Screening, Identification and Culture Condition Optimization of Antagonistic Endophytic Bacteria Against Gloeosporium theae-sinensis Miyake

ZHENG Shizhong; ZHOU Ziwei; CHEN Xiaohui; CAI Liewei; JIANG Shengtao; LIU Shengrong

doi:10.13305/j.cnki.jts.2023.02.006

2023 , Vol. 43 >Issue 2: 205 - 215

DOI: https://doi.org/10.13305/j.cnki.jts.2023.02.006

Research Paper

Screening, Identification and Culture Condition Optimization of Antagonistic Endophytic Bacteria Against Gloeosporium theae-sinensis Miyake

ZHENG Shizhong ,
ZHOU Ziwei ,
CHEN Xiaohui ,
CAI Liewei ,
JIANG Shengtao ,
LIU Shengrong

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College of Life Sciences College, Ningde Normal University, Ningde 352100, China

Received date: 2022-09-18

Revised date: 2022-12-06

Online published: 2023-05-05

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Abstract

To screen the antagonistic endophytic bacteria against Gloeosporium theae-sinensis Miyake, the healthy leaves of tea plants were utilized as materials, and the plate antagonistic method was used. The isolated bacteria were identified and evaluated for antimicrobial efficacy. The culture conditions were optimized using response surface methodology. One antagonistic bacterium X13 was screened from 162 strains of endophytic bacteria isolated, which had a good inhibitory effect on Colletotrichum gloeosporioides. The strain X13 was identified as Bacillus subtilis through morphological observation, physiological and biochemical tests, and 16S rDNA phylogenetic analysis. The inhibitory rate of strain X13 on the mycelial growth of C. gloeosporioides reached 61.6%. The logarithmic growth was between 2-14 h. The optimal culture conditions were 4.0% (mass percentage concentration) corn flour, 1.0% (mass percentage concentration) NaNO₃, and 3.5% (volume fraction) inoculation. This study laid a key theoretical foundation for the prevention of the tea pathogen G. theae-sinensis and the development of its biocontrol agents.

Key words： tea plant; Gloeosporium theae-sinensis; endophytic bacteria; Bacillus subtilis; culture condition

Cite this article

ZHENG Shizhong , ZHOU Ziwei , CHEN Xiaohui , CAI Liewei , JIANG Shengtao , LIU Shengrong . Screening, Identification and Culture Condition Optimization of Antagonistic Endophytic Bacteria Against Gloeosporium theae-sinensis Miyake[J]. Journal of Tea Science, 2023 , 43(2) : 205 -215 . DOI: 10.13305/j.cnki.jts.2023.02.006

References

[1] 张永乐. 山东省茶树主要叶部病害病原鉴定及生物学特性研究[D]. 泰安: 山东农业大学, 2018.
Zhang Y L.Identificantion and biological characteristics of tea leaf diseases in Shandong province [D]. Tai'an: Shandong Agricultural University, 2018.
[2] 张瑾, 邢玉娴, 韩涛, 等. 茶树诱导抗虫性的研究进展[J]. 昆虫学报, 2022, 65(3): 399-408.
Zhang J, Xing Y X, Han T, et al.Research progress of induced defense against insect pests in tea plant (Camellia sinensis)[J]. Acta Entomologica Sinica, 2022, 65(3): 399-408.
[3] Guo M, Pan Y M, Dai Y L, et al.First report of brown blight disease caused by Colletotrichum gloeosporioides on Camellia sinensis in Anhui Province, China[J]. Plant Disease, 2014, 98(2): 284. doi: 10.1094/PDIS-08-13-0896-PDN.
[4] 刘威, 袁丁, 尹鹏, 等. 茶树炭疽病的研究进展[J]. 热带农业科学, 2016, 36(11): 20-26.
Liu W, Yuan D, Yin P, et al.Research progress on anthracnose of tea plant[J]. Chinese Journal of Tropical Agriculture, 2016, 36(11): 20-26.
[5] 贡长怡, 刘姣姣, 邓强, 等. 茶树炭疽病病原菌鉴定及其致病性分析[J]. 园艺学报, 2022, 49(5): 1092-1101.
Gong C Y,Liu J J,Deng Q, et al.Identification and pathogenicity of Colletotrichum species causing anthracnose on Camellia sinensis[J]. Acta Horticulturae Sinica, 2022, 49(5): 1092-1101.
[6] 刘荣, 姚玉仙, 周爽爽, 等. 茶炭疽病的症状识别与防治研究[J]. 福建茶叶, 2022, 44(2): 24-26.
Liu R, Yao Y X, Zhou S S, et al.Identification and control of Gloeosporium theae-sinensis Miyake[J]. Tea in Fujian, 2022, 44(2): 24-26.
[7] 刘守安, 韩宝瑜, 付建玉, 等. 茶炭疽病菌毒素的致病活性及理化性质初探[J]. 茶叶科学, 2007, 27(2): 153-158.
Liu S A, Han B Y, Fu J Y, et al.Preliminary studies on pathogenic activity and physical and chemical characteristics of toxins from Gloeosporium theae-sinensis[J]. Journal of Tea Science, 2007, 27(2): 153-158.
[8] 陈雪芬. 我国茶树病害的发生趋势与绿色防控[J]. 中国茶叶, 2022, 44(6): 7-14.
Chen X F.The occurrence trend and green control of tea diseases in China[J]. China Tea, 2022, 44(6): 7-14.
[9] 朱咏珊, 罗晓欣, 梁浩然, 等. 一株茶树根际细菌的鉴定与生防效果研究[J]. 茶叶科学, 2022, 42(1): 87-100.
Zhu Y S, Luo X X, Liang H R, et al.Identification of a tea rhizosphere bacterium and its biocontrol of tea anthracnose disease[J]. Journal of Tea Science, 2022, 42(1): 87-100.
[10] Thakur R.Use of Trichoderma spp. as biocontrol for disease management[J]. Indian Farmer, 2021, 8(1): 108-115.
[11] 李聪. 放线菌TR19040的次级代谢产物及其生物活性研究[D]. 兰州: 兰州交通大学, 2021.
Li C.Secondary metabolites of actinomycete TR19040 and its biological activity [D]. Lanzhou: Lanzhou Jiaotong University, 2021.
[12] Christopher Y, Aguilar C, Gálvez D, et al.Interactions among Escovopsis, antagonistic microfungi associated with the fungus-growing antsymbiosis[J]. Journal of Fungi, 2021, 7(12): 1007. doi: 10.3390/jof7121007.
[13] Strobel G.Harnessing endophytes for industrial microbiology[J]. Current Opinion in Microbiology, 2006, 9(3): 240-244.
[14] 尚笑男, 刘君昂, 冯福山, 等. 油茶内生拮抗细菌的筛选、鉴定及防效[J]. 中国生物防治学报, 2021, 37(3): 575-583.
Shang X N, Liu J A, Feng F S, et al.Screening, identification and antagonistic effect of endophytic antagonistic bacterial strains from Camellia oleifera[J]. Chinese Journal of Biological Control, 2021, 37(3): 575-583.
[15] Deka B, Babu A, Peter A, et al.Beauveria bassiana: as a potential microbial biocontrol agent for tea mosquito bug, Helopeltis theivora waterhouse (Hemiptera: Miridae) in Dooars and Darjeeling, India[J]. Egyptian Journal of Biological Pest Control, 2021, 31(1): 134. doi: 10.1186/s41938-021-00478-3.
[16] Alviz M L, Pérez G A, Pérez-Cordero A.Inhibitory effect of compound type metabolites of endophytes bacteria against Colletotrichum gloeosporioides Burkholderia glumae[J]. Revista Colombiana de Ciencia Animal Recia, 2017, 9(5): 18-25.
[17] 布坎南, 吉本斯. 伯杰氏细菌鉴定手册[M]. 8版. 北京: 科学出版社, 1984: 661-870.
Buchanan R E, Gibbons N E.Bergey's manual of determinative bacteriology [M]. 8th ed. Beijing: Science Press, 1984: 661-870.
[18] 易有金, 罗宽, 刘二明. 内生细菌在植物病害生物防治中的作用[J]. 核农学报, 2007, 21(5): 474-477.
Yi Y J, Luo K, Liu E M.Effect of endophytic bacteria on biological control of plant disease[J]. Journal of Nuclear Agricultural Sciences, 2007, 21(5): 474-477.
[19] Trung D Q, Hang N T, Van D M, et al.Antagonic activity of endophytic bacteria isolated from man trau grass on stem rot disease of Pitaya (Hylocereus Undatus)[J]. Vietnam National University Journal of Science: Natural Sciences and Technology, 2021, 37(2): 60-69.
[20] 何玲敏, 叶建仁. 植物内生细菌及其生防作用研究进展[J]. 南京林业大学学报(自然科学版), 2014, 38(6): 153-159.
He L M, Ye J R.Endophytic bacteria: research advances and biocontrol applications[J]. Journal of Nanjing Forestry University (Natural Sciences Edition), 2014, 38(6): 153-159.
[21] Wu W, Chen W, Liu S, et al.Beneficial relationships between endophytic bacteria and medicinal plants[J]. Frontiers in Plant Science, 2021, 12: 646146. doi: 10.3389/fpls.2021.646146.
[22] Pinto-Carbó M, Gademann K, Eberl L, et al.Leaf nodule symbiosis: function and transmission of obligate bacterial endophytes[J]. Current Opinion in Plant Biology, 2018, 44(4): 23-31.
[23] 黄晓琴, 张丽霞, 刘会香, 等. 抗茶树冰核细菌内生菌的筛选及鉴定[J]. 茶叶科学, 2015, 35(1): 97-102.
Huang X Q, Zhang L X, Liu H X, et al.Screeining and identification of the endophytic bacterial strain against ice nucleation active bacteria of tea plant[J]. Journal of Tea Science, 2015, 35(1): 97-102.
[24] 王桥美, 严亮, 杨瑞娟, 等. 普洱地区茶叶内生细菌与根际土壤细菌群落结构分析[J]. 微生物学报, 2022, 62(2): 703-714.
Wang Q M, Yan L, Yang R J, et al.Analysis on the community structure of bacteria in tea leaves and rhizosphere soil in Pu’er[J]. Acta Microbiologica Sinica, 2022, 62(2): 703-714.
[25] 吴孔阳, 娄亚芳, 杨同香, 等. 枯草芽孢杆菌功能及相关机制研究进展[J]. 黑龙江畜牧兽医, 2020, 63(23): 55-58.
Wu K Y, Lou Y F, Yang T X, et al.Research advances in function and related mechanism of Bacillus subtilis[J]. Heilongjiang Animal Science and Veterinary Medicine, 2020, 63(23): 55-58.
[26] 马佳, 李颖, 胡栋, 等. 芽胞杆菌生物防治作用机理与应用研究进展[J]. 中国生物防治学报, 2018, 34(4): 639-648.
Ma J, Li Y, Hu D, et al.Progress on mechanism and applications of bacillus as a biocontrol microbe[J]. Chinese Journal of Biological Control, 2018, 34(4): 639-648.
[27] 陈百文, 刘伟, 赵恬欢, 等. 对茶树炭疽病菌具拮抗作用的茶树内生细菌的筛选[J]. 福建农林大学学报(自然科学版), 2010, 39(4): 341-346.
Chen B W, Liu W, Zhao K H, et al.Screening of endophytic strains to control Gloeosporium theae-sinensis Miyake[J]. Journal of Fujian Agriculture and Forestry University (Natural Science Edition), 2010, 39(4): 341-346.
[28] 代亚锋, 吴楠楠, 郜振, 等. 茶树炭疽病拮抗内生细菌贝莱斯芽胞杆菌的筛选与鉴定[J]. 信阳师范学院学报(自然科学版), 2021, 34(2): 201-207.
Dai Y F, Wu N N, Gao Z, et al.Screening and identificationof the endophytic bacteria Bacillus velezensis against tea anthracnose[J]. Journal of Xinyang Normal University (Natural Science Edition), 2021, 34(2): 201-207.
[29] 郝金辉, 包慧芳, 王宁, 等. 库尔勒香梨黑斑病菌拮抗菌筛选及其抑菌机理[J]. 微生物学通报, 2022, 49(9): 3709-3721.
Hao J H, Bao H F, Wang N, et al.Screening and possible mechanism of strains against black spot of Korla pear[J]. Microbiology China, 2022, 49(9): 3709-3721.
[30] Xu T, Zhu T H, Li S J.β-1,3-1,4-glucanase gene from Bacillus velezensis ZJ20 exerts antifungal effect on plant pathogenic fungi[J]. World Journal of Microbiology & Biotechnol, 2016, 32(2): 26-34.

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