猕猴桃AcNCED的克隆与表达分析
Cloning and Expression Analysis AcNCEDs in Kiwifruit
刘珂祎 1张灵婉 1黄婷 1夏惠 1梁东1
作者信息
- 1. 四川农业大学园艺学院,成都 611130
- 折叠
摘要
[目的]9-顺式环氧类胡萝卜素双加氧酶(NCED)是类胡萝卜素降解酶,同时是脱落酸(ABA)合成的关键酶.为探明AcNCED基因家族各成员在猕猴桃果实发育期的表达特性.[方法]利用RT-PCR技术从猕猴桃中分离4个NCED基因,分别命名为AcNCED1-4,并用qRT-PCR技术分析它们在猕猴桃不同组织和果实发育期的表达.[结果]AcNCED1、AcNCED2、AcNCED3和AcNCED4的开放阅读框(ORF)分别为1 755、1 830、1 797和1 788 bp,它们分别编码585、610、599和596个氨基酸.经预测,AcNCED1、AcNCED3和AcNCED4蛋白定位于叶绿体,AcNCED2蛋白定位于细胞质,均无信号肽,属于非分泌蛋白.qRT-PCR分析表明,AcNCEDs在茎中表达量最高,根中最低,在果实发育前期及成熟期高表达.[结论]4个基因编码的蛋白都较为保守且表达量均具有组织特异性,为进一步探明AcNCED的生物学功能奠定基础.
Abstract
[Objective]9-cis-epoxy carotenoid dioxygenase(NCED)is an enzyme involving in carot-enoid degrading and a key enzyme in the synthesis of abscisic acid(ABA).To explore the expression characteristics of all members of the AcNCED gene family during kiwifruit development.[Method]Four NCED genes were cloned from kiwifruit by RT-PCR,named AcNCED1-4,and their expression in vari-ousduring tissues and fruit developmental stages and in different tissues of both varieties was analyzed by qRT-PCR.[Result]The open reading frames(ORF)of AcNCED1,AcNCED2,AcNCED3 and AcNCED4 are 1 755,1 830,1 797 and 1 788 bp,respectively,which encode 585,610,599 and 596 amino acids,respectively.It was predicted that AcNCED1,AcNCED3 and AcNCED4 proteins are localized in chloro-plast,while AcNCED2 protein in cytoplasm,all of which lacking signal peptide and belong to non-secreted proteins.The qRT-PCR analysis showed that AcNCEDs has the highest expression level in the stem,the lowest in the roots,and high expression during early fruit development and maturity.[Conclusion]The proteins encoded by all four genes are highly conserved and exhibit tissue-specific ex-pression,which lays the foundation for further exploring the biological function of AcNCED.
关键词
猕猴桃/类胡萝卜素/9-顺式环氧类胡萝卜素双加氧酶/NCED/基因克隆Key words
kiwifruit/carotenoids/9-cis-epoxy carotenoid dioxygenase/NCED/gene cloning引用本文复制引用
基金项目
四川省科技厅项目(2023YFH0003)
四川省科技厅项目(2023ZHCG0098)
雅安市科技项目(23CGZH0007)
出版年
2024
国家科技期刊平台
NSTL
万方数据