Screening and Identification of the Antagonistic Bacteria Against Potato Scab Streptomyces galilaeus
YANG Xin1, FAN Wu-Jing1, LI Li-Shu1,*, TANG Xiu-Hua1, FENG Zhi-Wen2
1 Commercial Crop Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, China; 2 Potato Engineering and Technology Research Center, Inner Mongolia University, Hohhot 010021, China
Abstract：Antagonistic endophytic bacteria play an important role in the biological control of potato scab disease. Using healthy tubers of disease resistant potato (Solanum tuberosum) varieties ('Yunshu 304', 'Zhongshu 43', 'Zhongshu 566') as test materials and Streptomyces galilaeus as target bacteria, antagonistic bacteria were screened and identified using methods such as plate confrontation, Oxford cup method, strain morphology, physiological and biochemical characteristics, and bacterial 16S rRNA gene and gyrB gene sequencing. Their growth characteristics were analyzed and environmental adaptability was evaluated. The effectiveness of antagonistic bacteria in preventing potato scab disease was verified through pot experiments, in order to obtain efficient biocontrol strains for potato scab disease prevention and control. The results indicated that 3 strains YS304, ZS43 and ZS567 with significant antagonistic effect against pathogenic bacteria were obtained, The average diameters of bacteriostatic zones of primary and secondary screening were 18.71 and 31.57 mm, Gram staining of the strains was positive, It was identified as Bacillus velezensis combined with physiological and biochemical characteristics and 16S rRNA, gryB gene sequence analysis. The 3 strains still maintained certain antibacterial activity under pH 3~13 and 40~80 ℃, It was insensitive to common bactericides such as pentachloronitrobenzene, enoylmorpholine, mancozeb and haloperidine. In pot experiment of potato scab, the relative control effects were 60.84%, 69.15% and 75.76% respectively. The strains YS304, ZS43 and ZS567 have good biological control effects, strong environmental adaptability and stability, they can be used as a candidate strain to control potato scab.
[1] 崔凌霄, 杨成德, 杨丽萍, 等. 2021. 致病链霉菌毒素多样性及马铃薯疮痂病综合防治研究进展[J]. 西北农业学报, 30(4): 469-480. (Cui L X, Yang C D, Yang L P, et al.2021. Advance of research in diversity of pathogenic toxins from streptomyces and integrated control of potato scab[J]. Acta Agriculturae Borealioccidentalis Sinica, 30(4): 469-480) [2] 陈利达, 李磊, 谢学文, 等. 2020. 山东高密地区马铃薯疮痂病菌种类及致病性鉴定[J]. 华北农学报, 35(增刊): 347-354. (Chen L D, Li L, Xie X W.et al.2020. Species and pathogenicity identification of streptomyces species causing potato common scab in gaomi city, shangdong province[J]. Acta Agriculturae Boreali-sinica, 35(增刊): 347-354.) [3] 陈志垚, 王鹏, 王微, 等. 2021. 马铃薯疮痂病菌Stretomyces scabies拮抗细菌的筛选及BKS104鉴定[J]. 微生物学通报, 48(11): 4145-4155. (Chen Z Y, Wang P, Wang H, et al.2021. Screening of antagonistic bacteria against Streptomyces scabies and identification of BKS104[J].Microbiology China, 48(11): 4145-4155.) [4] 迟惠荣, 张亚惠, 曾欣, 等. 2019. 多花黄精内生贝莱斯芽孢杆菌的分离鉴定及其抗菌与促生作用分析[J].植物保护, 45(4): 122-131. (Chi H R, Zhang Y H, Zen Y, et al.2019. Isolation and identification of antagonistic endophytic Bacillus velezensis from polygonatum cyrtonema Hua and analysis of its antimicrobial and growth-promoting activity[J].Journal of Plant Protection, 45(4): 122-131) [5] 东秀珠, 蔡妙英. 2001. 常见细菌系统鉴定手册[M]. 北京: 科学出版社, pp. 362-373. (Dong X Z, Cai M Y.2001. Handbook of Identification of Common Bacterial Systems[M]. BeiJing:Science Press, pp. 362-373) [6] 方园, 彭勇政, 廖长贵, 等. 2022. 一株具有防病促生功能的贝莱斯芽孢杆菌SF327[J]. 微生物学报, 62(10): 4071-4088. (Fang Y, Peng Y Z, Liao C G, et al.2022. Bacillus velezensis SF327, a potential biocontrol agent with the functions of preventing plant diseases and promoting plant growth[J]. Acta Microbiologica Sinica, 62(10): 4071-4088.) [7] 傅奇, 林俊杰, 甘美裕, 等. 2020. 16S与gyrB基因联合构建树快速鉴定海水中的一株地衣芽胞杆菌[J]. 海洋科学, 44(4): 90-95. (Fu Q, Lin J J, Gan M Y, et al.2020. Identification of a Bacillus licheniformis strain from seawater using a phylogenetic tree based on 16S and gyrB gene sequences[J].Marine Sciences, 44(4): 90-95.) [8] 贾慧慧, 谢心悦, 潘园园, 等.2020. 芽孢杆菌BJ-6的鉴定及对甜瓜细菌性果斑病的防治[J]. 微生物学报, 60(5): 982-991. (Jia H H, Xie X Y, Pan Y Y, et al.2020. Identification of Bacillus sp.BJ-6 for biocontrol of bacterial fruit blotch of melon[J]. Microbiology China, 60(5): 982-991.) [9] 糜芳, 吴紫燕, 王承芳, 等. 2021. 1株解淀粉芽孢杆菌的分离、鉴定及在马铃薯疮痂病防治上的应用[J]. 江苏农业科学, 49(18): 122-127. (Mi F, Wu Z Y, Wang C F, et al.2021. Isolation and identification of a Bacillus amylolyticus strain and its application in the control of potato scab[J]. Jiangsu Agricultural Sciences, 49(18): 122-127.) [10] 马佳, 李颖, 胡栋, 等. 2018. 芽孢杆菌生物防治作用机理与应用研究进展[J]. 中国生物防治学报, 34(4): 639-648. (Ma J, Li Y, Hu D, et al.2018. Progress on mechanism and applications of Bacillus as a biocontrol microbe[J].Chinese Journal of Biological Control, 34(4): 639-648.) [11] 石莹莹, 赵盼, 宋双伟, 等. 2020. 马铃薯疮痂病拮抗菌YN-2-2的分离与鉴定[J]. 微生物学通报, 47(8): 2425-2435. (Shi Y Y, Zhao P, Song S W, et al.2020. Isolation and characterization of the antagonistic bacterium YN-2-2 against potato common scab[J].Microbiology China, 47(8): 2425-2435.) [12] 申永瑞, 向君亮, 王佳琦, 等. 2019. 疮痂链霉菌拮抗菌株BU396的分子鉴定与抗菌性质分析[J]. 微生物学通报, 46(10): 2601-2611. (Shen Y R, Xiang J L, Wang J Q, et al.2019. Isolation, identification and antimicrobial properties of a biocontrol strain BU396 against Streptomyces scabies[J]. Microbiology China, 46(10): 2601-2611.) [13] 台莲梅, 陈志垚, 王鹏, 等. 2021. 马铃薯疮痂病Streptomyces scabies拮抗细菌筛选及鉴定[J]. 河南农业科学, 50(11): 108-116. (Tai L M, Chen Z Y, Wang P, et al.2021. Screening and identification of antagonistic bacteria of Strepomyces scabies[J].Journal of Henan Agricultural Sciences, 50(11): 108-116.) [14] 陶永梅, 潘洪吉, 张小利, 等. 2020. 贝莱斯芽孢杆菌NSZ-YBGJ001的鉴定及其生物学特性[J]. 植物保护学报, 47(2): 324-331. (Tao Y M, Pan H J, Zhang X L, et al.2020. Identification and biological characteristics of Bacillus velezensis NSZ-YBGJ001[J]. Journal of Plant Protection, 47(2): 324-331.) [15] 徐李娟, 陈勇, 王则玉. 2023. 新疆拜城县马铃薯疮痂病病原菌的分离鉴定及生物学特性分析[J]. 新疆农业科学, 60(9): 2258-2265. (Xu L J, Chen Y, Wang Z Y, et al.2023. Isolation, identification and biological characteristics of potato scab pathogen in Baichen, Xinjiang[J]. Xingjiang Agricultual Sciences, 60(9): 2258-2265. [16] 徐淑琴, 贺曦, 龚紫凤, 等. 2022. 贝莱斯芽孢杆菌的生物学特性及其农业应用现状[J].饲料研究, 45(09): 143-147. (Xu S Q, He X, Gong Z F, et al.2022. Biological characteristics of Bacillus velezensis and its agricultural application[J]. Feed desearch, 45(09): 143-147.) [17] 杨鑫, 赖振光, 樊吴静, 等. 2023. 马铃薯疮痂病不同抗性品种发病与健康块茎内生细菌群落结构及多样性[J]. 江苏农业科学, 51(14): 134-140. (Yang X, Lai ZG, Fang W J, et al.2023. Community structure and diversity of endophytic bacteria in healthy and morbidity tubers with different resistance to potato scab[J]. Jiangsu Agricultural Sciences Journal, 51(14): 134-140.) [18] 严婉荣, 肖敏, 陈圆, 等. 2017. 芽孢杆菌基本特征、16S rRNA比对分析及特异性基因挖掘[J]. 基因组学与应用生物学, 36(11): 4686-4692. (Yan W R, Xiao M, Chen Y, et al.2017. Comparative analysis between basic characteristics and 16S rRNA,and specific gene discovery of Bacillus[J]. Genomics and Applied Biology, 36(11): 4686-4692.) [19] 朱展鹏. 2020. 马铃薯疮痂病病原菌多样性分析及抗病种质筛选[D]. 硕士学位论文, 华中农业大学, 导师: 蔡兴奎, pp. 44-45. (Zhu Z P.2020. Diversity analysis of potato common scab pathogens and disease-resistant germplasm screening[D]. Thesis for M.S., Huazhong Agricultural University, Supervisor: Cai X K, pp. 44-45.) [20] 赵永龙, 赵盼, 曹晶晶, 等. 2022. 疮痂链霉菌拮抗菌定向筛选及其功能评价[J]. 微生物学报, 62(7): 2624-2641. (Zhao Y L, Zhao P, Cao J J, et al.2022. Targeted screening and functional evaluation of the bacterial antagonistics to Streptomyces scabies[J]. Acta Microbiologica S, 62(7): 2624-2641) [21] 张铉哲, 赵雪, 陈苏慧, 等. 2022. 黑龙江省马铃薯疮痂病菌种群结构及 PAI致病基因分析[J]. 中国蔬菜, (5): 53-59. (Zhang X Z, Zhao X, Chen S H, et al. 2022. Analysis of potato common scab population structure and PAI pathogenic gene in heilongjiang province[J]. China Vegetables, (5): 53-59.) [22] 张铭铄. 2016. 一株马铃薯疮痂病病原菌的系统鉴定及生物防治的研究[D]. 硕士学位论文, 山东农业大学, 导师: 周波, pp. 26-37. (Zhang M S, 2016. System identification of a potato common scab pathogen and its biological control[D]. Thesis for M.S., Shangdong Agriculture university, Supervisor: Zhou B, pp. 26-37.) [23] Cui L X, Yang C D, Wei L J, et.al.2020. Isolation and identification of an endophytic bacteria Bacillus velezensis 8-4 exhibiting biocontrol activity against potato scab[J]. Biological Control, 141: 104156-104156. [24] Chen L, HengJ, Qin S.2018. A comprehensive understanding of the biocontrol potential of Bacillus velezensis LM2303 against fusariun head blight[J]. PLOS ONE, 13(6): e0198560 [25] Dees M W, Wanner L A.2013. In search of better management of potato common scab lesions in Norway[J]. Plant Pathology, 62(1): 217-225. [26] Hao J J, Meng Q X,Yin J F,et al.2009. Characterization of a new Streptomyces strain,DS3024,that causes potato common scab[J].Plant Disease, 93(12): 1329-1334. [27] Li C P, Shi W C, Wu D, et al.2020. Biocontrol of potato common scab by Brevibacillus laterosporus BL12 is related to the reduction of pathogen and changes in soil bacterial community[J]. Biological Control, 153: 104496. [28] Zhang X Y, Li C, Hao J J, et al.A novel Streptomyces sp. strain PBSH9 for controlling potato common scab caused by Streptomyces galiaeus[J]. Plant Disease, 104(7): 1986-1993.
