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金线兰茎腐病拮抗木霉菌鉴定及抑菌和促生效果评价

叶炜 颜沛沛 王培育 李尊文 郭莺 张剑亮 林敏水 江金兰

叶炜,颜沛沛,王培育,等. 金线兰茎腐病拮抗木霉菌鉴定及抑菌和促生效果评价 [J]. 福建农业学报,2024,39(X):1−9
引用本文: 叶炜,颜沛沛,王培育,等. 金线兰茎腐病拮抗木霉菌鉴定及抑菌和促生效果评价 [J]. 福建农业学报,2024,39(X):1−9
YE W, YAN P P, WAN P Y, et al. Identification and Evaluation of Stem Rot Antagonistic and Growth promotion Effects of Trichoderma on Anoectochilus roxburghii [J]. Fujian Journal of Agricultural Sciences,2024,39(X):1−9
Citation: YE W, YAN P P, WAN P Y, et al. Identification and Evaluation of Stem Rot Antagonistic and Growth promotion Effects of Trichoderma on Anoectochilus roxburghii [J]. Fujian Journal of Agricultural Sciences,2024,39(X):1−9

金线兰茎腐病拮抗木霉菌鉴定及抑菌和促生效果评价

基金项目: 福建省省级财政林业专项资金(闽财资环指〔2020〕10号);福建省科技计划项目(2023S0018、2023N0047);三明市科技计划项目(2023-N-6、2023-N-19)
详细信息
    作者简介:

    叶炜(1980 —),男,博士,副研究员,主要从事园艺植物生物技术研究,E-mail:yewei922@qq.com

    通讯作者:

    江金兰(1973 —),女,高级农艺师,主要从事园艺植物生物技术研究,E-mail: jjl75@qq.com

  • 中图分类号: S435.672

Identification and Evaluation of Stem Rot Antagonistic and Growth promotion Effects of Trichoderma on Anoectochilus roxburghii

  • 摘要:   目的  分离具有金线兰茎腐病拮抗作用的木霉菌,为生防菌的开发提供理论基础。  方法  以金线兰仿野生种植植株为材料,利用组织分离法分离木霉菌,利用形态特征与ITS和rpb2序列同源性分析鉴定其分类,利用平板对峙法鉴定其抗茎腐病能力,并对不同木霉菌的促生长作用进行评价。  结果  利用组织分离法分离3株木霉菌A21B-1、A21B-2和A21E。经鉴定,3株木霉株分别为哈茨木霉拟康宁木霉及Trichoderma longifialidicum。对峙生长表明,3种木霉菌株均对茎腐病病原菌尖孢镰刀菌ASP01表现较强的抑制作用,其抑制率分别达75.29%、73.55%和 60.02%。室内防效结果表明,A21B-1菌株对茎腐病有较强的抑制作用,接种15 d后病情抑制率达91.9%,可作为该病的生物防治候选菌株。促进生长试验表明,种植6个月后,施用3个木霉菌的金线兰植株的单株重、株高、茎粗、叶面积及SPAD值较对照均显著提高,其中A21B-2与A21E处理的植株单株重比对照分别提高了58.6%与58.9%,叶面积分别提高66.8%与59.7%,可作为金线兰促进生长的候选菌株。同时,施用木霉菌可有效提高金线兰多糖及金线莲苷的含量,其中A21B-2菌株效果最佳,其多糖及金线莲苷含量均较对照提高89.6%与11.8%,可作为促进金线兰药用成分积累的候选菌株。  结论  3种不同类型的木霉菌在金线兰对抗茎腐病、促进生长及提高多糖含量方面有显著作用。
  • 图  1  木霉菌形态特征

    a~d,e~h,i~l分别为木霉菌菌株A21B-1, A21B-2与A21E接种于PDA培养7 d后的菌落形态;a,e,i为正面;b,f, j为背面;c,g,k为分生孢子梗与瓶梗形态,分生孢子形态。

    Figure  1.  Morphological characterization of Trichoderma strains

    a-d,e-h,i-l:Colony morphologyof Trichodermastrains A21B-1, A21B-2 and A21E grown on PDA for 7 d; a, e, i are front; b, f, j are back; c,g,k are conidia stem and bottle stem, conidia morphology.

    图  2  基于序列ITS与rpb2多基因进化树分析

    标记T的菌株为模式菌,其余为参考菌株。

    Figure  2.  Phylogentic tree analysis based on ITS and rpb2 sequences

    The strains marked with T are type strains, and the rest are reference strains.

    图  3  不同木霉菌与尖孢镰刀菌菌株ASP01对峙培养

    a,e:尖孢镰刀菌ASP01;b,f:A21B-1与ASP01对峙培养;c,g:A21B-2与ASP01对峙培养;d,h:A21E与ASP01对峙培养;箭头所指为木霉菌菌丝侵入ASP01菌丝并导致尖孢镰刀菌孢子不能正常发育。

    Figure  3.  Confrontation culture of different Trichoderma strains against F. oxysporum f. sp. opponiarumASP01

    a, e: F. oxysporum f. sp. opponiarum ASP01; b,f: confrontation culture of A21B-1 and ASP01; c, g: confrontation culture of A21B-2 and ASP01; d, h: confrontation culture of A21E and ASP01; the arrow points to the invasion of Trichoderma hyphae into ASP01 hyphae, resulting in the inability of ASP01 spores to develop normally.

    表  1  木霉菌序列登录号及种名

    Table  1.   Trichoderma sequences accssion number and species

    菌株
    Strains
    登录号
    Accession NO.
    种名
    Species
    ITS rpb2
    A21B-1 ON209377 OR161371 T. harzianum
    A21B-2 ON209378 OR161372 T. konginggipsis
    A21E ON209384 OR161373 T. longifialidiucum
    下载: 导出CSV

    表  2  不同木霉菌对尖孢镰刀菌株ASP01抑制效果

    Table  2.   Inhibition effects of different Trichoderma strains against F. oxysporum f. sp. opponiarumASP01

    处理
    Treatment
    病原真菌菌落直径
    Colony diameter of pathogens/cm
    抑制率/%
    Inhibition rates
    CK 5.41±0.08 a 0±0 c
    A21B-1 1.33±0.32 c 75.29±6.45a
    A21B-2 1.43±0.40 c 73.55±7.38 a
    A21E 2.17±0.16 b 60.02±2.87 b
    数据为平均值±标准误,不同小字母表示0.05水平上差异显著。
    Dates were presented as mean ± SD, data with different lowercase letters indicated significant difference at 0.05 level.
    下载: 导出CSV

    表  3  木霉菌对茎腐病室内防效

    Table  3.   Indoor control effects of Trichoderma strains on stem rot disease

    处理
    Treatment
    7 d15 d
    感病面积
    Infected areas/mm2
    抑制率
    Inhibition rates/%
    感病面积
    Infected areas/mm2
    抑制率
    Inhibition rates/%
    CK354.0±90.70c995.3±55.60 d
    A21B-15.33±0.9498.5±0.26a79.9±14.791.9±1.47 a
    A21B-223.3±5.7393.4±1.62 b242.1±38.575.6±3.78 b
    A21E11.8±4.0996.7±1.15 b334.2±65.866.4±6.61c
    数据为平均值±标准误,不同字母代表在0.05水平存在显著差异。
    Date were presented as mean±SD, Different lowercase letters indicated significant differenceat 0.05 level.
    下载: 导出CSV

    表  4  木霉菌对金线兰促生效果

    Table  4.   Growth promotion effects of Trichoderma strains on A.roxburghii

    处理
    Treatment 
    株高
    Plant highth/
    cm
    茎粗
    Stem diameter/
    mm
    叶数
    Number of
    leaves
    叶长
    Leaf Lenth/
    mm
    叶宽
    Leaf Width/
    mm
    叶面积
    Leaf area/
    mm2
    叶厚
    Leaf thickth/
    mm
    根长
    Root lenth/
    cm
    根粗
    Root diameter/
    mm
    根数
    Root number
    SPAD 单株重
    Single plant
    weight/g
    CK 15.23±1.81 b 2.62±0.30c 6.67±1.37b 29.10±3.02b 26.78±1.84b 784.69±141.95 b 0.33±0.05 b 7.19±1.08b 2.02±0.26b 2.83±0.37 a 46.83±3.84c 3.17±0.44c
    A21B-1 19.15±3.26 a 3.05±0.40b 7.67±1.37ab 29.85±2.41 b 28.02±2.99b 984.27±151.95b 0.42±0.07 a 6.68±0.68b 1.95±0.21b 2.83±0.90 a 56.65±4.18 a 3.25±0.45 b
    A21B-2 21.02±2.05 a 3.08±0.19 b 8.33±0.47a 39.55±3.08 a 32.95±2.41a 1309.01±177.04a 0.40±0a 8.93±1.20 a 2.53±0.09a 2.33±0.47a 50.53±4.83b 5.03±0.62a
    A21E 19.65±2.00a 3.65±0.29 a 6.00±2.45b 38.10±0.73 a 32.85±1.67a 1253.43±87.91a 0.40±0 a 9.14±1.36a 2.10±0.24 b 3.00±0 a 56.95±2.16a 5.04±0.20a
    数据为平均值±标准误,不同字母代表在0.05水平存在显著差异。
    Date were presented as mean±SD, data with different lowercase letters indicated significant difference at 0.05 level.
    下载: 导出CSV

    表  5  木霉菌对金线兰促进次生代谢物积累效果

    Table  5.   Sencondary metabolite accumulated promotion effects of Trichoderma strains on A.roxburghii

    处理
    Treatment
    折干率
    Drying rate/%
    多糖含量
    Content of polysaccharide/(mg·g−1)
    黄酮含量
    Content of flavone/(mg·g−1
    金线莲苷含量
    Content of kinsenoside/(mg·g−1
    CK 11.25±0.71d 6.07±0.14 d 8.15±0.43 a (14.292±0.24) b
    A21B-1 12.88±0.73 cd 7.4±0.68 c 7.2±0.14c (14.6±0.45) b
    A21B-2 13.91±0.59b 11.51±0.35a 7.43±0.24bc (15.98±0.97) a
    A21E 14.58±0.01 a 8.05±0.99 b 7.76±0.78ab (14.96±1.26) b
    数据为平均值±标准误,不同字母代表在0.05水平存在显著差异。
    Date were presented as mean±SD, data with different lowercase letters indicated significant difference at 0.05 level.
    下载: 导出CSV
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  • 收稿日期:  2024-04-28
  • 修回日期:  2024-05-29
  • 网络出版日期:  2024-11-11

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