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园艺学报 ›› 2018, Vol. 45 ›› Issue (2): 205-216.doi: 10.16420/j.issn.0513-353x.2017-0179

• 研究论文 •    下一篇

苹果BR信号转录因子基因MdBZR1的克隆及表达分析

郑立伟,赵才德,宋春晖,马娟娟,张  东,樊  胜,张丽之,韩明玉   

  1. 西北农林科技大学园艺学院,陕西杨凌 712100
  • 出版日期:2018-02-25 发布日期:2018-02-25
  • 基金资助:

    国家现代农业产业技术体系建设专项资金项目(CARS-28);陕西省科技统筹项目(2015NY114;2016KTZDNY01-10);中央高校基本科研业务费项目(2452015291);国家苹果改良中心杨凌分中心项目;陕西省果业发展协同中心项目;中国博士后基金项目 (2015K3080215822)

Cloning and Expression of Transcription Factor Gene BZR1 in Malus × domestica

ZHENG Liwei,ZHAO Caide,SONG Chunhui,MA Juanjuan,ZHANG Dong,FAN Sheng,ZHANG Lizhi,and HAN Mingyu   

  1. College of HorticultureNorthwest A & F UniversityYanglingShaanxi 712100China
  • Online:2018-02-25 Published:2018-02-25

摘要:

从矮化苹果砧木Malling 9(M9)中分离1个BR信号转录因子BRASSINAZOLE-RESISTANT 1(BZR1)基因,命名为MdBZR1,其开放阅读框为888 bp,编码295个氨基酸,有5个保守结构域:核定位信号结构域NLS(MARRKPSWRERENNRRTERRR)、氮(N)端结构域(NLPKHCDNNEVLKALCLQ AGWTVEDDGT)、BRASSINOSTEROID-INSENSITIVE 2(BIN2)磷酸化结构域(STKISPYSSLNPSPIPS YOVSPSSSSYPSPTR)、PEST结构域(PTAATIPECDESDASTVDSGQ)和碳(C)端保守结构域(VKPWIGEK IHEVGLDDLELTLGNGKA)。系统进化树分析显示,MdBZR1与美国National Center for Biotechnology Information(NCBI)数据库中注释的苹果BZR1基因亲缘关系最近。MdBZR1在M9不同组织均有表达,在顶梢中的表达水平最高;外源BR和生长素处理促进苹果幼苗(M9和平邑甜茶)株高增长,MdBZR1表达量增加;GA处理虽也促进株高增长,但对MdBZR1的表达影响不显著。此外,半矮化砧穗组合长富2号/MM106和乔化砧穗组合长富2号/长富2号接穗中MdBZR1的表达量明显高于矮化砧穗组合长富2号/M9。因此,MdBZR1很可能对苹果树株高具有重要调控作用。

关键词: 苹果, MdBZR1, 基因克隆, 表达分析, 油菜素甾醇类

Abstract:

In order to study the regulation function of MdBZR1 on apple plant height,a BR signal transcription factorgene BRASSINAZOLE-RESISTANT 1BZR1)was cloned from apple dwarf rootstock Malling 9(M9),which was named as MdBZR1. MdBZR1contains an open reading frame(888 bp)and encodes a protein of 295 amino acids. Bioinformatics analysis showed that MdBZR1 shared five conserved domains:nuclear localization signal(NLS)(MARRKPSWRERENNRRTERRR),nitrogen terminal domain(N)(NLPKHCDNNEVLKALCLQAGWTVEDDGT),BIN2 phosphorylation domain (STKISPY 

SSLNPSPIPSYOVSPSSSSYPSPTR),PEST motif(PTAATIPECDESDASTVDSGQ)and a carboxyl- terminal(C(VKPWIGEKIHEVGLDDLELTLGNGKA). A phylogenetic analysis of BZR1 genes indicated that MdBZR1 shared high homologous amino acid sequences with apple BZR1 gene annotated in National Center for Biotechnology Information(NCBI)database. The expression pattern of MdBZR1 gene in different tissues of M9 was characterized. Results indicated that MdBZR1 transcript was detected in shoot tip,leaf,root,xylem and phloem of main stem,and the highest level appeared in shoot tip. Exogenous brassinosteroid(BR)and auxin increased primary shoot length of M9as well as Pingyitiancha(Malus hupehensis)and significantly induced the expression of MdBZR1. Primary shoot length was also increased by GA treatment,while MdBZR1 was neither induced nor repressed by GA obviously. In addition,MdBZR1 exhibited higher expression level in Nagafu/MM106 and Nagafu/Nagafu than Nagafu/M9. These results suggest that MdBZR1 may be involved in the regulation of apple tree size.

Key words: apple, MdBZR1, gene clone, expression analysis, brassinosteroids

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