2024年7月26日 星期五
黑莓1-氨基环丙烷-1-羧酸氧化酶基因RuACO1a和RuACO1b的克隆及功能鉴定
Cloning and functional identification of 1-aminocyclopropane-1-carboxylic acid oxidase genes RuACO1a and RuACO1b from blackberry (Rubus spp.)
2024年 第33卷 第3期 页码[14-26]    下载全文[21.6MB]  
摘要

基于前期黑莓(Rubus spp.)品种‘Navaho’果实转录组测序结果,通过反转录PCR克隆获得2个1-氨基环丙烷-1-羧酸氧化酶(ACO)基因,命名为RuACO1a和RuACO1b。结果表明:RuACO1a和RuACO1b的开放阅读框(ORF)长度分别为939和918 bp,分别含有4和3个外显子。系统进化分析结果显示RuACO1a和RuACO1b与月季(Rosa chinensis Jacq.)、蕨麻〔Argentina anserina (Linn.) Rydb.〕和野草莓(Fragaria vesca Linn.)ACO1蛋白的亲缘关系均较近,且分别属于蔷薇科(Rosaceae)植物中2类ACO1蛋白。RuACO1a在果实着色后28 d响应乙烯利诱导,其相对表达量急剧升高,而RuACO1b在果实着色后21 d响应乙烯利诱导,早于RuACO1a。脱落酸处理后,RuACO1a和RuACO1b的相对表达量在果实着色后14和28 d较高。RuACO1a和RuACO1b具有组织表达特异性,RuACO1a在幼果、花蕾和花中的相对表达量较高,RuACO1b在幼果和幼根中的相对表达量较高。与野生型相比,RuACO1a和RuACO1b过量表达转基因拟南芥〔Arabidopsis thaliana (Linn.) Heynh.〕植株均表现为叶片中叶绿素相对含量和氮含量升高、开花和角果成熟提前、ACO含量升高。综上所述,黑莓RuACO1a和RuACO1b基因均促进拟南芥角果提前成熟。

 

Abstract

Based on the previous transcriptome sequencing results of cultivar ‘Navaho’ of blackberry (Rubus spp.) fruits, two 1aminocyclopropane-1-carboxylic acid oxidase (ACO) genes were cloned via reverse transcription PCR and named as RuACO1a and RuACO1b. The results show that the open reading frames (ORFs) of RuACO1a and RuACO1b are 939 and 918 bp, respectively, and contain 4 and 3 exons, respectively. The phylogenetic analysis result shows that RuACO1a and RuACO1b both have close relationships with ACO1 proteins from Rosa chinensis Jacq., Argentina anserina (Linn.) Rydb., and Fragaria vesca Linn., and they belong to two categories of ACO1 proteins of Rosaceae plants. RuACO1a responds to ethephon induction at 28 d after fruit coloring, its relative expression increases sharply, while RuACO1b responds to ethephon induction at 21 d after fruit coloring, which is earlier than RuACO1a. After abscisic acid treatment, the relative expression of RuACO1a and RuACO1b is higher at 14 and 28 d after fruit coloring. RuACO1a and RuACO1b exhibit tissue-specific expression patterns, and the relative expression of RuACO1a is relatively high in young fruits, flower buds, and flowers, while those of RuACO1b are relatively high in young fruits and young roots. Compared with the wild type, transgenic Arabidopsis thaliana (Linn.) Heynh. plants overexpressing RuACO1a and RuACO1b exhibit increased chlorophyll relative content and nitrogen content in leaves, flowering and silique maturation in advance, and increased ACO content. In conclusion, both RuACO1a and RuACO1b genes from blackberry promote the early maturation of A. thaliana siliques.

 

关键词黑莓; 1-氨基环丙烷-1-羧酸氧化酶(ACO); 果实成熟; 乙烯; 脱落酸; 克隆; 功能鉴定
Key wordsblackberry (Rubus spp.); 1-aminocyclopropane-1-carboxylic acid oxidase (ACO); fruit ripening; ethylene; abscisic acid; cloning; functional identification
作者李洁1, 董金彦1, 闾连飞1, 吴文龙1, 李维林2a,2b, 张春红1
所在单位1. 江苏省中国科学院植物研究所(南京中山植物园) 江苏省植物资源研究与利用重点实验室, 江苏 南京 210014; 2. 南京林业大学林草学院、水土保持学院: a. 林木遗传育种全国重点实验室; b. 南方现代林业协同创新中心, 江苏 南京 210037
点击量675
下载次数719
基金项目江苏省种业振兴揭榜挂帅项目(JBGS[2021]021); 江苏省农业科技自主创新项目〔CX(22)2014〕