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白术根腐病病原菌分离鉴定、生物学特性及植物源农药筛选

谷清义 张耀洲 吴晓亚 黄雅琴 乔新荣 申君

谷清义,张耀洲,吴晓亚,等. 白术根腐病病原菌分离鉴定、生物学特性及植物源农药筛选 [J]. 福建农业学报,2024,39(X):1−9
引用本文: 谷清义,张耀洲,吴晓亚,等. 白术根腐病病原菌分离鉴定、生物学特性及植物源农药筛选 [J]. 福建农业学报,2024,39(X):1−9
GU Q Y, ZHANG Y Z, Wu X Y, et al. Isolation, identification, biological characteristics, and indoor drug screening of the pathogenic bacteria of Atractylodes macrocephala root rot disease [J]. Fujian Journal of Agricultural Sciences,2024,39(X):1−9
Citation: GU Q Y, ZHANG Y Z, Wu X Y, et al. Isolation, identification, biological characteristics, and indoor drug screening of the pathogenic bacteria of Atractylodes macrocephala root rot disease [J]. Fujian Journal of Agricultural Sciences,2024,39(X):1−9

白术根腐病病原菌分离鉴定、生物学特性及植物源农药筛选

基金项目: 河南省高等学校重点科研项目(21B210009);信阳农林学院青年教师科研基金(QN2021060);河南省科技攻关项目(222102110247)
详细信息
    作者简介:

    谷清义(1982 — ),男,硕士,助教,主要从事农药研究,E-mail:271490029@qq.com

    通讯作者:

    申君(1984 — ),女,博士,副教授,主要从事农药研究,E-mail:shenjun996@163.com

  • 中图分类号: S436

Isolation, identification, biological characteristics, and indoor drug screening of the pathogenic bacteria of Atractylodes macrocephala root rot disease

  • 摘要:   目的  明确河南信阳地区白术根腐病病原菌种属,研究其生物学特性并筛选可用于防控其病原菌的植物源农药。  方法  从种植区采集病株,室内分离病原菌并纯化,利用形态学和多基因联合分析对病原菌进行鉴定,并对病原菌的生物学特性进行研究,利用菌丝生长速率法评价3种植物源杀菌剂对其菌落的抑制作用。  结果  该病原菌菌丝白色,孢子呈卵圆形,两端稍尖,大型分生孢子3~5隔,大小为(7~10) μm×(3~4) μm,小型分生孢子2隔或无格,大小为(3~5) μm×(1~2) μm,多基因联合分析表明该病原菌与藤仓镰刀菌(Fusarium fujikuroi)同源性高,结合形态学特征与分子生物学分析将信阳白术根腐病病原菌鉴定为藤仓镰刀菌(Fusarium fujikuroi),这是藤仓镰刀菌引起白术根腐病的首次报道。生物学特性研究显示,该病原菌最适培养条件为温度28 ℃、pH为7,最适生长碳源为蔗糖,最适生长氮源为硝酸钾。3种植物源杀菌剂中,0.3%丁子香酚可溶液剂的EC50为6.906 mg·L−1,高于其他供试药剂,对病原菌的毒力最强。  结论  河南信阳地区白术根腐病病原菌为藤仓镰刀菌,最适培养条件为温度28 ℃、pH为7,最适生长碳源为蔗糖,最适生长氮源为硝酸钾。该研究结果为信阳地区白术根腐病科学防控提供了依据。
  • 图  1  白术正常植物与患根腐病植株

    注:图中1-6分别为正常植株、回接后患病植株、田间患病植株、正常根茎、回接后发病根茎、田间患病根茎。

    Figure  1.  Normal and the root rot disease of A. macrocephala

    Note: 1-6 in the figures represent normal plants, diseased plants, normal rhizomes, and diseased rhizomes, respectively

    图  2  病原菌的形态特征

    注:A-1为菌落正面;A-2为菌落反面;B-1为菌丝;B-2为大、小型分生孢子。

    Figure  2.  Morphological characteristics of pathogenic bacteria

    Note: A-1 represents the positive side of the colony; A-2 represents the negative side of the colony; B-1 mycelium; B-2 macroconidium and microconidium.

    图  3  基于ITS序列构建的系统发育树

    Figure  3.  Phylogenetic tree constructed based on ITS sequence

    图  4  基于TUB2序列构建的系统发育树

    Figure  4.  Phylogenetic tree constructed based on TUB2 sequence

    图  5  基于GAPDH序列构建的系统发育树

    Figure  5.  Phylogenetic tree constructed based on GAPDH sequence

    图  6  基于ACT序列构建的系统发育树

    Figure  6.  Phylogenetic tree constructed based on ACT sequence

    图  7  不同温度对病原菌生长的影响

    注: 图A:10、15、25、28、30 ℃条件下培养5 d的菌丝长度;不同字母表示处理间差异达显著水平(P<0.05),图810同。图B: 数字1-5分别为10、15、25、28、30 ℃条件下培养5 d的菌落大小。

    Figure  7.  Effect of Different Temperatures on the Growth of Pathogens

    Note: Fig.A:the mycelial length cultured at 10 °C, 15 °C, 25 °C, 28 °C and 30 °C for 5 d. Different letters indicated that the difference between treatments was significant ( P < 0.05 ), Same for Figs 8-10.Fig.B:Num.1-5 show colonies cultured at 10 ℃, 15 ℃, 25 ℃, 28 ℃, and 30 ℃ for 5 d, respectively.

    图  8  不同pH对病原菌生长的影响

    注: 图A:pH 5、6、7、8、9条件下培养5 d的菌丝长度;图B: 数字1-5分别为pH 5、6、7、8、9条件下培养5 d的菌落大小。

    Figure  8.  Effect of different pH on the growth of pathogenic bacteria

    Note: Fig.A:the mycelial length cultured at pH 5, 6, 7, 8, 9 for 5 d. Fig.B:Num.1-5 show colonies cultured at pH 5, 6, 7, 8, 9 for 5 d.

    图  9  不同碳源对病原菌生长的影响

    注: 图A:淀粉、乳糖、果糖、蔗糖、葡萄糖、麦芽糖条件下培养5 d的菌丝长度;图B: 数字1-5分别为淀粉、乳糖、果糖、蔗糖、葡萄糖、麦芽糖条件下培养5 d的菌落大小。

    Figure  9.  Effect of Different Carbon Sources on the Growth of Pathogens

    Note: Fig.A:the mycelial length cultured at Soluble starch, Lactose, Fructose, Sucrose, Glucose, Maltose for 5 d. Fig.B:Num.1-5 show colonies cultured at Soluble starch, Lactose, Fructose, Sucrose, Glucose, Maltose for 5 d.

    图  10  不同氮源对病原菌生长的影响

    注: 图A:硫酸铵、尿素、硝酸钾、硝酸铵、谷氨酸、丙氨酸条件下培养5 d的菌丝长度;图B: 数字1-5分别为硫酸铵、尿素、硝酸钾、硝酸铵、谷氨酸、丙氨酸条件下培养5 d的菌落大小。

    Figure  10.  Effect of different nitrogen sources on the growth of pathogenic bacteria

    Note: Fig.A:the mycelial length cultured at Sodium nitrate, Urea, Potassium nitrate, Ammonium nitrate, Glutamice, Alanine for 5 d. Fig.B:Num.1-5 show colonies cultured at Sodium nitrate, Urea, Potassium nitrate, Ammonium nitrate, Glutamice, Alanine for 5 d.

    表  1  引物信息

    Table  1.   Primer information

    基因
    Gene
    引物
    Primer
    序列
    Sequence
    ITS ITS1 5'-TCCGTAGGTGAACCTGCGG-3′
    ITS4 5′-TCCTCCGCTTATTGATATGC-3′
    ACT ACT-512F 5′-ATGTGCAAGGCCGGTTTCGC-3′
    ACT-783R 5′-TACGAGTCCTTCTGGCCCAT-3′
    TUB-2 T1 5′-GCCGTCAACGACCCCTTCATTGA-3′
    Bt2 5′-ACCCTCAGTGTAGTGACCCTTGGC-3′
    GAPDH GDF1 5′-GCCGTCAACGACCCCTTCATTGA-3′
    GDR1 5′-GGGTGGAGTCGTACTTGAGCATGT-3′
    下载: 导出CSV

    表  2  3种植物源农药室内毒力测定浓度设置

    Table  2.   Concentration settings for indoor toxicity determination of three plant-based pesticides

    供试药剂
    Pesticide
    药剂浓度
    Pesticide concentration/(μg·mL−1
    1%蛇床子素EW
    1% osthole EW
    200、 100、 50、 25、 12.5
    0.5%小檗碱AS
    0.5% berberine AS
    250、 125、 62.5、 31.25、15.625
    0.3%丁子香酚SL
    0.3% eugenol SL
    60、 30、 15、 7.5、 3.75
    CK
    下载: 导出CSV

    表  3  3种植物源杀菌剂对白术根腐病病原菌的室内毒力测定

    Table  3.   Indoor toxicity test results of plant-based fungicides on Fusarium fujikuroi

    药剂名称
    Pesticide
    毒力回归方程
    Toxicity regression equation
    R2 EC50
    EC50 value/(mg·L−1)
    0.3% 丁子香酚SL
    0.3% eugenol SL
    y=1.2062x+3.9877 0.9919 6.906
    0.5% 小檗碱AS
    0.5% berberine AS
    y=1.0629x+2.7199 0.9907 139.637
    1% 蛇床子素EW
    1% osthole EW
    y=1.2181x+3.5096 0.9909 16.749
    下载: 导出CSV
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  • 收稿日期:  2023-10-25
  • 修回日期:  2024-01-12
  • 网络出版日期:  2024-03-28

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