烟草疫霉拮抗细菌B44R-1的筛选及其生防效果

doi:10.16409/j.cnki.2095-039x.2022.06.012

摘要/Abstract

摘要： 为获得烟草疫霉Phytophthora nicotiana的优良拮抗菌，从烟草根际土壤和根系中分离对烟草疫霉具有较好拮抗作用的菌群，并从中筛选具有较强抑菌效果的拮抗菌株。通过形态学、生理生化试验、16S rDNA基因测序分析鉴定拮抗菌株种属，评价其防病效果及对烟草的促生作用。结果表明，从烟草根际土壤和根系分离得到拮抗菌群39个，并从中筛选得到拮抗效果较好的菌株B44R-1，初步鉴定其为皮特不动杆菌Acinetobater pittii。菌株B44R-1对烟草黑胫病的防效达到55.37%，对烟株根系生长发育影响明显，烟株鲜重、干重、株高、茎围、叶数和最大叶面积相比对照均有提高。本研究筛选获得对烟草黑胫病菌的拮抗菌不动杆菌，对烟草黑胫病防治效果和促生作用显著，具有较好的生防潜力。

关键词: 烟草黑胫病, 拮抗菌群, 不动杆菌, 防治效果, 促生作用

Abstract: To obtain excellent antagonistic bacteria of Phytophthora nicotiana in tobacco, the antagonistic strains of P. nicotiana were isolated from the rhizosphere soil and root of tobacco, and the antagonistic strains with strong inhibitory effects were screened. Afterwards, the antagonistic strains were further identified by morphological analysis, physiological and biochemical test, as well as 16S rDNA gene sequencing analysis. Besides, the effects of disease control and growth promotion on tobacco were also evaluated. The results showed that 39 antagonistic bacteria were isolated, and the strain B44R-1 with the best antagonistic effect was screened and preliminarily identified as Acinetobater pittii. The control efficacy of strain B44R-1 on tobacco black shank disease reached 55.37%. Moreover, this strain had obvious effect on the growth and development of tobacco root. Compared with the control, the agronomic traits of the treated tobacco plants were significantly improved, including fresh weight, dry weight, plant height, stem circumference, leaf number and maximum leaf area. In conclusion, the antagonistic bacterium A. pittii has significant control effect on tobacco black shank disease and has good biocontrol potential.

Key words: tobacco black shank, antagonistic microflora, Acinetobater pittii, control effect, growth promoting effect

中图分类号:

S476

刘天波, 赵誉强, 滕凯, 周向平, 肖艳松, 蔡海林, 肖志鹏, 陈武, 唐前君. 烟草疫霉拮抗细菌B44R-1的筛选及其生防效果[J]. 中国生物防治学报, 2022, 38(6): 1545-1552.

LIU Tianbo, Zhao Yuqiang, TENG Kai, ZHOU Xiangping, XIAO Yansong, CAI Hailin, XIAO Zhipeng, CHEN Wu, TANG Qianjun. Screening and Biocontrol Effect of Antagonistic Bacterium B44R-1 against Phytophthora parasitica[J]. Chinese Journal of Biological Control, 2022, 38(6): 1545-1552.

参考文献

[1] 陈瑞泰, 朱贤朝, 王智发, 等. 全国16个主产烟省(区)烟草侵染性病害调研报告[J]. 中国烟草科学, 1997, (4): 1-7.
[2] Debasish D, Ankita S, Maiti I B, et al. Recombinant promoter (MUASCsV8CP) driven totiviral killer protein 4 (KP4) imparts resistance against fungal pathogens in transgenic tobacco[J]. Frontiers in Plant Science, 2018, 9: 278.
[3] Niu J, Chao J, Xiao Y, et al. Insight into the effects of different cropping systems on soil bacterial community and tobacco bacterial wilt rate[J]. Journal of Basic Microbiology, 2017, 57(1): 3-11.
[4] You C, Zhang C, Kong F, et al. Comparison of the effects of biocontrol agent Bacillus subtilis and fungicide metalaxyl-mancozeb on bacterial communities in tobacco rhizospheric soil[J]. Ecological Engineering, 2016, 91:119-125.
[5] 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.
[6] Han T, You C, Zhang L, et al. Biocontrol potential of antagonist Bacillus subtilis Tpb55 against tobacco black shank[J]. Biocontrol, 2016, 61(2): 195-205.
[7] 王静, 孔凡玉, 张成省, 等. 放线菌F8对烟草黑胫病的拮抗作用及其产酶活性[J]. 中国烟草科学, 2013(2): 49-53.
[8] 张良, 刘好宝, 顾金刚, 等. 长柄木霉和泾阳链霉菌复配对烟苗生长及其抗病性的影响[J]. 应用生态学报, 2013, 10: 2961-2969.
[9] Arruda L, Beneduzi A, Martins A, et al. Screening of rhizobacteria isolated from maize (Zea mays L.) in rio grande do sul state (south brazil) and analysis of their potential to improve plant growth[J]. Applied Soil Ecology, 2013, 63: 15-22.
[10] Hayat R, Ali S, Amara U, et al. Soil beneficial bacteria and their role in plant growth promotion: a review[J]. Annals of Microbiology, 2010, 60(4): 579-598.
[11] Li S P, Tan J, Yang X, et al. Niche and fitness differences determine invasion success and impact in laboratory bacterial communities[J]. ISME Journal, 2019, 13: 402-412.
[12] Hu J, Wei Z, Friman V, et al. Probiotic diversity enhances rhizosphere microbiome function and plant disease suppression[J]. mBio, 2016,7: e01790.
[13] Liu T, Xiao Y, Yin J, et al. Effects of cultured root and soil microbial communities on the disease of Nicotiana tabacum caused by Phytophthora nicotianae[J]. Frontiers in Microbiology, 2020, 11: 929.
[14] Stumbriene K, Gudiukaite R, Semaskiene R, et al.Screening of new bacterial isolates with antifungal activity and application of selected Bacillus sp. cultures for biocontrol of Fusarium graminearum under field conditions[J]. Crop Protection, 2018, 113: 22-28.
[15] 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.
[16] 布坎南, 吉本斯. 伯杰细菌鉴定手册[M].中国科学院微生物研究所,译. 8版.北京:科学出版社,1984: 729-758.
[17] Paul N C, Ji S H, Jian X D, et al. Assemblages of endophytic bacteria in chili pepper (Capsicum annuum L.) and their antifungal activity against phytopathogens in vitro[J]. Plant Omics, 2013, 6(6): 441-448.
[18] 王万能, 全学军, 肖崇刚. 烟草疫霉的产孢和接种方法研究[J]. 植物保护学报, 2005, 32(1): 18-22.
[19] 烟草病虫害分级及调查方法(GB/T 23222-2008)[S]. 北京: 中国标准出版社, 2008.
[20] 郭建荣, 郑聪聪, 李艳迪, 等. NaCl处理对真盐生植物盐地碱蓬根系特征及活力的影响[J]. 植物生理学报, 2017, 53(1): 63-70.
[21] 白宝璋, 金锦子, 白菘, 等. 玉米根系活力TTC测定法的改良[J]. 玉米科学, 1994(2): 44-47.
[22] 胡伟. 土壤容重对烟草生长及植烟土化学性质影响的研究[D]. 昆明: 昆明理工大学, 2013.
[23] Qin C, Tao J, Liu T, et al. Responses of phyllosphere microbiota and plant health to application of two different biocontrol agents[J]. AMB Express, 2019, 9(1): 42.
[24] 郭桥燕. 烟草青枯病与黑胫病拮抗菌筛选及防治效果研究[D]. 郑州: 河南农业大学, 2007.
[25] 蔺经, 杨青松, 李晓刚, 等. 一株不动杆菌次生代谢活性产物的抑菌作用及对果树病害的防治效果[J]. 江西农业学报, 2010, 22(10): 78-79.
[26] 陈志超. 一株降解纤维素的皮特不动杆菌的分离鉴定,纤维素降解条件优化及转录组分析[D]. 柳州: 广西科技大学, 2019.
[27] 李静, 张瀚能, 赵翀, 等. 高效纤维素降解菌分离筛选、复合菌系构建及秸秆降解效果分析[J]. 应用与环境生物学报, 2016, 22(4): 689-696.
[28] 刘广超, 叶青, 车永梅, 等. 烟草根际高效解磷菌的筛选鉴定及促生作用研究[J]. 生物技术通报, 2022, 38(9): 84-91.
[29] El-Sharkawy H, Rashad Y M, Ibrahim S A. Biocontrol of stem rust disease of wheat using arbuscular mycorrhizal fungi and Trichoderma spp.[J]. Physiological & Molecular Plant Pathology, 2018, 103: 84-91.
[30] Vos C, Daphné Van Den Broucke, Lombi F M, et al. Mycorrhiza-induced resistance in banana acts on nematode host location and penetration[J]. Soil Biology & Biochemistry, 2012, 47: 60-66.
[31] Saini I, Yadav K, Aggarwal A. Response of arbuscular mycorrhizal fungi along with Trichoderma viride and Pseudomonas fluorescens on the growth, biochemical attributes and vase life of Chrysanthemum indicum[J]. Journal of Environmental Biology, 2019, 40(2): 183-191.
[32] Harrier L A, Watson C A.The potential role of arbuscular mycorrhizal (AM) fungi in the bioprotection of plants against soil-borne pathogens in organic and/or other sustainable farming systems[J]. Pest Management Science, 2010, 60(2): 149-157.
[33] 袁梦思, 李建文, 刘梦华, 等. 1株不动杆菌发酵液对水稻纹枯病菌的抑菌作用[J]. 江苏农业科学, 2016, 44(2): 164-165.
[34] 朱彤彤, 陈达川, 袁梦思, 等. 井冈霉素、多菌灵和不动杆菌A2混用对水稻纹枯病病菌的抑制作用[J]. 江苏农业科学, 2019, 47(12): 146-149.
[35] 吴俊林, 张正杨, 李翔, 等. 一株高效解钾菌的筛选鉴定及其对烟草吸收钾磷的影响[J]．江苏农业科学, 2020, 48(5): 276-280.
[36] 王鹏, 车永梅, 李雅华, 等. 多功能皮特不动杆菌3K32菌株的发酵优化[J]. 青岛农业大学学报(自然科学版), 2022, 39(2): 114-121.