Selection of reference genes for real-time RT-PCR normalization in brown alga Undaria pinnatifida

被引：0
作者：
Jing Li
Haishu Huang
Tifeng Shan
Shaojun Pang
机构：
[1] Chinese Academy of Sciences,CAS Key Laboratory of Experimental Marine Biology, Institute of Oceanology
[2] Qingdao National Laboratory for Marine Science and Technology,Laboratory for Marine Biology and Biotechnology
[3] University of Chinese Academy of Sciences,undefined
来源：
Journal of Applied Phycology | 2019年 / 31卷
关键词：
Reference genes; qRT-PCR; geNorm; NormFinder; Phaeophyta;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
Quantitative real-time PCR (qRT-PCR) has become widely used method for detecting gene expression of an organism at different developmental stages or under various stressful environmental conditions. Finding reference genes is a prerequisite in this process. In this research, we investigated the expression stability of eight candidate genes in the brown alga Undaria pinnatifida, including EF1α (elongation factor 1-alpha), eEF1β (eukaryotic elongation factor-1 B beta), RPL35 (60S ribosomal protein L35-3), GAPDH (glyceraldehyde-3-phosphatedehydrogenase), RPS17 (40S ribosomal protein S17), LHCP (light-harvesting complex protein), ribH (6,7-dimethyl-8-ribityllumazine synthase), and ilvC (ketol-acid reductoisomerase). The haploid male and female gametophytes at different stages, as well as the diploid sporophytes treated with different abiotic factors were analyzed by geNorm and NormFinder software. It was found that eEF1β and ribH could be used as the most stable reference genes in studies of gender or developmental gene expression in the gametophytes. As for the detection of gene expression in the sporophytes, eEF1β and ribH were recommended as the most suitable reference genes for irradiance treatment, eEF1β and RPL35 for temperature treatment, while EF1α and LHCP for nutrition treatment. These results will enable us to go further for quantifying gene expression study at transcript level in U. pinnatifida.
引用
收藏
页码：787 / 793
页数：6
相关论文
共 107 条
[1]  
Alves-Lima C(2017)Reference genes for transcript quantification in J Appl Phycol 29 731-740
[2]  
Cavacana N(2004) under drought stress Cancer Res 64 5245-5250
[3]  
Teixeira Chaves GA(1995)Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets Mol Gen Genet 246 455-464
[4]  
de Lima NO(2010)The gene family encoding the fucoxanthin chlorophyll proteins from the brown alga BMC Plant Biol 10 49-601
[5]  
Stefanello E(2005)Identification and evaluation of new reference genes in J Molec Endocrinol 34 597-28
[6]  
Colepicolo P(2008) for accurate normalization of real-time quantitative RT-PCR data J Mar Biol Assoc U K 88 21-17
[7]  
Hotta CT(2005)Quantitative real-time RT-PCR—a perspective Plant Physiol 139 5-924
[8]  
Andersen CL(2008)Population features of the invasive kelp J Appl Phycol 20 915-13
[9]  
Jensen JL(2009) (Phaeophyceae: Laminariales) in Nuevo Gulf (Patagonia, Argentina) Mar Ecol Prog Ser 386 1-5250
[10]  
Orntoft TF(2004)Genome-wide identification and testing of superior reference genes for transcript normalization in Cancer Res 64 5245-264
12345678910