Physical and Chemical Properties and Inhibitory Effect of Crude Antifungal Protein Produced by Bacillus tequilensis JN-369

doi:10.16409/j.cnki.2095-039x.2020.02.006

Abstract

Abstract: Bacillus tequilensis strain JN-369 is a biocontrol strain with antagonism on Magnaporthe oryzae. The antifungal protein crude extraction was precipitated by ammonium sulphate after fermentation. To determine the antifungal mechanism of the crude protein the inhibition spectrum, thermostability, acid-base stability, UV stability and enzyme stability were investigated, the inhibition effect to M. oryzae hyphae and spores was observed, and the antagonistic effects of 40% Isoprothiolane EC, 25% Tricyclazole WP and the crude antifungal protein against M. oryzae in rice were detected by tissue inoculation in vitro. Results indicated that the crude antifungal protein of the strain JN-369 had a broad inhibition spectrum, with inhibition activity on 13 plant pathogenic fungi and oomycetes, such as B. cinerea, M. oryzae, but no inhibition on S. sclerotiorum. The crude proteinstill had antifungal activity after heat treatment, pH treatment, UV treatment and enzyme treatment, indicating it has potential application value in biocontrol of food crops. After treatment with crude protein, M. oryzae hyphae became enlargement and could inhibit conidiospores germination. The in vitro efficacy experiments showed that all treatments had significant antagonistic effects on rice blast, and the crude antifungal protein at 50 ug/mL to 100 ug/mL had the best antagonistic effect on rice blast.The strain JN-369 was expected to be developed as a Biocontrol Strain for the control of rice blast.

Key words: Magnaporthe oryzae, Bacillus tequilensis, crude antifungal protein, biocontrol

CLC Number:

S476

ZHOU Hu, HU Ling, YU Xiyue, ZHU Huajun, REN Zuohua, LIU Erming. Physical and Chemical Properties and Inhibitory Effect of Crude Antifungal Protein Produced by Bacillus tequilensis JN-369[J]. Chinese Journal Of Biological Control, 2020, 36(2): 211-219.

References

[1] 杨华, 雷平, 郭照辉, 等. 细菌防治水稻稻瘟病研究进展[J]. 江苏农业科学, 2019, 47(7):99-104.
[2] Liu X, Fang Y, Xie F, et al. Synthesis and in vivo fungicidal activity of some new quinoline derivatives against rice blast[J]. Pest Management Science, 2017, 73(9):1900-1907.
[3] Wu L, Xiao W, Chen G, et al. Identification of Pseudomonas mosselii BS011 gene clusters required for suppression of rice blast fungus Magnaporthe oryzae[J]. Journal of Biotechnology, 2018, 16(4):1-9.
[4] 谢梓语, 郭恩辉, 孙宇波, 等. 枯草芽胞杆菌B1409对番茄和辣椒的防病促生作用[J]. 植物保护学报, 2018, 45(3):520-527.
[5] Johansson A H, Bejai S, Niazi A, et al. Studies of plant colonisation by closely related Bacillus amyloliquefaciens biocontrol agents using strain specific quantitative PCR assays[J]. Antonie van Leeuwenhoek, 2014, 106(6):1247-1257.
[6] Gatson J W, Benz B F, Chandrasekaran C, et al. Bacillus tequilensis sp nov. isolated from a 2000-year-old Mexican shaft-tomb, is closely related to Bacillus subtilis[J]. International Journal of Systematic and Evolutionary Microbiology, 2006, 56(7):1475-1484.
[7] Gholami M, Khakvar R, Aliasgarzad N. Application of endophytic bacteria for controlling anthracnose disease (Colletotrichum lindemuthianum) on bean plants[J]. Archives Phytopathology and Plant Protection, 2013, 46(15):1831-1838.
[8] 袁雪, 侯旭, 胡晓, 等. 桃褐腐病拮抗细菌的筛选、鉴定及生防作用[J]. 北京农学院学报, 2018, 33(4):11-17.
[9] 张猛, 王琼, 冯发运, 等. 植物内生特基拉芽胞杆菌的分离、鉴定及防治西瓜枯萎病效果[J]. 中国生物防治学报, 2017, 33(3):371-377.
[10] 谢洁, 任慧爽, 唐翠明, 等. 一株桑树内生细菌的鉴定和对桑椹核地杖菌的拮抗作用[J]. 蚕业科学, 2016, 41(5):815-824.
[11] Adenekan M K, Fadimu G J, Odunmbaku L A, et al. Effect of isolation techniques on the characteristics of pigeon pea (Cajanus cajan) protein isolates[J]. Food Science and Nutrition, 2018, 6(1):146-152.
[12] 周瑚, 邹秋霞, 胡玲, 等. 特基拉芽胞杆菌JN-369的分离鉴定及其抑菌物质分析[J]. 农药学学报, 2019, 21(1):52-58.
[13] 邹秋霞, 任佐华, 高诗涵, 等. 枯草芽胞杆菌YN145分离鉴定及抑菌活性[J]. 中国生物防治学报, 2017, 33(3):421-426.
[14] Gotor-Vila A, Teixidó N, Sisquella M, et al. Biological characterization of the biocontrol agent Bacillus amyloliquefaciens CPA-8:the effect of temperature, pH and water activity on growth, susceptibility to antibiotics and detection of enterotoxic genes[J]. Current Microbiology, 2017, 74:1089-1099.
[15] 周瑚, 任佐华, 虞选杰, 等. 湖南桃江病圃稻瘟病菌对24个水稻抗稻瘟基因的毒性分析[J]. 植物保护, 2017, 43(3):165-170.
[16] Ma J, Lei C, Xu X, et al. Pi64, encoding a novel CC-NBS-LRR protein, confers resistance to leaf and neck blast in rice[J]. Molecular Plant-Microbe Interaction, 2015, 28(5):558-568.
[17] 马晨燕, 袁正杰, 杨海河, 等. 水稻离体叶片抗纹枯病接种方法的研究[J]. 浙江农业学报, 2016, 28(10):1730-1737.
[18] Oberoi R, Beg Q K, Puri S, et al. Characterization and wash performance analysis of an SDS-stable alkaline protease from a Bacillus sp.[J]. World Journal of Microbiology and Biotechnology, 2001, 17(5):493-497.
[19] 隋好林, 王仁山, 王义华, 等. 土壤酸碱度对作物生长的影响及改良剂应用效果初探[J]. 山东农业科学, 2007(2):90-91.
[20] 周盈, 陈琳, 柴鑫莉, 等. 魔芋内生拮抗细菌的分离及其抗菌物质特性研究[J]. 微生物学报, 2007, 47(6):1076-1079.
[21] 王青华, 孙晓晖, 唐旭, 等. 深海贝莱斯芽胞杆菌DH82的筛选、鉴定及其抗菌粗蛋白性质分析[J]. 海洋通报, 2019(1):63-69.
[22] Thiel T, Wolk C P. Autoinhibition of spore germination in Nostoc spongiaeforme (Cyanophyceae)[J]. Journal of Phycology, 2010, 18(2):305-306.
[23] 孙平平, 贾晓辉, 崔建潮, 等. 梨灰霉病拮抗放线菌L-30的筛选、鉴定及作用机制研究[J]. 园艺学报, 2016, 43(12):2335-2346.
[24] 秦楠, 郝林, 李鑫. 解淀粉芽胞杆菌HRH317抗菌蛋白的分离纯化及其抗菌作用[J]. 植物保护学报, 2015, 42(5):813-819.
[25] 王奕丁, 姜婷婷, 王全. 棉花黄萎病拮抗细菌Bacillus subtilis ZL2-70抗菌蛋白的理化性质和抑菌机理[J]. 棉花学报, 2017, 29(6):560-569.
[26] 杨胜清, 张帆, 马贵龙. 贝莱斯芽胞杆菌S6拮抗物质分离纯化及抑菌机理[J]. 农药, 2017, 56(9):645-648.
[27] 杨定祥, 林巧玲, 卢乃会, 等. 拮抗辣椒疫霉菌海洋细菌菌株SH-27的筛选鉴定及其防病促生作用[J]. 微生物学通报, 2018, 45(1):54-63.