梨花芽休眠进程中microRNA实时定量PCR内参基因的筛选  

Selection of microRNA Reference Genes for Real-time Quantitative PCR in Pear Flower Bud during Dormant Phase

作　　者：马鑫瑞 李亮[1] 杨梦洁 廖正萍 王文婷[1] 刘靖媛[1] 吴少华[1] 李永裕[1] Ma Xinrui;Li Liang;Yang Mengjie;Liao Zhengping;Wang Wenting;Liu Jingyuan;Wu Shaohua;Li Yongyu(College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou, 35000)

机构地区：[1]福建农林大学园艺学院,福州350002

出　　处：《分子植物育种》2018年第14期4696-4704,共9页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31501694);高等学校博士学科点专项科研基金(2011351511011)共同资助

摘　　要：本研究通过分析候选内参基因的表达稳定性,筛选出适合梨花芽休眠进程中microRNA实时荧光定量PCR(qRT-PCR)的内参基因。本研究以休眠不同时期的砂梨花芽为材料,挑选了15个候选内参基因,用qRT-PCR技术检测候选基因在休眠不同时期的表达量,利用geNorm、Norm Finder和Best Keeper软件进行表达稳定性综合分析。研究结果表明,梨花芽休眠不同时期中表达最稳定的单个内参基因是miR3.0_22782,最稳定的内参基因组合是miR3.0_22782和miR171。本研究为准确探究梨花芽休眠进程中miRNAs的表达及进一步研究提供科学依据。The objective of the study was to select appropriate reference genes for qRT-PCR of miRNAs during dormant phase of pear flower bud by analyzing the stabilities of candidate reference genes.In this study,15 candidate reference genes were selected from the flower bud of pear at different periods of dormancy,and the expression of candidate genes in different periods of dormancy was detected by qRT-PCR technique.The stabilities were comprehensively analyzed by geNorm,Norm Finder and Best Keeper.The results showed that miR3.0_22782 was the most stable single reference gene in different dormancy periods of pear flower bud,and the most stable combination of reference genes was miR3.0_22782 and miR171.This study provided a scientific basis for the accurate detection and further study on the expression of miRNAs during dormant phase of pear flower bud.

关 键 词：梨 花芽 miRNA 内参基因 QRT-PCR 

分 类 号：Q943.2[生物学—植物学] S661.2[农业科学—果树学]