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福建省一种蓝莓枝枯病病原鉴定

张小艳 陈细红 谢丽雪 张立杰 郑姗 李韬

张小艳,陈细红,谢丽雪,等. 福建省一种蓝莓枝枯病病原鉴定 [J]. 福建农业学报,2023,38(10):1214−1219 doi: 10.19303/j.issn.1008-0384.2023.10.010
引用本文: 张小艳,陈细红,谢丽雪,等. 福建省一种蓝莓枝枯病病原鉴定 [J]. 福建农业学报,2023,38(10):1214−1219 doi: 10.19303/j.issn.1008-0384.2023.10.010
ZHANG X Y, CHEN X H, XIE L X, et al. Pathogen of Branch Blight on Blueberry Plants in Fujian [J]. Fujian Journal of Agricultural Sciences,2023,38(10):1214−1219 doi: 10.19303/j.issn.1008-0384.2023.10.010
Citation: ZHANG X Y, CHEN X H, XIE L X, et al. Pathogen of Branch Blight on Blueberry Plants in Fujian [J]. Fujian Journal of Agricultural Sciences,2023,38(10):1214−1219 doi: 10.19303/j.issn.1008-0384.2023.10.010

福建省一种蓝莓枝枯病病原鉴定

doi: 10.19303/j.issn.1008-0384.2023.10.010
基金项目: 福建省科技计划公益类专项(2020R10280011)
详细信息
    作者简介:

    张小艳(1982 —),女,硕士,助理研究员,主要从事果树病虫害综合治理研究,E-mail:zxyan1204@163.com

    通讯作者:

    李韬(1969 —),男,学士,副研究员,主要从事果树遗传育种与病虫害研究,E-mail: leetao06@163.com

  • 中图分类号: S436.63

Pathogen of Branch Blight on Blueberry Plants in Fujian

  • 摘要:   目的  明确福建省三明地区蓝莓枝枯病的病原菌,为该病害的有效防治提供理论依据。  方法  采集受害蓝莓枝条样品,通过常规组织分离法进行病原菌分离纯化,采用针刺法进行致病性测定完成柯赫氏法则验证,利用形态学特征并联合分子生物学,明确致病菌的分类地位。  结果  从病组织中分离获得菌株KW1-4,将其回接至健康蓝莓枝条后,接种部位出现干枯病斑,再次分离获得的菌株与接种菌株一致,表明该菌株是引起蓝莓枝枯病的病原菌。菌株KW1-4菌落圆形,白色至灰色,有2种类型分生孢子,α型分生孢子长椭圆形至梭形,具2个明显油球,(4.77~7.63)µm×(1.55~2.71)µm,β型分生孢子线形,无油球,(11.95~19.65)µm×(1.05~1.94)µm。采用ITSTEF1-ɑβ-tubulin3段基因联合构建系统发育树,供试菌株KW1-4与Diaporthe australiana聚在同一分支。对比形态学特征,确定菌株KW1-4为间座壳属真菌Diaporthe australiana  结论  引起福建省三明市蓝莓枝枯病的病原菌是Diaporthe australiana,本研究首次报道Diaporthe australiana能够侵染蓝莓枝条。
  • 图  1  蓝莓枝枯病田间症状

    Figure  1.  Symptoms of blueberry branch blight in the field

    图  2  KW1-4的接种症状

    Figure  2.  Pathogenicity of KW1-4

    图  3  KW1-4 菌落形态及其分生孢子

    A:3 d ;B:9 d ;C:25 d ;D:α型分生孢子; E:β型分生孢子。

    Figure  3.  Colony morphology and conidia of KW1-4

    A: 3 d; B: 9 d; C: 25 d; D: alpha conidia; E: beta conidia.

    图  4  基于rDNA-ITSTEF1-ɑβ-tubulin基因序列构建的间座壳菌系统发育树

    Figure  4.  Phylogenetic tree of Diaporthe species constructed based on concatenated sequences of ITS, TEF1-ɑ, and β-tubulin

    表  1  PCR扩增ITSTEF1-ɑβ-tubulin片段的引物

    Table  1.   Primers for PCR amplification of fragments of ITS, TEF1-ɑ, and β-tubulin

    基因
    Gene
    引物
    Primer
    序列(5′-3′)
    Sequence(5′-3′)
    参考文献
    Reference
    ITSITS1TCCGTAGGTGAACCTGCGG[21]
    ITS4TCCTCCGCTTATTGATATGC
    TEF1-ɑEF-728FCATYGAGAAGTTCGAGAAGG[22]
    EF2GGARGTACCAGTSATCATGTT[23]
    β-tubulinBt2aGGTAACCAAATCGGTGCTGCTTTC[24]
    Bt2bACCCTCAGTGTAGTGACCCTTGGC
    下载: 导出CSV

    表  2  ITSTEF1-ɑβ-tubulin引物的PCR反应条件

    Table  2.   PCR reaction conditions for fragments of ITS, TEF1-ɑ, and β-tubulin

    扩增片段
    Amplified fragment
    预变性
    Pre-denaturation
    变性
    Denaturation
    退火
    Annealing
    延伸
    Extension
    循环次数
    Cycle number
    延伸
    Extension
    ITS94 ℃,3 min94 ℃,45 s52 ℃,45 s72 ℃,60 s3072 ℃,7 min
    TEF1-ɑ95 ℃,5 min95 ℃,60 s55 ℃,80 s72 ℃,90 s3572 ℃,5 min
    β-tubulin95 ℃,5 min95 ℃,60 s45 ℃,80 s72 ℃,90 s3572 ℃,5 min
    下载: 导出CSV
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出版历程
  • 收稿日期:  2023-06-30
  • 修回日期:  2023-07-26
  • 网络出版日期:  2023-10-25
  • 刊出日期:  2023-10-28

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