RT-qPCR normalization of reference genes in different lifehistory stages of Gracilaria vermiculophylla(Rhodophyta)  

作　　者：Yingyue ZHANG Jinxin YANG Ze YANG Cong QI Di XU 

机构地区：[1]Key Laboratory of Marine Genetics and Breeding,Ministry of Education,College of Marine Life Sciences,Ocean University of China,Qingdao 266003,China

出　　处：《Journal of Oceanology and Limnology》2023年第5期1910-1917,共8页海洋湖沼学报（英文）

基　　金：Supported by the Key Program of Science and Technology Innovation in Ningbo (No.2019B10009);the National Natural Science Foundation of China (No.41476111)。

摘　　要：The quantitative realtime polymerase chain reaction(RT-qPCR)is a powerful and sensitive method to measure expression of targeted gene but it highly relies on the use of suitable reference genes for data normalization.We evaluated the expressions of 8 housekeeping genes:18S ribosomal rDNA(18S rDNA),28S ribosomal r DNA(28S rDNA),rubisco large subunit(rbc-L),β-actin(ACT),glyceraldehyde-3-phosphate dehydrogenase(GAPDH),elongation factor 1(EF1),β-tubulin(Tub B),and P-phycoerythrin B(PEB),to select the suitable reference genes for different life-history stages(tetrasporophyte,carposporophyte,and male/female gametophyte)of Gracilaria vermiculophylla by absolute quantitative method.Softwares geNorm and BestKeeper were used to verify the results acquired from copy number analysis.Results show that the expression of identified reference genes varied in comparing groups composed of different type of life stages.It is suggested that 18S rDNA and TubB could be used for highly complex samples composed of mixed ploidy and phases.18S rDNA and 28S rDNA were also preferred for using among the matured isomorphic samples.But for samples with different maturities,TubB and ACT were recommended for tetrasporophytes and gametophytes respectively.

关 键 词：reference gene Gracilaria vermiculophylla life-history stage quantitative real-time polymerase chain reaction(RT-qPCR) red algae 

分 类 号：Q943[生物学—植物学]