黄芪根腐病拮抗芽孢杆菌的筛选鉴定及其对根围细菌群落的影响
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摘要
根腐病是严重危害黄芪产业化生产的一类土传病害。该文以山西省黄芪根腐病的优势致病菌Fusarium solani和F. acuminatum为靶标,筛选具有良好抑菌防病效果的生防芽孢杆菌,并测定其对黄芪根围土壤细菌群落的影响,以期为生防制剂的后续开发提供物质基础和理论依据。试验采用平皿对峙拮抗、发酵液抑菌活性测定、菌丝生长抑制、以及离体防病效果测定的方法,对分离自黄芪根围土壤中的芽孢杆菌进行多重筛选,并对获得的目标拮抗芽孢杆菌SXKF16-1进行形态学、生理生化特性、16S r DNA序列鉴定,同时采用DGGE技术分析其对土壤细菌群落的影响。结果显示,菌株SXKF16-1抑菌效果显著,其无菌发酵滤液对F. solani,F. acuminatum的抑菌圈直径分别为(25. 90±1. 18),(25. 86±1. 85) mm,且持效性良好;离体防病试验中,F. solani,F. acuminatum侵染后保护性处理和治疗性处理的病情指数分别是(37. 50±8. 58),(41. 67±4. 90)和(25. 00±8. 33),(38. 89±9. 62),与对照相比均差异显著(P<0. 05)。该菌株鉴定为萎缩芽孢杆菌Bacillus atrophaeus,其发酵液施入土壤后,明显提高了土壤中的细菌多样性。B. atrophaeus SXKF16-1能够有效拮抗根腐病病原菌,并且提高黄芪根围土壤的细菌多样性,具有开发为专用生防制剂的潜力。
The Astragalus membranaceus root rot disease,a soil-borne disease,has become increasingly severe in Shanxi province.This study was aimed at getting antagonistic Bacillus with excellent bio-control effects,and determining its effects on bacterial communities in root zone soil. With Fusarium solani and F. acuminatum as the target,antagonistic Bacillus was selected through such tests as living body dual culture,antifungal effect of bacteria-free filtrate,mycelia growth inhibition in vitro and control effect in detached roots,and identified with morphology,physio-biochemical characteristics and 16 S r DNA sequence analysis. The results showed that the Bacillus strain SXKF16-1 had obvious antifungal effect. The diameter of inhibition zone of its bacteria-free filtrate to F. solani and F. acuminatum was( 25. 90±1. 18) mm and( 25. 86±1. 85) mm respectively,and showed a lasting inhibition effect to mycelia growth. The disease index of the protective treatment and that of the cure treatment in detached roots test to F. solani and F. acuminatum were( 37. 50±8. 58),( 41. 67±4. 90) and( 25. 00±8. 33),( 38. 89±9. 62) respectively,both being significantly different( P<0. 05) from that of the control. The strain SXKF16-1 was identified as Bacillus atrophaeus. The B. atrophaeus SXKF16-1 showed significantly inhibition effect to pathogen causing root rot and could increase the bacterial diversity in root zone soil. It has potential to be developed as a special biocontrol agent.
The Astragalus membranaceus root rot disease,a soil-borne disease,has become increasingly severe in Shanxi province.This study was aimed at getting antagonistic Bacillus with excellent bio-control effects,and determining its effects on bacterial communities in root zone soil. With Fusarium solani and F. acuminatum as the target,antagonistic Bacillus was selected through such tests as living body dual culture,antifungal effect of bacteria-free filtrate,mycelia growth inhibition in vitro and control effect in detached roots,and identified with morphology,physio-biochemical characteristics and 16 S r DNA sequence analysis. The results showed that the Bacillus strain SXKF16-1 had obvious antifungal effect. The diameter of inhibition zone of its bacteria-free filtrate to F. solani and F. acuminatum was( 25. 90±1. 18) mm and( 25. 86±1. 85) mm respectively,and showed a lasting inhibition effect to mycelia growth. The disease index of the protective treatment and that of the cure treatment in detached roots test to F. solani and F. acuminatum were( 37. 50±8. 58),( 41. 67±4. 90) and( 25. 00±8. 33),( 38. 89±9. 62) respectively,both being significantly different( P<0. 05) from that of the control. The strain SXKF16-1 was identified as Bacillus atrophaeus. The B. atrophaeus SXKF16-1 showed significantly inhibition effect to pathogen causing root rot and could increase the bacterial diversity in root zone soil. It has potential to be developed as a special biocontrol agent.
引文
[1]高芬,赵晓霞,秦雪梅,等.山西省蒙古黄芪根腐病优势致病菌群分析[J].植物保护学报,2018,45(4):878.
[2]高芬,任小霞,王梦亮,等.中草药根腐病及其微生物防治研究进展[J].中国中药杂志,2015,40(21):4122.
[3] Zobel S,Kumpfmüller J,Süssmuth R D,et al. Bacillus subtilis as heterologous host for the secretory production of the non-ribosomal cyclodepsipeptide enniatin[J]. Appl Microbiol Biotechnol,2015,99(2)):681.
[4]刘丹,张丽萍,史延茂,等.生防细菌对植物根围微生态效应的研究进展[J].中国农学通报,2014,30(7):260.
[5]任小霞,秦雪梅,王梦亮,等.山西黄芪根腐病优势病原菌鉴定及其拮抗菌的多重筛选[J].中药材,2016,39(10):2173.
[6]陈垣,朱蕾,郭凤霞,等.甘肃渭源蒙古黄芪根腐病病原菌的分离与鉴定[J].植物病理学报,2011,41(4):428.
[7]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.
[8]布坎南R E,吉本斯N E.伯杰细菌鉴定手册[M].北京:科学出版社,1984.
[9]尹丹韩,高观朋,夏飞,等.生防菌哈茨木霉T4对黄瓜根围土壤细菌群落的影响[J].中国农业科学,2012,45(2):246.
[10]刘绍雄,王明月,王娟,等.基于PCR-DGGE技术的剑湖湿地湖滨带土壤微生物群落结构多样性分析[J].农业环境科学学报,2013,32(7):1405.
[11]高芬,郝锐,秦雪梅,等.拮抗黄芪根腐病菌的根际促生菌的室内筛选与鉴定[J].中国中药杂志,2016,41(22):4188.
[12] Mwita L,Chan W Y,Pretorius T,et al. Gene expression regulation in the plant growth promoting Bacillus atrophaeus UCMB-5137 stimulated by maize root exudates[J]. Gene,2016,590(1):18.
[13]蔡燕飞,廖宗文,章家恩,等.生态有机肥对蕃茄青枯病及土壤微生物多样性的研究[J].应用生态学报,2003,14(3):349.
[14] Ellis R J,Morgan P,Weightman A J,et al.Cultivation dependent and independent approaches for determining bacterial diversity inheavy-metal-contaminated soil[J]. Appl Environ Microbiol,2003,69(6):3223.
[15]朱伟杰,王楠,郁雪平,等.生防菌Pseudomonas fluorescens 2P24对甜瓜根围土壤微生物的影响[J].中国农业科学,2010,43(7):1389.
[16]范文艳,陈瑾,姜述君,等.复合生防菌群对连作大豆根际土壤可培养微生物区系的影响[J].中国油料作物学报,2012,34(3):286.
[2]高芬,任小霞,王梦亮,等.中草药根腐病及其微生物防治研究进展[J].中国中药杂志,2015,40(21):4122.
[3] Zobel S,Kumpfmüller J,Süssmuth R D,et al. Bacillus subtilis as heterologous host for the secretory production of the non-ribosomal cyclodepsipeptide enniatin[J]. Appl Microbiol Biotechnol,2015,99(2)):681.
[4]刘丹,张丽萍,史延茂,等.生防细菌对植物根围微生态效应的研究进展[J].中国农学通报,2014,30(7):260.
[5]任小霞,秦雪梅,王梦亮,等.山西黄芪根腐病优势病原菌鉴定及其拮抗菌的多重筛选[J].中药材,2016,39(10):2173.
[6]陈垣,朱蕾,郭凤霞,等.甘肃渭源蒙古黄芪根腐病病原菌的分离与鉴定[J].植物病理学报,2011,41(4):428.
[7]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.
[8]布坎南R E,吉本斯N E.伯杰细菌鉴定手册[M].北京:科学出版社,1984.
[9]尹丹韩,高观朋,夏飞,等.生防菌哈茨木霉T4对黄瓜根围土壤细菌群落的影响[J].中国农业科学,2012,45(2):246.
[10]刘绍雄,王明月,王娟,等.基于PCR-DGGE技术的剑湖湿地湖滨带土壤微生物群落结构多样性分析[J].农业环境科学学报,2013,32(7):1405.
[11]高芬,郝锐,秦雪梅,等.拮抗黄芪根腐病菌的根际促生菌的室内筛选与鉴定[J].中国中药杂志,2016,41(22):4188.
[12] Mwita L,Chan W Y,Pretorius T,et al. Gene expression regulation in the plant growth promoting Bacillus atrophaeus UCMB-5137 stimulated by maize root exudates[J]. Gene,2016,590(1):18.
[13]蔡燕飞,廖宗文,章家恩,等.生态有机肥对蕃茄青枯病及土壤微生物多样性的研究[J].应用生态学报,2003,14(3):349.
[14] Ellis R J,Morgan P,Weightman A J,et al.Cultivation dependent and independent approaches for determining bacterial diversity inheavy-metal-contaminated soil[J]. Appl Environ Microbiol,2003,69(6):3223.
[15]朱伟杰,王楠,郁雪平,等.生防菌Pseudomonas fluorescens 2P24对甜瓜根围土壤微生物的影响[J].中国农业科学,2010,43(7):1389.
[16]范文艳,陈瑾,姜述君,等.复合生防菌群对连作大豆根际土壤可培养微生物区系的影响[J].中国油料作物学报,2012,34(3):286.
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