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草地早熟禾及其矮化突变材料锈病病原菌鉴定及抗病机制初探

甘露 苏浩天 凌欣闻 尹淑霞

甘露, 苏浩天, 凌欣闻, 尹淑霞. 草地早熟禾及其矮化突变材料锈病病原菌鉴定及抗病机制初探[J]. 北京林业大学学报, 2017, 39(3): 87-92. doi: 10.13332/j.1000-1522.20160315
引用本文: 甘露, 苏浩天, 凌欣闻, 尹淑霞. 草地早熟禾及其矮化突变材料锈病病原菌鉴定及抗病机制初探[J]. 北京林业大学学报, 2017, 39(3): 87-92. doi: 10.13332/j.1000-1522.20160315
GAN Lu, SU Hao-tian, LING Xin-wen, YIN Shu-xia. Rust pathogen identification and mechanism of disease-resistance research on Kentucky bluegrass dwarf mutant[J]. Journal of Beijing Forestry University, 2017, 39(3): 87-92. doi: 10.13332/j.1000-1522.20160315
Citation: GAN Lu, SU Hao-tian, LING Xin-wen, YIN Shu-xia. Rust pathogen identification and mechanism of disease-resistance research on Kentucky bluegrass dwarf mutant[J]. Journal of Beijing Forestry University, 2017, 39(3): 87-92. doi: 10.13332/j.1000-1522.20160315

草地早熟禾及其矮化突变材料锈病病原菌鉴定及抗病机制初探

doi: 10.13332/j.1000-1522.20160315
基金项目: 

国家自然科学基金项目 31302016

深圳市科技计划项目 JSGG20160229155434792

详细信息
    作者简介:

    甘露,博士生。主要研究方向:草坪草分子生理。Email:ganlu2016@bjfu.edu.cn  地址:100083  北京市海淀区清华东路35号北京林业大学草坪研究所

    责任作者:

    尹淑霞,教授,博士生导师。主要研究方向:草坪科学与管理。Email: yinsx369@bjfu.edu.cn  地址:同上

  • 中图分类号: S688.4

Rust pathogen identification and mechanism of disease-resistance research on Kentucky bluegrass dwarf mutant

  • 摘要: 锈病是草坪草的重要病害,且病原菌种类多,鉴定困难。本研究以草地早熟禾野生型和矮化突变材料中的感病株系为对象,通过观察锈菌的形态特征,并结合ITS和β-tubulin基因序列分析,其中ITS序列的同源比对分析和β-tubulin基因的分子进化分析结果表明,该菌种与小麦禾柄锈菌的同源关系接近,所以初步认定该菌株属于禾柄锈菌,这是国内对禾柄锈菌引起草地早熟禾锈病的首次报道。同时,我们对病原菌诱导后草地早熟禾PR1LNPR1L基因的表达变化和PRs蛋白表达进行了研究,发现PR1LNPR1L基因在病菌诱导12 h时的表达量达到了峰值,且在矮化突变植株(A16)中的相对表达量分别达到了8.8-fold、4.5-fold,均大于在对照植株(WT)中的表达量。另外植物的PRs蛋白在禾柄锈菌侵染植物第8天后的表达量明显高于未侵染的植株。初步对草地早熟禾锈病的防御机制进行了探讨,为今后开展草地早熟禾锈病的预防及抗病育种研究奠定基础。

     

  • 图  1  感病植株的发病症状和锈菌菌株夏孢子形态的光学显微镜观察

    Figure  1.  Symptoms of Kentucky bluegrass leaves infected by urediospore of rust and morphological investigation of the urediospore under optical microscope

    图  2  供试菌株及Puccinia相关菌株β-tubulin基因的系统发育树

    候选锈菌菌株为供试材料的锈病病原菌。进化树右端的字母和数字代表GenBank中的序列号,分支上的数字代表自举的可信度,0.02为序列分歧度。

    Figure  2.  Phylogenetic tree of isolate strain and their relatives in Puccinia

    Putative Puccinia is the isolate rust strain. Numbers in parentheses represent the sequences' accession number in Gen Bank. The number at each branch points is the percentage supported by bootstrap. Bar, 2% sequence divergence.

    图  3  草地早熟禾及其矮化突变材料受禾柄锈菌侵染后PR1LNPR1L基因的表达变化情况

    Figure  3.  Expression pattern of PR1L, NPR1L induced by Puccinia graminis in Kentucky bluegrass wild type (WT) and its dwarf mutants (A16)

    图  4  草地早熟禾野生型(WT)及其矮化突变(A16)植株接种禾柄锈菌前后的病程相关蛋白电泳分析

    1.蛋白marker;2.未接种的WT样品;3.未接种的A16样品;4.接种后的A16样品;5.接种后的WT样品。

    Figure  4.  12% SDS-PAGE analysis of pathogenesis-related proteins extracted from leaves of WT and A16 inoculated with/without Puccinia graminis

    1, protein marker; 2, the A16 sample inoculated without Puccinia graminis; 3, the WT sample inoculated without Puccinia graminis; 4, the A16 sample inoculated with Puccinia graminis; 5, the WT sample inoculated with Puccinia graminis.

    表  1  实验所需引物的序列

    Table  1.   List of primer sequences used in this study

    引物名称
    Primer name
    引物序列
    Primer sequence(5′—3′)
    ITS1TCCGTAGGTGAACCTGCGG
    ITS4TCCTCCGCTTATTGATATGC
    Tubulin-FATGGAYTCGGTYCGATCTGGCG
    Tubulin-RCYTCTCCRGTGTACCAATGCAGG
    PR1L-qFCGCTACGCCCGCTCCC
    PR1L-qRGCCCCTCGTCCACCCA
    NPR1L-qFCAAGGAAGGGCAGACTAA
    NPR1L-qRGCAGCGATGTGAAGAACA
    Actin-qFTTGACTGAGAGGGGCT
    Actin-qRTCATACGGTCTGCGAT
    下载: 导出CSV

    表  2  草地早熟禾的抗病相关的候选基因筛选

    Table  2.   List of candidate disease-resistant gene studied in Kentucky bluegrass clones

    基因名称
    Gene ID
    候选基因注释
    Candidate gene annotation
    开放阅读框序列长度
    ORF length/bp
    同源基因编号
    GenBank accession of homologs
    同源物种
    Allied species
    PR1Lpathogenesis-related protein 1A-like552XM_004975168Setaria italica
    NPR1LNPR1-like 1 protein1 758JX424315硬粒小麦Triticum durum
    下载: 导出CSV
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出版历程
  • 收稿日期:  2016-09-29
  • 修回日期:  2016-11-19
  • 刊出日期:  2017-03-01

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