Journal of Nuclear Agricultural Sciences ›› 2021, Vol. 35 ›› Issue (6): 1273-1280.DOI: 10.11869/j.issn.100-8551.2021.06.1273

• Induced Mutations for Plant Breeding·Agricultural Biotechnology • Previous Articles     Next Articles

Cloning of Peach PpNAC19 and Its Regulation on PpCCD4 Promoter Activity

QIN Juan1,2, YU Fan1,2, LIU Lu1, ZHU Tingting1, CHEN Wei1, CAO Shifeng1, YANG Zhenfeng1, SHI Liyu1,*   

  1. 1Zhejiang Wanli University, Ningbo, Zhejiang 315100;
    2Shanghai Ocean University, Shanghai 201306
  • Received:2020-03-09 Online:2021-06-10 Published:2021-04-16

PpNAC19的克隆及其对PpCCD4启动子活性的调节分析

秦娟1,2, 余凡1,2, 刘璐1, 朱婷婷1, 陈伟1, 曹士锋1, 杨震峰1, 施丽愉1,*   

  1. 1浙江万里学院,浙江 宁波 315100;
    2上海海洋大学,上海 201306
  • 通讯作者: *施丽愉,女,讲师,主要从事农产品贮藏与加工方面研究。E-mail:shiliyu@163.com
  • 作者简介:秦娟,女,主要从事农产品贮藏与加工方面研究。E-mail:1005068921@qq.com
  • 基金资助:
    宁波市自然科学基金项目(2019A610432),浙江省“生物工程”一流学科自设课题项目(ZS2019007),水蜜桃保鲜贮运与深加工关键技术研究与示范(2019C10069),水蜜桃保鲜贮运与深加工关键技术研究与示范(2019C10069)

Abstract: In order to explore the regulation effect of NAC gene on carotenoid accumulation, this study cloned PpNAC19 gene from peach. Sequence analysis showed that the open reading frame sequence of PpNAC19 (XP_007223324.1) was 867 bp in length, encoding 288 amino acids with a NAM domain. The predicted molecular weight of the protein was 33.2 kDa, and the average hydrophobic coefficient was-0.719, which was a hydrophilic protein. Subcellular localization predicted that PpNAC19 protein was localized in the nucleus. Amino acid sequence alignment and cluster analysis showed that PpNAC19 had a close relationship with Prunus dulcis (VVA20514.1). The results of qRT-PCR revealed that PpNAC19 was expressed in different organs of peach, and the expression was the highest in hard-core fruit. During the whole fruit ripening process, the carotenoid content in yellow fleshed peach was obviously higher than that in white fleshed peach. The expression of PpNAC19 gene was higher in yellow fleshed peach, while the expression of carotenoid degradation gene PpCCD4 in yellow fleshed peach is significantly lower than that in white fleshed peach. The results of double luciferase test showed that PpNAC19 could transcriptionally inhibit the activity of PpCCD4 promoter. This experiment provides a theoretical basis for further research on the function of PpNAC19 gene in the carotenoid metabolism of peach fruits.

Key words: peach, NAC, expression analysis, carotenoids

摘要: 为探究桃NAC基因对类胡萝卜素积累的调控作用,本研究从桃中克隆得到PpNAC19基因。序列分析表明,PpNAC19(XP_007223324.1)开放阅读框为867 bp,编码288个氨基酸,预测蛋白质分子量为33.2 kDa,平均疏水系数为-0.719,属于亲水性蛋白,含有一个典型的NAM结构域。亚细胞定位预测PpNAC19蛋白定位在细胞核中。氨基酸序列比对与聚类分析表明,PpNAC19与巴旦杏(Prunus dulcis,VVA20514.1)亲缘关系较近。荧光定量PCR结果表明,PpNAC19在桃不同组织中均有表达,且在硬核期果实中表达量最高。在果实成熟过程中,黄肉桃果实中类胡萝卜素含量显著高于白肉桃果实。PpNAC19基因在黄肉桃中表达量较高,类胡萝卜素降解基因PpCCD4在黄肉桃中表达量显著低于白肉桃。双荧光素酶试验结果表明,PpNAC19能够转录抑制PpCCD4启动子活性。本试验为进一步研究PpNAC19基因在桃果实类胡萝卜素代谢过程中的功能提供了理论依据。

关键词: 桃, NAC, 表达分析, 类胡萝卜素