解淀粉芽孢杆菌HZ-6-3的筛选鉴定及其防治番茄灰霉病效果的评价

doi:10.11686/cyxb2019213

草业学报 ›› 2020, Vol. 29 ›› Issue (2): 31-41.DOI: 10.11686/cyxb2019213

解淀粉芽孢杆菌HZ-6-3的筛选鉴定及其防治番茄灰霉病效果的评价

荆卓琼^1,2, 郭致杰^1,2,*, 徐生军^1,2,*, 何苏琴^1,2

1.甘肃省农业科学院植物保护研究所,甘肃兰州730070;
2.农业部天水作物有害生物科学观测实验站,甘肃天水 741200

收稿日期:2019-03-26 修回日期:2019-07-16 出版日期:2020-02-20 发布日期:2020-02-20
通讯作者: E-mail: guozhijie@gsagr.ac.cn, xusj1001@aliyun.com
作者简介:荆卓琼(1984-),女,甘肃徽县人,助理研究员,硕士。E-mail: jingzhuoqiong@qq.com
基金资助:
国家重点研发计划(2017YFD0200608)和甘肃省农业科学院农业科技创新专项(2014GAAS24; 2016GAAS26; 2017GAAS23; 2019GAAS09; 2019GAAS23)资助

Screening, identification as Bacillus amyloliquefaciens strain HZ-6-3 and evaluation of inhibitory activity against tomato gray mold, of a bacterial isolate

JING Zhuo-qiong^1,2, GUO Zhi-jie^1,2,*, XU Sheng-jun^1,2,*, HE Su-qin^1,2

1.Institute of Plant Protection, Gansu Academy of Agricultural Sciences, Lanzhou 730070, China;
2.Scientific Observing and Experimental Station of Crop Pests in Tianshui, Ministry of Agriculture, Tianshui 741200, China

Received:2019-03-26 Revised:2019-07-16 Online:2020-02-20 Published:2020-02-20
Contact: E-mail: guozhijie@gsagr.ac.cn, xusj1001@aliyun.com

摘要/Abstract

摘要： 采用平板对峙法,从银杏组织分离的内生细菌菌株中筛选对灰葡萄孢菌抑菌活性较好的菌株,其中菌株HZ-6-3的抑制作用最强,抑制率为79.06%。通过菌落形态观察、生理生化特征、16S rRNA基因序列和gyrA基因分析将菌株HZ-6-3鉴定为解淀粉芽孢杆菌。抑菌谱测定结果显示,该菌株对其他8种植物病原真菌均具有不同程度的拮抗作用。该菌株可产生蛋白酶、果胶酶、β-1,3-葡聚糖酶和淀粉酶;能够显著抑制病原菌的菌丝生长,使菌丝生长扭曲,细胞膨大、形成串状泡囊结构等畸变。利用盆栽试验测定了该菌株对番茄灰霉病的防控效果,结果显示1×10⁸ CFU·mL^-1的预防效果和治疗效果最高,可达81.12%和70.45%。上述结果显示菌株HZ-6-3具有作为生物制剂的潜能。

关键词: 解淀粉芽孢杆菌, 银杏, 内生细菌, 番茄灰霉病菌, 防控效果

Abstract: Bacterial strains exhibiting antifungal activity against Botrytis cinerea, which causes tomato gray mold, were screened from endophytic bacterial strains isolated from tissue of Ginkgo biloba by dual culture. The strain HZ-6-3 showed strong antagonistic activity against B. cinerea with an inhibition efficiency of 79.06%. Based on cultural and morphological characterization, physiological and biochemical characteristics, 16S rRNA gene sequence and gyrA gene sequence, strain HZ-6-3 was identified as Bacillus amyloliquefaciens. The results of antimicrobial spectrum determination showed that strain HZ-6-3 inhibited the growth of eight plant pathogenic fungal species. It could produce proteinase, pectinase, β-1,3-glucanase and amylase. The strain HZ-6-3 exhibited a strong antagonistic activity, resulting in abnormal mycelial growth of pathogenic fungi, including twist, cell-expansion and bubble deformation structures. The efficiency of strain HZ-6-3 for prevention and control of gray mold in tomato plant was evaluated by pot experiment. An application of 1×10⁸ CFU·mL^-1 was highly effective against gray mold, and which showed a prevention efficiency of 81.12% and a control efficiency of 70.45%, respectively. These results indicate that B. amyloliquefaciens strain HZ-6-3 may have potential as biological control agent.

Key words: Bacillus amyloliquefaciens, Ginkgo biloba, endophytic bacteria, Botrytis cinerea, prevention and control efficiency

荆卓琼, 郭致杰, 徐生军, 何苏琴. 解淀粉芽孢杆菌HZ-6-3的筛选鉴定及其防治番茄灰霉病效果的评价[J]. 草业学报, 2020, 29(2): 31-41.

JING Zhuo-qiong, GUO Zhi-jie, XU Sheng-jun, HE Su-qin. Screening, identification as Bacillus amyloliquefaciens strain HZ-6-3 and evaluation of inhibitory activity against tomato gray mold, of a bacterial isolate[J]. Acta Prataculturae Sinica, 2020, 29(2): 31-41.

参考文献

[1] Chen F P, Han P, Zhang Z Z, et al. Studies on antifungal activity of fungicide 5-(4-chlorophenyl)-2, 3-dimethyl -3-(pyridine-3)-oxazoline against Botrytis cinerea. Chinese Journal of Pesticide Science, 2010, 12(1): 42-48.
陈凤平, 韩平, 张真真, 等. 啶菌噁唑对番茄灰霉病菌的抑菌作用研究. 农药学学报, 2010, 12(1): 42-48.
[2] Li B J, Zhu G R, Guan T S, et al. Studies on the epidemic regularities of tomato grey mould disease in energy-saving greenhouse. Plant Protection, 2003, 2(29): 313-325.
李宝聚, 朱国仁, 关天舒, 等. 节能日光温室中番茄灰霉病发生规律的研究. 植物保护, 2003, 2(29): 313-325.
[3] Ji J J, Zhang X F, Wang W Q, et al. Research progress on control of tomato gray mold. Chinese Agricultural Science Bulletin. 2012, 28(31): 109-113.
纪军建, 张小风, 王文桥, 等. 番茄灰霉病防治研究进展. 中国农学通报, 2012, 28(31): 109-113.
[4] Chen C, Cui T B, Yu P R. Identification of an anti-fungal strain of Amyloliquefaciens Bacillus and the properties of the antifungal substance. Modern Food Science and Technology, 2011, 27(1): 36-39.
陈成, 崔堂兵, 于平儒. 一株抗真菌的解淀粉芽孢杆菌的鉴定及其抗菌性研究. 现代食品科技, 2011, 27(1): 36-39.
[5] Zhang S, Wang Y, Meng L, et al. Isolation and characterization of antifungal lipopeptides produced by endophytic Bacillus amyloliquefaciens TF28. African Journal of Microbiology Research, 2012, 6(8): 1747-1755.
[6] Chen Z, Huang J, Zhao J, et al. Isolation and identification of pathogenic fungus of Botrytis cinerea and screening of antagonistic bacteria against tomato gray mold. Biotechnology Bulletin, 2017, 33(8): 81-87.
陈哲, 黄静, 赵佳, 等. 番茄灰霉病病原菌分离鉴定及拮抗菌筛选. 生物技术通报, 2017, 33(8): 81-87.
[7] Li H X, Zhang D P, Zhao H X, et al. Physical and chemical properties of antimicrobial from bacillus amyloliquefaciens strain MH71 and its inhibition activities against Botrytis cinerea. Chinese Journal of Biological Control, 2016, 32(4): 485-492.
李红晓, 张殿朋, 赵洪新, 等. 解淀粉芽胞杆菌MH71抗菌物质理化特性及对番茄灰霉病菌的抑菌活性. 中国生物防治学报, 2016, 32(4): 485-492.
[8] Wang J L, Lu C G, Liu W C, et al. Identification and biological characteristics analysis of bacillus strain QD-10. Chinese Journal of Biological Control, 2014, 30(4): 564-572.
王俊丽, 卢彩鸽, 刘伟成, 等. 一株芽胞杆菌QD-10的鉴定及生防特性分析. 中国生物防治学报, 2014, 30(4): 564-572.
[9] Chen X, Koumoutsi A, Scholz R, et al. Comparatuve analysis of the complete genome sequence of the plant growth-promoting bacterium Bacillus amyloliquefaciens FZB42. Nature Biotechnology, 2007, 25(9): 1007-1014.
[10] Yang R X, Ji J H, Wang H Z, et al. Isolation, indentification and inhibitory activity of lipopeptides of endophytic bacteria from the root of Paeonia suffruticosa. Microbiology China, 2015, 42(6): 1081-1088.
杨瑞先, 姬俊华, 王华祖, 等. 牡丹根部内生细菌的分离鉴定及脂肽类物质的拮抗活性研究. 微生物学通报, 2015, 42(6): 1081-1088.
[11] Guo J P, Ma G, Wang Z J, et al. Identification and analysis of biocontrol proteins from a strain of Bacillus amyloliquefaciens. Biotechnology Bulletin, 2018, 34(1): 202-207.
郭继平, 马光, 王志杰, 等. 一株解淀粉芽孢杆菌生防蛋白的鉴定及分析. 生物技术通报, 2018, 34(1): 202-207.
[12] Yang R X, Tao Y F, Song M X, et al. Inhibition of endophytic bacterial strains isolated from Ginkgo biloba against phytophthora blight of pepper. Chinese Journal of Biological Control, 2012, 28(4): 552-559.
杨瑞先, 陶玉凤, 宋美仙, 等. 银杏内生细菌防治辣椒疫病研究. 中国生物防治学报, 2012, 28(4): 552-559.
[13] Yang R X, Song M X, Qiu S X, et al. Screening, identification of biocontrol bacterial strain Hy11 against white radish soft rot and its colonization characteristics. Plant Protection, 2013, 40(2): 109-114.
杨瑞先, 宋美仙, 邱思鑫, 等. 萝卜软腐病生防菌株Hy11的筛选、鉴定及定殖特性. 植物保护学报, 2013, 40(2): 109-114.
[14] Fan X J, Huang W, Qiu S X, et al. Isolation and identification of endophytic bacterial strains from Ginkgo biloba and their antimicrobial activities. Microbiology China, 2013, 40(9): 1638-1648.
范晓静, 黄未, 邱思鑫, 等. 银杏内生细菌的分离鉴定及抑菌活性初探. 微生物学通报, 2013, 40(9): 1638-1648.
[15] Li C G, Cao C L, Qin S, et al. Identification of an endophytic bacterium XZNUM 033 from Ginkgo biloba L. and its physicochemical characteristic of anti-sapstain fungus activity. Forest Research, 2010, 23(5): 708-712.
李长根, 曹成亮, 秦盛, 等. 银杏内生细菌XZNUM 033的鉴定及其抗杨树变色真菌活性物质的理化性质. 林业科学研究, 2010, 23(5): 708-712.
[16] Xu S J, Kim B S. Biocontrol of Fusarium crown and root rot and promotion of growth of tomato by Paenibacillus strains isolated from soil. Mycobiology, 2014, 42: 158-166.
[17] Kumar R S, Ayyadurai N, Pandiaraja P, et al. Characterization of antifungal metabolite produced by a new strain Pseudomonas aeruginosa PUPa3 that exhibits broad-spectrum antifungal activity and biofertilizing traits. Journal of Applied Microbiology, 2005, 98(1): 45-54.
[18] Dong X Z, Cai M Y. Common bacterial systematic identification manual. Beijing: Science Press, 2001.
东秀珠, 蔡妙英. 常见细菌系统鉴定手册. 北京: 科学出版社, 2001.
[19] Pan H Y, Jin W Y, Zhang X M, et al. Inhibition of antifungal substances from Bacillus amyloquefaciens B15 against Botrytis Cinerea—the agent of “gray mold” of grape. Acta Microbiologica Sinica, 2018, 58(7): 1245-1254.
潘虹余, 金玮鋆, 张晓蒙, 等. 解淀粉芽孢杆菌 B15抑菌物质对葡萄灰霉病灰葡萄孢的抑菌机理. 微生物学报, 2018, 58(7): 1245-1254.
[20] Liu C, Liu H W, Wang B Q, et al. Isolation of antifungal substances from Bacillus amyloliquefaciens BA-26 and its antifungal activity against Botrytis cinerea. Biotechnology Bulletin, 2019, 35(7): 22-28.
刘超, 刘洪伟, 汪步青, 等. 解淀粉芽孢杆菌BA-26抗菌物质分离及对灰葡萄孢抑菌作用研究. 生物技术通报, 2019, 35(7): 22-28.
[21] Asraful S M, Math R K, Kim J M, et al. Effect of plant age on endophytic bacterial diversity of balloon flower (Platycodon grandiflorum) root and their antimicrobial activities. Current Microbiology, 2010, 61(4): 346-356.
[22] Danti R, Sieberr T N, Sanguineti G, et al. Decline in diversity and abundance of endophytic fungi in twigs of Fagus sylvatica L. after experimental long-term exposure to sodium dodecylbenzene sulphonate (SDBS) aerosol. Environmental Microbiology, 2002, 4(11): 696-702.
[23] Chun J, Bae K S. Phylogenetic analysis of Bacillus subtilis and related taxa based on partial gyrA gene sequence. Antonievan Leeywenhoek, 2000, 78(2): 123-127.
[24] Wang L T, Lee F L, Tai C J, et al. Compariosn of gyrB gene sequences, 16S rRNA gene sequences and DNA-DNA hybridization in the Bacillus subtilis group. International Journal of Systematic and Evolutionary Microbiology, 2007, 57: 1846-1850.
[25] Yamamoto S, Harayama S. Phylogenetic relationships of Pseudomonas putida strains deduced from the nucleotide sequences of gyrB, rpoD and 16S rRNA genes. Interntional Journal of Systematic Bacteriology, 1998, 48: 813-819.
[26] Klein T A, Burgess L W, Ellison F W. The incidence and spatial patterns of wheat plants infectd by Fusariym graminearum group land the effect of crown roton yield. Australian Journal of Agricultural Research, 1991, 42(3): 399-407.
[27] Yang L M, Tong Z H, Guo L H, et al. Screening, identification and biocontrol effects of antagonistic bacterial strain CQ against Botrytis cinerea from tomato. Chinese Journal of Biological Control, 2015, 31(6): 956-960.
杨利敏, 仝赞华, 郭立华, 等. 番茄灰霉生防菌CQ的分子鉴定及其生防效果研究. 中国生物防治学报, 2015, 31(6): 956-960.
[28] Zhang Y X, Li G, Zhang G M. Colonization of antagonistic strains of tomato leaf in green house and their effects on gray mold (Botrytis cinerea). Acta Phytopathologica Sinica, 2000, 30(1): 91.
张玉勋, 李光, 张光明. 拮抗细菌在大棚温室番茄叶片定殖及对灰霉病害的控制效果. 植物病理学报, 2000, 30(1): 91.

解淀粉芽孢杆菌HZ-6-3的筛选鉴定及其防治番茄灰霉病效果的评价

Screening, identification as Bacillus amyloliquefaciens strain HZ-6-3 and evaluation of inhibitory activity against tomato gray mold, of a bacterial isolate

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 12

编辑推荐

Metrics

[1]	杨成德, 崔月贞, 冯中红, 薛莉, 金梦军. 内生枯草芽孢杆菌265ZY4对温度和紫外光胁迫下紫花针茅生化特征的影响[J]. 草业学报, 2019, 28(6): 101-108.
[2]	江绪文, 李贺勤, 谭勇. 藿香内生细菌HX-2的鉴定、耐性及对宿主植物的促生作用[J]. 草业学报, 2018, 27(1): 161-168.
[3]	胡进玲, 汪治刚, 徐娜, 党淑钟, 李彦忠. 防治紫花苜蓿真菌病害的生防菌筛选[J]. 草业学报, 2017, 26(6): 145-152.
[4]	李红亚, 李文, 李术娜, 王树香, 李猛, 田苗苗, 朱宝成. 解淀粉芽孢杆菌复合菌剂对玉米秸秆的降解作用及表征[J]. 草业学报, 2017, 26(6): 153-167.
[5]	柏玉晶, 姚玉玲, 张振粉, 杨成德, 薛莉. 紫花苜蓿根腐病原——厚垣镰刀菌的鉴定及其拮抗菌的筛选[J]. 草业学报, 2017, 26(2): 78-87.
[6]	杨成德, 王玉琴, 陈秀蓉, 张振粉, 薛莉, 王颖, 姚玉玲. 2种嵩草属牧草休眠期内生细菌多样性研究[J]. 草业学报, 2016, 25(8): 136-144.
[7]	冯中红, 王玉琴, 杨成德, 薛莉, 陈秀蓉. 番茄细菌性叶斑病菌的拮抗菌筛选、鉴定及其拮抗性能评价[J]. 草业学报, 2015, 24(8): 166-173.
[8]	杨成德,王颖,王玉琴,姚玉玲,薛莉,徐长林,陈秀蓉. 东祁连山高寒草地几株醉马草内生细菌的生物功能评价及鉴定[J]. 草业学报, 2014, 23(5): 249-255.
[9]	王颖,王玉琴,杨成德,姚玉玲,陈秀蓉,薛莉. 马铃薯贮藏期主要病害拮抗内生细菌的筛选、鉴定及功能评价[J]. 草业学报, 2014, 23(3): 269-275.
[10]	畅涛,王涵琦,杨成德,王颖,杨小利,薛莉,陈秀蓉,徐长林. 高寒草地禾草内生细菌B-401的鉴定及生物防治潜力评价[J]. 草业学报, 2014, 23(3): 282-289.
[11]	高晓星,满百膺,陈秀蓉,杨成德. 东祁连山线叶嵩草内生细菌X4的产吲哚乙酸、解磷、抗菌和耐盐特性研究及分子鉴定[J]. 草业学报, 2013, 22(4): 137-146.
[12]	车建美,刘波,张彦,胡桂萍,黄勤楼,陈忠钿,翁伯琦. 几种禾本科牧草内生细菌的分布特性[J]. 草业学报, 2010, 19(3): 124-131.