基于实时荧光定量PCR筛选巨菌草内参基因  

Screening of reference genes of Pennisetum giganteum based on real-time fluorescence quantitative PCR

作　　者：李苏涛 李妍 张磊 陈思齐 韩雪林 张娟 苏德伟[2] 罗海凌[2] 周晶 LI Sutao;LI Yan;ZHANG Lei;CHEN Siqi;HAN Xueling;ZHANG Juan;SU Dewei;LUO Hailing;ZHOU Jing(College of Animal Sciences(College of Bee Science),Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China;National Engineering Research Center of Juncao,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China)

机构地区：[1]福建农林大学动物科学学院(蜂学学院),福建福州350002 [2]福建农林大学国家菌草工程技术研究中心,福建福州350002

出　　处：《草业科学》2022年第5期907-921,共15页Pratacultural Science

基　　金：国家自然科学基金项目(32101418、42075116);福建省重大专项(2021NZ0101)。

摘　　要：以巨菌草(Pennisetum giganteum)的叶片、茎、根为试验材料,通过分析8个基因18s rRNA、Actin、GAPDH、ACTB、EF-1α、UBQ、CYP、TUB在正常生长、干旱以及盐碱胁迫下荧光定量PCR中稳定性表达情况,筛选巨菌草的稳定内参基因。利用geNorm、NormFinder和BestKeeper软件计算内参基因的表达稳定值并进行排序,最终通过赋值法综合排序确定稳定内参基因。结果表明,内参基因的稳定性在不同处理下存在差异。其中,ACTB稳定性好,为本研究筛选出巨菌草的内参基因。本研究筛选出的内参基因,为后续巨菌草功能基因表达分析奠定了基础。Using the leaves,stems,and roots of giant juncao as experimental materials,eight genes(18s rRNA,Actin,GAPDH,ACTB,EF-1α,UBQ,CYP,TUB)were analyzed using fluorescence quantitative Polymerase Chain Reaction(PCR)to select stable reference genes under normal,drought,and salt-alkali growth conditions.We used geNorm,NormFinder,and BestKeeper software to calculate the expression stability of genes and sorted the genes by the comprehensive ranking of the assignment method.The results showed that the stability of the genes differed under different treatments.In contrast,ACTB was stably expressed and used as the reference gene in this experiment.The experimental results will lay a foundation for the study of functional genes in giant juncao.

关 键 词：巨菌草 实时荧光定量PCR 内参基因 稳定性评价 干旱胁迫 盐碱胁迫 赋值法综合排序 

分 类 号：S543.9[农业科学—作物学]