烟草黑胫病生防菌的筛选、鉴定及其防病促生作用

doi:10.16409/j.cnki.2095-039x.2025.02.048

摘要/Abstract

摘要： 由烟草疫霉引起的黑胫病是威胁烟草生产的最具破坏性和最广泛的土传病害之一。本研究通过梯度稀释涂布法和平板对峙法从云南省曲靖市烟草黑胫病重灾区根际土壤中分离得到一株高效拮抗菌株t1.1。经形态学、生理生化以及16S rRNA和gyrB基因联合建树鉴定为植物乳植杆菌Lactiplantibacillus plantarum。进一步研究表明，菌株t1.1具有产嗜铁素、1-氨基环丙烷-1-羧酸（ACC）脱氨酶、溶解有机磷和无机磷能力等多种促生、防病能力。盆栽试验结果表明，菌株t1.1发酵液对烟草黑胫病的防效为67.14%，显著高于对照药剂（58%甲霜·锰锌） 57.14%的防效，且对烟草的株高、根长、鲜重及叶片表面积等指标具有较好的促生效果。综上，菌株t1.1对烟草黑胫病具有良好的的生物防治潜力，且具有较好的促生效果，丰富了烟草黑胫病的生防菌株资源，为乳酸菌防治烟草黑胫病提供了一定的借鉴。

关键词: 烟草黑胫病, 植物乳植杆菌, 生物防治, 盆栽试验

Abstract: Tobacco black shank, caused by Phytophthora nicotianae, is one of the most destructive and widespread soil-borne diseases of tobacco production. In this study, an efficient antagonistic strain, t1.1, was isolated from the rhizosphere soil of tobacco black shank-affected fields in Qujing, Yunnan Province, using the dilution coating plate method and the plate confrontation method. Based on morphological, physiological and biochemical characterization, as well as the phylogenetic trees constructed from the 16S rRNA and gyrB sequences, the strain was identified and named as Lactiplantibacillus plantarum t1.1. Further studies revealed that strain t1.1 producing siderophores, 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity and the ability to solubilize both organic and inorganic phosphates are capable of stimulating plant growth and disease-suppressing capabilities. Pot experiment revealed that the strain t1.1 provided achieved 67.14% a control efficacy of tobacco black shank, significantly surpassing the positive control (58% metalaxyl-mancozeb at 57.14% efficacy). Concurrently, it exhibited significantly plant growth-promoting effects, markedly enhancing tobacco growth metrics including plant height, root length, fresh weight, and leaf surface area. In summary, strain t1.1 demonstrates promising biocontrol effects against tobacco black shank and exhibits notable plant growth-promoting effects. This study enriched the biological strain resources of this disease and provided a theoretical basis support for the control of lactic acid bacteria for tobacco black shank.

Key words: tobacco black shank, Lactobacillus plantarum, biological control, pot experiment

中图分类号:

S476

郭健杰, 易晗, 唐家昊, 刘颜玉, 韦金妮, 杨咖儿, 张红岩, 申乃坤, 陈伯昌. 烟草黑胫病生防菌的筛选、鉴定及其防病促生作用[J]. 中国生物防治学报, 2025, 41(4): 927-938.

GUO Jianjie, YI Han, TANG Jiahao, LIU Yanyu, WEI Jinni, YANG Kaer, ZHANG Hongyan, SHEN Naikun, CHEN Bochang. Screening and Identification of Biocontrol Bacterium and Its Effect on Tobacco Black Shank Disease and Plant Growth Promotion[J]. Chinese Journal of Biological Control, 2025, 41(4): 927-938.

参考文献

[1] Liang F, Chai Y X, Li Y Y, et al. The effect of mixed microbial agents on tobacco black shank disease[J]. Physiological and Molecular Plant Pathology, 2024, 134: 102442.
[2] 赵辉, 王喜英, 刘国权, 等. 烟草黑胫病发生因素及综合防治研究进展[J]. 湖南农业科学, 2020(11): 99-103.
[3] 高文华, 李江美. 烟草黑胫病防治研究进展[J]. 热带农业科技, 2020, 43(1): 50-54.
[4] Ma Y N, Gu Y L, Liu J, et al. Deciphering the rhizosphere bacteriome associated with biological control of tobacco black shank disease[J]. Frontiers in Plant Science, 2023, 14: 1152639.
[5] Guo D S, Yuan C H, Luo Y Y, et al. Biocontrol of tobacco black shank disease (Phytophthora nicotianae) by Bacillus velezensis Ba168[J]. Pesticide Biochemistry and Physiology, 2020, 165: 104523.
[6] Zhu Q X, Chen Y, Jia M. et al. Managing tobacco black shank disease using biochar: direct toxicity and indirect ecological mechanisms[J]. Microbiology Spectrum, 2024, 12(10): e0014924.
[7] Zhou F F, Pan Y H, Zhang X L, et al. Accumulation patterns of tobacco root allelopathicals across different cropping durations and their correlation with continuous cropping challenges[J]. Frontiers in Plant Science, 2024, 15: 1326942.
[8] Sullivan M J, Melton T A, Shew H D. Managing the race structure of Phytophthora parasitica var. nicotianae with cultivar rotation[J]. Plant Disease, 2005, 89(12): 1285-1294.
[9] Pandey R. Controlling tobacco diseases: an overview of black shank and fusarium wilt[J]. International Journal of Applied Sciences and Biotechnology, 2023, 11(1): 1-7.
[10] Li X X, Wang J, Lv Y, et al. Screening and identification of Paenibacillus polymyxa GRY-11 and its biological control potential against apple replant disease[J]. Folia Microbiologica, 2024, 70(2): 475-487.
[11] 何明川, 王志江, 谢永辉, 等. 烟草黑胫病拮抗菌的筛选、鉴定及发酵条件优化[J]. 中国生物防治学报, 2022, 38(2): 428-439.
[12] 千慧敏, 文艺, 赵辉, 等. 烟草黑胫病和根黑腐病生防假单胞杆菌的筛选与鉴定[J]. 中国生物防治学报, 2019, 35(6): 940-948.
[13] Ying H, Ma L, Fang D H, et al. Isolation and characterization of rhizosphere bacteria active against Meloidogyne incognita, Phytophthora nicotianae and the root knot-black shank complex in tobacco[J]. Pest Management Science, 2015, 71(3): 415-422.
[14] 彭海莹, 田叶韩, 李晓芳, 等. 生防真菌与氨基寡糖素组合对烟草黑胫病和根黑腐病的防治[J]. 中国烟草科学, 2021, 42(1): 47-53.
[15] 刘天波, 赵誉强, 滕凯, 等. 烟草疫霉拮抗细菌B44R-1的筛选及其生防效果[J]. 中国生物防治学报, 2022, 38(6): 1545-1552.
[16] 韩瑞华, 江凯, 康业斌. 玫瑰黄链霉菌联合杀菌剂对烟草疫霉的防治效果[J]. 中国生物防治学报, 2024, 40(3): 710-716.
[17] 张幸博, 郭小红, 刘玉珍, 等. 烟草黑胫病生防菌LG-3的分离鉴定及防效研究[J]. 湖北农业科学, 2021, 60(2): 82-84, 89.
[18] Chen D M, Yang H J, Huang J G, et al. Lysobacter enzymogenes LE16 autolysates have potential as biocontrol agents-Lysobacter sp. autolysates as biofungicide[J]. Journal of Applied Microbiology, 2020,129(6): 1684-1692.
[19] Liu Y L, He P B, He P F, et al. Potential biocontrol efficiency of Trichoderma species against oomycete pathogens[J]. Frontiers in Microbiology, 2022, 13: 974024.
[20] Volynchikova E, Kin K D. Biological control of oomycete soilborne diseases caused by Phytophthora capsici, Phytophthora infestans, and Phytophthora nicotianae in solanaceous crops[J]. Microbiology, 2022, 50(5): 269-293.
[21] Siedler S, Balti R, Neves R A. Bioprotective mechanisms of lactic acid bacteria against fungal spoilage of food[J]. Current Opinion in Biotechnology, 2019, 56: 138-146.
[22] 王晓宇, 吴梦娜, 于巧如, 等. 植物乳杆菌ST3.5的分离鉴定及其对霉菌的抑制作用[J]. 食品工业科技, 2023, 44(13): 141-149.
[23] 黄若琪, 李瑜玲, 杨恩. 乳酸菌对真菌毒素的脱毒作用研究进展[J]. 微生物学通报, 2023, 50(9): 4260-4274.
[24] Lv J, Jiang W T, Xu Z H, et al. Comparative analysis of the effects of crude metabolic extracts of three biocontrol bacteria on microbial community structure provides a new strategy for the biological control of apple replant disease[J]. Horticulture, 2024, 10(10): 1035.
[25] Yi Z Q, Yue Y S, Kan J Q, et al. Biocontrol of Fusarium oxysporum-infested Gastrodia elata Bl. by Lactobacillus curvatus 2768-VOCs and mechanism of inhibition[J]. Food Bioscience, 2024, 62: 105174.
[26] 曹然, 杨青青, 崔美林, 等. 玉米青贮中优良产酸乳酸菌筛选及生物学特性分析[J]. 中国酿造, 2024, 43(8): 144-152.
[27] 李枢妍. 香蕉枯萎病高效生防菌株的筛选、鉴定及抑菌机理研究[D]. 南宁: 广西民族大学, 2024.
[28] 许佳露, 张平, 李美芳, 等. 产铁载体菌株的分离、培养条件优化及初步应用[J]. 微生物学通报, 2022, 49(03): 1004-1016.
[29] 朱滕滕, 何学莲, 勉小娟, 等. 硒砂瓜根际促生菌筛选鉴定及其促生效应研究[J]. 干旱地区农业研究, 2023, 41(2): 221-229.
[30] 胡美娟, 周星辰, 王丽, 等. 苦豆子内生放线菌产酶特性分析及活性菌株鉴定[J]. 北方园艺, 2013(16): 98-102.
[31] 张奇, 王一兵, 申乃坤, 等. 产几丁质酶菌株GXUN-20的筛选、鉴定及其产酶条件优化[J]. 食品工业科技, 2021, 42(24): 119-127.
[32] 殷豆豆, 黄宏智, 宋超东, 等. 广西小花老鼠簕可培养细菌多样性及促生、防病、产酶活性分析[J]. 南方农业学报, 2024, 55(4): 973-984.
[33] 沈萍, 陈向东. 微生物学实验[M]. 第4 版. 北京: 高等教育出版社, 2017．
[34] 陆君铭, 吉春喜, 郭健杰, 等. 一株耐镉促生细菌Achromobacter sp. A81的分离鉴定及特性[J]. 微生物学报, 2025, 65(1): 90-105.
[35] 布坎南, 吉本斯. 伯杰细菌鉴定手册[M]. 第8 版. 北京: 科学出版社, 1984.
[36] Reyes-Escogido L, Balam-Chi M, Rodríguez-Buenfil I, et al. Purification of bacterial genomic DNA in less than 20 min using chelex-100 microwave: examples from strains of lactic acid bacteria isolated from soil samples[J]. Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology, 2010, 98(4): 465-474.
[37] GB/T23222—2008. 烟草病虫害分级及调查方法[S].
[38] 韦国敏, 秦健. 7种杀菌剂对樟树溃疡病菌的室内抑菌效果[J]. 广西植保, 2017, 30(4): 39-41.
[39] 巫锡奎, 刘志均, 陈君志, 等. 广西香蕉叶斑病病原菌研究[J]. 广西农业科学, 1990(4): 24-27.
[40] 韦绍龙, 霍秀娟, 李朝生. 广西香蕉枯萎病菌4 号生理小种对桂蕉6 号的致病力分化研究[J]. 湖北农业科学, 2016, 55(11): 2796-2798,2801.
[41] 蔡毅, 鄢敏, 胡万波, 等. 3种烟草根茎病害防治药剂的筛选[J]. 湖南农业科学, 2022(6): 53-56.
[42] 徐丹阳. 生防菌株DY-6-6的筛选、鉴定及其对烟草黑胫病的生物防治研究[D]. 杨凌: 西北农林科技大学, 2023.
[43] 陈梦多, 胡春艳, 马肖静, 等. 植物根际细菌HQ1-2的根际定殖与土壤微生态调节及枯萎病防治[J]. 中国生物防治学报, 2023, 39(4): 924-932.
[44] Akhtar M, Nosheen A, Keyani R, et al. Biocontrol of Rhizoctonia solani in basmati rice by the application of Lactobacillus and Weissella spp.[J]. Scientific Reports, 2023, 13(1): 13855.
[45] Pruthviraj, Naik K M, Ekabote D S, et al. Evaluation of Lactiplantibacillus argentoratensis MYSJK8 from jackfruit for its multifarious antibacterial, probiotic, plant growth promoting and bio-control attributes[J]. Journal of Crop Health, 2024, 76(6): 1-16.
[46] 刘邮洲, 沈佳慧, 乔俊卿, 等. 芽孢杆菌嗜铁素研究进展[J]. 江苏农业学报, 2023, 39(1): 266-276.
[47] 李枢妍, 何艳丽, 宋超东, 等. 铜绿假单胞菌Gxun-2产铁载体发酵条件优化及其抑菌效果[J]. 食品工业科技, 2024, 45(10): 118-125.
[48] 王慧楠, 朱静, 谢文文, 等. 一株海洋杆菌属新菌种XAAS-72T的植物促生功能分析及ACC脱氨酶蛋白结构预测[J]. 新疆农业科学, 2024, 61(07): 1778-1785.
[49] Herpell J B, Alickovic A, Diallo B, et al. Phyllosphere symbiont promotes plant growth through ACC deaminase production[J]. The ISME Journal, 2023, 17(8): 1267-1277.
[50] Kowalska B, Szczech M, Lisek A. Inhibition of Botrytis cinerea and Escherichia coli by lactic acid bacteria on leafy vegetables[J]. Agriculture, 2024, 14(8): 1228.
[51] Yan F, Ma J C, Peng M J, et al. Lactic acid induced defense responses in tobacco against Phytophthora nicotianae[J]. Scientific Reports, 2024, 14(1): 9338.
[52] 田所会, 刘涛, 刘俊, 等. 烟草黑胫病的病原鉴定及诱抗剂处理效果[J]. 中国植保导刊, 2022, 42(10): 20-24, 66.