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玉米咖啡酸O-甲基转移酶的全基因组鉴定与功能分析

张璐 赵月强 付家锋 张权 马春业 李英壮 张兰 王改革

张璐,赵月强,付家锋,等. 玉米咖啡酸O-甲基转移酶的全基因组鉴定与功能分析 [J]. 福建农业学报,2023,38(11):1259−1266 doi: 10.19303/j.issn.1008-0384.2023.11.001
引用本文: 张璐,赵月强,付家锋,等. 玉米咖啡酸O-甲基转移酶的全基因组鉴定与功能分析 [J]. 福建农业学报,2023,38(11):1259−1266 doi: 10.19303/j.issn.1008-0384.2023.11.001
ZHANG L, ZHAO Y Q, FU J F, et al. Identification and Functions of Caffeic Acid O-methyltransferase Genes in Maize [J]. Fujian Journal of Agricultural Sciences,2023,38(11):1259−1266 doi: 10.19303/j.issn.1008-0384.2023.11.001
Citation: ZHANG L, ZHAO Y Q, FU J F, et al. Identification and Functions of Caffeic Acid O-methyltransferase Genes in Maize [J]. Fujian Journal of Agricultural Sciences,2023,38(11):1259−1266 doi: 10.19303/j.issn.1008-0384.2023.11.001

玉米咖啡酸O-甲基转移酶的全基因组鉴定与功能分析

doi: 10.19303/j.issn.1008-0384.2023.11.001
基金项目: 国家玉米产业技术体系建设专项(nycytx-02)
详细信息
    作者简介:

    张璐(1986 —),女,硕士,助理研究员,主要从事作物遗传育种相关研究,E-mail: zhanglu19860320@163.com。张璐和赵月强对本研究具有同等贡献

    通讯作者:

    付家锋(1981 —),男,硕士,助理研究员,主要从事作物遗传育种相关研究,E-mail:fujiafeng11@126.com

  • 中图分类号: S513

Identification and Functions of Caffeic Acid O-methyltransferase Genes in Maize

  • 摘要:   目的  咖啡酸O-甲基转移酶(Caffeic acid O-methyltransferase,COMT)是褪黑素生物合成中的关键酶,在植物的生长、发育和抵御胁迫中发挥着重要的作用。本研究旨在探究COMT在玉米全基因组中的分布与盐胁迫响应情况,鉴定玉米咖啡酸O-甲基转移酶(COMT)基因。  方法  通过分析玉米COMT基因家族成员的结构特征、系统发育关系、基因结构、表达模式等,对ZmCOMTs基因进行系统分析,并通过过表达ZmCOMT12拟南芥对ZmCOMT的功能进行初步验证。  结果  在玉米全基因组中共鉴定出了28个COMT基因,根据系统发育分析分为2个分支(分支Ⅰ~II),大部分的COMT成员具有相似的motif组成和基因结构特征。利用玉米数据库分析ZmCOMTs的表达模式发现部分基因存在组织特异性,实时荧光定量PCR发现ZmCOMTs被盐胁迫诱导表达,说明ZmCOMTs成员参与了盐胁迫的调控,过表达ZmCOMT12拟南芥提高了褪黑素的含量和耐盐性。  结论  通过序列比对、亲缘关系、蛋白结构分析得到了假定的褪黑素合成基因ZmCOMT12,过表达拟南芥发现该基因与褪黑素的合成和耐盐性相关。
  • 图  1  28个玉米ZmCOMTs的系统发育分析

    A:28个玉米ZmCOMTs的系统发育分析;B:拟南芥AtCOMT与玉米ZmCOMTs的系统发育分析。

    Figure  1.  Phylogenetic analysis on 28 ZmCOMTs

    A: Phylogenetic analysis on 28 ZmCOMTs; B: phylogenetic analyses on AtCOMT and ZmCOMTs.

    图  2  玉米ZmCOMTs的进化关系及motif组成与基因结构的关联分析

    A:ZmCOMTs的系统发育分析;B:ZmCOMTs的蛋白motif组成;C:ZmCOMTs的基因结构。

    Figure  2.  Evolutionary relationships and association between motif composition and gene structure of ZmCOMTs in maize

    A: Phylogenetic analysis on ZmCOMTs; B: protein motif composition of ZmCOMTs; C: gene structure of ZmCOMTs.

    图  3  ZmCOMTs的染色体定位与基因复制

    Figure  3.  Chromosomal localization and gene duplication of ZmCOMTs

    图  4  ZmCOMTs在不同组织中的表达

    a:节间6~7;b:节间7~8;c:叶片区1;d:叶片区2;e:叶片区3;f:成熟叶片8;g:果皮糊粉27 d;h:初生根5 d;i:根皮层5 d;j:根伸长区5 d;k:根分生区5 d;l:次生根7~8 d。

    Figure  4.  Expressions of ZmCOMTs in different tissues

    a: internode 6–7; b: internode 7–8; c: Leaf zone 1; d: Leaf zone 2; e: Leaf zone 3; f: Mature leaf 8; g: Pericarp aleurone 27 d; h: Primary root 5 d; i: Root cortex 5 d; j: Root elongation zone 5 d; k: Root meristem zone 5 d; l: Secondary root 7–8 d.

    图  5  ZmCOMTs在盐胁迫下的表达

    *、**分别表示与对照相比差异显著(P<0.05)和极显著(P<0.01)。

    Figure  5.  Expression of ZmCOMTs under salt stress

    * and ** mean significant difference (P<0.05) and extremely signigifant difference (P<0.01) with control.

    图  6  ZmCOMT12的功能研究

    A:ZmCOMT12过表达植株的表型;B:ZmCOMT12的相对表达水平;C:褪黑素含量检测;D:POD酶活检测;E:SOD酶活检测。**P < 0.01。

    Figure  6.  Functions of ZmCOMT12

    A: Phenotypes of ZmCOMT12 overexpressed plants; B: relative expression of ZmCOMT12; C: melatonin content determination; D: POD enzyme activity determination; E: SOD enzyme activity assay. **: P<0.01.

    表  1  引物信息

    Table  1.   Information on primer

    基因
    Gene
    上游引物 5′-3′
    Forward primer 5′-3′
    下游引物 5′-3′
    Reverse primer 5′-3′
    ZmCOMT4TACCGTCTCCAGCATCTTCGTCCTTGAACCACTCTT
    ZmCOMT6TCTGTTCCACGAGAGCATTGTCATCACCGAAGTTACC
    ZmCOMT13ATGTTGGCGGAGATATGTTGATAGATGGCTCGGAAGG
    ZmCOMT22GCTGATGCCGTTCTTCTAAGCCGATACCATTCTTCCTC
    ZmCOMT23GGACAGCCACTTCATCATGCAGTTCCTCAGTATCTTCA
    ZmCOMT24CACGAACGCCATACTGAAGACTCATCATATCCAACTACCA
    ZmCOMT25GGCTGATGAACCTGAACACGCTCGGAATGTACTCAA
    Actin1GCAGGTATTGTGATGGATTCCATTAGGTGGTCGGTGAG
    下载: 导出CSV

    表  2  ZmCOMT家族成员的基本信息

    Table  2.   Basic information on ZmCOMTs

    基因
    Gene
    编号
    ID
    氨基酸数量
    Amino
    acid amount
    等电点
    Isoelectric
    point
    分子质量
    Molecular
    weight/Da
    ZmCOMT1Zm00001eb0186303656.0439538.69
    ZmCOMT2Zm00001eb0186403755.7240568.66
    ZmCOMT3Zm00001eb0186603727.2740187.3
    ZmCOMT4Zm00001eb0405703724.9740596.44
    ZmCOMT5Zm00001eb0902303656.3740018.77
    ZmCOMT6Zm00001eb0908303635.6540252.51
    ZmCOMT7Zm00001eb0927703665.5339785.78
    ZmCOMT8Zm00001eb1647503545.5239183.45
    ZmCOMT9Zm00001eb1682903645.5439700.6
    ZmCOMT10Zm00001eb1695203915.4142526.25
    ZmCOMT11Zm00001eb1705903715.6640477.9
    ZmCOMT12Zm00001eb1724203645.4839553.44
    ZmCOMT13Zm00001eb1909203745.1539590.18
    ZmCOMT14Zm00001eb1915904175.0745060.54
    ZmCOMT15Zm00001eb1981403645.5739200.14
    ZmCOMT16Zm00001eb1981503795.7741100.43
    ZmCOMT17Zm00001eb2022003655.2339345.35
    ZmCOMT18Zm00001eb2928403595.8338600.51
    ZmCOMT19Zm00001eb2928503555.3738002.74
    ZmCOMT20Zm00001eb2928803565.4238880.9
    ZmCOMT21Zm00001eb2928903585.4638816.89
    ZmCOMT22Zm00001eb3067003654.8739122.97
    ZmCOMT23Zm00001eb3531103645.6139605.53
    ZmCOMT24Zm00001eb3923403625.5439456.21
    ZmCOMT25Zm00001eb4001903655.6639687.64
    ZmCOMT26Zm00001eb4051604055.7643159.01
    ZmCOMT27Zm00001eb4051803975.7642617.41
    ZmCOMT28Zm00001eb4122603645.4439950.98
    下载: 导出CSV
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出版历程
  • 收稿日期:  2023-03-13
  • 修回日期:  2023-07-30
  • 网络出版日期:  2023-12-21
  • 刊出日期:  2023-11-28

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