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园艺学报 ›› 2015, Vol. 42 ›› Issue (11): 2133-2143.doi: 10.16420/j.issn.0513-353x.2015-0269

• 果树 • 上一篇    下一篇

桃果实PpOAT 和PpP5CS 的克隆及其对外源#br# GABA 处理的响应表达

宋春波1,2,晁青青2,梁敏华2,邵佳蓉2,陈 伟2,杨震峰2,*   

  1. 1 上海海洋大学食品学院,上海 201306;2 浙江万里学院生物与环境学院,浙江宁波 315100
  • 出版日期:2015-11-25 发布日期:2015-11-25
  • 基金资助:

    国家自然科学基金项目(31371866);浙江省自然科学基金项目(LQ15C00004);浙江省教育厅攀登计划项目(PD2013328)

Molecular Cloning of PpOAT and PpP5CS and Their Expression Profile in#br# Responses to Exogenous GABA in Postharvest Peach Fruit

SONG Chun-bo1,2,CHAO Qing-qing2,LIANG Min-hua2,SHAO Jia-rong2,CHEN Wei2,and YANG#br# Zhen-feng2,*   

  1. 1College of Food Science and Technology,Shanghai Ocean University,Shanghai 201306,China;2College of Biological
    and Environmental Sciences,Zhejiang Wanli University,Ningbo,Zhejiang 315100,China
  • Online:2015-11-25 Published:2015-11-25

摘要:

利用RT-PCR 结合RACE 技术从‘玉露’桃果实中克隆得到Δ1–吡咯啉–5–羧酸合成酶
(P5CS)和鸟氨酸转氨酶(OAT)基因的全长cDNA 序列,分别命名为PpP5CS 和PpOAT(GenBank 登
录号分别为KP973954 和KP973956)。PpP5CS 全长2 511 bp,开放阅读框为2 151 bp,编码717 个氨基酸
组成的蛋白质多肽,5′-UTR 长度为123 bp,3′-UTR 序列长度为235 bp;PpOAT 全长1 686 bp,开放阅读
框1 416 bp,编码472 个氨基酸,5′-UTR 长度为151 bp,3′-UTR 序列长度为119 bp。进化树分析发现,
PpOAT 和PpP5CS 与湖北海棠的同源性最高,与MhOAT 和MhP5CS 的相似性分别达到88%和91%。采
用荧光定量PCR 分析了外源5 mmol · L-1 GABA 处理对桃果实0 ℃贮藏期间果实冷害发生和内源脯氨酸
合成关键基因PpOAT 和PpP5CS 表达的影响。结果表明,GABA 处理能显著抑制‘玉露’桃果实0 ℃贮
藏期间果肉出汁率的下降,减轻冷害。通过上调果实中PpOAT 和PpP5CS 的表达,提高内源脯氨酸的合
成和积累,进而增强了果实抵抗低温胁迫的能力,可能是外源GABA 处理减轻桃果实冷害的重要原因。

关键词: 桃, 果实, PpP5CS, PpOAT, 脯氨酸, 冷害

Abstract:

Full-length cDNAs of Δ1-pyrroline-5-carboxylate synthetase(PpP5CS)and ornithine
aminotransferase(PpOAT)were isolated from peach fruit using degenerate RT-PCR and RACE(rapid
amplification of cDNA ends)method. The sequences of these two genes were then deposited in GenBank
database with the accession number KP973954 and KP973956,respectively. PpP5CS was 2 511 bp in full
length and encoded a predicted protein of 717 amino acids(ORF length 2 151 bp),and flanked by 123
nucleotides at the 5′-UTR and 235 nucleotides at the 3′-UTR. The full length of PpOAT was 1 686 bp with
ORF 1 416 bp,5′-UTR 151 bp and 3′-UTR 119 bp,which encoded a deduced polypeptide of 472 amino
acids. Phylogenetic analysis indicated that PpOAT and PpP5CS shared high similarity with otherplants with 88% and 91% homology with MhOAT and MhP5CS,respectively. Furthermore,the effect of
5 mmol · L-1 γ-aminobutyric acid treatment on chilling injury incidence and PpOAT and PpP5CS
expression,which were involved in proline biosynthesis,in postharvest peach fruit stored at 0 ℃ was
investigated. The results showed that 5 mmol · L-1 γ-aminobutyric acid treatment significantly inhibited the
decline of extractable juice,and alleviated chilling injury in peach fruit during cold storage. Meanwhile,
the expression of these two genes in peach fruit was induced by the treatment during cold storage. These
results suggested that γ-aminobutyric acid treatment could increase proline biosynthesis and accumulation
by up-regulating the expression levels of PpOAT and PpP5CS thereby promote chilling tolerance in
cold-stored peach fruit.

Key words: peach, fruit, PpP5CS, PpOAT, proline, chilling injury

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