不同株系球形棕囊藻qRT-PCR内参基因的筛选  

Screening Suitable Reference Genes for qRT-PCR in Different Strains of Phaeocystis globosa

作　　者：许歆 王锦荣[2] 许欣红 徐肖 李靖 杨杰 XU Xin;WANG Jinrong;XU Xinhong;XU Xiao;LI Jing;YANG Jie(Fujian Key Laboratory on Conservation and Sustainable Utilization of Marine Biodiversity,College of Geography and Oceanography,Minjiang University,Fuzhou,Fujian 350108,China;The Second Geological Exploration Institute of China Metallurgical Geology Bureau,Fuzhou,Fujian 350108,China)

机构地区：[1]闽江学院地理与海洋学院,福建省海洋生物多样性保护与永续利用重点实验室,福建福州350108 [2]中国冶金地质总局第二地质勘查院,福建福州350108

出　　处：《闽江学院学报》2024年第5期67-79,共13页Journal of Minjiang University

基　　金：国家自然科学基金(42306179);福建省中青年教师教育科研项目(JAT200433);中国冶金地质总局二局科研项目(CMGB02KY202302);闽江学院发树慈善基金会捐赠资金研究专项(MFK23015)。

摘　　要：研究了7个候选内参基因(α-tubulin、β-tubulin、Actin、Ubiquitin、18 S rRNA、EF-1α和GAPDH),在3株成囊能力不同的球形棕囊藻(成囊株系PG-1和PG-2以及不成囊的单细胞株系PG-3)不同生长阶段下的表达稳定性。结果表明:PG-1的最佳内参基因组合为Ubiquitin、β-tubulin和18 S rRNA;PG-2的最佳内参基因组合为α-tubulin、18 S rRNA、β-tubulin和EF-1α,可见在成囊株系中表达较为稳定的内参基因为β-tubulin和18 S rRNA;在单细胞株系PG-3中表达最稳定的内参基因为Ubiquitin和α-tubulin;对3个株系球形棕囊藻的基因表达差异进行标准化分析可使用EF-1α、Ubiquitin和18 S rRNA为内参基因组合。所得结论为更准确地研究不同株系球形棕囊藻在不同生长时期的基因表达、验证成囊的关键基因奠定基础。Expression stability of seven candidate reference genes in different growth stages of three Phaeocystis globosa strains with different colony-formation ability was studied.Results showed that the optimal reference genes in colony-forming strain PG-1 are Ubiquitin,β-tubulin and 18 S rRNA.α-tubulin,18 S rRNA,β-tubulin and EF-1αare the best combination for another colony-forming strain PG-2.Therefore,β-tubulin and 18 S rRNA were stably expressed in the colony-forming strains.The most stable reference genes in non-colony-forming strain PG-3 were Ubiquitin andα-tubulin.EF-1α,Ubiquitin and 18 S rRNA could be used for the standardized analysis of gene expression level among three P.globosa strains.This study provides a basis for a more accurate study on the gene expression levels of P.globosa in different strains and different growth stages,especially verifying the key genes of colony formation.

关 键 词：球形棕囊藻 不同株系 内参基因 实时荧光定量PCR 

分 类 号：X171[环境科学与工程—环境科学]