猕猴桃多胺氧化酶基因家族的鉴定及表达分析

doi:10.15933/j.cnki.1004-3268.2023.10.012

摘要/Abstract

摘要： 通过鉴定猕猴桃多胺氧化酶（PAO）基因家族成员，分析其在猕猴桃果实生长发育、成熟及储藏过程中的表达变化，发掘与猕猴桃果实成熟、软化相关的重要候选基因，为调控猕猴桃鲜果的发育及采后保鲜提供参考。以拟南芥AtPAO、水稻OsPAO蛋白序列作为参考序列，利用BLAST程序分别从中华猕猴桃、毛花猕猴桃、软枣猕猴桃基因组或转录组中筛选和鉴定PAO家族成员；利用Compute pI/Mw分析蛋白质分子质量、理论等电点，利用SignalP-4.1、TMHMM-2.0预测信号肽和跨膜区，利用WoLF PSORT进行亚细胞定位预测；利用MEGA 6构建系统发育进化树，利用MEME进行保守基序分析，利用TBtools分析PAO基因的共线性关系；基于转录组数据和荧光定量PCR技术，分析PAO基因的组织表达情况及其在猕猴桃果实生长发育、成熟及储藏过程中的表达变化。结果表明，中华猕猴桃、毛花猕猴桃、软枣猕猴桃中各存在4、4、5个PAO基因，分别是AcPAO1—4、AePAO1—4、AaPAO1—5。AcPAO、AePAO 位于中华猕猴桃、毛花猕猴桃的8条染色体上，基因长度为1 722~9 837 bp。AcPAO、AePAO、AaPAO 编码的氨基酸数目为420~496个，分子质量为45.74~55.92 ku，等电点为5.29~5.79，均不含有信号肽和跨膜区；AcPAO、AaPAO主要定位在过氧化物酶体，而AePAO分布范围较广；依据系统进化关系，13个猕猴桃PAO蛋白分属于3个类群，同一类群含有相似的motif分布及保守结构域；AcPAO、AePAO 间存在4对共线性基因，它们在进化中经历了纯化选择。AcPAO3的表达量随猕猴桃果实成熟、软化呈现先升高后降低趋势，且能够响应不同储藏温度；AaPAO为组成型表达，多数成员的表达量在果实储藏过程中亦呈现先升高后降低的趋势。

关键词: 猕猴桃, 多胺氧化酶（PAO）, 基因家族, 生物信息学分析, 表达分析

Abstract: The aim of this study was to identify members of polyamine oxidase（PAO） family from kiwifruit，to analyze their expression changes during growth，development，maturation and storage of kiwifruit，to mine important candidate genes involved in fruit maturation and softening，and to provide reference for regulating development and post‑harvest preservation of kiwifruit.The PAO proteins of Arabidopsis thaliana and Oryza sativa were used as probes，and the family members in Actinidia chinensis，Actinidia eriantha and Actinidia arguta were screened out by BLAST.Compute pI/Mw，SignalP‑4.1，TMHMM‑2.0，WoLF PSORT，MEGA 6，MEME and TBtools were used for analyzing and predicting the molecular weight，theoretical isoelectric point，signal peptide，transmembrane region，subcellular localization，phylogenetic relationship，conserved motifs and syntenic relationship，respectively.Expression analysis of PAO genes among different tissues and biological processes of kiwifruit were analyzed based on transcriptome data and qPCR.The results showed that four，four and five PAO genes were identified in Actinidia chinensis，Actinidia eriantha and Actinidia arguta，named AcPAO1—4，AePAO1—4，AaPAO1—5，respectively.AcPAO and AePAO were distributed on eight chromosomes，with the length of 1 722—9 837 bp.The amino acid number of PAO proteins ranged from 420 aa to 496 aa，the molecular weight was 45.74—55.92 ku，and the isoelectric point was 5.29—5.79.Neither signal peptides nor transmembrane regions were included in PAO proteins. AcPAO and AaPAO were mainly located in peroxisome，while AePAO widely distributed in cells.Based on phylogenetic analysis，thirteen kiwifruit PAO proteins were divided into three groups，and there were similar motif distribution patterns and conservative domains in members which were clustered into the same group. Four syntenic gene pairs were identified among AcPAO and AePAO genes.The Ka/Ks values of syntenic gene pairs were less than 1，suggesting that they underwent purifying pressure during the evolution process.The expression of AcPAO3 showed a trend of increase at first and then decrease with maturation and ripening of kiwifruit.In addition，AcPAO3 could respond to different storage temperature to some extent.AaPAO genes were expressed in all the tested tissues，the expression during storage of which mostly also showed the same trend as that of AcPAO3.

Key words: Kiwifruit, Polyamine oxidase（PAO）, Gene family, Bioinformatic analysis, Expression analysis

中图分类号:

S663.4

谢羽恬, 张新业, 苏彦苹, 吴双, 张泽银, 褚卓栋, 冯雪, 孙艳香. 猕猴桃多胺氧化酶基因家族的鉴定及表达分析[J]. 河南农业科学, 2023, 52(10): 109-119.

XIE Yutian, ZHANG Xinye, SU Yanping, WU Shuang, ZHANG Zeyin, CHU Zhuodong, FENG Xue, SUN Yanxiang. Identification and Expression Analysis of PAO Gene Family in Kiwifruit[J]. Journal of Henan Agricultural Sciences, 2023, 52(10): 109-119.

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