Application of Pseudomonas chlororaphis subsp. aurantiaca against Gray Leaf Spot of Tomato

doi:10.16409/j.cnki.2095-039x.2021.06.018

Abstract

Abstract: In order to obtain the antagonistic strain with stronger inhibition and biocontrol effect on gray leaf spot of tomato. In this study, the biocontrol bacterium YS05 isolated from tomato field soil samples had significant inhibitory effect on Stemphylium lycopersici (Enjoji) Yamamoto by gradient dilution plate method. The strain YS05 was identified as Pseudomonas chlororaphis subsp. aurantiaca based on biological characteristics, physiological and biochemical characteristics, and multilocus sequence analysis. The results of biological characters assay show that strain YS05 could produce protease, HCN, phenazine-1-carboxilic acid and pyrrolnitrin during the metabolic process, and exhibited broad inhibitory spectrum against nine pathogenic fungi and three pathogenic bacteria. Under the condition of in vitro leaves and potted plants in greenhouse, the results of biocontrol study show that the disease incidence and index of tomato inoculated with strain YS05 were significantly reduced. The control efficacy was 71.40% and 58.27%, respectively. In summary, the strain YS05 has excellent biocontrol characteristics and potential in biocontrol of vegetable diseases.

Key words: Pseudomonas chlororaphis subsp. aurantiaca, gray leaf spot of tomato, Stemphylium lycopersici (Enjoji) Yamamoto, biological control

CLC Number:

S476

HUANG Yishuo, XIE Xuewen, SHI Yanxia, CHAI Ali, LI Lei, LI Baoju. Application of Pseudomonas chlororaphis subsp. aurantiaca against Gray Leaf Spot of Tomato[J]. Chinese Journal of Biological Control, 2021, 37(6): 1265-1275.

References

[1] 谢学文.我国蔬菜匍柄霉叶斑病的病原学及对啶酰菌胺抗药性研究[D].沈阳:沈阳农业大学, 2017.
[2] 李宝聚,周艳芳,李金萍,等.李宝聚博士诊病手记(三十)番茄匍柄霉叶斑病(灰叶斑病)的诊断与防治[J].中国蔬菜, 2010(23):24-26.
[3] 邱德文.我国植物病害生物防治的现状及发展策略[J].植物保护, 2010, 36(4):15-18.
[4] 章茂林.拮抗细菌SU8抑菌活性与抑菌物质提取及其理化性质初步研究[D].长沙:湖南农业大学, 2014.
[5] 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, 24(3):9-11.
[6] Ilhan K, Karabulut O A. Efficacy and population monitoring of bacterial antagonists for gray mold (Botrytis cinerea Pers. ex. Fr.) infecting strawberries[J]. BioControl, 2013, 58(4):457-470.
[7] 孙广正,姚拓,赵桂琴,等.荧光假单胞菌防治植物病害研究现状与展望[J].草业学报, 2015, 24(4):174-190.
[8] 李永斌,李云龙,关国华,等.植物根际促生菌的筛选、鉴定及其对小麦的减肥增产效果[J].农业生物技术学报, 2020, 28(8):145-150.
[9] 刘程程,王媛花,张雯婷,等.草莓炭疽病拮抗菌SKL108的分离、鉴定及拮抗活性[J].江苏农业科学, 2020, 48(10):125-130.
[10] 李静,冯淑杰,肖晶,等.小白菜内生假单胞菌XBC-PS的生防作用[J].中国蔬菜, 2007, 1(5):21-23.
[11] 胡菁.恶臭假单胞菌A1在番茄青枯病生物防治中的应用[D].福州:福建农林大学, 2017.
[12] Jiao Z, Wu N, Hale L, et al. Characterisation of Pseudomonas chlororaphis subsp. aurantiaca strain Pa40 with the ability to control wheat sharp eyespot disease[J]. Annals of Applied Biology, 2013, 163(3):444-453.
[13] 许帅,谢学文,张昀,等.马铃薯枯萎病生防芽胞杆菌筛选及生防效果研究[J].中国生物防治学报, 2020, 36(5):761-770.
[14] 黄艺烁,谢学文,石延霞,等.多粘类芽胞杆菌ZF197对白菜茎基腐病防治效果[J].园艺学报, 2020, 47(6):1059-1071.
[15] 李新宇,李磊,陈利达,等.番茄匍柄霉叶斑病拮抗细菌的筛选与鉴定[J].园艺学报, 2020, 47(4):741-748.
[16] 郑斐,李磊,石延霞,等.水生拉恩氏菌ZF7分离鉴定及其促生防病效果研究[J].植物病理学报, 2019, 49(6):836-845.
[17] 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.
[18] Buchanan R E, Bergey N E. Bergey's manual of determinative bacteriology, 9th edition[M]. Baltimore:Williams and Wilkins Company, 1994.
[19] Lalucat J, Mulet M, Gomila M, et al. Genomics in bacterial taxonomy:impact on the genus pseudomonas[J]. Genes, 2020, 11(2):139.
[20] Hesse C, Schulz F, Bull C T, et al. Genome-based evolutionary history of Pseudomonas spp.[J]. Environmental Microbiology, 2018, 20(6):2142-2159.
[21] Stonier T. Agrobacterium tumefaciens conn. II. production of an antibiotic substance[J]. Journal of Bacteriology, 1960, 79:889.
[22] 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.
[23] 李昕,曾洁,王岱,等.细菌耐药耐受性机制的最新研究进展[J].中国抗生素杂志, 2020, 45(2):113-121.
[24] Zhang Y, Fernando W G D, Kievit T R D E, et al. Detection of antibiotic-related genes from bacterial biocontrol agents with polymerase chain reaction[J]. Canadian Journal of Microbiology, 2006, 52(5):476-481.
[25] Delaney S M, Mavrodi D V, Bonsall R F, et al. PhzO, a gene for biosynthesis of 2-hydroxylated phenazine compounds in Pseudomonas aureofaciens 30-84[J]. Journal of Bacteriology, 2001, 183(1):318-327.
[26] 魏海雷,王烨,张力群,等.生防菌株2P24与CPF-10的鉴定及其生防相关性状的初步分析[J].植物病理学报, 2004(1):80-85.
[27] Briard B, Heddergott C, Latgé J P, et al. Volatile compounds emitted by Pseudomonas aeruginosa stimulate growth of the fungal pathogen Aspergillus fumigatus.[J]. Mbio, 2016, 7(2):e00219-16.
[28] Li B J, Yuan H Z, Fang J C, et al. Recent progress of highly efficient in vivo biological screening for novel agrochemicals in China[J]. Pest Management Science, 2010, 66(3):238-247.
[29] 农业部农药检定所生测室.农药田间药效实验准则[M].北京:中国标准出版社, 1993, 45-51.
[30] Peix A, Valverde A, Rivas R, et al. Reclassification of Pseudomonas aurantiaca as a synonym of Pseudomonas chlororaphis and proposal of three subspecies, P. chlororaphis subsp. chlororaphis subsp. nov., P. chlororaphis subsp. aureofaciens subsp. nov., comb. nov. and P. chlororaphis subsp. aurantiaca subsp. nov., comb. nov.[J]. International Journal of Systematic and Evolutionary Microbiology, 2007, 57(6):1286-1290.
[31] Rovera M, Pastor N, Niederhauser M, et al. Evaluation of Pseudomonas chlororaphis subsp. aurantiaca SR1 for growth promotion of soybean and for control of Macrophomina phaseolina[J]. Biocontrol Science and Technology, 2014, 24(9):1012-1025.
[32] Hu W, Gao Q, Hamada M S, et al. Potential of Pseudomonas chlororaphis subsp aurantiaca strain Pcho10 as a biocontrol agent against Fusarium graminearum[J]. Phytopathology, 2014, 104(12):1289-1297.
[33] 张莹莹.绿针假单胞菌HL5-4对番茄灰霉病的抑制活性及其初步研究[D].郑州:河南农业大学, 2013.
[34] 年洪娟.荧光假单胞菌的分子生物学[M].云南:科学技术出版社, 2010, 5-7.
[35] 王灿华,祝新德,许煜泉,等.假单胞菌株M18分泌羧基吩嗪抑制黄瓜枯萎病害[J].上海交通大学学报, 2000(11):1574-1578.
[36] 郭洁心,张育铭,朱洪磊,等.解淀粉芽胞杆菌XJ5挥发性物质抑菌活性及对苹果褐腐病防效测定[J].中国生物防治学报, 2020, 36(4):575-580.
[37] 杨艺炜,黎妍妍,张安盛,等.烟草黑胫病拮抗菌XF10的筛选与鉴定[J].烟草科技, 2018, 51(4):20-27.
[38] 冯福山,刘君昂,胡廉成,等.枯草芽胞杆菌Y13挥发性物质的分析及抑菌活性[J].中国生物防治学报, 2019, 35(4):597-604.
[39] 陈田硕.脱落酸对番茄防御灰叶斑病的功能研究[D].泰安:山东农业大学, 2020.
[40] 李敏.建立高效的人工接种实验体系筛选抗灰叶斑病的番茄种质[D].泰安:山东农业大学, 2019.