Isolation and Identification of Antagonistic Fungus against Albizzia Julibrissin Wilt and Pot Control Experiment

doi:10.16409/j.cnki.2095-039x.2023.02.021

Abstract

Abstract: Albizzia julibrissin wilt is one of the major factors that trigger the weakness and death of A. julibrissin, and there are no efficient approaches for preventing this disease at present. In this study, plate confrontation and toxic medium method have been used to isolate potential antagonists from the rhizosphere soil of A. Julibrissin. The antagonistic fungus was identified based on morphological characteristics and combined ITS-TEF1 phylogenetic analysis, and the pot experiment was conducted to verify the efficacy of the antagonistic strains against A. Julibrissin wilt. The result showed that 11 potential antagonists were collected from rhizosphere soil, and 5-2 has the highest inhibitory effect. After six days of confrontation, the inhibition rate of 5-2 on Fusarium oxysporum and Fusarium solani were 76.88% and 84.08%, respectively, and the two pathogens can hardly grow on the plate containing 6.67% concentration of 5-2 spore suspension (1.0×10⁷ spore/mL). Microscopic observation showed that 5-2 could entangle the mycelium of A. Julibrissin wilt pathogen and parasitic growth. Pot experiment showed that strain 5-2 had good antagonistic effect on F. oxysporum and F. solani with the control efficacies of 78.26% and 57.89%, respectively. Combined with morphological characteristics and ITS-TEF1 phylogenetic analysis, strain 5-2 was identified as Trichoderma koningii. These results indicate that T. koningii (strain no. 5-2) has positive effect on preventing and treating Fusarium wilt, and could be used as a candidate strain for biological control of A. Julibrissin wilt disease.

Key words: Albizzia julibrissin wilt, antagonistic fungus, Trichoderma koningii, control efficacy

CLC Number:

S476

CAI Yue, TANG Xiaofei, HU Ting, GE Chunmei, RUAN Banfeng. Isolation and Identification of Antagonistic Fungus against Albizzia Julibrissin Wilt and Pot Control Experiment[J]. Chinese Journal of Biological Control, 2023, 39(2): 397-406.

References

[1] 曹杰, 柏超. 合欢枯萎病的综合防控技术研究[J]. 北京园林, 2015, 31(4):55-62.
[2] 吴玲, 李绍凯. 合欢枯萎病研究初报[J]. 森林病虫通讯, 1990(4):13-14.
[3] 蔡悦, 王储炎, 黄明华, 等. 合欢枯萎病菌的分离鉴定和培养特性[J]. 浙江农业学报, 2019, 31(4):588-599.
[4] 冯雪. 合欢枯萎病病原菌生物学特性及防治药剂筛选[D]. 武汉:华中农业大学, 2014.
[5] Nelson P E. Life cycle and epidemiology of Fusarium oxysporum[M]. Fungal Wilt Diseases of Plants. New York:Academic Press, 1981:51-80
[6] Bubici G, Kaushal M, Prigigallo M I, et al. Biological control agents against Fusarium wilt of banana[J]. Frontiers in Microbiology, 2019, 10(616):1-33.
[7] 余小兰, 邹立飞, 邹雨坤, 等. 甜瓜枯萎病拮抗菌的筛选及鉴定[J]. 南方农业学报, 2018, 49(6):1118-1124.
[8] 李枢妍, 阳黎恒, 肖雪婷, 等. 一株香蕉枯萎病拮抗菌的筛选、鉴定及生防效果研究[J]. 南方农业学报, 2021, 52(7):1826-1834.
[9] 高苇, 杨利娟, 刘春艳, 等. 西瓜枯萎病拮抗菌株的筛选与鉴定[J]. 中国瓜菜, 2021, 34(1):19-23.
[10] 刘侃诚, 许伟, 郁世军, 等. 防治榉树枯萎病室内杀菌剂的筛选[J]. 林业科技开发, 2015, 29(2):127-129.
[11] 吕娟, 亓玉昆, 季延平, 等. 山东省黑松枯萎病病原鉴定、致病性及其生长适应性研究[J]. 山东农业大学学报(自然科学版), 2022, 53(1):77-84.
[12] 彭昀, 肖宇明, 刘衍超, 等. 橡胶树枯萎病病原菌室内毒力测定[J]. 热带生物学报, 2019, 10(3):272-275.
[13] 胡忠亮, 邹东霞, 黄敏仁, 等. 镰刀菌毒素滤液对转基因香石竹抗枯萎病性能的测定[J]. 科学技术与工程, 2016, 16(27):132-136.
[14] 王嘉, 梁晓洁, 高暝, 等. 千年桐根部黄酮类化合物生物合成对枯萎病菌侵染的响应[J]. 林业科学, 2022, 58(2):159-170.
[15] 韩长志. 植物病害生防菌的研究现状及发展趋势[J]. 中国森林病虫, 2015, 34(1):33-37, 25.
[16] 刘新月, 李凡, 陈海如, 等. 致病性尖孢镰刀菌生物防治研究进展[J]. 云南大学学报, 2008, 30(S1):89-93.
[17] 杨辉辉, 王坦, 黄思良, 等. 一株芝麻枯萎病菌的鉴定及其生物学特性[J]. 中国油料作物学报, 2014, 36(3):385-392.
[18] Crous P W. Fusarium:more than a node or a foot-shaped basal cell[J]. Studies in Mycology, 2021, 98:100116.
[19] 梁晓洁, 刘志华, 张平, 等. 抗油桐枯萎病木霉菌的分离鉴定及抑菌作用研究[J]. 菌物学报, 2020, 39(5):795-805.
[20] 高智谋, 曹君, 潘月敏, 等. 哈茨木霉TH-1对棉花枯萎病菌和黄萎病菌的拮抗机制研究[J]. 棉花学报, 2007, 19(3):168-172.
[21] 张胜利, 李增智. 球孢白僵菌对两种植物病原镰孢的拮抗作用[J]. 中国生物防治学报, 2011, 27(3):349-356.
[22] 刘彩云, 季洪亮, 王瑞, 等. 近缘毛壳菌生防菌株LB-1对几种常见植物病原真菌的拮抗作用及其生长适应性分析[J]. 植物保护学报, 2018, 45(2):332-339.
[23] 田桢, 曹传旺, 季世达, 等. 新疆野果林木霉分离鉴定、抑菌作用及固态发酵配方优化[J]. 微生物学通报, 2021, 48(12):4696-4709.
[24] 方中达. 植病研究方法(第三版)[M]. 北京:中国农业出版社, 1998
[25] 倪方方, 徐红梅, 宋腾蛟, 等. 生防菌对白术根腐病菌的拮抗作用及盆栽防治效果[J]. 浙江中医药大学学报, 2017, 41(3):179-185.
[26] 纪兆林, 谈彬, 朱薇, 等. 我国不同产区桃褐腐病病原鉴定与分析[J]. 微生物学通报, 2019, 46(4):869-878.
[27] Samuels G J, Dodd S L, Lu B S, et al. The Trichoderma koningii aggregate species[J]. Studies in Mycology, 2006, 56:67-133.
[28] 周玉宝. 分离自安徽土壤绿僵菌物种和遗传多样性的研究[D]. 合肥:安徽农业大学, 2011.
[29] White T J, Bruns T, Lee S, et al. PCR protocols a guide to methods and applications[M]. New York:Academic Press, 1990, 315-322.
[30] Bustamante D E, Calderon M S, Leiva S, et al. Three new species of Trichoderma in the Harzianum and Longibrachiatum lineages from Peruvian cacao crop soils based on an integrative approach[J]. Mycologia, 2021, 113(5):1056-1072.
[31] Swofford D L. PAUP*. Phylogenetic analysis using parsimony (*and other methods). version 4.0b10[M]. Sunderland, Massachusetts:Sinauer Associates, 2002.
[32] Ronquist F, Huelsenbeck JP. MrBayes 3:Bayesian phylogenetic inference under mixed models[J]. Bioinformatics, 2003, 19(12):1572-1574.
[33] Cai Y, Nie Y, Gao Y, et al. Natural populations of the entomopathogenic fungus Beauveria bassiana in Chinese forest ecosystems are diverse and reveal equal frequencies of mating types within phylogenetic species[J]. Fungal Ecology, 2022, 56:101139.
[34] 魏萍, 王恒玺, 李际会. 临沂地区合欢枯萎病的发生调查与综合防治技术[J]. 农业与技术, 2020, 40(21):117-119.
[35] Loc N H, Huy N D, Quang H T, et al. Characterisation and antifungal activity of extracellular chitinase from a biocontrol fungus, Trichoderma asperellum PQ34[J]. Mycology, 2020, 11(1):38-48.
[36] 叶旻硕, 俞键烽, 马艳, 等. 不同微生物菌剂对辣椒疫病的防控效果及对土壤性状的影响[J]. 江苏农业学报, 2019, 35(4):811-817.
[37] Wu Q, Sun R, Ni M, et al. Identification of a novel fungus, Trichoderma asperellum GDFS1009, and comprehensive evaluation of its biocontrol efficacy[J]. PLoS ONE, 2017, 12:e0179957.
[38] Gava C A T, Pinto J M. Biocontrol of melon wilt caused by Fusarium oxysporum Schlect f. sp. melonis using seed treatment with Trichoderma spp. and liquid compost[J]. Biological Control, 2016, 97:13-20.
[39] 张小杰, 周天旺, 王春明, 等. 长枝木霉TS-1的分离鉴定、拮抗作用及固体发酵条件初探[J]. 中国农学通报, 2021, 37(27):105-111.
[40] 刘佳, 张悦, 沈志彦, 等. 长枝木霉T6菌株对美洲南瓜枯萎病菌的抑制作用[J]. 西北农业学报, 2020, 29(12):1891-1897.
[41] 张玉博, 庄文颖. 对峙培养条件下木霉拮抗植物病原真菌能力的评价[J]. 菌物学报, 2017, 36(7):1-9.
[42] 覃柳燕, 郭成林, 黄素梅, 等. 棘孢木霉菌株PZ6对香蕉促生效应及枯萎病室内防效的影响[J]. 南方农业学报, 2017, 48(2):277-283.
[43] 李淼, 产祝龙, 檀根甲, 等. 木霉菌防治植物真菌病害研究进展[J]. 生物技术通讯, 2009, 20(2):286-290.
[44] 邢咏梅, 李向东, 李莉, 等. 木霉属真菌拮抗金钗石斛病原菌的研究[J]. 中国医药生物技术, 2017, 12(1):35-39.