|
|
|
| Antimicrobial Stability of Bacillus subtilis Strain Rbac1 and Its Biocontrol Effect on Plant Anthracnose |
| HU Qian1,*, JIANG Hao2,*, LIU Yang3, WANG Yi1, HE Lu-Qian1, KONG Li-Ya1, WANG Zhan-Qi1, ZHANG Li-Qin1, MIN Li-Jing1, TANG Zhao-Yang1,** |
1 College of Life Sciences/Zhejiang Key Laboratory of Biology and Ecological Regulation of Crop Pathogens and Insects, Huzhou Normal University, Huzhou 313000, China;
2 National Key Laboratory for Tea Plant Germplasm Innovation and Resource Utilization, Anhui Agricultural University, Hefei 230036, China;
3 Qujiang District Bureau of Agriculture and Rural Affairs of Quzhou City, Quzhou 324000, China |
|
|
|
|
Abstract Anthracnose poses a severe threat to the green and sustainable development of the agricultural industry. To explore novel biological control technologies, a Bacillus subtilis strain Rbac1 was isolated and purified in this study. The control efficacy of strain Rbac1 against various important diseases, especially anthracnose, was investigated. Results showed that Rbac1 strain significantly inhibited multiple pathogens, including Colletotrichum camelliae, Co. fructicola, Fusarium solani, Aspergillus flavus, and Phytophthora sojae. The conidial germination test indicated that the fermentation filtrate of strain Rbac1 remarkably suppressed the conidial germination of Co. camelliae and Co. fructicola. The antibacterial stability test revealed that the fermentation broth of this strain exhibited favorable thermal stability. There was no significant difference in inhibition rates at pH 3~9 and no obvious change following a 60 min ultraviolet irradiation treatment. The detached leaf biocontrol test demonstrated that the fermentation filtrate of strain Rbac1 effectively controlled anthracnose in 'Baiye 1' and 'Huang jinya' tea(Camellia sinensis) varieties, with control efficiencies of 58.03% and 70.99% respectively. In conclusion, B. subtilis Rbac1 showed great potential for biological control of tea anthracnose. This study provides theoretical basis and strain resources for the development of novel biological control agents.
|
|
Received: 24 October 2025
|
|
|
|
Corresponding Authors:
**zytang@zjhu.edu.cn
|
|
|
|
[1] 曹阳, 孙平平, 刘彬. 2022. 一株枯草芽孢杆菌发酵培养基和培养条件的优化[J]. 绿色科技, 24(24): 237-240+257.
(Cao Y, Sun P P, Liu B. 2022. Optimization of fermentation medium and culture conditions for a Bacillus subtilis strain[J]. Journal of Green Science and Technology, 24(24): 237-240+257.)
[2] 贡长怡, 刘姣姣, 邓强, 等. 2022. 茶树炭疽病病原菌鉴定及其致病性分析[J]. 园艺学报, 49(05): 1092-1101.
(Gong C Y, Liu J J, Deng Q, et al.2022. Identification and pathogenicity of Colletotrichum species causing anthracnose on Camellia sinensis[J]. Acta Horticulturae Sinica, 49(05): 1092-1101.)
[3] 纪宏伟, 王小敏, 庞宏伟, 等. 2015. 枯草芽孢杆菌与巨大芽孢杆菌对土壤有效态Cd的影响研究[J]. 水土保持学报, 29(03): 325-329.
(Ji H W, Wang X M, Pang H W, et al.2015. Effect of Bacillus subtilis and Bacillus megaterium on soil available Cd[J]. Journal of Soil and Water Conservation, 29(03): 325-329.)
[4] 李娟, 宋洁, 姜路路, 等. 2015. 草莓炭疽病拮抗芽孢杆菌TJX-012的筛选与鉴定[J]. 辽东学院学报(自然科学版), 22(01): 23-26+47.
(Li J, Song J, Jiang L L, et al. 2015. Screening and identification of antagonistic Bacillus DTJ-012 on strawberry anthracnose[J]. Journal of Liaodong University(Natural Science Edition), 22(01): 23-26+47.)
[5] 李欣, 黄月玲, 韦俊峰. 2019. 布央侗寨茶产业助推乡村振兴的路径与模式[J]. 茶叶通讯, 46(01): 125-128.
(Li X, Hunag Y L, Wei J F.2019. The path and model of buyang's tea industry promoting rural revitalization[J]. Journal of Tea Communication, 46(01): 125-128.)
[6] 李颖, 金鑫, 沙海天, 等. 2016. 枯草芽孢杆菌SH-1发酵条件优化及抑菌活性物质稳定性研究[J]. 食品科技, 41(09): 25-29.
(Li Y, Jin X, Sha H T, et al.2016. Optimization of fermentation conditions of Bacillus subtilis SH-1 and stability of antimicrobial substances[J]. Food Science and Technology, 41(09): 25-29.)
[7] 凌光汉, 陈小媛. 2015. 新昌县茶树炭疽病的发生与防治对策[J]. 中国茶叶, 37(12): 19.
(Lin G H, Chen X Y. 2015. Occurrence and control strategies of tea anthracnose in Xinchang County[J]. China Tea, 37(12): 19.)
[8] 刘丽萍, 高洁, 李玉. 2020. 植物炭疽菌属Colletotrichum真菌研究进展[J]. 菌物研究, 18(04): 266-281.
(Liu L P, Gao J, Li Y.2020. Advances in knowledge of the fungi referred to the genus Colletotrichum[J]. Journal of Fungal Research, 18(04): 266-281.)
[9] 刘荣, 姚玉仙, 周爽爽, 等. 2022. 茶炭疽病的症状识别与防治研究[J]. 福建茶叶, 44(02): 24-26.
(Liu R, Yao Y X, Zhou S S, et al.2022. Symptom identification and control research on tea anthracnose[J]. Tea in Fujian, 44(02): 24-26.)
[10] 潘朝勃, 李其利, 莫贱友, 等. 2013. 枯草芽孢杆菌9A对杧果炭疽菌的抑菌作用及防病效果[J]. 果树学报, 30(06): 1023-1029.
(Pan Z B, Li Q L, Mo J Y, et al.2013. Inhibitory effects on Colletotrichum gloeosporioides and control efficacy against mango anthracnose by Bacillus subtilis 9A[J]. Journal of Fruit Science, 30(06): 1023-1029.)
[11] 宋璐, 张花, 李保珍, 等. 2022. 枯草芽孢杆菌对镉胁迫下连翘生长生理的影响. 森林与环境学报[J]. 42(06): 617-622.
(Song L, Zhang H, Li B Z, et al.2022. Effects of Bacillus subtilis on growth physiology and cadmium[J]. Journal of Forest and Environment, 42(06): 617-622.)
[12] 唐朝阳, 孔丽娅, 胡骞, 等. 2024. 贝莱斯芽孢杆菌YJK1鉴定及其对茶炭疽病的拮抗效果[J]. 茶叶科学, 44(03): 443-452.
(Tang Z Y, Kong L Y, Hu Q, et al.2024. Antagonistic activity and utilization of Bacillus velezensis strain YJK1 as a biocontrol agent against anthracnose on Camellia sinensis[J]. Journal of Tea Science, 44(03): 443-452.)
[13] 王玉春, 刘守安, 卢秦华, 等. 2019. 中国茶树炭疽菌属病害研究进展及展望[J]. 植物保护学报, 46(05): 954-963.
(Wang Y C, Liu S A, Lu Q H, et al.2019. Research progress and prospects of Colletotrichum species causing tea plant diseases in China[J]. Journal of Plant Protection, 46(05): 954-963.)
[14] 吴艳清, 王游游, 王畅, 等. 2018. 枯草芽孢杆菌WL2脂肽粗提物对致病疫霉的抑制作用及其分离鉴定[J]. 河北大学学报(自然科学版), 38(06): 632-639.
(Wu Y Q, Wang Y Y, Wang C, et al.2018. Inhibitory effect of lipopeptide crude extract produced by Bacillus subtilis WL2 on Phytophthora infestans and its isolation and identification[J]. Journal of Hebei University(Natural Science Edition), 38(06): 632-639.)
[15] 肖小露. 2017. 枯草芽孢杆菌BS193对辣椒疫病的生防作用及其抗菌机制初探[D].硕士学位论文, 福建农林大学, 导师: 陈庆河, pp. 32-33.
(Xiao X L.2017. Biological control and mechanism of Bacillus subtilis BS193 against pepper phytophthora blight(Phytophthora capsici)[D]. Thesis for M.S., Fujian Agriculture and Forestry University, Suppervisor: Chen Q H, pp. 32-33.)
[16] 徐小文, 王义勋, 张子一, 等. 2022. 山茶炭疽菌CcSnf1基因参与调控山茶炭疽菌的生长发育和致病力[J]. 植物病理学报, 52(06): 927-939.
(Xu X W, Wang Y X, Zhang Z Y, et al.2022. CcSnf1 is involved in regulating development and pathogenecity in Colletotrichum camelliae[J]. Acta Phytopathologica Sinica, 52(06): 927-939.)
[17] 尹杨燕, 彭昊, 李小宁, 等. 2022. 枯草芽孢杆菌体外抑菌试验及其抑菌蛋白的鉴定[J]. 畜牧与兽医, 54(12): 57-63.
(Yin Y Y, Peng H, Li X N, et al.2022. Antibacterial test in vitro of Bacillus subtilis and identification of its antibacterial proteins[J]. Animal Husbandry & Veterinary Medicine, 54(12): 57-63.)
[18] 余贤美, 侯长明, 王洁, 等. 2017. 枯草芽孢杆菌Bs-15对柿树炭疽病的离体防治效果[J]. 山东农业科学, 49(07): 125-127.
(Yu X M, Hou C M, Wang J, et al.2017. In vitro control effect of Bacillus subtilis Bs-15 on persimmon anthracnose[J]. Shandong Agricultural Sciences, 49(07): 125-127.)
[19] 曾大兴, 张晓阳, 贾书娟, 等. 2015. 枯草芽孢杆菌C-D6对辣椒炭疽菌附着胞形成的抑制作用研究[J]. 微生物学通报, 42(12): 2377-2385.
(Zeng D X, Zhang X Y, Jia S J, et al.2015. Bacillus subtilis C-D6 as a potential biocontrol agent against appressorium formation of Colletotrichum capsica[J]. Microbiology China, 42(12): 2377-2385.)
[20] 张凯歌, 魏琛, 余璐, 等. 2024. 枯草芽孢杆菌PW2挥发性产物对黄曲霉的抑制作用[J]. 食品与生物技术学报, 43(05): 122-129.
(Zhang K G, Wei C, Yu L, et al.2024. Inhibition of Aspergillus flavus by volatile organic compounds produced by Bacillus subtilis PW2[J]. Journal of Food Science and Biotechnology, 43(05): 122-129.)
[21] 张淘崴, 乌日娜, 纪帅奇, 等. 2024. 豆酱源枯草芽孢杆菌S1-2产细菌素分离鉴定及其稳定性研究[J]. 食品工业科技, 45(08): 158-164.
(Zhang T W, Wu R N, Ji S Q, et al.2024. Isolation and identification of bacteriocin-producing Bacillus subtilis S1-2 from soybean paste and its stability study[J]. Science and Technology of Food Industry, 45(08): 158-164.)
[22] 赵金, 高子晴, 丛丽娜. 2023. 贝莱斯芽孢杆菌 BA-300 的鉴定、发酵条件优化及应用[J]. 大连工业大学学报, 42(3): 176-180.
(Zhao J, Gao Z Q, Cong L N.2023. Identification, optimal fermentation conditions and application of Bacillus velezensis BA-300[J]. Journal of Dalian Polytechnic University, 42(3): 176-180.)
[23] 祖雪, 周瑚, 朱华珺, 等. 2022. 枯草芽孢杆菌K-268的分离鉴定及对水稻稻瘟病的防病效果[J]. 生物技术通报, 38(06): 136-146.
(Zu X, Zhou H, Zhu H J, et al.2022. Isolation and identification of Bacillus subtilis K-268 and its biological control effect on rice blast[J]. Biotechnology Bulletin, 38(06): 136-146.)
[24] Da Silva L L, Moreno H L A, Correia H L N, et al.2020. Colletotrichum: Species complexes, lifestyle, and peculiarities of some sources of genetic variability[J]. Applied Microbiology Biotechnology, 104(5): 1891-1904.
[25] ElNogoumy Baher A, Salem Mohamed A, ElKot Gabr A, et al.2022. Evaluation of the impacts of potassium bicarbonate, Moringa oleifera seed extract, and Bacillus subtilis on sugar beet powdery mildew[J]. Plants, 11(23): 3258-3258.
[26] Nguyen A T V, Nguyen D V, Tran M T, et al.2015. Isolation and characterization of Bacillus subtilis CH16 strain from chicken gastrointestinal tracts for use as a feed supplement to promote weight gain in broilers[J]. Letters in Applied Microbiology, 60(6): 580-588.
[27] Cannon P F, Damm U, Johnston P R, et al.2012. Colletotrichum-current status and future directions[J]. Studies in Mycology, 73: 181-213.
[28] Tang Z Y, Lou J, He L Q, et al.2022. First report of Colletotrichum fructicola causing anthracnose on cherry(Prunus avium) in China[J]. Plant Disease, 106(1): 317.
[29] Tian Y, Ji S H, Zhang E R, et al.2023. Complete genome analysis of Bacillus subtilis TY-1 reveals its biocontrol potential against tobacco bacterial wilt[J]. Marine Genomics, 68: 101018.
[30] Wang W L, Li X, Li Z Z, et al.2025. Revisiting causal organisms of tea anthracnose: Pathogen isolation and pathogenicity identification[J]. Beverage Plant Research, 5: e016. |
|
|
|
