基于根系分泌物的根际强定殖生防细菌筛选及其在黄瓜枯萎病防治中的应用

doi:10.7668/hbnxb.20195721

华北农学报 ›› 2025, Vol. 40 ›› Issue (6): 168-176. doi: 10.7668/hbnxb.20195721

所属专题： 黄瓜；植物保护；蔬菜专题；

• 资源环境·植物保护 • 上一篇下一篇

基于根系分泌物的根际强定殖生防细菌筛选及其在黄瓜枯萎病防治中的应用

曲远航¹^,², 苏振贺², 董丽红², 张晓云², 李社增², 郭庆港², 马平²

¹ 河北农业大学植物保护学院,河北保定 071000
² 河北省农林科学院植物保护研究所,农业农村部华北北部作物有害生物综合治理重点实验室,河北省农业有害生物综合防治技术创新中心,河北省作物有害生物综合防治国际科技联合研究中心,河北保定 071000

收稿日期:2024-12-27 出版日期:2025-12-31
通讯作者:
马平(1964—),男,河北保定人,研究员,博士,博士生导师,主要从事植物病害生物防治研究。
郭庆港(1976—),男,河北沧州人,研究员,博士,博士生导师,主要从事植物病害生物防治研究。
作者简介:
曲远航(1990—),男,山东泰安人,助理研究员,博士,主要从事植物病害生物防治研究。
基金资助:
河北省农林科学院基本科研业务费青年项目(2023120102); 河北省农林科学院科技创新专项(2022KJCXZX-ZBS-1)

Screening of Strong Rhizosphere-colonizing Biocontrol Bacteria Based on Root Exudates and Their Application in Cucumber Fusarium Wilt Disease Management

QU Yuanhang¹^,², SU Zhenhe², DONG Lihong², ZHANG Xiaoyun², LI Shezeng², GUO Qinggang², MA Ping²

¹ College of Plant Protection,Hebei Agricultural University,Baoding 071000,China
² Plant Protection Institute,Hebei Academy of Agriculture and Forestry Sciences,Key Laboratory of Integrated Pest Management on Crops in Northern Region of North China,Ministry of Agriculture and Rural Affairs,P.R.China,IPM Innovation Center of Hebei Province,International Science and Technology Joint Research Center on IPM of Hebei Province,Baoding 071000,China

Received:2024-12-27 Published:2025-12-31

摘要/Abstract

摘要：

旨在建立一种基于根系分泌物的生防菌筛选体系,筛选具有根际强定殖能力的生防细菌,并利用该体系筛选与菌糠复配防治黄瓜枯萎病的细菌菌株。通过病原菌胁迫和菌糠诱导,调控黄瓜根系分泌物的化学组成,以待测菌株对根系分泌物的趋化性、代谢增殖和生物膜形成等定殖相关因素为定量指标,采用毛细管试验、96孔板培养和生物膜形成测定,实现根际强定殖细菌菌株的高效筛选。结果表明,从黄瓜根际分离的200株细菌对根系分泌物的趋化性、代谢增殖能力和生物膜形成能力存在显著差异。利用该筛选体系,筛选出的贝莱斯芽孢杆菌MTC-5菌株在所有筛选指标中均表现优异。温室试验结果表明,MTC-5菌株与香菇菌糠复配具有更强的根际定殖能力,并在防病和促生上表现出显著的协同增效作用。MTC-5和菌糠的复配菌剂对黄瓜枯萎病的田间防病效果为74.5%,比单独施用菌糠和MTC-5菌株的防效分别超出10.7,24.6百分点。建立了一个基于根系分泌物的高通量生防菌筛选体系,揭示了根系分泌物在细菌招募中的关键作用。基于该体系创制的MTC-5菌株与菌糠的复配菌剂在防治黄瓜枯萎病中展现出显著的协同增效,为生防菌筛选和应用提供了新的思路和技术支持。

关键词: 生防菌, 筛选, 根系分泌物, 菌糠, 黄瓜枯萎病, 芽孢杆菌

Abstract:

This study aims to establish a biocontrol bacteria screening system based on root exudates to screen biocontrol strains with strong rhizosphere colonization ability.Using this system,bacterial strains capable of synergistic control of cucumber Fusarium wilt in combination with spent mushroom substrate(SMS)were screened.The chemical composition of cucumber root exudates was modulated by pathogen stress and SMS induction.Chemotaxis,metabolic proliferation,and biofilm formation,which were colonization-related factors,were used as quantitative indicators for the tested strains.Efficient screening of rhizosphere-colonizing bacteria was achieved using capillary assays,96-well plate cultures,and biofilm formation tests.Among 200 bacterial strains isolated from the cucumber rhizosphere,significant differences were observed in chemotaxis,metabolic proliferation,and biofilm formation in response to root exudates.Using this screening system,Bacillus velezensis MTC-5 was screened as an outstanding strain across all screening indicators.Greenhouse experiments demonstrated that MTC-5 strain combined with SMS exhibited enhanced rhizosphere colonization ability and showed significant synergistic effects in disease suppression and plant growth promotion.Field trials revealed that the combined application of MTC-5 strain and SMS achieved a disease control efficacy of 74.5% against cucumber Fusarium wilt,which was 10.7,24.6 percent points higher than the efficacy of SMS or MTC-5 strain alone,respectively.It established a high-throughput biocontrol bacteria screening system based on root exudates,highlighting the critical role of root exudates in bacterial recruitment.The MTC-5 and SMS combination developed through this system demonstrated significant synergistic effects in controlling cucumber Fusarium wilt,providing new insights and technical support for biocontrol bacteria screening and application.

Key words: Biocontrol bacteria, Screening, Root exudates, Spent mushroom substrate, Cucumber Fusarium wilt, Bacillus

中图分类号:

曲远航, 苏振贺, 董丽红, 张晓云, 李社增, 郭庆港, 马平. 基于根系分泌物的根际强定殖生防细菌筛选及其在黄瓜枯萎病防治中的应用[J]. 华北农学报, 2025, 40(6): 168-176. doi: 10.7668/hbnxb.20195721.

QU Yuanhang, SU Zhenhe, DONG Lihong, ZHANG Xiaoyun, LI Shezeng, GUO Qinggang, MA Ping. Screening of Strong Rhizosphere-colonizing Biocontrol Bacteria Based on Root Exudates and Their Application in Cucumber Fusarium Wilt Disease Management[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(6): 168-176. doi: 10.7668/hbnxb.20195721.

http://www.hbnxb.net/CN/Y2025/V40/I6/168

图/表 7

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菌株 Strains	细胞密度(OD₆₀₀)Cell density		增幅/% Increase rate
菌株 Strains	CK-CRE	SMS-CRE	增幅/% Increase rate
MTC-5	0.049±0.007 3b	0.177±0.009 0a	261.22
GY2-27	0.080±0.011 1b	0.221±0.011 9a	176.25
MTS-35	0.060±0.008 0b	0.135±0.007 2a	125.00
MTS-29	0.067±0.007 7b	0.148±0.010 6a	120.90
R-10	0.064±0.007 2b	0.135±0.011 0a	110.94
MTC-3	0.063±0.009 2b	0.129±0.010 3a	104.76
LY2-15	0.077±0.009 3b	0.154±0.011 9a	100.00
GYS-17	0.071±0.006 9b	0.137±0.006 0a	92.96
LYS-28	0.072±0.008 3b	0.135±0.009 3a	87.50
MTC-2	0.063±0.008 2b	0.114±0.006 9a	80.95
R-8	0.075±0.006 1b	0.133±0.010 0a	77.33
MTS-13	0.067±0.007 5b	0.115±0.007 7a	71.64
GY2-16	0.068±0.009 8b	0.111±0.007 3a	63.24
MTC-20	0.069±0.007 1b	0.112±0.012 0a	62.32
R-9	0.181±0.009 1b	0.289±0.006 3a	59.67
GY2-28	0.080±0.011 9b	0.126±0.008 2a	57.50
MTC-6	0.078±0.010 4b	0.121±0.011 0a	55.13
LYS-14	0.093±0.010 1b	0.140±0.010 8a	50.54
LY2-5	0.105±0.009 3b	0.158±0.008 1a	50.48
MTC-1	0.073±0.010 1b	0.109±0.011 4a	49.32
GYS-3	0.086±0.008 3b	0.127±0.008 4a	47.67
GYS-34	0.097±0.007 2b	0.141±0.009 2a	45.36
GYS-19	0.075±0.008 9b	0.108±0.010 6a	44.00
GYS-22	0.077±0.008 3b	0.108±0.011 4a	40.26
MTC-25	0.081±0.006 6b	0.113±0.011 5a	39.51
LY2-2	0.103±0.009 7b	0.143±0.007 0a	38.83
R-1	0.110±0.007 5b	0.152±0.010 2a	38.18
GYS-21	0.115±0.009 6b	0.155±0.007 3a	34.78
GYS-28	0.106±0.009 5a	0.118±0.007 1a	11.32
GYS-32	0.104±0.008 7a	0.113±0.010 1a	8.65
GY2-4	0.107±0.006 9a	0.111±0.007 9a	3.74
LY2-1	0.104±0.010 9a	0.106±0.011 0a	1.92
GYS-5	0.117±0.009 8a	0.118±0.009 4a	0.85
MTS-3	0.137±0.009 8a	0.134±0.009 3a	-2.19
GYS-31	0.149±0.006 4a	0.142±0.010 1a	-4.70
LY2-19	0.120±0.009 4a	0.109±0.011 1a	-9.17
MTC-18	0.120±0.010 3a	0.107±0.008 3a	-10.83
LY2-16	0.127±0.011 4a	0.110±0.010 9a	-13.39
GYS-36	0.129±0.007 5a	0.109±0.008 8a	-15.50
MTC-29	0.123±0.008 0a	0.103±0.009 1a	-16.26
GYS-12	0.133±0.006 3a	0.109±0.010 6a	-18.05
MTS-33	0.129±0.009 1a	0.104±0.010 9b	-19.38
MTS-24	0.129±0.011 5a	0.100±0.007 1b	-22.48
GYS-2	0.139±0.009 5a	0.102±0.007 4b	-26.62
GY2-24	0.137±0.008 0a	0.100±0.008 5b	-27.01
MTC-22	0.142±0.011 4a	0.102±0.007 2b	-28.17
LYS-24	0.146±0.006 8a	0.101±0.011 0b	-30.82

菌株 Strains	细胞密度(OD₆₀₀)Cell density		增幅/% Increase rate
菌株 Strains	CK-CRE	SMS-CRE	增幅/% Increase rate
MTC-5	0.049±0.007 3b	0.177±0.009 0a	261.22
GY2-27	0.080±0.011 1b	0.221±0.011 9a	176.25
MTS-35	0.060±0.008 0b	0.135±0.007 2a	125.00
MTS-29	0.067±0.007 7b	0.148±0.010 6a	120.90
R-10	0.064±0.007 2b	0.135±0.011 0a	110.94
MTC-3	0.063±0.009 2b	0.129±0.010 3a	104.76
LY2-15	0.077±0.009 3b	0.154±0.011 9a	100.00
GYS-17	0.071±0.006 9b	0.137±0.006 0a	92.96
LYS-28	0.072±0.008 3b	0.135±0.009 3a	87.50
MTC-2	0.063±0.008 2b	0.114±0.006 9a	80.95
R-8	0.075±0.006 1b	0.133±0.010 0a	77.33
MTS-13	0.067±0.007 5b	0.115±0.007 7a	71.64
GY2-16	0.068±0.009 8b	0.111±0.007 3a	63.24
MTC-20	0.069±0.007 1b	0.112±0.012 0a	62.32
R-9	0.181±0.009 1b	0.289±0.006 3a	59.67
GY2-28	0.080±0.011 9b	0.126±0.008 2a	57.50
MTC-6	0.078±0.010 4b	0.121±0.011 0a	55.13
LYS-14	0.093±0.010 1b	0.140±0.010 8a	50.54
LY2-5	0.105±0.009 3b	0.158±0.008 1a	50.48
MTC-1	0.073±0.010 1b	0.109±0.011 4a	49.32
GYS-3	0.086±0.008 3b	0.127±0.008 4a	47.67
GYS-34	0.097±0.007 2b	0.141±0.009 2a	45.36
GYS-19	0.075±0.008 9b	0.108±0.010 6a	44.00
GYS-22	0.077±0.008 3b	0.108±0.011 4a	40.26
MTC-25	0.081±0.006 6b	0.113±0.011 5a	39.51
LY2-2	0.103±0.009 7b	0.143±0.007 0a	38.83
R-1	0.110±0.007 5b	0.152±0.010 2a	38.18
GYS-21	0.115±0.009 6b	0.155±0.007 3a	34.78
GYS-28	0.106±0.009 5a	0.118±0.007 1a	11.32
GYS-32	0.104±0.008 7a	0.113±0.010 1a	8.65
GY2-4	0.107±0.006 9a	0.111±0.007 9a	3.74
LY2-1	0.104±0.010 9a	0.106±0.011 0a	1.92
GYS-5	0.117±0.009 8a	0.118±0.009 4a	0.85
MTS-3	0.137±0.009 8a	0.134±0.009 3a	-2.19
GYS-31	0.149±0.006 4a	0.142±0.010 1a	-4.70
LY2-19	0.120±0.009 4a	0.109±0.011 1a	-9.17
MTC-18	0.120±0.010 3a	0.107±0.008 3a	-10.83
LY2-16	0.127±0.011 4a	0.110±0.010 9a	-13.39
GYS-36	0.129±0.007 5a	0.109±0.008 8a	-15.50
MTC-29	0.123±0.008 0a	0.103±0.009 1a	-16.26
GYS-12	0.133±0.006 3a	0.109±0.010 6a	-18.05
MTS-33	0.129±0.009 1a	0.104±0.010 9b	-19.38
MTS-24	0.129±0.011 5a	0.100±0.007 1b	-22.48
GYS-2	0.139±0.009 5a	0.102±0.007 4b	-26.62
GY2-24	0.137±0.008 0a	0.100±0.008 5b	-27.01
MTC-22	0.142±0.011 4a	0.102±0.007 2b	-28.17
LYS-24	0.146±0.006 8a	0.101±0.011 0b	-30.82

菌株 Strains	定殖量/(×10⁵ cfu/g) Colonization quality		增幅/% Increase rate
菌株 Strains	BAC	SMS-BAC	增幅/% Increase rate
MTC-5	8.61±0.11b	13.51±0.12a	57.20
GYS-21	8.28±0.42b	12.65±0.86a	52.30
MTC-25	8.22±0.62b	12.03±0.85a	46.30
MTC-20	9.54±0.67b	11.91±0.56a	24.40
LY2-5	7.41±0.47b	9.95±0.36a	34.30
GY2-16	8.87±0.67a	9.26±0.35a	4.40
R-10	6.39±0.87b	8.70±0.12a	36.20
GY2-27	5.71±0.89b	8.40±0.24a	47.30
GYS-17	7.63±0.53a	8.10±0.72a	6.20
GY2-28	6.20±0.38a	6.46±0.17a	4.10
MTS-13	5.13±0.22a	5.29±0.37a	3.10
GYS-19	2.89±0.04b	4.41±0.76a	52.40
R-9	1.66±0.12b	2.18±0.11a	31.00
LYS-14	0.69±0.02a	7.25±0.33a	4.40

菌株 Strains	定殖量/(×10⁵ cfu/g) Colonization quality		增幅/% Increase rate
菌株 Strains	BAC	SMS-BAC	增幅/% Increase rate
MTC-5	8.61±0.11b	13.51±0.12a	57.20
GYS-21	8.28±0.42b	12.65±0.86a	52.30
MTC-25	8.22±0.62b	12.03±0.85a	46.30
MTC-20	9.54±0.67b	11.91±0.56a	24.40
LY2-5	7.41±0.47b	9.95±0.36a	34.30
GY2-16	8.87±0.67a	9.26±0.35a	4.40
R-10	6.39±0.87b	8.70±0.12a	36.20
GY2-27	5.71±0.89b	8.40±0.24a	47.30
GYS-17	7.63±0.53a	8.10±0.72a	6.20
GY2-28	6.20±0.38a	6.46±0.17a	4.10
MTS-13	5.13±0.22a	5.29±0.37a	3.10
GYS-19	2.89±0.04b	4.41±0.76a	52.40
R-9	1.66±0.12b	2.18±0.11a	31.00
LYS-14	0.69±0.02a	7.25±0.33a	4.40

菌株 Strains	BAC		SMS-BAC		防效增幅/百分点 Efficiency enhancement
菌株 Strains	发病率/% Disease incidence	防病效果/% Control effect	发病率/% Disease incidence	防病效果/% Control effect	防效增幅/百分点 Efficiency enhancement
CK	87.60±7.02a	0.00	87.60±7.02a	0.00	0.00
LY2-13	70.41±5.43cde	19.62	59.05±5.51d	32.59	12.97
GY2-11	72.98±5.41bcd	16.69	57.01±6.39de	34.92	18.23
GYS-32	83.98±7.23ab	4.13	78.32±6.10ab	10.59	6.46
GYS-21	61.82±7.88def	29.43	32.38±6.19g	63.04	33.61
GYS-11	58.73±6.00ef	32.96	32.13±5.12g	63.32	30.36
GY2-1	79.63±5.64abc	9.10	63.67±5.06cd	27.32	18.22
LYS-17	51.08±6.51fg	41.69	19.87±4.80h	77.32	35.63
MTS-27	66.96±5.66cde	23.56	45.34±6.87ef	48.24	24.68
GYS-13	85.47±5.21a	2.43	72.52±4.41bc	17.21	14.78
MTC-5	46.51±4.13g	46.91	13.41±2.80h	84.69	37.78
LY2-3	65.79±6.38cde	24.90	40.83±3.17fg	53.39	28.49
MTC-28	71.19±7.35bcde	18.73	56.46±4.51de	35.55	16.82
LYS-13	86.65±7.12a	1.08	76.04±4.95ab	13.20	12.12
GY2-3	59.71±6.26ef	31.84	45.77±6.64ef	47.75	15.91
SMS	81.32±9.81abc	7.17	—	—	—

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基于根系分泌物的根际强定殖生防细菌筛选及其在黄瓜枯萎病防治中的应用

Screening of Strong Rhizosphere-colonizing Biocontrol Bacteria Based on Root Exudates and Their Application in Cucumber Fusarium Wilt Disease Management

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菌株 Strains	株高/mm Height	干质量/g Dry weight	SPAD
MTC-25	186.67±10.65bcd	2.13±0.22a	50.96±5.74abc
MTC-5	221.67±19.56a	2.15±0.18a	46.08±5.55cd
GYS-21	219.08±17.10a	2.02±0.20ab	50.14±5.95abc
MTC-20	204.33±15.98ab	2.23±0.18a	49.41±4.93bc
LY2-5	203.25±28.45abc	1.89±0.22abc	45.38±4.79cd
GY2-16	197.83±20.53abc	1.57±0.14c	46.15±5.62cd
R-10	197.17±13.00abc	1.74±0.15bc	48.51±2.98c
GY2-27	191.75±15.87abc	2.18±0.24a	52.12±7.35abc
GYS-17	183.55±18.84bcd	1.55±0.13c	49.82±5.29bc
GY2-28	183.00±24.65bcd	2.26±0.18a	53.28±6.42abc
MTS-13	171.67±16.37cd	1.28±0.13d	54.09±3.65abc
GYS-19	163.17±13.99cd	1.25±0.11d	60.54±4.13a
R-9	156.08±17.80de	1.10±0.12d	55.82±5.74abc
LYS-14	151.82±24.84de	1.20±0.12d	59.34±5.70ab
CK	127.45±13.62e	0.85±0.09e	37.66±3.50d

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基于根系分泌物的根际强定殖生防细菌筛选及其在黄瓜枯萎病防治中的应用

Screening of Strong Rhizosphere-colonizing Biocontrol Bacteria Based on Root Exudates and Their Application in Cucumber Fusarium Wilt Disease Management

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