猕猴桃褪黑素合成相关基因AcTDC、AcT5H和AcSNAT的克隆与表达分析

doi:10.16036/j.issn.1000-2650.202202052

摘要/Abstract

摘要： 【目的】探明猕猴桃中褪黑素合成的相关基因对非生物逆境的响应机制。【方法】利用RT-PCR技术从猕猴桃中克隆了3个褪黑素合成相关基因AcTDC、AcT5H和AcSNAT,并用qRT-PCR技术分析了它们在不同组织和逆境胁迫下的表达。【结果】 AcTDC、AcT5H和AcSNAT的ORF区长度分别为1 506、618和579 bp,编码501、205和192个氨基酸,亚细胞定位预测分别在线粒体、内质网和叶绿体中。AcTDC仅在幼果中表达,在根、茎和叶中不表达;AcT5H在幼果中表达量相对较高,茎次之,根和叶较低;AcSNAT在叶中表达量极高,是其他部位表达量的几十至几百倍。测定逆境胁迫下叶片中各基因的表达量,AcTDC仍不表达,AcT5H表达量在脱落酸处理后下调幅度较大,AcSNAT表达量在各处理后下调幅度均较为明显。【结论】 3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,为进一步研究褪黑素在猕猴桃中的抗逆机制提供了理论基础。

关键词: 猕猴桃, 褪黑素, 基因克隆, 表达分析

Abstract: 【Objective】 In order to deeply explore genes related to melatonin synthesis in kiwifruit and reveal melatonin function in kiwifruit under abiotic stress. 【Method】 AcTDC、AcT5H and AcSNAT genes were cloned using RT-PCR and their expression in different tissues and under abiotic stress conditions were measured by qRT-PCR. 【Result】 The ORF regions of AcTDC、AcT5H and AcSNAT are 1 506、618 and 579 bp, coding 501、205 and 192 amino acid and the prediction of their subcellular localization are in mitochondria、endoplasmic reticulum and chloroplast,respectively. AcTDC only had expression in young fruits but not in roots、stems and leaves;the expression of AcT5H was comparatively high in young fruits,followed by stems, roots and leaves;the expression of AcSNAT was extremely high that dozens to hundreds of times higher than that of other tissues. When measuring their expression in leaves under abiotic stress,AcTDC still not expressed. AcT5H expression was down-regulated significantly after abscisic acid treatment and AcSNAT expression was down-regulated significantly after all treatments. 【Conclusion】 Proteins encoded by three genes are conservative and tissue-specific,providing theoretic basis for further study of melatonin resistance mechanism in kiwifruit.

Key words: kiwifruit, melatonin, gene cloning, expression analysis

中图分类号:

S663.4

吕晓雨, 刘芯伶, 张雪峰, 梁东, 夏惠. 猕猴桃褪黑素合成相关基因AcTDC、AcT5H和AcSNAT的克隆与表达分析[J]. 四川农业大学学报, 2022, 40(2): 191-197.

LYU Xiaoyu, LIU Xinling, ZHANG Xuefeng, LIANG Dong, XIA Hui. Cloning and Expression Analysis of Melatonin Biosynthesis Gene AcTDC、AcT5H and AcSNAT in Kiwifruit[J]. Journal of Sichuan Agricultural University, 2022, 40(2): 191-197.

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