Isolation, identification, biocontrol effect and its colonization in organic substrate of antagonistic bacterium strain LCQ1801 against fusarium wilt

ZHANG Yaoyao; ZHENG Yuhua; SUN Jianbo; ZHU Baibi; XIONG Guoru; LI Shuxia; PENG Ming; LI Chunqiang

doi:10.15886/j.cnki.rdswxb.20230023

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Volume 16 Issue 1

Mar. 2025

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Article Navigation > Journal of Tropical Biology > 2025 > 16(1): 71-78

ZHANG Yaoyao, ZHENG Yuhua, SUN Jianbo, ZHU Baibi, XIONG Guoru, LI Shuxia, PENG Ming, LI Chunqiang. Isolation, identification, biocontrol effect and its colonization in organic substrate of antagonistic bacterium strain LCQ1801 against fusarium wilt[J]. Journal of Tropical Biology, 2025, 16(1): 71-78. doi: 10.15886/j.cnki.rdswxb.20230023

Citation:

ZHANG Yaoyao, ZHENG Yuhua, SUN Jianbo, ZHU Baibi, XIONG Guoru, LI Shuxia, PENG Ming, LI Chunqiang. Isolation, identification, biocontrol effect and its colonization in organic substrate of antagonistic bacterium strain LCQ1801 against fusarium wilt[J]. Journal of Tropical Biology, 2025, 16(1): 71-78. doi: 10.15886/j.cnki.rdswxb.20230023

Isolation, identification, biocontrol effect and its colonization in organic substrate of antagonistic bacterium strain LCQ1801 against fusarium wilt

doi: 10.15886/j.cnki.rdswxb.20230023

1. School of Tropical Crops, Hainan University, Haikou, Hainan 570228, China;
2. Sanya Research Institute/Institute of Tropical Bioscience and Biotechnology/Ministry of Agriculture and Rural Affairs Key Laboratory of Biology and Genetic Resources of Tropical Crops, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China;
3. Institute of Vegetables/Hainan Key Laboratory of Vegetable Biology/Hainan Engineering Center for Breeding of Vegetables, Hainan Academy of Agricultural Sciences, Haikou, Hainan 571100, China

Received Date: 2023-02-28
Rev Recd Date: 2023-12-09
Publish Date: 2025-03-15

Abstract

To develop biological organic fertilizer for preventing and controling Fusarium wilt of banana, a bio-control bacterial strain LCQ1801 was isolated from the soil of rubber plantation in Chengmai, Hainan and its inhibition activity against Fusarium oxysporum f. sp. cubense race 4（Foc4）, the causal agent of banana Fusarium wilt was determined by using plate dual culture. The results indicated that LCQ1801 had obviously antagonistic effect on Foc4,and the inhibition rate was 65.9%. The LCQ1801 was identified as Bacillus velezensis by morphological observation,physiological and biochemical tests, combined with 16S rDNA sequencing analysis. Foc4 was cocultured with LCQ1801 bacterial liquid and filtrate, respectively. The result found that LCQ1801 bacterial fluid and filtrate caused abnormal growth or rupture of Foc4 hyphae and affected spore germination. At the same time, LCQ1801 was cultured on different substrates by using plate assay and culture flask culture. The plate assay indicated that the growth of LCQ1801 in the animal substrates of sheep dung, pig dung and cow dung was better than that in chicken dung. The growth of LCQ1801 in the plant substrates of wheat bran, corn flour, soybean meal and rice straw was better than that in the plant substrates of bagasse and coconut coir. Culture flask culture test showed that the number of viable bacteria of LCQ1801 in cow dung and sheep dung reached 16.8 billions·g^-1 and 24.0 billions·g^-1, respectively. The number of viable bacteria of LCQ1801 in rice straw, soybean meal and corn flour reached 32.2 billions·g^-1, 24.9 billions·g^-1 and 21.7 billions·g^-1, respectively. LCQ1801 hardly grew in the substrate of coconut coir in both culture styles. This study provides strain materials and technical support for biological control of Fusarium wilt of banana.
- biocontrol bacteria,
- fusarium wilt,
- organic substrate,
- biological fertilizer

References

[1]	BENIZRI E, PIUTTI S, VERGER S, et al. Replant diseases:bacterial community structure and diversity in peach rhizosphere as determined by metabolic and genetic fingerprinting[J]. Soil Biology and Biochemistry, 2005,37(9):1738-1746.
[2]	郝晓娟,刘波,谢关林.植物枯萎病生物防治研究进展[J]. 中国农学通报, 2005, 21(7):319-322.
[3]	郝晓娟.作物枯萎病生防细菌的研究[D]. 杭州:浙江大学, 2006.
[4]	PLOETZ R C. Fusarium wilt of banana is caused by several pathogens referred to as Fusarium oxysporum f. sp.cubense[J]. Phytopathology, 2006, 96(6):653-656.
[5]	邓晓.香蕉枯萎病区土壤微生物多样性研究[D]. 海口:海南大学, 2012.
[6]	宋荣浩,戴富明,杨红娟,等.西瓜品种资源对枯萎病和蔓枯病的抗性鉴定[J]. 植物保护, 2009, 35(1):117-120.
[7]	DONG H, FAN H, LEI Z, et al. Histological and gene expression analyses in banana reveals the pathogenic differences between races 1 and 4 of banana Fusarium wilt pathogen[J]. Phytopathology, 2019, 109(6):1029-1042.
[8]	孙茜茜,李春强,易小平.几种生防细菌对尖孢镰刀菌的抑菌效果及其相互间包容性生长的快速检测[J]. 热带农业科学, 2019, 39(12):36-41.
[9]	彭埃天,宋晓兵,凌金锋,等.香蕉枯萎病菌4号生理小种分子检测与枯萎病生物防治研究进展[J]. 果树学报,2009, 26(1):77-81.
[10]	王超,郭坚华,席运官,等.拮抗细菌在植物病害生物防治中应用的研究进展[J]. 江苏农业科学, 2017,45(18):1-6.
[11]	HWANG S C, KO W H. Cavendish banana cultivars resistant to Fusarium wilt acquired through somaclonal variation in Taiwan[J]. Plant Disease, 2004, 88(6):580-588.
[12]	TRIPATHI L, MWANGI M, ABELE S, et al. Xanthomonas wilt:a threat to banana production in east and central Africa[J]. Plant Disease, 2009, 93(5):440-451.
[13]	杨颂,杨利民.微生物菌剂在农业生产上的应用[J]. 吉林农业, 2015(12):86-87.
[14]	孙雪丽,郝向阳,王天池,等.香蕉枯萎病防控和抗病育种研究进展[J]. 果树学报, 2018, 35(7):870-879.
[15]	陆娇娇,畅瑛,李波,等.两株灰黄霉素产生菌的18S rDNA和ITS序列测定与分析[J]. 福建农业科技,2019(10):10-15.
[16]	YU T, LI H Y, ZHENG X D. Synergistic effect of chitosan and Cryptococcus laurentii on inhibition of Penicillum expansum infections[J]. International Journal of Food Microbiology, 2007, 114(3):261-266.
[17]	BOURQUE S N, VALERO J R, LAVOIE M C, et al.Comparative analysis of the 16S to 23S ribosomal intergenic spacer sequences of Bacillus thuringiensis strains and subspecies and of closely related species[J]. Applied and Environmental Microbiology, 1995, 61(7):2811.
[18]	姬彦飞,董欣欣,田野,等.根际促生菌的生防机理及用作生防制剂的潜能[J]. 中国农学通报, 2021,37(14):141-149.
[19]	BLOEMBERG G V, LUGTENBERG B J J. Molecular basis of plant growth promotion and biocontrol by rhizobacteria[J]. Current Opinion in Plant Biology, 2001,4(4):343-350.
[20]	HAN S H, SONG M H, KEUM Y S. Effects of azole fungicides on secreted metabolomes of Botrytis cinerea[J]. Journal of Agricultural and Food Chemistry, 2020, 68(19):5309-5317.
[21]	MORIMURA H, ITO M, YOSHIDA S, et al. In vitro assessment of biocontrol effects on Fusarium head blight and deoxynivalenol(DON) accumulation by DONdegrading bacteria[J]. Toxins, 2020, 12(6):339.
[22]	王松.贝莱斯芽孢杆菌YQ1菌株对香蕉枯萎病菌的拮抗效果研究[D]. 昆明:云南大学, 2020.
[23]	万青,包晓东,李文彬,等.一株贝莱斯芽孢杆菌B18的分离纯化与鉴定及其对香蕉枯萎病的防效[J]. 基因组学与应用生物学, 2021, 40(Z3):3209-3215.
[24]	王小慧,张国漪,李蕊,等.拮抗菌强化的生物有机肥对西瓜枯萎病的防治作用[J]. 植物营养与肥料学报,2013, 19(1):223-231.
[25]	王敏,梁振深,肖日新,等.海南甜瓜椰糠基质栽培技术规程[J]. 广东农业科学, 2010, 37(4):56-58.
[26]	SATYANARAYANA K G, PILLAI C K S, SUKUMARAN K, et al. Structure property studies of fibres from various parts of the coconut tree[J]. Journal of Materials Science,1982,17(8):2453-2462.

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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

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Isolation, identification, biocontrol effect and its colonization in organic substrate of antagonistic bacterium strain LCQ1801 against fusarium wilt

1. School of Tropical Crops, Hainan University, Haikou, Hainan 570228, China;

2. Sanya Research Institute/Institute of Tropical Bioscience and Biotechnology/Ministry of Agriculture and Rural Affairs Key Laboratory of Biology and Genetic Resources of Tropical Crops, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China;

3. Institute of Vegetables/Hainan Key Laboratory of Vegetable Biology/Hainan Engineering Center for Breeding of Vegetables, Hainan Academy of Agricultural Sciences, Haikou, Hainan 571100, China

Received Date: 2023-02-28

Rev Recd Date: 2023-12-09

Publish Date: 2025-03-15

Key words:

Abstract: To develop biological organic fertilizer for preventing and controling Fusarium wilt of banana, a bio-control bacterial strain LCQ1801 was isolated from the soil of rubber plantation in Chengmai, Hainan and its inhibition activity against Fusarium oxysporum f. sp. cubense race 4（Foc4）, the causal agent of banana Fusarium wilt was determined by using plate dual culture. The results indicated that LCQ1801 had obviously antagonistic effect on Foc4,and the inhibition rate was 65.9%. The LCQ1801 was identified as Bacillus velezensis by morphological observation,physiological and biochemical tests, combined with 16S rDNA sequencing analysis. Foc4 was cocultured with LCQ1801 bacterial liquid and filtrate, respectively. The result found that LCQ1801 bacterial fluid and filtrate caused abnormal growth or rupture of Foc4 hyphae and affected spore germination. At the same time, LCQ1801 was cultured on different substrates by using plate assay and culture flask culture. The plate assay indicated that the growth of LCQ1801 in the animal substrates of sheep dung, pig dung and cow dung was better than that in chicken dung. The growth of LCQ1801 in the plant substrates of wheat bran, corn flour, soybean meal and rice straw was better than that in the plant substrates of bagasse and coconut coir. Culture flask culture test showed that the number of viable bacteria of LCQ1801 in cow dung and sheep dung reached 16.8 billions·g^-1 and 24.0 billions·g^-1, respectively. The number of viable bacteria of LCQ1801 in rice straw, soybean meal and corn flour reached 32.2 billions·g^-1, 24.9 billions·g^-1 and 21.7 billions·g^-1, respectively. LCQ1801 hardly grew in the substrate of coconut coir in both culture styles. This study provides strain materials and technical support for biological control of Fusarium wilt of banana.

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Reference (26)

[1]	BENIZRI E, PIUTTI S, VERGER S, et al. Replant diseases:bacterial community structure and diversity in peach rhizosphere as determined by metabolic and genetic fingerprinting[J]. Soil Biology and Biochemistry, 2005,37(9):1738-1746.
[2]	郝晓娟,刘波,谢关林.植物枯萎病生物防治研究进展[J]. 中国农学通报, 2005, 21(7):319-322.
[3]	郝晓娟.作物枯萎病生防细菌的研究[D]. 杭州:浙江大学, 2006.
[4]	PLOETZ R C. Fusarium wilt of banana is caused by several pathogens referred to as Fusarium oxysporum f. sp.cubense[J]. Phytopathology, 2006, 96(6):653-656.
[5]	邓晓.香蕉枯萎病区土壤微生物多样性研究[D]. 海口:海南大学, 2012.
[6]	宋荣浩,戴富明,杨红娟,等.西瓜品种资源对枯萎病和蔓枯病的抗性鉴定[J]. 植物保护, 2009, 35(1):117-120.
[7]	DONG H, FAN H, LEI Z, et al. Histological and gene expression analyses in banana reveals the pathogenic differences between races 1 and 4 of banana Fusarium wilt pathogen[J]. Phytopathology, 2019, 109(6):1029-1042.
[8]	孙茜茜,李春强,易小平.几种生防细菌对尖孢镰刀菌的抑菌效果及其相互间包容性生长的快速检测[J]. 热带农业科学, 2019, 39(12):36-41.
[9]	彭埃天,宋晓兵,凌金锋,等.香蕉枯萎病菌4号生理小种分子检测与枯萎病生物防治研究进展[J]. 果树学报,2009, 26(1):77-81.
[10]	王超,郭坚华,席运官,等.拮抗细菌在植物病害生物防治中应用的研究进展[J]. 江苏农业科学, 2017,45(18):1-6.
[11]	HWANG S C, KO W H. Cavendish banana cultivars resistant to Fusarium wilt acquired through somaclonal variation in Taiwan[J]. Plant Disease, 2004, 88(6):580-588.
[12]	TRIPATHI L, MWANGI M, ABELE S, et al. Xanthomonas wilt:a threat to banana production in east and central Africa[J]. Plant Disease, 2009, 93(5):440-451.
[13]	杨颂,杨利民.微生物菌剂在农业生产上的应用[J]. 吉林农业, 2015(12):86-87.
[14]	孙雪丽,郝向阳,王天池,等.香蕉枯萎病防控和抗病育种研究进展[J]. 果树学报, 2018, 35(7):870-879.
[15]	陆娇娇,畅瑛,李波,等.两株灰黄霉素产生菌的18S rDNA和ITS序列测定与分析[J]. 福建农业科技,2019(10):10-15.
[16]	YU T, LI H Y, ZHENG X D. Synergistic effect of chitosan and Cryptococcus laurentii on inhibition of Penicillum expansum infections[J]. International Journal of Food Microbiology, 2007, 114(3):261-266.
[17]	BOURQUE S N, VALERO J R, LAVOIE M C, et al.Comparative analysis of the 16S to 23S ribosomal intergenic spacer sequences of Bacillus thuringiensis strains and subspecies and of closely related species[J]. Applied and Environmental Microbiology, 1995, 61(7):2811.
[18]	姬彦飞,董欣欣,田野,等.根际促生菌的生防机理及用作生防制剂的潜能[J]. 中国农学通报, 2021,37(14):141-149.
[19]	BLOEMBERG G V, LUGTENBERG B J J. Molecular basis of plant growth promotion and biocontrol by rhizobacteria[J]. Current Opinion in Plant Biology, 2001,4(4):343-350.
[20]	HAN S H, SONG M H, KEUM Y S. Effects of azole fungicides on secreted metabolomes of Botrytis cinerea[J]. Journal of Agricultural and Food Chemistry, 2020, 68(19):5309-5317.
[21]	MORIMURA H, ITO M, YOSHIDA S, et al. In vitro assessment of biocontrol effects on Fusarium head blight and deoxynivalenol(DON) accumulation by DONdegrading bacteria[J]. Toxins, 2020, 12(6):339.
[22]	王松.贝莱斯芽孢杆菌YQ1菌株对香蕉枯萎病菌的拮抗效果研究[D]. 昆明:云南大学, 2020.
[23]	万青,包晓东,李文彬,等.一株贝莱斯芽孢杆菌B18的分离纯化与鉴定及其对香蕉枯萎病的防效[J]. 基因组学与应用生物学, 2021, 40(Z3):3209-3215.
[24]	王小慧,张国漪,李蕊,等.拮抗菌强化的生物有机肥对西瓜枯萎病的防治作用[J]. 植物营养与肥料学报,2013, 19(1):223-231.
[25]	王敏,梁振深,肖日新,等.海南甜瓜椰糠基质栽培技术规程[J]. 广东农业科学, 2010, 37(4):56-58.
[26]	SATYANARAYANA K G, PILLAI C K S, SUKUMARAN K, et al. Structure property studies of fibres from various parts of the coconut tree[J]. Journal of Materials Science,1982,17(8):2453-2462.

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Isolation, identification, biocontrol effect and its colonization in organic substrate of antagonistic bacterium strain LCQ1801 against fusarium wilt

doi: 10.15886/j.cnki.rdswxb.20230023

Abstract

References

Proportional views

通讯作者: 陈斌, bchen63@163.com

Proportional views

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