洋葱伯克霍尔德氏菌KRS634对棉花红腐病的生防效果

doi:10.13560/j.cnki.biotech.bull.1985.2025-0497

生物技术通报 ›› 2025, Vol. 41 ›› Issue (12): 294-303.doi: 10.13560/j.cnki.biotech.bull.1985.2025-0497

洋葱伯克霍尔德氏菌KRS634对棉花红腐病的生防效果

吴琼¹^,²(), 陈德勇³, 朱鹤⁴, 王丹⁵, 张晓军¹(), 陈捷胤²()

^1.牡丹江师范学院生命科学与技术学院，牡丹江 157000
^2.中国农业科学院植物保护研究所植物病虫害综合治理全国重点实验室，北京 100193
^3.中国农业科学院西部农业研究中心，昌吉 831100
^4.辽宁省经济作物研究所，辽阳 111000
^5.浙江农林大学林业与生物技术学院，杭州 311300

收稿日期:2025-05-14 出版日期:2025-12-26 发布日期:2026-01-06
通讯作者: 陈捷胤，男，博士，研究员，研究方向：植物病理学；E-mail: chenjieyin@caas.cn
张晓军，男，硕士，教授，研究方向：植物保护；E-mail: swxzxj@126.com
作者简介:吴琼，女，硕士，研究方向：植物学；E-mail: 875031242@qq.com
基金资助:
国家重点研发子课题(2022YFD1401202)

Biocontrol Effect of Burkholderia contaminans KRS634 on Cotton Red Rot Disease

WU Qiong¹^,²(), CHEN De-yong³, ZHU He⁴, WANG Dan⁵, ZHANG Xiao-jun¹(), CHEN Jie-yin²()

^1.College of Life Science and Technology, Mudanjiang Normal University, Mudanjiang 157000
^2.National Key Laboratory of Integrated Management of Plant Diseases and Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193
^3.Western Agricultural Research Center, Chinese Academy of Agricultural Sciences, Changji 831100
^4.Liaoning Institute of Cash Crops, Liaoyang 111000
^5.College of Forestry and Biotechnology, Zhejiang A&F University, Hangzhou 311300

Received:2025-05-14 Published:2025-12-26 Online:2026-01-06

摘要/Abstract

摘要：

目的获得绿色、安全、高效的对棉花红腐病具有良好防效的生防菌剂。方法以拟轮枝镰孢霉Fusarium verticillioides为靶标菌，通过平板对峙法筛选拮抗菌株，通过形态观察、16S rRNA序列的系统发育分析及促生特性对菌株进行鉴定分析；采用盆栽试验进一步测定菌株生防及促生效果；采用对峙培养法和对扣熏蒸法测试菌株对多种真菌的抑制作用。结果洋葱伯克霍尔德氏菌（Burkholderia contaminans）KRS634对F. verticillioides的抑菌率为73.80%；该菌株具有解磷、解钾、固氮及产嗜铁素的促生特性；室内防效试验表明，KRS634能够有效降低棉花苗期红腐病的发生，经KRS634处理的棉花红腐病病株率为85.71%，病情指数为36.31，显著低于对照组；KRS634发酵液还能够提高棉花出苗率，促进棉花生长；另外，KRS634对大丽轮枝菌（Verticillium dahliae）、稻瘟病菌（Magnaporthe oryzae）、胶孢炭疽菌（Colletotrichum gloeosporioides）等多种病原真菌均具有良好的拮抗效果。结论洋葱伯克霍尔德氏菌KRS634菌株特性优异，在防治棉花红腐病等真菌病害方面具有较好的应用潜力，可作为开发棉花红腐病生防菌剂的候选菌株。

关键词: 棉花红腐病, 拟轮枝镰孢霉, 生物防治, 洋葱伯克霍尔德氏菌

Abstract:

Objective To develop an environmentally-friendly, safe, and highly effective biocontrol agent against cotton red rot disease caused by Fusarium verticillioides. Method F. verticillioides was used as the target fungus, antagonistic strains were screened by the plate confrontation method to obtain a strong antagonistic activity strain. The strain was identified and analyzed through morphological observation, phylogenetic analysis of 16S rRNA sequences, and plant growth-promoting characteristics. Pot experiments were further carried out to determine the biocontrol and plant growth-promoting effects of the strain. The inhibitory effects of the strain on various fungi were tested by the confrontation culture method and the sealed fumigation method. Result The strain KRS634 exhibiting strong antagonistic activity against F. verticillioides was screened through plate confrontation assays, and was identified as Burkholderia contaminans based in 16S rRNA sequence analysis. KRS634 had the plant growth-promoting characteristics including phosphorus solubilization, potassium solubilization, nitrogen fixation, and siderophore production. The indoor experiments showed that KRS634 effectively reduced the occurrence of cotton red rot disease at the seedling stage. The disease plant rate of cotton red rot disease treated with KRS634 was 85.71%, and the disease index was 36.31, which was significantly lower than that of the control group. The fermentation broth of KRS634 also increased the emergence rate of cotton and promoted the growth of cotton. In addition, KRS634 had a broad-spectrum antagonistic effect on various pathogenic fungi such as Verticillium dahliae, Magnaporthe oryzae, and Colletotrichum gloeosporioides. Conclusion The strain B. contaminans KRS634 has a good application potential in the prevention and control of fungal diseases such as cotton red rot disease. It can be used as a candidate strain for the development of biocontrol agents against cotton red rot disease.

Key words: cotton red rot disease, Fusarium verticillioides, biological control, Burkholderia contaminans

吴琼, 陈德勇, 朱鹤, 王丹, 张晓军, 陈捷胤. 洋葱伯克霍尔德氏菌KRS634对棉花红腐病的生防效果[J]. 生物技术通报, 2025, 41(12): 294-303.

WU Qiong, CHEN De-yong, ZHU He, WANG Dan, ZHANG Xiao-jun, CHEN Jie-yin. Biocontrol Effect of Burkholderia contaminans KRS634 on Cotton Red Rot Disease[J]. Biotechnology Bulletin, 2025, 41(12): 294-303.

图/表 10

表1 棉花红腐病的病情分级标准

Table 1 Grading standards for cotton red rot disease

级别 Grade	分级标准 Grading standard	级值 Value
0级	无病状，健康植株	0
1级	下胚轴有少数小的病斑（褐色凹陷斑或红肿斑纹），病斑横向直径<下胚轴直径的1/4	1
2级	下胚轴病斑稍大，下胚轴直径的1/4≤病斑直径<下胚轴直径的1/2	2
3级	下胚轴病斑较大，下胚轴直径的1/2≤病斑直径<下胚轴直径的3/4，或下胚轴缢缩	3
4级	病斑直径>下胚轴直径的3/4，或整株萎蔫或死亡	4

图1 不同拮抗菌株对拟轮枝镰孢霉的抑制效果

Fig. 1 Inhibitory effect of different antagonistic strains against F. verticillioides

表2 不同菌株对拟轮枝镰孢霉的抑制效果

Table 2 Inhibitory effect of different antagonistic strains against F. verticillioides

编号 Number	菌落直径平均值 Average colony diameter （mm）	抑菌率 Inhibition rate （%）
CK	68.03±0.52	-
KRS504	（35.90±0.97） a	（47.23±0.01） g
KRS507	（32.38±0.47） b	（52.40±0.00） f
KRS515	（31.91±0.20） b	（53.09±0.01） f
KRS516	（21.37±0.56） cde	（68.59±0.01） cde
KRS523	（20.02±0.37） ef	（70.57±0.00） bc
KRS524	（22.44±1.06） c	（67.01±0.02） e
KRS620	（17.99±0.14） g	（73.56±0.00） a
KRS625	（19.36±0.75） f	（71.54±0.01） b
KRS631	（21.71±0.79） cd	（68.09±0.01） de
KRS634	（17.83±0.21） g	（73.80±0.00） a
KRS638	（20.11±1.41） ef	（70.44±0.02） bc
KRS642	（32.86±0.95） b	（51.70±0.01） f
KRS666	（20.38±0.62） def	（70.04±0.01） bcd
KRS671	（19.98±0.93） ef	（70.63±0.02） bc
KRS684	（22.73±0.78） c	（66.59±0.01） e

图2 菌株KRS634在 LB培养基上的菌落形态

Fig. 2 Colony morphology of strain KRS634 on LB plate

图3 基于16S rRNA序列构建的菌株KRS634和相关菌株系统发育树

Fig. 3 Phylogenetic tree of strain KRS634 and related strains constructed based on 16S rRNA sequences

图4 促生特性测定

Fig. 4 Determination of plant growth-promoting characteristics

图5 拮抗细菌KRS634对棉花红腐病的防治效果A：棉花表型； a-d：棉花正视图；e-h：棉花俯视图；i-l：棉花根部表型；B：棉花病株率；C：棉花病情指数；*表示对照组和处理组在P<0.05水平上差异显著（t检验）， ***表示对照组和处理组在P<0.001水平上差异显著（t检验），下同

Fig. 5 Biocontrol effect of antagonistic bacteria KRS634 on cotton red rot diseaseA: Cotton phenotype. a-d: Front view of cotton; e-h: top view of cotton; i-l: root phenotype of cotton. B: Rate of diseased cotton plants. C: Disease index of cotton; * indicates a significant difference between the control group and the treatment group at the P<0.05 level (t-test), and *** indicates a significant difference between the control group and the treatment group at the P<0.001 level (t-test). The same below

表3 KRS634对棉花生长情况的影响

Table 3 Impact of KRS634 on the growth situation of cotton plants

处理

Treatment

株高

Plant height （mm）

茎粗

Stem diameter （cm）

干重

Dry weight （g）

鲜重

Fresh weight （g）

氮含量

Nitrogen content

菌液浇灌 Irrigationn with bacterial liquid

图6 KRS634对7种病原真菌生长抑制的平板对峙结果A：菌落生长表型； B：菌落直径； C：菌落抑制率； ****表示对照组和处理组在P<0.000 1水平上差异显著（t检验），下同

Fig. 6 Results of KRS634 against seven pathogenic fungi using the plate confrontation methodA: Phenotype of colony growth. B: Colony diameter. C: Bacterial colony inhibition rate. **** indicates the significant differences between control group and treatment group at P<0.000 1 level (t test), the same below

图7 KRS634挥发性有机化合物对7种病原真菌的广谱抑制作用

Fig. 7 Broad-spectrum inhibitory effect of volatile organic compounds released from KRS634 against seven pathogenic fungi

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洋葱伯克霍尔德氏菌KRS634对棉花红腐病的生防效果

Biocontrol Effect of Burkholderia contaminans KRS634 on Cotton Red Rot Disease

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