQ、TUA、EF1和CYP，利用实时荧光定量PCR技术，结合GeNorm、NormFinder和RefFinder软件对其在喀西茄不同组织、不同植物生长调节剂处理、非生物和生物胁迫下的表达稳定性进行评价。结果表明，TUA和18sRNA在喀西茄不同组织中的表达稳定性最好，18sRNA和ACTIN在不同植物生长调节剂处理条件下表达水平最稳定，EF1和GAPDH在干旱和盐条件下表达稳定性最好，TUA和EF1在所有喀西茄测试样品组中（包括不同组织、不同植物生长调节剂处理、非生物及生物胁迫）的表达稳定性最高，而CYP和ACTIN在喀西茄不同试验条件下的表达稳定性较差。

关键词: 喀西茄, 实时荧光定量PCR, 内参基因

Abstract: The present study aimed to select stable and optimal reference genes for gene expression analysis in Solanum aculeatissimum. A total of seven traditional reference genes including GAPDH，ACTIN，18sRNA，UBQ，TUA，EF1 and CYP were screened and compared by the real-time quantitative PCR. The mRNA levels of these candidate genes were evaluated under different experimental conditions，including different tissues，different hormone treatments，abiotic and biotic stress using three common statistical softwares，GeNorm，NormFinder and RefFinder. The data showed that TUA and 18sRNA were the most stable genes in different tissues. 18sRNA and ACTIN showed high stability in different hormone treatments sample pool. EF1 and GAPDH were the most stably expressed reference genes under salt and drought stress while TUA and EF1 exhibited the most stable expression across all of the tested S. aculeatissimum samples. CYP and ACTIN exhibited the least stable expression under most of the experimental conditions which are not recommended.

Key words: Solanum aculeatissimum, real-time quantitative PCR, reference gene

中图分类号:

S 641.1

周晓慧刘军庄勇. 喀西茄内参基因实时荧光定量PCR表达稳定性评价[J]. 园艺学报, 2014, 41(8): 1731-1738.

ZHOU Xiao-hui，LIU Jun，and ZHUANG Yong*. Selection of Appropriate Reference Genes in Solanum aculeatissimum for Quantitative Gene Expression Studies Under Different Experimental Conditions[J]. ACTA HORTICULTURAE SINICA, 2014, 41(8): 1731-1738.

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