水产学报2017，Vol.41Issue(7)：1064-1072,9.DOI:10.11964/jfc.20160810518
高光胁迫下坛紫菜定量PCR内参基因的筛选
Selection of the internal control gene for expression analyses of Pyropia haitanensis under high light stress by quantitative real-time PCR
摘要
Abstract
Pyropia haitanensis is a sessile marine macroalgae that inhabits the intertidal zone. High light was the main environmental stressor of P. haitanensis during low tides and the key factor affecting yield. To explore the molecular mechanisms of high light tolerance, gene expression analysis was the first step. Gene expression studies using quantitative real-time PCR should start by selecting an appropriate internal control gene; therefore, the absolute expression levels of six housekeeping genes [18S rRNA (18S), ubiquitin-conjugating enzyme (UBC), actin (ACT), β-tubulin (TUB), elongation factors 2 (EF2), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH)] were examined by the quantitative real-time PCR in samples corresponding to different high light stress treatments. Their expression stabilities were assessed by the comparative cycle threshold (Ct) method and by three different software packages: ge Norm, Bestkeeper and Norm Finder. The results showed that the Ct value of UBC had the least variation, while that of 18S obviously changed under different light intensities treatments. The analysis of ge Norm also exhibited that the most stable housekeeping genes were UBC and EF2. The analysis of both Bestkeeper and Norm Finder exhibited that stability measure M of UBC was the lowest (0.044 and 0.80, respectively) among six candidate genes. Thus the most stable housekeeping gene is UBC. Based on the above results, it is proposed that the most appropriate internal control gene for expression analyses under high light stress of P. haitanensis is UBC.关键词
坛紫菜/高光胁迫/内参基因/qRT-PCRKey words
Pyropia haitanensis/high light stress/housekeeping genes/qRT-PCR分类
生物科学引用本文复制引用
昌晶,陈陆丹,徐燕,纪德华,陈昌生,谢潮添..高光胁迫下坛紫菜定量PCR内参基因的筛选[J].水产学报,2017,41(7):1064-1072,9.基金项目
国家"八六三"高技术研究发展计划(2012AA10A411) （2012AA10A411）
国家自然科学基金(41276177) （41276177）
福建省自然科学基金(2014J05041, 2014J07006) National High Technology Research and Development Program 863 (2012AA10A411) （2014J05041, 2014J07006）
National Natural Science Foundation of China (41276177) （41276177）
Natural Science Foundation of Fujian, China (2014J05041, 2014J07006) （2014J05041, 2014J07006）
