摘要
RT-qPCR是目前应用最广泛的基因表达分析方法,选择合适的内参基因对数据进行标准化至关重要。结合geNorm、Normfinder和RefFinder 3个软件分析了8个候选miRNA内参基因在核桃不同分化期花芽与叶芽、不同组织及不同品种中的表达稳定性。结果显示,jre-miR394a,jre-miR159a和jre-miR159c可作为不同分化期花芽中基因表达分析的内参;5.8S rRNA和jre-miRn3可作为不同分化期叶芽中基因表达分析的内参;5.8S rRNA,jre-miRn3、jre-miR159b-3p和jre-miR159c可作为不同组织中基因表达分析的内参;jre-miR159b-3p,jre-miR159c和jre-miR159a可作为不同品种中基因表达分析的内参。5.8S rRNA不适合作为不同分化期花芽和不同品种中基因表达分析的内参。本研究为核桃miRNA表达分析中的数据标准化提供数据支持和理论参考。
Reverse transcription quantitative real-time PCR (RT-qPCR) is the most widely used gene expression analysis method, and the selection of appropriate reference genes is crucial for data normalization. Eight candidate miRNA reference genes were screened to evaluate their expression stability in flower buds and leaf buds at different differentiation phase, different tissues and cultivars of walnut (Juglans regia L.) based on three softwares (geNorm, Normfinder and RefFinder). The results demonstrated that jre-miR394a, jre-miR159a and jre-miR159c could be used as reference genes for gene expression analysis in flower buds at different stages of differentiation;5.8S rRNA and jre-miRn3 could be used as reference genes for gene expression analysis in leaf buds at different stages of differentiation;5.8S rRNA, jre-miRn3, jre-miR159b-3p and jre-miR159c could be used as reference genes for gene expression analysis in different tissues;jre-miR159b-3p, jre-miR159c and jre-miR159a could be used as reference genes for gene expression analysis in different cultivars. Moreover, 5.8S rRNA was not a suitable reference gene for gene expression analysis in flower buds at different stages of differentiation and different cultivars. The study could provide data support and theoretical reference for data standardization in miRNA expression analysis of walnut.
作者
周丽
全绍文
马丽
徐航
牛建新
Zhou Li;Quan Shaowen;Ma Li;Xu Hang;Niu Jianxin(Key Laboratory of Special Fruits and Vegetables Cultivation Physiology and Germplasm Resources Utilization of Xinjiang Production and Construction Corps, Department of Horticulture, College of Agriculture, Shihezi University, Shihezi, 832003)
出处
《分子植物育种》
CAS
CSCD
北大核心
2019年第7期2270-2278,共9页
Molecular Plant Breeding
基金
国家自然科学基金项目(30560090)
新疆重大科技专项(201130102-1-4)共同资助
