芒果乙烯信号转导关键基因MiCTR1的克隆与序列分析  

Cloning and Sequence Analysis of Key Ethylene Signal Transduction Gene MiCTR1 in Mango

作　　者：袁芳[1] 王春艳 李丽 许丹妮[1] 李志红 甘婷 隆宇涵 YUAN Fang;WANG Chunyan;LI Li;XU Danni;LI Zhihong;GAN Ting;LONG Yuhan(College of Chemistry and Biological Engineering,Guangxi Minzu Normal University,Chongzuo,Guangxi 532200,China;Agro-food Science and Technology Research Institute,Guangxi Academy of Agricultural Sciences,Nanning,Guangxi 530007,China;Guangxi Key Laboratory of Fruits and Vegetables Storage-processing Technology,Nanning,Guangxi 530007,China)

机构地区：[1]广西民族师范学院化学与生物工程学院,广西崇左532200 [2]广西壮族自治区农业科学院农产品加工研究所,广西南宁530007 [3]广西果蔬贮藏与加工新技术重点实验室,广西南宁530007

出　　处：《热带作物学报》2024年第10期2035-2043,共9页Chinese Journal of Tropical Crops

基　　金：国家重点研发计划专项(No.2021YFD1600100);国家自然科学基金项目(No.3230181455);广西重点研发计划项目(桂科AB23075095)。

摘　　要：芒果属于典型的呼吸跃变型果实,采后代谢旺盛且对乙烯非常敏感,CTR1是乙烯信号转导途径的负调控因子,在乙烯信号通路中发挥着核心作用。为了研究芒果CTR1在芒果采后贮藏过程中的表达模式及可能的作用,本研究从台农1号芒果转录组数据库中筛选1个CTR基因(MiCTR1),利用生物学方法对其编码的蛋白质进行测序分析,并对MiCTR1基因在芒果后熟过程中经乙烯抑制剂1-MCP处理后的表达模式进行分析。结果表明:MiCTR1的开放阅读框(ORF)长度为1551 bp,编码516个氨基酸,预测蛋白的分子式为C_(2503)H_(3932)N_(712)O_(797)S_(26),总原子数为7970,分子量为57.58 kDa,理论等电点(pI)为5.72,负电荷和正电荷的氨基酸残基总数分别为64和50个,脂肪系数为70.78,亲水性总平均值为–0.596,有111个磷酸化位点,且以丝氨酸(Ser)的磷酸化修饰为主,苏氨酸(Thr)为辅,无跨膜结构,无信号肽,亚细胞预测分析显示其定位于细胞核。蛋白质结构域预测发现,MiCTR1蛋白含有1个保守的PB1结构域,位于氨基酸序列189~285处。系统进化分析表明,MiCTR1与扁桃(Prunus dulcis)、桃(Prunus persica)、甜樱桃(Prunus avium)、杏(Prunus armeniaca)、梅(Prunus mume)的亲缘关系较近。荧光定量PCR结果显示,MiCTR1的相对表达量在芒果后熟过程中升高,且经1-MCP处理后其表达量下调。本研究结果为芒果后熟的分子机制提供基础。Mango is a typical respiration leap fruit,with vigorous postharvest metabolism and very sensitive to ethylene.CTR1 is a negative regulator of the ethylene signaling pathway and plays a central role in the ethylene signaling pathway.In order to study the possible role of mango CTR1 expression pattern in mango postharvest storage,a CTR gene(MiCTR1)was screened from the Tainung No.1)mango transcriptome database,and its encoded gene was used using biological methods.The protein was sequenced and analyzed,and the expression pattern of MiCTR1 gene during mango ripening and under 1-MCP treatment was analyzed.The results showed that the length of the open reading frame of MiCTR1 was 1551 bp,encoding 516 amino acids,The molecular formula of the predicted protein was C_(2503)H_(3932)N_(712)O_(797)S_(26),the total atomic number was 7970,and the weight of the protein was 57.58 kDa,the theoretical isoelectric point(pI)was 5.72,the fat coefficient was 70.78,and the overall average hydrophilicity was-0.596,with 111 phosphorylation sites,and the phosphorylation modification of serine was the main one,supplemented by threonine,had no transmembrane structure and no signal peptide.Subcellular prediction analysis showed that it was located in the nucleus.Protein domain prediction MiCTR1 protein contained a conserved PB1 domain located at amino acid sequence 189-285.Phylogenetic analysis showed that MiCTR1 was closely related to Prunus dulcis,Prunus persica,Prunus avium,Prunus armeniaca,Prunus mume.The results of fluorescence quantitative PCR showed that the relative expression of MiCTR1 increased during the ripening process of mango,and 1-MCP down-regulated its expression.In this study,the MiCTR1 gene was cloned in mango and its biological analysis was analyzed,and its expression pattern during postharvest storage was analyzed,which would provide a basis for the molecular mechanism of mango ripening.

关 键 词：芒果 乙烯响应转录因子 CTR 克隆 表达分析 

分 类 号：S667[农业科学—果树学]