黑土区大豆根际土壤放线菌的分离与功能研究

doi:10.13560/j.cnki.biotech.bull.1985.2024-0994

生物技术通报 ›› 2025, Vol. 41 ›› Issue (5): 255-266.doi: 10.13560/j.cnki.biotech.bull.1985.2024-0994

• 研究报告 • 上一篇

黑土区大豆根际土壤放线菌的分离与功能研究

陈永旗¹^,²(), 李志文¹^,², 李鑫¹, 原若曦¹, 王春旭¹^,², 韩毅强¹^,²^,³, 高亚梅¹^,²^,⁴()

^1.黑龙江八一农垦大学生命科学技术学院，大庆 163319
^2.寒区环境微生物与农业废弃物资源化利用黑龙江省重点实验室，大庆 163319
^3.国家杂粮工程中心，大庆 163319
^4.农业农村部东北平原农业绿色低碳重点实验室，大庆 163319

收稿日期:2024-10-12 出版日期:2025-05-26 发布日期:2025-06-05
通讯作者: 高亚梅，女，教授，研究方向：微生物资源开发与功能研究；E-mail: gaoym800@126.com
作者简介:陈永旗，女，硕士，研究方向：寒区环境微生物与农业废弃物资源化利用；E-mail: 3065408202@qq.com
基金资助:
黑龙江省面上基金项目(LH2021C065);黑龙江八一农垦大学大学生创新训练项目(XS2024031)

Isolation and Function Study of Actinomycetes from Rhizosphere Soil of Soybean in the Black Soil Region

CHEN Yong-qi¹^,²(), LI Zhi-wen¹^,², LI Xin¹, YUAN Ruo-xi¹, WANG Chun-xu¹^,², HAN Yi-qiang¹^,²^,³, GAO Ya-mei¹^,²^,⁴()

^1.College of Life Science and Biotechnology, Heilongjiang Bayi Agricultural University, Daqing 163319
^2.Heilongjiang Provincial Key Laboratory of Environmental Microbiology and Resource Utilization of Agricultural Waste in Cold Regions, Daqing 163319
^3.National Coarse Cereals Engineering Research Center, Daqing 163319
^4.Key Laboratory of Low-carbon Green Agriculture in Northeastern China, Ministry of Agriculture and Rural Affairs, Daqing 163319

Received:2024-10-12 Published:2025-05-26 Online:2025-06-05

摘要/Abstract

摘要：

目的大豆疫霉根腐病是由大豆疫霉引起的严重土传病害，从黑土区大豆根际土壤中筛选并鉴定对大豆疫霉具有拮抗作用的放线菌资源。方法选择4种分离培养基，利用稀释划线法从健康和患病大豆植株根际土壤中进行菌株分离，通过平板对峙法筛选对大豆疫霉有抑制活性的菌株，并对候选菌株进行培养特征、生理生化特性测定和基于16S rRNA的分子鉴定。结果从健康植株根际土壤中分离了87株放线菌，患病植株根际土壤中分离了43株放线菌。通过平板对峙实验在130株菌中筛选出了11株对大豆疫霉有较好抑制效果的菌株。来自患病根际土的HV-HDN-12对大豆疫霉抑制率为45.48%，健康土壤中分离的LSV-JDN-1对大豆疫霉抑制率达45.84%。HV-HDN-12和LSV-JDN-1具有ACC脱氨酶活性和促进植物生长的吲哚乙酸（IAA），LSV-JDN-1可产铁载体。根据其形态和生理生化结果以及16S rRNA序列分析，HV-HDN-12鉴定为Streptomyces albidoflavus，LSV-JDN-1鉴定为Streptomyces cavourensis。结论 HV-HDN-12和LSV-JDN-1来自黑土大豆种植区大豆根际，具有广谱抗真菌和植物促生的优良特性，是有较好生防潜力的放线菌资源。

关键词: 放线菌, 拮抗菌, 筛选, 拮抗作用, 大豆疫霉

Abstract:

Objective Soybean root rot, caused by Phytophthora sojae, is a serious soil-borne disease. The actinomycetes resources with antagonistic effect on P. sojae in the black soil region were explored. Method Actinomycete from the rhizosphere soil of healthy and diseased soybean plants were isolated by the dilution plate scribing method using four isolation media, and strains with inhibitory activity against P. sojae were screened by the plate confrontation method, then the cultivation characteristics, physiological and biochemical properties of candidate strains were measured, and molecular identification based on 16S rRNA was performed. Result The total 87 strains of actinomycetes were isolated from rhizosphere soil of healthy plants and total 43 strains from rhizosphere soil of diseased plants. Among the 130 strains, eleven strains showed obvious antifungal activity. HV-HDN-12 from the diseased root soil significantly inhibited P. sojae with an inhibition rate of 45.48%, and LSV-JDN-1 from the healthy root soil significantly inhibited P. sojae with an inhibition rate of 45.84%. HV-HDN-12 and LSV-JDN-1 have ACC deaminase activity and produce plant growth-promoting indoleacetic acid (IAA). LSV-JDN-1 produced siderophore. HVNDN-12 showed broad-spectrum antifungal activity and was identified as Streptomyces albidoflavus based on its morphological, physiological and biochemical results and 16S rRNA sequence analysis, and LSV-JDN-1 was identified as Streptomyces cavourensis. Conclusion S. alboflavus HVNDN-12 and S. cavourensis LSV-JDN-1 are actinomycete resource with great biocontrol potential against the root rot of soybean with comprehensive anti-fungi and plant growth promotion from soybean rhizosphere in black soil region.

Key words: actinomycete, antagonistic bacteria, screening, antagonism, Phytophthora sojae

陈永旗, 李志文, 李鑫, 原若曦, 王春旭, 韩毅强, 高亚梅. 黑土区大豆根际土壤放线菌的分离与功能研究[J]. 生物技术通报, 2025, 41(5): 255-266.

CHEN Yong-qi, LI Zhi-wen, LI Xin, YUAN Ruo-xi, WANG Chun-xu, HAN Yi-qiang, GAO Ya-mei. Isolation and Function Study of Actinomycetes from Rhizosphere Soil of Soybean in the Black Soil Region[J]. Biotechnology Bulletin, 2025, 41(5): 255-266.

图/表 11

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菌株 Strain	大豆疫霉 P. sojae	尖孢镰刀菌 F. oxysporum	小麦赤霉菌 G. triticum	燕麦镰刀菌 F. oat	玉米大斑病菌 J. spot	核盘菌 S. sclerotiorum	芹菜灰霉菌 Celery cinerine mold	麦根镰刀菌 F. rhizome	稻瘟病菌 M. oryzae
LSV-JDN-1	45.84	30.67	14.74	11.16	17.88	17.53	19.55	30.98	26.98
LSV-JDN-3	44.67	30.79	27.99	20.48	20.78	19.35	17.46	17.69	27.38
GS2-JDN-7	40.27	32.03	20.61	19.52	12.48	16.35	15.20	18.34	25.56
LSV-JDN-8	42.56	27.80	15.05	14.62	14.45	18.26	24.55	27.38	35.95
LSV-JDN-10	41.58	20.74	14.70	12.64	20.75	17.68	25.64	26.10	28.39
GS-JDN-13	45.64	22.56	无	无	8.56	18.91	18.58	25.87	34.10
HV-HDN-5-1	42.87	30.69	13.26	8.64	16.58	15.24	23.13	19.35	30.64
LSV-HDN-6	40.77	20.14	12.84	18.76	13.26	19.21	20.68	25.67	31.49
HV-HDN-12	45.48	34.59	10.19	21.15	20.18	18.18	22.87	24.87	28.76
GS-HDN-12-1	43.44	34.98	20.69	25.05	22.14	16.54	15.74	11.43	23.98
GS-HDN-12-2	43.64	36.39	无	12.38	23.67	12.36	21.35	18.64	29.86

菌株 Strain	大豆疫霉 P. sojae	尖孢镰刀菌 F. oxysporum	小麦赤霉菌 G. triticum	燕麦镰刀菌 F. oat	玉米大斑病菌 J. spot	核盘菌 S. sclerotiorum	芹菜灰霉菌 Celery cinerine mold	麦根镰刀菌 F. rhizome	稻瘟病菌 M. oryzae
LSV-JDN-1	45.84	30.67	14.74	11.16	17.88	17.53	19.55	30.98	26.98
LSV-JDN-3	44.67	30.79	27.99	20.48	20.78	19.35	17.46	17.69	27.38
GS2-JDN-7	40.27	32.03	20.61	19.52	12.48	16.35	15.20	18.34	25.56
LSV-JDN-8	42.56	27.80	15.05	14.62	14.45	18.26	24.55	27.38	35.95
LSV-JDN-10	41.58	20.74	14.70	12.64	20.75	17.68	25.64	26.10	28.39
GS-JDN-13	45.64	22.56	无	无	8.56	18.91	18.58	25.87	34.10
HV-HDN-5-1	42.87	30.69	13.26	8.64	16.58	15.24	23.13	19.35	30.64
LSV-HDN-6	40.77	20.14	12.84	18.76	13.26	19.21	20.68	25.67	31.49
HV-HDN-12	45.48	34.59	10.19	21.15	20.18	18.18	22.87	24.87	28.76
GS-HDN-12-1	43.44	34.98	20.69	25.05	22.14	16.54	15.74	11.43	23.98
GS-HDN-12-2	43.64	36.39	无	12.38	23.67	12.36	21.35	18.64	29.86

实验项目 Test	LSV-JDN-1	HV-HDN-12	实验项目 Test item	LSV- JDN-1	HV-HDN-12
乳糖 Lactose	+	+	苏氨酸 Threonine	+	+
鼠李糖 Rhamnose	+	-	谷氨酰胺 Glutamine	+	+
棉子糖 Marshmallow	+	-	酪氨酸 Tyrosine	-	+
甘露醇 Mannitol	+	-	精氨酸 Argnine	+	+
葡萄糖 Glucose	+	+	丙氨酸 Alanine	+	+
木糖 Xylose	-	+	牛奶凝固与胨化 Milk peptonization	+	+
甘露糖 Mannose	+	+	明胶液化 Gelatin Liquefaction	+	+
阿拉伯糖 Arabinose	-	+	淀粉水解 Starch hydrolysis	+	+
肌醇 Inositol	+	-	硫化氢产生 Hydrogen Sulfide Generation	-	-
半乳糖 Galactose	+	+	硝酸盐还原 Nitrate Reduction	+	+
蔗糖 Fructose	+	-	纤维素水解 Urease	-	-
天冬氨酸 Aspartic acid	-	-	脲酶 Lipase	-	-
肌酸 Creatine	+	+

实验项目 Test	LSV-JDN-1	HV-HDN-12	实验项目 Test item	LSV- JDN-1	HV-HDN-12
乳糖 Lactose	+	+	苏氨酸 Threonine	+	+
鼠李糖 Rhamnose	+	-	谷氨酰胺 Glutamine	+	+
棉子糖 Marshmallow	+	-	酪氨酸 Tyrosine	-	+
甘露醇 Mannitol	+	-	精氨酸 Argnine	+	+
葡萄糖 Glucose	+	+	丙氨酸 Alanine	+	+
木糖 Xylose	-	+	牛奶凝固与胨化 Milk peptonization	+	+
甘露糖 Mannose	+	+	明胶液化 Gelatin Liquefaction	+	+
阿拉伯糖 Arabinose	-	+	淀粉水解 Starch hydrolysis	+	+
肌醇 Inositol	+	-	硫化氢产生 Hydrogen Sulfide Generation	-	-
半乳糖 Galactose	+	+	硝酸盐还原 Nitrate Reduction	+	+
蔗糖 Fructose	+	-	纤维素水解 Urease	-	-
天冬氨酸 Aspartic acid	-	-	脲酶 Lipase	-	-
肌酸 Creatine	+	+

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黑土区大豆根际土壤放线菌的分离与功能研究

Isolation and Function Study of Actinomycetes from Rhizosphere Soil of Soybean in the Black Soil Region

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