文献类型：期刊文献

中文题名：‘麦缘锦楸’叶色表型qRT-PCR内参基因筛选及验证

英文题名：Selection and Validation of Reference Genes for Leaf Color Phenotype in'Maiyuanjinqiu',a Catalpa fargesii Variety,by qRT-PCR

作者：杨英英[1,2] 赵林姣[3,2] 杨桂娟[2] 张玉[2] 付鹏跃[2,4] 胡继文[2] 刘莹[2] 王楠[2]

第一作者：杨英英

机构：[1]三峡大学生物技术研究中心,湖北宜昌443002;[2]林木遗传育种国家重点实验室,中国林业科学研究院林业研究所,国家林业和草原局林木培育重点实验室,楸树国家创新联盟,北京100091;[3]西南林业大学林学院,云南昆明650224;[4]东北林业大学林木遗传育种国家重点实验室,黑龙江哈尔滨150040

年份：2022

卷号：35

期号：1

起止页码：123-131

中文期刊名：林业科学研究

外文期刊名：Forest Research

收录：CSTPCD;;Scopus;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：中央级公益性科研院所基本科研业务费专项资金项目(CAFYBB2020SZ003)省部级。

语种：中文

中文关键词：‘麦缘锦楸’;叶片;内参基因;qRT-PCR

外文关键词：Catalpa fargesii;'Maiyuanjinqiu';leaf;reference gene;qRT-PCR

分类号：S792.99

摘要：[目的]筛选不同叶色部位稳定表达的内参基因,为后期开展‘麦缘锦楸’叶色形成的分子机制奠定基础。[方法]以‘麦缘锦楸’不同叶色部位及灰楸对应部位叶片为材料,利用实时荧光定量PCR(qRT-PCR)技术检测了7个候选基因(CfUBC、CfActin11、CfPP2A、CfMADH、CfGADPH、CfEF-1及CbuActin)的相对表达量,利用GeNorm、NormFinder和BestKeeper等分析软件对内参基因进行了稳定性评价。分析了‘麦缘锦楸’和灰楸不同叶色部位中萜类合成相关基因(CfGES)的表达模式,验证了上述稳定性评价结果的可靠性。[结果]7个候选内参基因在‘麦缘锦楸’和灰楸叶片中均具有作为内参基因的性能,其中,CfMADH和CfEF-1在不同叶色部位的表达量最稳定,CfGADPH和CfActin11次之,CfUBC最差。以萜类合成相关基因CfGES验证内参稳定性发现,单独或组合使用CfEF-1及CfMADH为内参基因时,CfGES的表达差异与转录组数据趋势一致。[结论]单独或组合使用CfMADH和CfEF-1来校准‘麦缘锦楸’不同叶色部位的基因表达量,可显著提高实验结果的可靠性。
[Objective]To select the housekeeping genes stably expressed in leaves in order to support the study of the molecular mechanism of leaf color formation on'Maiyuanjinqiu',a Catalpa fargesii variety.[Method]The relative expression of seven candidate genes,including CfUBC,CfActin11,CfPP2A,CfMADH,CfGADPH,CfEF-1 and CbuActin were detected by real-time quantitative PCR(qRT-PCR)technology using leaves of different leaf color sectors of'Maiyuanjinqiu'and the corresponding sectors of Catalpa fargesii Bur.The stability of the seven candidate reference genes was evaluated by using internal reference analysis software such as GeNorm,NormFinder and Best-Keeper.Furthermore,the expression patterns of terpenoid biosynthesis related gene(CfGES)in the leaves of different leaf color parts of'Maiyuanjinqiu'and the corresponding sectors of C.fargesii were analyzed respectively.The results verified the stability evaluation results mentioned above.[Result]All the seven candidate reference genes can be used as internal reference genes in the leaves of'Maiyuanjinqiu'and C.fargesii.The expression levels of CfMADH and CfEF-1 were the most stable in different color tissues,followed by CfGADPH and CfActin11,while the CfUBC was the worst.The stability of internal reference gene was verified by terpenoid biosynthesis related gene(CfGES).It was found that the expressions of CfGES gene were consistent with the trend of transcriptome data,when CfEF-1 and CfMADH genes were used as internal reference for qRT-PCR alone or in combination.[Conclusion]Using CfMADH and CfEF-1 individually or in combination to calibrate the gene expression levels of different leaf color parts in'Maiyuanjinqiu'can greatly improve the reliability of experimental results.