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植物研究 ›› 2020, Vol. 40 ›› Issue (6): 913-922.doi: 10.7525/j.issn.1673-5102.2020.06.014

• 研究报告 • 上一篇    

澳洲坚果MiMYB2基因克隆及结构与功能分析

王文林1, 陈海生1(), 郑树芳1, 樊松乐2,3, 王立丰3, 谭秋锦1, 覃振师1, 黄锡云1, 贺鹏1, 汤秀华1, 许鹏1   

  1. 1.广西南亚热带农业科学研究所,龙州 532400
    2.海南大学热带作物学院,海南省热带生物资源可持续利用重点实验室,海口 570228
    3.农业农村部橡胶树生物学与遗传资源利用重点实验室;省部共建国家重点实验室培育基地-海南省热带作物栽培生理学重点实验室;农业农村部儋州热带作物科学观测实验站;中国热带农业科学院橡胶研究所,海口 571101
  • 收稿日期:2019-12-05 出版日期:2020-11-20 发布日期:2020-11-04
  • 通讯作者: 陈海生 E-mail:chenhs6179@163.com
  • 作者简介:王文林(1980—),高级农艺师,主要从事热带果树遗传育种方面的研究。
  • 基金资助:
    广西科技基地和人才专项(桂科AD18281087);崇左市科技计划项目(崇科攻2019027);广西农业科学院科技发展基金(桂农科2019ZX2015);广西农科院基本科研业务费项目(桂农科2020YM46)

Cloning,Structure and Function Analysis of MiMYB2 Gene from Macadamia integrifolia

Wen-Lin WANG1, Hai-Sheng CHEN1(), Shu-Fang ZHENG1, Song-Le FAN2,3, Li-Feng WANG3, Qiu-Jin TAN1, Zhen-Shi QIN1, Xi-Yun HUANG1, Peng HE1, Xiu-hua TANG1, Peng XU1   

  1. 1.Guangxi South Subtropical Agricultural Research Institute,Longzhou 532400
    2.Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresource,College of Tropical Crop,Hainan University,Haikou 570228
    3.Key Laboratory of Biology and Genetic Resources of Rubber Tree,Ministry of Agriculture and Rural Affairs,P. R. China;State Key Laboratory Incubation Base for Cultivation & Physiology of Tropical Crops;Danzhou Investigation & Experiment Station of Tropical Crops,Ministry of Agriculture and Rural Affairs,P. R. China; Rubber Research Institute,Chinese Academy of Tropical Agricultural Sciences,Haikou 571101
  • Received:2019-12-05 Online:2020-11-20 Published:2020-11-04
  • Contact: Hai-Sheng CHEN E-mail:chenhs6179@163.com
  • About author:WANG Wen-Lin(1980—),senior agronomist, mainly engaged in the genetic and breeding research of tropical fruit trees.
  • Supported by:
    Guangxi Science and Technology Base and Talent Special Project(Guike AD18281087);Chongzuo Science and Technology Plan Project(Chongkegong(2019027);2019039;Guangxi Academy of Agricultural Sciences Science and Technology Development Fund (Guinongke 2019ZX2015,2019ZX17,2020YM148);Basic Scientific Research Fund of Guangxi Academy of Agricultural Sciences(Guinongke 2020YM46,2020YM141)

摘要:

为研究R2R3-MYB转录因子家族成员在澳洲坚果生长发育和产量形成中的作用机制,利用PCR技术从澳洲坚果品种“桂热一号”叶片中克隆MiMYB2基因并采用生物信息学对其结构及功能进行分析。结果表明,克隆获得的MiMYB2(NCBI登录号:MN254976)基因cDNA序列全长1 210 bp,开放阅读框(ORF)1 002 bp,编码333个氨基酸。MiMYB2编码一个无跨膜结构、无信号肽且定位于细胞核的不稳定亲水蛋白,含两个SANT保守结构域,属于R2R3-MYB家族。BLAST分析发现MiMYB2与荷花NnMYB3-like氨基酸序列同源性最高。系统进化分析将MiMYB2与AtMYB17、AtMYB106和AtMYB16聚类为S9亚族。通过转录组数据分析MiMYB2基因在“桂热一号”和“695”品种澳洲坚果枝条、花和叶片中的表达模式,表明MiMYB2在“桂热一号”品种花中的表达量最低,“695”品种花中的表达量最高。推测MiMYB2与澳洲坚果花生长发育密切相关。本研究为阐明MiMYB2基因在澳洲坚果生长发育和产量形成中的作用机制提供理论参考。

关键词: 澳洲坚果, MiMYB2, 基因克隆, 生物信息学分析

Abstract:

In order to study the mechanism of R2R3-MYB transcription factor gene family member in growth,development and yield formation in Macadamia integrifolia,the MiMYB2 gene was cloned from the leaves of M.integrifolia “GUIRE1” cultivar by PCR and its structure and function were analyzed by bioinformatics method. The cloned cDNA sequence of MiMYB2(NCBI accession number:MN254976) gene is 1 210 bp in length,has 1002 bp open reading frame(ORF),encoding 333 amino acids. MiMYB2,an unstable hydrophilic protein that has no transmembrane structure,no signal peptide and localizes in the nucleus,has two SANT conserved domains belonging to the R2R3-MYB family. BLAST analysis showed that the MiMYB2 share high amino acid sequence identity with Lotus NnMYB3-like protein. Phylogenetic analysis clustered MiMYB2 and AtMYB17,AtMYB106 and AtMYB16 into S9 subfamily. The transcriptome data were used to analyze the expression patterns of MiMYB2 gene in branch,flower and leave of M.integrifolia “GUIRE1” and “695” cultivar. Results indicated that MiMYB2 has the lowest expression in the flower of “GUIRE1”,and the expression level of “695” is the highest. It is speculated that MiMYB2 is closely related to M.integrifolia growth and development. This study would provide a theoretical reference for clarification the role MiMYB2 in growth,development and yield formation mechanism of M.integrifolia.

Key words: Macadamia integrifolia, MiMYB2, gene cloning, bioinformatics analysis

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