三株海洋木霉的应用潜力
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国家自然科学基金(31501693);国家海洋局重点实验室开放基金(MBSMAT-2014-03);国家高技术“863”研究发展计划(2011AA10A202-2)


Application potential of three marine Trichoderma spp.
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    摘要:

    [目的] 探索3株海洋生境木霉的应用潜力。[方法] 经过筛选和诱变,获得高抑菌活性及产孢量的木霉突变株;通过优化培养基、温度、初始pH考察其产孢量及最适培养条件;综合抑菌谱、重寄生及抑菌相关基因考察其抑菌活性;采用特殊培养基法考察其产纤维素酶、植酸酶、铁载体以及降解磷钾的能力,高效液相色谱法测定其产吲哚乙酸能力。[结果] 3株木霉菌的产孢量分别为3.45×108、3.10×108和2.55×108 CFU/cm2,与野生型相比分别提高了88.52%、63.16%和180.22%;且均可产生厚垣孢子,其中XG20-1厚垣孢子产量最高,达到3.56×108 CFU/mL。3株木霉菌具有较广抑菌谱及对番茄早疫病菌的重寄生作用,同时扩增得到Tex1、Nag1、Eg1基因,生物学测试显示其均具有产纤维素酶、几丁质酶以及铁载体的能力,证明其抑菌活性是多种机制共同作用的结果;菌株可以降解磷钾,且吲哚乙酸产量分别为2.61、1.57和1.92 mg/L,具有促进植物生长的潜力。[结论] 本文中3株木霉菌在开发为生防菌与生物肥料方面展现出良好的应用潜力。

    Abstract:

    [Objective] To explore the application potential of three marine Trichoderma spp. as biocontrol agents and bio-fertilizer. [Methods] Three Trichoderma spp. with high antimicrobial activity and spore germination were selected by isolating, screening and mutagenesis. Several indicators related with application potential of these strains were tested. Sporulation capacity was measured after optimizing culture medium, temperature and initial pH. Antimicrobial activity was evaluated based on inhibition spectrum, superparasitism and related antifungal genes. Special culture method was used to determine the secretion of cellulase, phytase and siderophore, and the ability to release phosphorus and potassium. High performance liquid chromatography was used to quantify indole-3-acetic acid production. [Results] We obtained three Trichoderma spp. with the spore germinations of 3.45×108, 3.10×108 and 2.55×108 CFU/cm2, respectively. Moreover, these mutants formed chlamydospores, and strain XG20-1 had the highest chlamydospore germination of 3.56×108 CFU/mL. All three strains showed a broad inhibition spectrum against 6 fungal phytopathogen and the superparasitism to Alternaria solani. Moreover, Tex1, Nag1 and Eg1 genes were found in all strains, with cellulose, phytase and siderophore detected. Indole-3-acetic acid productions in three strains were 2.61, 1.57 and 1.92 mg/L, respectively, and all three strains had the capacity of releasing phosphorus and potassium. [Conclusion] Three Trichoderma spp. had high antimicrobial activities against plant pathogenic fungi through integrated mechanism, high spore-bearing yields and growth-promoting effects on plants, which showed their potential application as biocontrol agents and bio-fertilizer.

    参考文献
    [1] Bissett J. A revision of the genus Trichoderma Ⅲ:section pachybasium. Canadian Journal of Botany, 1991, 69(11):2373-2417.
    [2] Reino JL, Guerrero RF, Collado IG, Hernández-Galán R. Secondary metabolites from species of the biocontrol agent Trichoderma. Phytochemistry Reviews, 2008, 7(1):89-123.
    [3] Daniel JF, Filho ER. Peptaibols of Trichoderma. Natural Product Reports, 2007, 24(5):1128-1141.
    [4] Ye XB, Zeng QC, Jiang XL. Research advances on enzyme for mycoparasitism of Trichoderma. Chinese Journal of Biological Control, 2009, 25(3):276-280. (in Chinese)叶小波, 曾千春, 蒋细良. 木霉菌重寄生过程中的酶学研究进展. 中国生物防治学报, 2009, 25(3):276-280.
    [5] Gray KA, Zhao L, Emptage M. Bioethanol. Current Opinion in Chemical Biology, 2006, 10(2):141-146.
    [6] 陈华癸. 土壤微生物学. 北京:高等教育出版社, 1957.
    [7] Contreras-Cornejo HA, Macías-Rodríguez L, Cortés-Penagos C, López-Bucio J. Trichoderma virens, a plant beneficial fungus, enhances biomass production and promotes lateral root growth through an auxin-dependent mechanism in Arabidopsis. Plant Physiology, 2009, 149(3):1579-1592.
    [8] Ratledge C, Dover LG. Iron metabolism in pathogenic bacteria. Annual Review of Microbiology, 2000, 54:881-941.
    [9] Haas H. Molecular genetics of fungal siderophore biosynthesis and uptake:the role of siderophores in iron uptake and storage. Applied Microbiology and Biotechnology, 2003, 62(4):316-330.
    [10] Szabó O, Farkas E. Characterization of Mn (Ⅱ) and Mn (Ⅲ) binding capability of natural siderophores desferrioxamine B and desferricoprogen as well as model hydroxamic acids. Inorganica Chimica Acta, 2011, 376(1):500-508.
    [11] Braud A, Jézéquel K, Lebeau T. Impact of substrates and cell immobilization on siderophore activity by Pseudomonads in Fe and/or Cr, Hg, Pb containing-medium. Journal of Hazardous Materials, 2007, 144(1/2):229-239.
    [12] Pócsi I, Jeney V, Kertai P, Pócsi I, Emri T, Gyémánt G, Fésüs L, Balla J, Balla G. Fungal siderophores function as protective agents of LDL oxidation and are promising anti-atherosclerotic metabolites in functional food. Molecular Nutrition and Food Research, 2008, 52(12):1434-1447.
    [13] Samuels GJ, Pardo-Schultheiss R, Hebbar KP, Lumsden RD, Bastos CN, Costa JC, Bezerra JL. Trichoderma stromaticum sp. nov., a parasite of thecacao witches broom pathogen. Mycologi Research, 2000, 104(6):760-764.
    [14] Sun M, Ren QY, Guan GQ, Liu ZJ, Li YQ, Ma ML, Liu AH, Yang JF. Optimization of cultural conditions of Beauveria bassiana pathogenic for ticks. Microbiology China, 2011, 38(7):1022-1030. (in Chinese)孙明, 任巧云, 关贵全, 刘志杰, 李有全, 马米玲, 刘爱红, 杨吉飞. 对蜱致病性球孢白僵菌培养条件的优化. 微生物学通报, 2011, 38(7):1022-1030.
    [15] Yang HT, Yu XY, Wei YL, Zhou HZ, Song LL, Hu JD, Li JS. Isolation, screening and identification of multifunctional Trichoderma spp. with antipathogenic biocontrol capacity. Shandong Science, 2009, 22(5):27-30. (in Chinese)杨合同, 于雪云, 魏艳丽, 周红姿, 宋莉璐, 扈进冬, 李纪顺. 多功能植病生防木霉的分离筛选与鉴定. 山东科学, 2009, 22(5):27-30.
    [16] Ye JY. A new differential medium for cellulose decomposing microorganisms. Microbiology China. 1997, 24(4):251-252. (in Chinese)叶姜瑜. 一种纤维素分解菌鉴别培养基. 微生物通报, 1997, 24(4):251-252.
    [17] Shi FC, Yin ZW, Jiang HM, Fan BQ. Screening, identification of P-dissolving fungus P83 strain and its effects on phosphate solubilization and plant growth promotion. Acta Microbiologica Sinica, 2014, 54(11):1333-1343. (in Chinese)史法超, 殷中伟, 江红梅, 范丙全. 一株溶磷真菌筛选鉴定及其溶磷促生效果. 微生物学报, 2014, 54(11):1333-1343.
    [18] Wang MY. Determination of indole acetic acid content in the fermentation broth using HPLC. Tropical Agricultural Engineering, 2009, 33(5):1-3. (in Chinese)王明月. 高效液相色谱法测定发酵液中的吲哚乙酸含量. 热带农业工程, 2009, 33(5):1-3.
    [19] Milagres AM, Machuca A, Napoleão D. Detection of siderophore production from several fungi and bacteria by a modification of chrome azurol S (CAS) agar plate assay. Journal of Microbiological Methods, 1999, 37:1-6.
    [20] Harman GE, Howell CR, Viterbo A, Chet I, Lorito M. Trichoderma species-opportunistic, avirulent plant symbionts. Nature Renew Microbiology, 2004, 2(2):43-56.
    [21] Lang BY, Li J, Xie J, Ren MR, Liu XZ, Chen YH. Screening of Trichoderma activity inhibiting five kinds of pathogens. Biological Disaster Science, 2013(2):142-147. (in Chinese)郎八一, 李靖, 谢津, 任梅蓉, 刘小烛, 陈玉慧. 木霉对5种病原菌抑菌活性筛选. 生物灾害科学, 2013(2):142-147.
    [22] Zhao J, Zhao L. Identification of an antagonistic Trichoderma spp. strain with plant growth-promoting activity. Shandong Agricultural Sciences, 2013, 45(4):86-89. (in Chinese)赵娇, 赵蕾. 一株促生拮抗木霉的鉴定. 山东农业科学, 2013, 45(4):86-89.
    [23] 滕安娜. 木霉菌对植物的促生效果及其机理的研究. 山东师范大学硕士学位论文. 2010.
    [24] 邹勇. 木霉厚垣孢子的产生、萌发和保存条件研究. 四川农业大学硕士学位论文. 2006.
    [25] Zhuang JH, Gao ZG, Liu X, Chen J, Yang Y, Huang YQ. Effect of fermentation factors on spore types of Trichoderma. Chinese Journal of Biological Control, 2005, 21(1):37-40. (in Chinese)庄敬华, 高增贵, 刘限, 陈捷, 杨宇, 黄艳青. 不同发酵条件对木霉产孢类型的影响. 中国生物防治学报, 2005, 21(1):37-40.
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李闯,张道敬,罗远婵,张久明,田黎. 三株海洋木霉的应用潜力[J]. 微生物学报, 2017, 57(5): 690-700

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  • 收稿日期:2016-09-28
  • 最后修改日期:2016-12-07
  • 在线发布日期: 2017-05-02
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