Studies on Antibacterial and Plant-Growth-Promoting Characterization in Biocontrol Bacterial Strains PA2101 and PG3402

doi:10.16409/j.cnki.2095-039x.2020.01.009

Abstract

Abstract: We investigated the antimicrobial activity, inhibition and plant-growth-promoting characterization of Pseudomonas aeruginosa PA2101 and Pseudomonas granadensis PG3402 with great biocontrol potential for tobacco in our laboratory. The results showed that PA2101 and PG3402 greatly inhibited Fusarium oxysporum, Alternaria alternata, Rhizoctonia cerealis, Macrophomina phaseolina and Phoma herbarum. The fermentation filtrates of the two bacteria caused the abnormal hyphal growth of Phytophthora nicotianae and Thielaviopsis basicola, as well as inhibited spore germination of T. basicola. They could produce volatile substances with inhibitory activity against P. nicotianae and T. basicola. Both bacteria were detected to be able to produce protease, cellulose and β-1,3-glucanase. Meanwhile, PA2101 was detected to be able to secrete chitinase. Both could synthesize HCN and harbored the genes for synthesis of phenazine-1-carboxylic acid (PCA). PG3402 had the genes for synthesis of 2,4-diacetylphloroglucinal (DAPG). It was found that PA2101 and PG3402 could produce siderophores and IAA, and that they had the characteristic of solubilizing phosphorus. To sum up, the two bacteria PA2101 and PG3402 have a broad antimicrobial spectrum, and can inhibit the growth of P. nicotianae and T. basicola. They can produce a variety of inhibition and plant growth promoting substances, which also contain specific antibiotic genes, indicating that PA2101 and PG3402 have multiple biocontrol functions and that they can be used for prevention of tobacco disease.

Key words: biocontrol bacteria, antimicrobial activity, inhibition characterization, plant-growth-promoting mechanism

CLC Number:

S476

QIAN Huimin, ZHAO Hui, LIU Xintao, NI Yunxia, QIU Rui, LI Xiaojie, ZHAO Xinbei, LI Shujun, LIU Hongyan, WEN Yi. Studies on Antibacterial and Plant-Growth-Promoting Characterization in Biocontrol Bacterial Strains PA2101 and PG3402[J]. Chinese Journal Of Biological Control, 2020, 36(1): 135-144.

References

[1] 杨春云, 邓渊钰, 李伟, 等. 小麦全蚀病生防细菌的筛选和鉴定[J]. 中国生物防治学报, 2018, 34(6):873-881.
[2] Patil S V, Jayamohan N S, Kumudini B S. Strategic assessment of multiple plant growth promotion traits for shortlisting of fluorescent Pseudomonas spp. and seed priming against ragi blast disease[J]. Plant Growth Regulation, 2016, 80(1):1-12.
[3] 唐佳频, 邵宗泽, 张智涛, 等. 南极土壤来源的恶臭假单胞菌1A00316抗南方根结线虫的机制[J]. 应用与环境生物学报, 2014, 20(6):1045-1051.
[4] 郭丛, 杨金广, 申莉莉, 等. 一株对TMV具有显著拮抗活性的恶臭假单胞菌的筛选与鉴定[J]. 华南农业大学学报, 2011, 32(3):57-60.
[5] Vassilev N, Vassileva M, Lopez A, et al. Unexploited potential of some biotechnological techniques for biofertilizer production and formulation[J]. Applied Microbiology and Biotechnology, 2015, 99(12):4983-4996.
[6] 方运玲, 孙爽, 申阅, 等. 微生物源农药申嗪霉素的研制与应用[J]. 农药学学报, 2014, 16(4):387-393.
[7] Ma L, Zhang H Y, Zhou X K, et al. Biological control tobacco bacterial wilt and black shank and root colonization by bio-organic fertilizer containing bacterium Pseudomonas aeruginosa NXHG29[J]. Applied Soil Ecology, 2018, 129:136-144.
[8] 朱流红, 安启菲, 吴轶凡, 等. 烟草黑胫病菌对7种杀菌剂的敏感性[J]. 山地农业生物学报, 2016, 35(1):18-22.
[9] 顾金刚, 方敦煌, 李天飞, 等. 两株荧光假单胞杆菌菌株对烟草黑胫病病原菌的抑制作用[J]. 中国生物防治, 2004, 23(1):76-78, 81.
[10] Gómez-Lama Cabanás C, Legarda G, Ruano-Rosa D, et al. Indigenous Pseudomonas spp. strains from the olive (Olea europaea L.) rhizosphere as effective biocontrol agents against Verticillium dahliae:from the host roots to the bacterial genomes[J]. Frontiers in Microbiology, 2018, 9:277.
[11] Redondo-nieto M, Barret M, Morrissey J, et al. Genome sequence reveals that Pseudomonas fluorescens F113 possesses a large and diverse array of systems for rhizosphere function and host interaction[J]. BMC Genomics, 2013, 14(54):1-17.
[12] Jitendra Mishra, Naveen Kumar Arora. Secondary metabolites of fluorescent pseudomonads in biocontrol of phytopathogens for sustainable agriculture[J]. Applied Soil Ecology, 2018, 125:35-45.
[13] Wallace R L, Hirkala D L, Nelson L M. Mechanisms of action of three isolates of Pseudomonas fluorescens active against postharvest grey mold decay of apple during commercial storage[J]. Biological Control, 2018, 117:13-20.
[14] 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, 34(1):80-85.
[15] 余贤美, 郑服丛. 嗜铁素在促进植物生长及病害防治等方面的应用[J]. 中国农学通报, 2007(8):507-510.
[16] Kucey R M N, Janzen H H, Leggett M E. Microbially mediated increases in plant-available phosphorus[J]. Advances in Agronomy, 1989, 42:199-228.
[17] 王欢, 王敬敬, 徐松, 等. 有机磷降解菌的筛选及其促生特性[J]. 微生物学报, 2017, 57(5):667-680.
[18] 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.
[19] 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.
[20] Roberts W K, Selitrennikoff C P. Plant and bacterial chitinases differ in antifungal activity[J]. Microbiology, 1988, 134(1):169-176.
[21] Ghose T K. Measurement of cellulase activities[J]. Pure and Applied Chemistry, 1987, 59(2):257-268.
[22] 王明江, 章如意, 林多多, 等. 棉花黄萎病不同抗性品种内生菌数量调查与拮抗菌筛选[J]. 江苏农业科学, 2010, 38(2):102-104.
[23] 何碧珀, 郝学政, 刘红彦, 等. 解淀粉芽孢杆菌B10-26对芝麻的促生防病效果及其定殖能力分析[J]. 河南农业科学, 2018, 47(12):78-83.
[24] Castric K F, Castric P A. Method for rapid detection of cyanogenic bacteria[J]. Applied and Environmental Microbiology, 1983, 45(2):701-702.
[25] Prabhukarthikeyan S R, Keerthana U, Raguchander T. Antibiotic-producing Pseudomonas fluorescens mediates rhizome rot disease resistance and promotes plant growth in turmeric plants[J]. Microbiological Research, 2018, 210:65-73.
[26] 李洪林. 荧光假单胞菌生防菌株的筛选及对烟草赤星病菌的抑制作用[D]. 大庆:黑龙江八一农垦大学, 2008.
[27] 杨常娥, 鲁艳莉, 倪捍成, 等. 创伤弧菌产铁载体菌株的筛选及其诱导条件的响应面优化[J]. 食品工业科技, 2017, 38(3):159-165.
[28] 杨顺, 杨婷, 林斌, 等. 两株溶磷真菌的筛选、鉴定及溶磷效果的评价[J]. 微生物学报, 2018, 58(2):264-273.
[29] Patten C L, Glick B R. Role of Pseudomonas putida indoleacetic acid in development of the host plant root system[J]. Applied and Environmental Microbiology, 2002, 68(8):3795-3801.
[30] 易龙, 马冠华, 肖崇刚. 烟草根黑腐病拮抗内生细菌的筛选及其抑菌作用[J]. 微生物学通报, 2012, 39(10):1464-1470.
[31] 刘烈花. 4种生防菌对烟草黑胫病的防治作用[J]. 植物医生, 2018, 31(4):42-44.
[32] 檀建新, 陈忠义, 张杰, 等. 产几丁质酶菌的分离鉴定及其抑菌作用的初步研究[J]. 植物保护, 2001, 27(2):1-3.
[33] 金志雄, 周围, 王娅, 等. 内生枯草芽孢杆菌SWB8菌株β-1,3-1,4-葡聚糖酶的抗菌作用及细胞毒性[J]. 微生物学报, 2011, 51(11):1527-1537.
[34] 蒋海霞, 周莲, 何亚文. 铜绿假单胞菌生防菌株抑菌代谢产物及其生防应用研究进展[J]. 微生物学通报, 2015, 42(7):1338-1349.
[35] Souza J T D, Arnould C, Deulvot C, et al. Effect of 2, 4-diacetylphloroglucinol on Pythium:cellular responses and variation in sensitivity among propagules and species[J]. Phytopathology, 2003, 93(8):966-975.
[36] GómezLama Cabanás C, Schilirò E, Valverdecorredor A, et al. The biocontrol endophytic bacterium Pseudomonas fluorescens PICF7 induces systemic defense responses in aerial tissues upon colonization of olive roots[J]. Frontiers in Microbiology, 2014, 5:427.
[37] Voisard C, Keel C, Haas D, et al. Cyanide production by Pseudomonas fluorescens helps suppress black root rot of tobacco under gnotobiotic conditions[J]. EMBO Journal, 1989, 8(2):351-358.
[38] 张琳, 陈彦宏, 王雪妍, 等. 三七植物内生细菌的促生作用[J]. 大连工业大学学报, 2018, 37(4):244-248.
[39] Arya N, Rana A, Rajwar A, et al. Biocontrol efficacy of siderophore producing indigenous pseudomonas strains against Fusarium wilt in tomato[J]. National Academy Science Letters, 2018, 41(3):133-136.
[40] 李静, 张金羽, 张琪, 等. 玉米根际无机磷溶解菌的筛选与促生特性[J/OL]. 应用与环境生物学报, 2019:1-11.
[41] Rijavec T, Lapanje A. Hydrogen cyanide in the rhizosphere:not suppressing plant pathogens, but rather regulating availability of phosphate[J]. Front Microbiology, 2016, 7:1785.