华北农学报 ›› 2024, Vol. 39 ›› Issue (4): 10-19. doi: 10.7668/hbnxb.20194777

所属专题: 玉米 抗旱节水 生物技术

• 作物遗传育种·种质资源·生物技术 • 上一篇    下一篇

玉米ZmPAP26b基因的克隆及其在干旱胁迫下表达模式分析

郭昭阳1, 殷宇航2, 刘钰1, 解一桐1, 裴玉贺2, 宋希云2, 赵美爱1,3   

  1. 1 青岛农业大学 生命科学学院,山东 青岛 266109
    2 青岛市主要农作物种质创新与应用重点实验室,青岛农业大学 农学院,山东 青岛 266109
    3 东营青农大盐碱地高效农业技术产业研究院,山东 东营 257000
  • 收稿日期:2024-05-10 出版日期:2024-08-27
  • 通讯作者:
    赵美爱(1973—),女,吉林磐石人,副教授,博士,硕士生导师,主要从事玉米分子育种研究。
  • 作者简介:

    郭昭阳(1999—),男,山东潍坊人,在读硕士,主要从事玉米分子育种研究。

  • 基金资助:
    山东省农业良种工程(2019LZGC002)

Cloning and Expression Pattern Analysis of ZmPAP26b Gene in Maize under Drought Stress

GUO Zhaoyang1, YIN Yuhang2, LIU Yu1, XIE Yitong1, PEI Yuhe2, SONG Xiyun2, ZHAO Meiai1,3   

  1. 1 College of Life Sciences,Qingdao Agricultural University,Qingdao 266109,China
    2 Qingdao Key Laboratory of Germplasm Innovation and Application of Main Crops,College of Agronomy,Qingdao Agricultural University,Qingdao 266109,China
    3 Academy of Dongying Efficient Agricultural Technology and Industry on Saline and Alkaline Land in Collaboration with Qingdao Agricultural University,Dongying 257000,China
  • Received:2024-05-10 Published:2024-08-27

摘要:

干旱胁迫对玉米生长发育造成严重影响,从而导致玉米产量降低。紫色酸性磷酸酶是一种参与植物多种生理生化功能的磷脂酶蛋白,为进一步深入研究紫色酸性磷酸酶家族基因在玉米抗逆过程中的作用,探究了ZmPAP26b基因在干旱胁迫下响应模式,利用实时荧光定量分析模拟干旱条件下不同玉米自交系中的基因相对表达量;从玉米中克隆ZmPAP26b(GenBank:NC_050104.1),并对玉米、拟南芥、水稻、小麦、高粱和二穗短柄草中的PAP基因进行鉴定和生物信息学分析;同时构建原核过表达菌株进行功能验证。结果表明,干旱胁迫下该基因在耐旱材料中表达量下降,干旱敏感材料中表达量上升。该基因CDS全长1 431 bp,编码476个氨基酸。在6个物种中共发现228个PAP基因,系统发育分析发现,共分成4个亚家族。玉米中的19个PAP基因分布在9条染色体上并且具有相似的保守结构域,启动子顺式作用元件分析显示含有响应干旱和激素等元件。原核表达试验发现,含有重组质粒pET28a-ZmPAP26b的菌株在10%PEG-6000和15%PEG-6000模拟干旱胁迫下的生长与空载菌株都受到了抑制。综上所述,推测ZmPAP26b在干旱胁迫下呈负调控模式。

关键词: 玉米, ZmPAP, 干旱胁迫, 原核表达, 生物信息学

Abstract:

Drought stress has a serious effect on the growth and development of maize,which leads to a decrease of maize yield.Purple acid phosphatase is a phospholipase protein involved in many physiological and biochemical functions of plants.In order to further study the role of purple acid phosphatase family genes in the process of stress resistance of maize,this paper explored the response mode of ZmPAP26b gene under drought stress,and Real-time fluorescence Quantitavive analysis was used to analyze the relative gene expression in different maize inbreeding lines under simulated drought conditions;ZmPAP26b(GenBank:NC_050104.1)was cloned from maize,and PAP genes in Zea mays,Arabidopsis thaliana,Oryza sativa L.,Triticum aestivum L.,Sorghum bicolor and Brachypodium distachyon were identified and bioinformatic analysis was performed.Meanwhile,prokaryotic overexpression strains were constructed for functional verification.The results showed that the expression of this gene decreased in drought tolerant materials and increased in drought sensitive materials under drought stress.The CDS length of this gene was 1 431 bp,encoding 476 amino acids.A total of 228 PAP genes were found in six species,divided into 4 subfamilies by phylogenetic analysis.The 19 PAP genes in maize were distributed on 9 chromosomes and had similar conserved domains.Analysis of promoter cis-acting elements showed that they contained elements responding to drought and hormones.Prokaryotic expression experiments showed that the growth of strains containing the recombinant plasmid pET28a-ZmPAP26b was inhibited compared with non-loaded strains under 10% PEG-6000 and 15%PEG-6000 simulated drought stress.In summary,it is speculated that ZmPAP26b is negatively regulated under drought stress.

Key words: Maize, ZmPAP, Drought stress, Prokaryotic expression, Bioinformatics

引用本文

郭昭阳, 殷宇航, 刘钰, 解一桐, 裴玉贺, 宋希云, 赵美爱. 玉米ZmPAP26b基因的克隆及其在干旱胁迫下表达模式分析[J]. 华北农学报, 2024, 39(4): 10-19. doi: 10.7668/hbnxb.20194777.

GUO Zhaoyang, YIN Yuhang, LIU Yu, XIE Yitong, PEI Yuhe, SONG Xiyun, ZHAO Meiai. Cloning and Expression Pattern Analysis of ZmPAP26b Gene in Maize under Drought Stress[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(4): 10-19. doi: 10.7668/hbnxb.20194777.