Screening and Identification of Antagonistic Strains from Endophytic Bacteria Isolated from Seeds of Stylosanthes and Exploration of Inhibitory Effects against Colletotrichum gloeosporioides

doi:10.16409/j.cnki.2095-039x.2025.02.003

Abstract

Abstract: Stylosanthes (stylo) is a leguminous forage widely planted in tropical regions of the world. Anthracnose caused by Colletotrichum is the major disease that greatly limits the growth of stylo. Currently, chemical pesticides are the major approaches to control stylo anthracnose, and there is no effective biological pesticides. In order to screen antagonistic bacteria against Colletotrichum, endophytic strains were isolated by co-culturing the seed extract from stylo resistant line with C. gloeosporioides strain. The plate confrontation test showed that 10 strains could significantly inhibit the mycelial growth of C. gloeosporioides, and the ZW21 strain had the strongest antagonistic effect, with the inhibition rates of 70.10%, 73.41%, and 76.86% on the C. gloeosporioides strains HK04, DZ19, and W2, respectively. In addition, the ZW21 strain could also effectively inhibit the mycelial growth of four different C. gloeosporioides strains isolated from major tropical plants. The scanning electron microscope examination showed that the strain ZW21 caused the swelling, distortion and deformity of mycelia. The extracellular enzyme detection showed that the ZW21 strain could produce cellulase, protease, cellulase, xylanase and etc., destroying the structure of pathogen and limiting its growth. Based on physiological, biochemical and molecular biological analysis, strain ZW21 was identified as Bacillus cereus. The inoculation experiments on the detached stylo leaves showed that the fermentation broth of ZW21 strain had a biocontrol efficiency of 89.02%. These results indicate that ZW21 is expected to be used to create bio-pesticides for the prevention and control of stylo anthracnose.

Key words: Stylosanthes, anthracnose, endophytic bacteria, Bacillus cereus, antagonism

CLC Number:

S476

XU Yunfeng, ZHANG Jichang, LUO Lijuan, JIANG Lingyan. Screening and Identification of Antagonistic Strains from Endophytic Bacteria Isolated from Seeds of Stylosanthes and Exploration of Inhibitory Effects against Colletotrichum gloeosporioides[J]. Chinese Journal of Biological Control, 2025, 41(1): 132-142.

References

[1] Luo J J, Liu Y X, Zhang H K, et al. Metabolic alterations provide insights into Stylosanthes roots responding to phosphorus deficiency[J]. BMC Plant Biology, 2020, 20(2): 3-22.
[2] 易克贤, 郑金龙, 习金根, 等. 柱花草炭疽病流行因子初探[J]. 热带农业科学, 2014, 34(2): 74-78.
[3] 贾艳星. 柱花草炭疽病新致病菌的鉴定及致病性研究[D]. 海口: 海南大学, 2017.
[4] 刘丽萍, 高洁, 李玉. 植物炭疽菌属Colletotrichum 真菌研究进展[J]. 菌物研究, 2020, 18(4): 266-281.
[5] Youssef M M A, Eissa M F M. Biofertilizers and their role in management of plant parasitic nematodes: A review[J]. E3 Journal of Biotechnology and Pharmaceutical Research, 2014, 5(1): 1-6.
[6] 徐秋红. 浅析生物防治在蔬菜病虫害防治中的应用[J]. 种子科技, 2022, 40(13): 72-74.
[7] 翟颖妍. 甲基营养型芽孢杆菌ZH-9的筛选、鉴定及其对炭疽病的生物防治研究[D]. 咸阳: 西北农林科技大学, 2020.
[8] 李坤, 洪秀杰, 王欣悦, 等. 贝莱斯芽孢杆菌TC-52的分离鉴定及其对水稻幼苗生长和立枯病的影响[J]. 江苏农业科学, 2024, 52(10): 129-137.
[9] 谢学文, 赵昱榕, 孙雪莹, 等. 番茄疫霉根腐病生防菌的分离鉴定及其防治效果[J]. 植物病理学报, 2020, 50(5): 610-617.
[10] 张宁, 张晶晶, 李雅淑, 等. 人参黑斑病生防菌的分离鉴定及防效[J]. 中国生物防治学报, 2022, 18(2): 1-8.
[11] 梁艳琼,唐文,吴伟怀,等. 甘蔗赤腐病菌拮抗细菌TWC2的分离鉴定及其防效研究[J]. 热带农业科学, 2018, 38(2): 61-68.
[12] 付晓云. 柱花草炭疽病生防菌筛选、抑菌物质分析及防病效果评价[D]. 南京: 南京农业大学, 2018.
[13] 梁艳琼, 吴伟怀, 习金根, 等. 生防菌JNC2对柱花草炭疽病的生防效果及其机理作用[J].基因组学与应用生物学, 2020, 39(12): 5567-5573.
[14] Wang B Z, Li L, Lin Y H, et al. Targeted isolation of biocontrol agents from plants through phytopathogen co-culture and pathogen enrichment[J]. Phytopathology Research, 2022, 4(19): 1-14.
[15] 邓振山, 赵龙飞, 张薇薇, 等. 银杏内生真菌的分离及其对苹果腐烂病病原菌的拮抗作用[J]. 西北植物学报, 2009, 29(3): 608-613.
[16] 东秀珠, 蔡妙英. 常见细菌系统鉴定手册[M]. 北京: 科学出版社, 2001.
[17] 王绍春, 冯淑芬. 粤西地区橡胶树炭疽病流行因素分析[J]. 热带作物学报, 2001, (01): 15-22.
[18] 代真林, 何洪磊, 姚秀英, 等. 抗生素溶杆菌13-6对玉米小斑病的生防作用机制初步研究[J]. 植物病理学报, 2021, 51(3): 393-402.
[19] Han S, Zheng Z L, Li S J, et al. Impact of root rot disease of Zanthoxylum armatum on rhizosphere soil microbes and screening of antagonistic bacteria[J]. Forests, 2023, 14(8): 1561.
[20] Li J L, Hu M, Xue Y, et al. Screening, identification and efficacy evaluation of antagonistic bacteria for biocontrol of soft rot disease caused by Dickeya zeae[J]. Microorganisms, 2020, 8(5): 697-697.
[21] Sun Z Z, Liu T T, Liu Z, et al. Screening of antagonistic bacteria against the blue mold of citrus fruit from soil by a new parallel screening method without prior isolation of single strains[J]. Biological Control, 2022, 176: 105066.
[22] 杨睿, 汪琳罗沙, 姚迪, 等. 猕猴桃根际土壤可培养细菌分离及溃疡病拮抗菌筛选[J]. 安徽农业科学, 2023, 51(22): 98-103.
[23] 张静雅, 李欣雨, 张成, 等. 木薯炭疽病拮抗木霉菌筛选与室内防效研究[J]. 中国生物防治学报, 2022, 38(1): 115-124.
[24] Miljakovi D, Marinkovi J, Baleevi-Tubi S. The significance of bacillus spp. in disease suppression and growth promotion of field and vegetable crops[J]. Microorganisms, 2020, 8(7): 1037.
[25] Rita, Grosch, Simone, et al. Biocontrol of Rhizoctonia solani: complex interaction of biocontrol strains, pathogen and indigenous microbial community in the rhizosphere of lettuce shown by molecular methods[J]. Plant and Soil, 2012, 361: 343-357.
[26] Abriouel H, Franz C M A P, Omar N B, et al. Diversity and applications of Bacillus bacteriocins[J]. FEMS Microbiology Reviews, 2011, 35(1): 201-232.
[27] 冯印印, 李斌, 杨洋, 等. 烟草青枯病菌拮抗细菌的筛选、鉴定和抑菌机制研究[J]. 中国生物防治学报, 2021, 37(2): 331-339.
[28] Bubici G, Kaushal M, Prigigallo M. Biological control agents against Fusarium wilt of banana[J]. Frontiers in Microbiology, 2019, 10: 616.
[29] Hu Y, Li Y Y, Yang X Q, et al. Effects of integrated biocontrol on bacterial wilt and rhizosphere bacterial community of tobacco[J]. Scientific Reports, 2021, 11(1): 2653-2653.
[30] Wang N, Wang L Y, Zhu K, et al. Plant root exudates are involved in Bacillus cereus Ar156 mediated biocontrol against Ralstonia solanacearum[J]. Frontiers in Microbiology, 2019, 10: 98.
[31] 刘任, 韩静君, 游春平, 等. 稻瘟病菌拮抗细菌bio-2的抑菌作用及其鉴定[J]. 植物保护学报, 2011, 38(1): 91-92.
[32] Li C Y, Wang L J, Chai S S, et al. Screening of Bacillus subtilis HAAS01 and its biocontrol effect on Fusarium wilt in sweet potato[J]. Phyton-International Journal of Expermental Botany, 2022, 8: 1779-1793.
[33] 张艳群, 来航线, 韦小敏, 等. 两株生防芽孢细菌筛选、鉴定及拮抗研究[J]. 微生物学通报, 2014, 41(2): 281-289.
[34] 章乐乐, 王冠, 柳凤, 等. 芒果炭疽病拮抗菌分离、鉴定及生防机制研究[J]. 生物技术通报, 2023, 39(4): 277-287.
[35] Richter B S, Ivors K, Shi W, et al. Cellulase activity as a mechanism for suppression of Phytophthora root rot in mulches[J]. Phytopathology, 2011, 101(2): 223-230.
[36] Sindhu S S, Dadarwal K R. Chitinolytic and cellulolytic Pseudomonas sp. antagonistic to fungal pathogens enhances nodulation by Mesorhizobium sp. cicer in chickpea[J]. Microbiological Research, 2001, 156(4): 353-358.
[37] Fan X, Ye T, Li Q, et al. Potential of a quorum quenching bacteria isolate Ochrobactrum intermedium D-2 against soft rot pathogen Pectobacterium carotovorum subsp. carotovorum[J]. Frontiers in Microbiology, 2020(11): 898.
[38] See-Too W S, Convey P, Pearce D A, et al. Characterization of a novel N-acylhomoserine lactonase, aidp, from antarctic Planococcus sp.[J]. Microbial Cell Factories, 2018, 17(1): 179.
[39] Zhang W P, Luo Q Q, Zhang Y Y, et al. Quorum quenching in a novel Acinetobacter sp. XN-10 bacterial strain against Pectobacterium carotovorum subsp. carotovorum[J]. Microorganisms, 2020, 8(8): 1100.
[40] 丁海霞. 生防蜡样芽胞杆菌905 PlcR-PapR群体感应途径研究[D]. 北京: 中国农业大学, 2016.