链霉菌XG40的鉴定及其对琯溪蜜柚黑斑病的防效

doi:10.16409/j.cnki.2095-039x.2022.02.020

摘要/Abstract

摘要： 为筛选出对琯溪蜜柚黑斑病菌有拮抗效果的生防菌株，本研究采用平板对峙法和牛津杯法分离筛选植物根际土壤放线菌，共分离纯化得到12株能够拮抗蜜柚黑斑病菌Phyllosticta citriasiana的放线菌菌株，其中菌株XG40对蜜柚黑斑病菌的拮抗能力最强，抑菌带宽度为16.3 mm。根据形态特征、生理生化特征以及16S rDNA、recA、rpoB、gyrB序列分析，将菌株XG40确定为小串链霉菌Streptomyces catenulae。抗菌谱试验结果表明，菌株XG40的发酵液对番茄叶霉病菌、蜜柚炭疽病菌、蜜柚黑点病菌等12种植物病原真菌和2种植物病原细菌也有较好的拮抗作用，抗菌性能广谱高效。菌株XG40对黑斑病菌的抑菌机制表现为受抑制菌丝发生畸变、粗细不均匀、大量断裂，其发酵液能使黑斑病菌孢子内含物消解并产生液泡。防效测定结果表明，菌株XG40发酵液对室内离体果实黑斑病的预防效果和治疗效果分别为83.52%和78.87%；对田间果实黑斑病的预防效果和治疗效果分别为78.91%和71.15%。因此，菌株XG40可作为防治蜜柚黑斑病的生防材料，具有良好的开发价值和应用前景。

关键词: 链霉菌, 抗菌活性, 鉴定, 琯溪蜜柚黑斑病, 生防效果

Abstract: To screen the strains with antagonistic effect against Phyllosticta citriasiana, the plant rhizosphere actinomyces strains were isolated and screened by plate-confrontation and oxford cup method. A total of 12 actinomyces strains had inhibition activity against P. citriasiana, and the actinomyces XG40 was the strongest, with inhibition zone diameter reaching to 16.3 mm. Based on the morphological, physiobiochemical characteristics and 16S rDNA、recA、rpoB、gyrB sequence analysis, XG40 was identified as Streptomyces catenulae. The antifungal spectrum experiment indicated that Streptomyces catenulae XG40 also showed broad-spectrum inhibition activity to 12 plant pathogenic fungi and 2 plant pathogenic bacteria, such as Fulvia fulva, Colletotrichum gloeosporioides and Diaporthe citri. The inhibition mechanism of XG40 against P. citriasiana was investigated by optical Microscope observation showed that the hyphae inhibited by the strain XG40 was distorted, with uneven thickness, and broken in large quantity. Fermentative liquid of XG40 could dissolve conidia, and produce vacuoles simultaneously. In addition, the prevention and cure efficacies of the fermented broth against P. citriasiana were up to 83.52% and 78.87% in vitro, 78.91% and 71.15% in field, respectively. Therefore, strain XG40 could be used for controlling the honey pomelo black spot disease, and has a wonderful potential for application.

Key words: Streptomyces, antimicrobial activity, identification, honey pomelo black spot disease, biocontrol efficacy

中图分类号:

S476

赖宝春, 戴瑞卿, 吴振强, 姚锦爱, 曾天宝, 王家瑞. 链霉菌XG40的鉴定及其对琯溪蜜柚黑斑病的防效[J]. 中国生物防治学报, 2022, 38(3): 662-672.

LAI Baochun, DAI Ruiqing, WU Zhenqiang, YAO Jinai, ZENG Tianbao, WANG Jiarui. Screening, Identification and Biological Control Activity of Streptomyces sp. XG40 against Black Spot Disease of Guanxi Honey Pomelo Cultivar[J]. Chinese Journal of Biological Control, 2022, 38(3): 662-672.

参考文献

[1] 赖宝春, 吴顺章, 郑春明, 等. 琯溪蜜柚炭疽病病原鉴定[J]. 果树学报, 2017, 34(9):1178-1184.
[2] 罗金水, 张汉荣, 卢松茂, 等. 琯溪蜜柚黑斑病菌分离鉴定及其致病性测定[J]. 福建热作科技, 2011, 6(4):1-5.
[3] 张晓杉, 王宏毅, 卢友民, 等. 蜜柚黑斑病症状及病原鉴定[J]. 福建农林大学学报(自然科学版), 2014, 43(3):245-250.
[4] 赖宝春, 戴瑞卿, 林明辉, 等. 一株拮抗放线菌的鉴定及其对香蕉枯萎病的生物防效[J]. 南方农业学报, 2020, 51(4):836-843.
[5] 郑域茹, 林秀香, 武英, 等. 琯溪蜜柚黑斑病拮抗细菌分离及鉴定[J]. 中国农学通报, 2015, 31(34):193-196.
[6] Lucon C M M, Guzzo S D, Jesus C O, et al. Postharvest harpin or Bacillus thuringiensis treatments suppress citrus black spot in ‘Valencia’ oranges[J]. Crop Protection, 2010, 29(7):766-772.
[7] Lombardo P, Guimaraens A, Franco J, et al. Effectiveness of essential oils for postharvest control of Phyllosticta citricarpa (citrus black spot) on citrus fruit[J]. Postharvest Biology and Technology, 2016, 121:1-8.
[8] Kupper K C, Moretto R K, Fujimoto A. Production of antifungal compounds by Bacillus spp. isolates and its capacity for controlling citrus black spot under field conditions[J]. World Journal of Microbiology and Biotechnology, 2020, 36(1):2-10.
[9] Mauricio B F, Leonardo T, Marcio P P, et al. Volatile organic compounds produced by Saccharomyces cerevisiae inhibit the in vitro development of Guignardia citricarpa, the causal agent of citrus black spot[J]. World Journal of Microbiology and Biotechnology, 2010, 26(5):925-932.
[10] 关统伟, 张小平, Rashey S G, 等. 放线菌系统分类技术[M]. 北京:化学工业出版社, 2016, 52-81.
[11] 方中达. 植病研究方法3版[M]. 北京:中国农业出版社, 1998, 243-250.
[12] 赖宝春, 吴振强, 戴瑞卿, 等. 琯溪蜜柚炭疽病拮抗放线菌的筛选和鉴定[J]. 热带作物学报, 2020, 41(9):1876-1882.
[13] 中国科学院微生物研究所放线菌分类组. 链霉菌鉴定手册[M]. 北京:科学出版社, 1975, 14-15.
[14] 宋利沙, 蒋妮, 缪剑华, 等. 肿节风炭疽病拮抗细菌的筛选与鉴定[J]. 植物保护, 2018, 44(6):61-65, 72.
[15] 侯艳娇, 罗欲承, 邵晨, 等. 快速检测肺炎链球菌recA、ply及16S rRNA基因的多重降落PCR方法建立及应用[J]. 江苏大学学报(医学版), 2014, 24(1):40-43, 47.
[16] Trantas E A, Sarris P F, Mpalantinaki E E, et al. A new genomovar of Pseudomonas cichorii, a causal agent of tomato pith necrosis[J]. European Journal of Plant Pathology, 2013, 137(3):477-493.
[17] Ruan H C, Shi N N, Du Y X, et al. First report of Pseudomonas cichorii tomato pith necrosis in Fujian Province, China[J]. Plant Disease, 2018, 103(1):145.
[18] 徐威主编. 微生物学实验(第二版)[M].北京:中国医药科技出版社, 2014, 227-229.
[19] 阎逊初编著. 放线菌的分类和鉴定[M]. 北京:科学出版社, 1992, 413-414.
[20] 卢松茂, 林秀香, 蔡坤秀, 等. 柚黑斑病菌的生长特性及寄主范围测定[J]. 果树学报, 2016, 33(12):1532-1541.
[21] 史婷婷, 马静静, 郭鑫, 等. 拮抗水稻白叶枯病菌链霉菌的筛选、鉴定和防效研究[J]. 植物病理学报, 2021, 51(3):403-412.
[22] 王燕, 宗兆锋, 程联社. 放线菌在植物病害生物防治中的应用[J]. 杨凌职业技术学院学报, 2005, 4(3):21-23.
[23] Yasuhiro I, Linkai Y, Megumi I, et al. Jomthonic acid a, a modifified amino acid from a soil-derived Streptomyces[J]. Journal of Natural Products, 2012, 75:986-990.
[24] Sabutskii Y E, Denisenko V A, Dmitrenok P S, et al. Development of approches to fibrostatin F, N-acetyl-L-cysteinyl-containing 1,4-naphthoquinone metabolite of Streptomyces catenulae[J]. Russian Chemical Bulletin, 2016, 65(3):774-778.
[25] Kwan H L, Harold W M. Partial synthesis of fibrostatins[J]. Journal of the Korean Chemical Society, 2003, 47(1):47-58.
[26] Brigitte R, Larry D, John P N. Biotransformations of tocopherols by Streptomyces catenulae[J]. American Oil Chemists Society, 1997, 32(1):79-84.
[27] Zeinat K, Mahmoud S, Noha E N. Biosynthesis, characterization, and antimicrobial activity of silver nanoparticles from actinomycetes[J]. Research Journal of Pharmaceutical, Biological and Chemical, 2016, 7(1):119-127.
[28] 蒋常德, 胡艳晖. 一种防治小麦全蚀病的复合微生物菌肥及其制备方法[P]. 中国专利, 2016, CN 106244490 A.
[29] Hoda H A, Atef F M, Safa S H. Effect of the actinomycetes and arbuscular mycorrhiae isolated from heavy metal polluted sites on tolerance of peanut plants grown in soil spiked with manganese, zinc and iron[J]. African Journal of Biological Sciences, 2010, 6(1):47-72.