Isolation and Identification of PseudomonasZF509 and Its Control Effect on Bacterial Fruit Blotch of Melon

doi:10.16409/j.cnki.2095-039x.2024.02.017

Abstract

Abstract: In order to obtain the biocontrol potential strain against bacterial fruit blotch (BFB), strain ZF509 isolated from potato rhizosphere soil in Heilongjiang Province was tested against Acidovorax citrulli. The stain ZF509 had biocontrol efficiency on BFB. Morphological and biochemical characterization tests were carried out on the stain ZF509 for identification. Phylogenetic analysis based on multilocus sequence revealed that stain ZF509 clustered with Pseudomonas protegens. Agar disk diffusion test showed that strain ZF509 had inhibitory effect on 4 pathogenic bacterial and 8 pathogenic fungi. Enzyme activity detection showed that strain ZF509 could dissolve phosphorus, secrete protease, synthesize biofilm and produce hydrocyanic acid. PCR amplification of genes of four bacteriostatic substances with specific primers showed that strain ZF509 had genes of producing 2-hydroxylated phenazine, 2,4-DAPG, pyrrolnitrin and pyoluteori. Pot experiments showed that the disease index significantly reduced after inoculation with ZF509 and the control efficacy was up to 73.46%, which was superior to that of benziothiazolinone. The strain ZF509 has excellent biocontrol and practical potential against BFB.

Key words: Pseudomonas protegens, bacterial fruit blotch, biocontrol efficiency, antibacterial substances

CLC Number:

S476

ZHAO Zixuan, LI Junhui, DU Gongfu, SHI Yanxia, XIE Xuewen, CHAI Ali, FAN Tengfei, LI Baoju, LI Lei. Isolation and Identification of PseudomonasZF509 and Its Control Effect on Bacterial Fruit Blotch of Melon[J]. Chinese Journal of Biological Control, 2024, 40(3): 690-700.

References

[1] 季苇芹, 叶云峰, 张爱萍, 等. 我国瓜类细菌性果斑病研究新进展[J]. 中国瓜菜, 2022, 35(9): 1-8.
[2] 阎莎莎, 王铁霖, 赵廷昌. 瓜类细菌性果斑病研究进展[J]. 植物检疫, 2011, 25(3): 71-76.
[3] 蔡馥宇, 关巍, 乔培, 等. 瓜类细菌性果斑病研究新进展[J]. 中国瓜菜, 2017, 30(11): 1-5.
[4] 王晓青, 杨扬, 赵朔, 等. 我国瓜类细菌性果斑病发生现状和综合防控策略[J]. 植物检疫, 2012, 26(5): 79-82.
[5] Fravel D R. Commercialization and implementation of biocontrol[J]. Annual Review of Phytopathology, 2005, 43: 337-359.
[6] 张伟琼, 聂明, 肖明. 荧光假单胞菌生防机理的研究进展[J]. 生物学杂志, 2007(03): 9-11.
[7] 杨珊. 绿针假单胞菌zm-1生物防治花生白绢病的作用机理研究[D]. 郑州: 河南大学, 2022.
[8] 江聪, 张雪, 徐代英, 等. 一种防治小麦赤霉病的假单胞菌CXZ8、生防菌剂及其应用[P]. 中国发明专利, 2022, CN115232776A.
[9] 金利容, 许冬, 王玲等. 恶臭假单胞菌HB3S-20在棉花中的定殖及诱导抗性研究[J]. 中国生物防治学报, 2024, 40(1): 117-125.
[10] 聂晨曦, 向涛, 张雪洪, 等. 防御假单胞菌H78中crp基因对抗生素合成的调控作用[J]. 生物学杂志, 2023, 40(3): 11-15.
[11] 潘静文, 王正, 张雪洪, 等. 防御假单胞菌H78中sRNA基因crcY/Z正调控藤黄绿菌素合成[J]. 基因组学与应用生物学, 2022, 41(4): 771-777.
[12] 常琳, 李倩, 童蕴慧, 等. 生防细菌FD6的鉴定及其对番茄灰霉病菌的作用机制[J]. 植物保护学报, 2011, 38(6): 487-492.
[13] Yu F, Jing X, Li X,et al. RecombineeringPseudomonas protegensCHA0: An innovative approach that improves nitrogen fixation with impressive bactericidal potency[J]. Microbiological Research, 2019, 218: 58-65.
[14] Henkels M D, Kidarsa T A, Shaffer B T,et al.Pseudomonas protegens Pf-5 causes discoloration and pitting of mushroom caps due to the production of antifungal metabolites[J]. Molecular Plant Microbe Interaction, 2014, 27(7): 733-746.
[15] 李磊, 李宝聚, 黄艺烁, 等. 一株绿针假单胞菌桔黄亚种在防治番茄匍柄霉叶斑病中的应用[J]. 中国生物防治学报, 2021, 37(6): 1265-1275.
[16] 东秀珠, 洪俊华. 原核微生物的多样性[J]. 生物多样性, 2001(1): 18-24.
[17] Buchanan R E. 伯杰细菌鉴定手册: 第八版[M]. 北京: 科学出版社, 1984.
[18] 赵子璇, 曾先锋, 覃诗扬, 等. 贝莱斯芽胞杆菌ZF438菌株的鉴定及其发酵上清液对辣椒炭疽病的抑菌作用[J]. 农业生物技术学报, 2023, 31(10): 2163-2175.
[19] Kasana R C, Salwan R, Dhar H,et al. A rapid and easy method for the detection of microbial cellulases on agar plates using gram's iodine[J]. Current Microbiology, 2008, 57(5): 503-507.
[20] Jiao Z, Wu N, Hale L,et al. Characterisation ofPseudomonas chlororaphissubsp.aurantiaca strain Pa40 with the ability to control wheat sharp eyespot disease[J]. Annals of Applied Biology, 2013, 163(3): 444-453.
[21] 李佳珣, 叶雨寒, 胡心怡等. 抑制铜绿假单胞菌和金黄色葡萄球菌生物膜形成的乳杆菌筛选及机制初探[J]. 微生物学杂志, 2023, 43(6): 89-96.
[22] 费诺亚, 陈华民, 杨玉文, 等. 瓜类细菌性果斑病国外研究新进展[J]. 中国瓜菜, 2022, 35(7): 1-5.
[23] Jousset A, Schuldes J, Keel C,et al. Full-Genome Sequence of the Plant Growth-Promoting BacteriumPseudomonas protegens CHA0[J]. Genome Announcements, 2014, 2(2): e00322-14.
[24] 吴丽媛, 刘宝玉, 王钰杰, 等. 甜瓜细菌性果斑病生防菌株BW-6的筛选和鉴定[J]. 植物保护, 2014, 40(1): 43-47.
[25] 王雪妍, 岳丹丹, 张志龙, 等. 一株贝莱斯芽孢杆菌HP-24及其在制备防治瓜类细菌性果斑病菌液中的应用[P]. 中国发明专利, 2022, CN113755393B.
[26] 梁靖聆, 朱勃, 聂文翰, 等. 生防假单胞菌及其应用[P]. 中国发明专利, 2020, CN111187741A.
[27] 钟小燕, 梁妙芬, 甄锡壮, 等. 假单胞菌对香蕉枯萎病菌的抑制作用[J]. 植物保护, 2009, 35(1): 86-89.
[28] 娄志英, 王云鹏, 袁启明, 等. 生防假单胞菌Pf-5对秀丽隐杆线虫的毒性及其作用机制[J]. 南京农业大学学报, 2019, 42(5): 827-834.
[29] Andreolli M, Zapparoli G, Angelini E,et al.Pseudomonas protegens MP12: A plant growth-promoting endophytic bacterium with broad-spectrum antifungal activity against grapevine phytopathogens[J]. Microbiological Research, 2019, 219: 123-131.
[30] Lai X, Niroula D, Burrows M,et al. Identification and characterization of bacteria-derived antibiotics for the biological control of pea aphanomyces root rot[J]. Microorganisms, 2022, 10(8): 1596.
[31] Harting R, Nagel A, Nesemannet al.Pseudomonasstrains induce transcriptional and morphological changes and reduce root colonization ofVerticillium spp.[J]. Frontiers in Microbiology, 2021, 12: 652468.
[32] 董秋伶. 荧光假单胞菌2P24中grxD调控2,4-DAPG产生机理及其功能分析[D]. 南宁: 广西大学, 2022.
[33] 陈韵, 汪晓雷, 黄显清, 等. 假单胞菌M18株las系统负调控藤黄绿脓菌素和吩嗪-1-羧酸的合成[J]. 中国科学(C辑: 生命科学), 2008(3): 230-237.
[34] 许煜泉, 黄显清. 假单胞菌M18中藤黄绿菌素生物合成调控因子的基因序列[P]. 中国发明专利, 2004, CN1523107.