不同组织及干旱胁迫下黄薇内参基因的筛选与验证  

Screening and validation of reference genes in Heimia myrtifolia in different tissues and under drought stress

作　　者：赵雨 林琳 王群[1,2,3] 张国哲 王杰 尚林雪 洪思丹[1,2,3] 马清清 顾翠花 ZHAO Yu;LIN Lin;WANG Qun;ZHANG Guozhe;WANG Jie;SHANG Linxue;HONG Sidan;MA Qingqing;GU Cuihua(College of Landscape and Architecture,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China;Zhejiang Provincial Key Laboratory of Germplasm Innovation and Utilization for Garden Plants,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China;Key Laboratory of National Forestry and Grassland Administration on Germplasm Innovation and Utilization for Southern Garden Plants,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China)

机构地区：[1]浙江农林大学风景园林与建筑学院,浙江杭州311300 [2]浙江农林大学浙江省园林植物种质创新与利用重点实验室,浙江杭州311300 [3]浙江农林大学南方园林植物种质创新与利用国家林业和草原局重点实验室,浙江杭州311300

出　　处：《浙江农林大学学报》2023年第3期665-672,共8页Journal of Zhejiang A&F University

基　　金：浙江省农业新品种重大专项花卉育种专项(2021C02071-4);浙江省自然科学基金资助项目(LY21C160001)。

摘　　要：【目的】为黄薇Heimia myrtifolia不同组织及不同干旱胁迫下基因表达分析筛选最适内参基因。【方法】选取黄薇盛花期的根、茎、叶、花,以及5种不同干旱处理的叶片作为实验材料,借助RT-qPCR技术对黄薇转录组数据筛选的9个候选内参基因进行分析,并利用软件geNorm、BestKeeper、NormFinder和RefFinder综合评价候选基因的表达稳定性。最后选取2个与胁迫相关的基因CSLD和SOD,对所选内参基因进行验证。【结果】geNorm、BestKeeper和NormFinder分析得出的候选内参基因排序存在一定差异。利用RefFinder对上述3个软件的结果综合分析得出:在不同组织中,最稳定的内参基因为GAPDH,最不稳定的内参基因为TUA;在干旱胁迫中,最稳定的内参基因为GAPDH,最不稳定的内参基因为TUB。在全部样本中,最稳定的内参基因为GAPDH,最不稳定的内参基因为18S RNA。对不同组织和干旱胁迫下的CSLD和SOD基因表达模式进行验证表明:上述2个基因与筛选所得内参基因的表达量和变化趋势均较为一致。【结论】在不同组织和干旱处理后,GAPDH是最适合黄薇基因表达的内参基因。图4表4参33。[Objective]The objective is to study the gene expression of Heimia myrtifolia in different tissues and under different drought stress.[Method]The root,stem,leaf,and flower of H.myrtifolia at full flowering stage and 5 leaves under different drought treatments were selected as experimental materials,and 9 candidate internal reference genes screened from the transcriptome data were analyzed by RT-qPCR.The software geNorm,BestKeeper,NormFinder,and RefFinder were used to comprehensively evaluate the expression stability of candidate reference genes.Finally,2 stress-related genes CSLD and SOD were selected to verify the selected internal reference genes.[Result]GeNorm,BestKeeper,and NormFinder showed some differences in the ranking of candidate internal parameters.By using the online software RefFinder to comprehensively analyze the results of the above 3 software,it was concluded that GAPDH was the most stable internal reference gene and TUA was the most unstable in different tissues.In drought stress,GAPDH was the most stable internal reference gene and TUB was the least stable.In all samples,the most stable reference gene was GAPDH,and the most unstable one was 18S RNA.The expression patterns of CSLD and SOD genes in different tissues and under drought stress were verified,which showed that the expression levels and change trends of the above 2 genes were consistent with those of the selected internal reference genes.[Conclusion]GAPDH is the best internal reference gene for gene expression of H.myrtifolia in different tissues and under drought treatment.[Ch,4 fig.4 tab.33 ref.]

关 键 词：黄薇 实时荧光定量PCR 内参基因 

分 类 号：S718.46[农业科学—林学]