绛红褐链霉菌CC2-6抑制大白菜根肿病的效果及机制分析

doi:10.16420/j.issn.0513-353x.2023-0146

园艺学报 ›› 2024, Vol. 51 ›› Issue (5): 1137-1150.doi: 10.16420/j.issn.0513-353x.2023-0146

绛红褐链霉菌CC2-6抑制大白菜根肿病的效果及机制分析

王青¹^,³^,^*, 胡燕²^,^*, 陈姗¹, 陆景伟², 郑阳², 陶伟林², 孙现超¹, 周娜²^,^**(), 陈国康¹^,^**()

¹ 西南大学植物保护学院，重庆 400716
² 重庆市农业科学院蔬菜花卉研究所，重庆 401329
³ 浙江省嘉善县林业技术推广站，浙江嘉善 314199

收稿日期:2023-10-08 修回日期:2024-02-10 出版日期:2024-05-25 发布日期:2024-05-29
通讯作者: ** E-mail：zhouna0214@qq.com，chenguokang@swu.edu.cn
作者简介:
^* 共同第一作者
基金资助:
国家现代农业产业技术体系建设专项(CARS-25-G-40); 重庆市技术创新与应用发展专项(CSTB2022TIAD-LDX0020)

Biocontrol Efficacy and Mechanisms of Streptomyces purpeofuscus CC2-6 Against Chinese Cabbage Clubroot Caused by Plasmodiophora brassicae

WANG Qing¹^,³, HU Yan², CHEN Shan¹, LU Jingwei², ZHENG Yang², TAO Weilin², SUN Xianchao¹, ZHOU Na²^,^**(), CHEN Guokang¹^,^**()

¹ College of Plant Protection，Southwest University，Chongqing 400716，China
² Institute of Vegetable and Flower Research，Chongqing Academy of Agricultural Sciences，Chongqing 400055，China
³ Forestry Technology Extension Station of Jiashan County，Jiashan，Zhejiang 314199，China

Received:2023-10-08 Revised:2024-02-10 Published:2024-05-25 Online:2024-05-29
Contact: ** E-mail：zhouna0214@qq.com，chenguokang@swu.edu.cn

摘要/Abstract

摘要：

为筛选可抑制芸薹根肿菌（Plasmodiophora brassicae）的生防菌株，以棉花枯萎病尖孢镰刀菌（Fusarium oxysporum）为指示菌，通过平板对峙法筛选出优良菌株CC2-6，其抑制率为50.03%；该菌株发酵液能抑制根肿菌休眠孢子萌发及根肿菌根毛侵染，提前7 d接菌CC2-6对大白菜根肿病的盆栽防效为53.20%。同时，该菌株CC2-6对包括烟草叶斑病菌（Pestalotiopsis nicotiae）等9种病原菌均有抑制效果，其抑菌率在47.20% ~ 76.19%，表明菌株CC2-6具有广谱抑菌活性。形态学观察及16S rDNA基因序列分析，初步鉴定菌株CC2-6为绛红褐链霉菌（Streptomyces purpeofuscus）。菌株CC2-6可产生蛋白酶、纤维素酶、几丁质酶，其发酵液可诱导大白菜中的PPO、POD、PAL活性增强，SA、JA含量升高，降低ET含量。GC-TOF/LC-QTOF/QE非靶标代谢组学检测结果表明，菌株CC2-6发酵液中含有潮霉素B（Hygromycin B）、喜树碱（Camptothecin）、2-苯乙醇（2-Phenylethanol）、柠檬酸（Citric acid）等抑菌活性物质，表明CC2-6可能通过产生上述化合物抑制根肿菌侵染寄主。

关键词: 根肿菌, 生防放线菌, 绛红褐链霉菌, 诱导抗性, 代谢产物

Abstract:

In order to screen out biocontrol strains that can effectively inhibit Plasmodiophora brassicae，Fusarium oxysporum was used as the indicator fungus，and the strain CC2-6 was screened by plate confrontation method. The inhibitory rate was 50.03%. The fermentation broth of strain CC2-6 could inhibit the germination of resting spores of P. brassicae and the infection of P. brassicae from root hairs，and and the control efficacy of Chinese cabbage clubroot was 53.20% in pot trials. In addition，CC2-6 had inhibitory effects on nine pathogens including Pestalotiopsis nicotiae，and its inhibition rate rangeed more than 47.20%-76.19%，indicating that CC2-6 had broad-spectrum bacteriostatic activity. The strain CC2-6 was preliminarily identified as Streptomyces purpeofuscus by morphological observation and 16S rDNA gene sequence analysis. The strain CC2-6 could produce protease，cellulase and chitinase，and its fermentation broth could induce the increase of PPO，POD and PAL activities，the increase of SA and JA contents and the decrease of ET content in Chinese cabbage. The results of GC-TOF/LC-QTOF/QE non-target metabolomics showed that the fermentation broth of strain CC2-6 contained antagnostic active substances such as hygromycin B，camptothecin，2-phenylethanol and citric acid，indicating that CC2-6 might inhibit the infection of P. brassicae by metabolizing these compounds.

Key words: Plasmodiophora brassicae, biocontrol actinomycetes, Streptomyces purpeofuscus, induced resistance, metabolites

王青, 胡燕, 陈姗, 陆景伟, 郑阳, 陶伟林, 孙现超, 周娜, 陈国康. 绛红褐链霉菌CC2-6抑制大白菜根肿病的效果及机制分析[J]. 园艺学报, 2024, 51(5): 1137-1150.

WANG Qing, HU Yan, CHEN Shan, LU Jingwei, ZHENG Yang, TAO Weilin, SUN Xianchao, ZHOU Na, CHEN Guokang. Biocontrol Efficacy and Mechanisms of Streptomyces purpeofuscus CC2-6 Against Chinese Cabbage Clubroot Caused by Plasmodiophora brassicae[J]. Acta Horticulturae Sinica, 2024, 51(5): 1137-1150.

图/表 10

参考文献 34

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病原菌 Target pathogen	抑制率 Inhibition rate	病原菌 Target pathogen	抑制率 Inhibition rate
棉花枯萎病菌Fusarium oxysporum	50.30 ± 1.50	油菜菌核病菌Sclerotinia sclerotiorum	76.19 ± 1.19
甘薯软腐病菌Rhizopus stolonifer	51.67 ± 0.78	棉花立枯病菌Thanatepehorus cucumeris	52.79 ± 1.51
茄褐纹病菌Phomopsis vexans	55.58 ± 1.24	辣椒疫霉病菌Phytophthora capsici	66.67 ± 1.67
烟草灰霉病菌Botrytis cinerea	53.95 ± 0.63	黄瓜猝倒病菌Pythium delicense	72.31 ± 0.96
烟草炭疽病菌Colletotrichum micotianae	47.20 ± 5.21	烟草叶斑病菌Pestalotiopsis nicotiae	59.71 ± 0.75

病原菌 Target pathogen	抑制率 Inhibition rate	病原菌 Target pathogen	抑制率 Inhibition rate
棉花枯萎病菌Fusarium oxysporum	50.30 ± 1.50	油菜菌核病菌Sclerotinia sclerotiorum	76.19 ± 1.19
甘薯软腐病菌Rhizopus stolonifer	51.67 ± 0.78	棉花立枯病菌Thanatepehorus cucumeris	52.79 ± 1.51
茄褐纹病菌Phomopsis vexans	55.58 ± 1.24	辣椒疫霉病菌Phytophthora capsici	66.67 ± 1.67
烟草灰霉病菌Botrytis cinerea	53.95 ± 0.63	黄瓜猝倒病菌Pythium delicense	72.31 ± 0.96
烟草炭疽病菌Colletotrichum micotianae	47.20 ± 5.21	烟草叶斑病菌Pestalotiopsis nicotiae	59.71 ± 0.75

处理 Treatment	防治效果/% Indoor control effect
同时接菌 Strain CC2-6 was simultaneously inoculated with P. brassicae	27.46 ± 2.37 b
提前7 d接种菌株CC2-6 Strain CC2-6 was inoculated 7 d in advance	53.20 ± 1.82 a

处理 Treatment	防治效果/% Indoor control effect
同时接菌 Strain CC2-6 was simultaneously inoculated with P. brassicae	27.46 ± 2.37 b
提前7 d接种菌株CC2-6 Strain CC2-6 was inoculated 7 d in advance	53.20 ± 1.82 a

编号	代谢物	归一化后的代谢物丰度 Metabolic abundance after normalization
Code	Metabolite components	CC2-6-1	CC2-6-2	CC2-6-3
pos_628	诺氟沙星 Norfloxacin	0.001539502	0.000906463	0.001681919
pos_695	四环素 Tetracycline	2.28324E-05	6.55759E-06	2.04925E-05
pos_690	潮霉素B Hygromycin B	3.22625E-05	1.30073E-07	3.03196E-05
pos_664	托普霉素 Tobramycin	0.000110993	6.0012E-05	6.48009E-05
pos_683	盐霉素 Salinomycin	0.023219173	0.022620707	0.022703959
pos_474	2-苯乙醇 2-Phenylethanol	6.18249E-06	5.00147E-06	4.44714E-06
pos_445	莽草酸 Shikimic acid	3.38127E-06	3.21205E-06	4.07632E-06
pos_708	紫杉醇 Paclitaxel	2.47289E-05	3.74363E-05	3.1205E-05
pos_482	熊果苷 Arbutin	0.000933082	0.00089494	0.00096134
pos_236	L-岩藻糖 L-Fucose	5.18191E-06	3.25384E-06	3.57858E-06
pos_26	麦芽四糖Maltotetraose	0.006656552	0.006144853	0.007064496
pos_345	乙酰丁香酮 Acetosyringone	0.000384261	0.001579729	0.000450847
pos_54	姜油酮 Zingerone	2.82372E-05	2.29699E-05	3.25111E-05
pos_250	香豆酸 Coutaric acid	0.000821003	0.001010641	0.000847733
pos_469	阿斯特酚苷 Astringin	5.47227E-05	5.21228E-06	4.92987E-05

绛红褐链霉菌CC2-6抑制大白菜根肿病的效果及机制分析

Biocontrol Efficacy and Mechanisms of Streptomyces purpeofuscus CC2-6 Against Chinese Cabbage Clubroot Caused by Plasmodiophora brassicae

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编号	代谢物	归一化后的代谢物丰度 Metabolic abundance after normalization
Code	metabolite components	CC2-6-1	CC2-6-2	CC2-6-3
neg_575	土霉素 Oxytetracycline	1.87E-05	6.75E-07	0.075964
neg_51	放线酰胺素 actinonin	0.023284	0.014191	0.000149
neg_611	氟康唑Fluconazole	0.00664	0.014538	0
neg_511	印楝素A Azadirachtin A	0.00039	0.000211	0.015373
neg_569	柚皮苷 Naringin	0.004988	0.005777	1.48E-05
neg_372	硫乳霉素 Thiolactomycin	2.64E-05	1.16E-05	0.003713
neg_518	银杏内酯B Ginkgolide B	0.001923	0.001084	0.000244
neg_233	4-羟基苯甲酸 4-Hydroxybenzoic acid	0.000101	0.000116	0.001317
neg_510	喜树碱 Camptothecin	0.000331	0.001183	1.92E-05
neg_552	潮霉素B Hygromycin B	0	0	0.001455
neg_159	苯乙胺 Phenylethylamine	0.000451	0.000677	0.000183
neg_9	柠檬酸 Citric acid	0.00079	0.000511	0
neg_152	二羟丙酮 Dihydroxyacetone	0.00047	0.000458	0.000222
neg_446	噻苯咪唑 Thiabendazole	0.00034	0.000636	0.000151
neg_648	淫羊藿苷 Icariin	0.000323	0.000301	0
neg_354	对羟基肉桂酸 4-Hydroxycinnamic acid	1.61E-05	1.5E-05	0.000363
neg_320	新生霉素 Novobiocin	0	0	0.000288
neg_617	多球壳菌素 Myriocin	0	0	0.00028
neg_513	四环素 Tetracycline	0.000129	0.000133	1.25E-06
neg_667	依诺沙星 Enoxacin	7.17E-06	5.7E-06	0.000149

编号	代谢物	归一化后的代谢物丰度 Metabolic abundance after normalization
Code	metabolite components	CC2-6-1	CC2-6-2	CC2-6-3
neg_575	土霉素 Oxytetracycline	1.87E-05	6.75E-07	0.075964
neg_51	放线酰胺素 actinonin	0.023284	0.014191	0.000149
neg_611	氟康唑Fluconazole	0.00664	0.014538	0
neg_511	印楝素A Azadirachtin A	0.00039	0.000211	0.015373
neg_569	柚皮苷 Naringin	0.004988	0.005777	1.48E-05
neg_372	硫乳霉素 Thiolactomycin	2.64E-05	1.16E-05	0.003713
neg_518	银杏内酯B Ginkgolide B	0.001923	0.001084	0.000244
neg_233	4-羟基苯甲酸 4-Hydroxybenzoic acid	0.000101	0.000116	0.001317
neg_510	喜树碱 Camptothecin	0.000331	0.001183	1.92E-05
neg_552	潮霉素B Hygromycin B	0	0	0.001455
neg_159	苯乙胺 Phenylethylamine	0.000451	0.000677	0.000183
neg_9	柠檬酸 Citric acid	0.00079	0.000511	0
neg_152	二羟丙酮 Dihydroxyacetone	0.00047	0.000458	0.000222
neg_446	噻苯咪唑 Thiabendazole	0.00034	0.000636	0.000151
neg_648	淫羊藿苷 Icariin	0.000323	0.000301	0
neg_354	对羟基肉桂酸 4-Hydroxycinnamic acid	1.61E-05	1.5E-05	0.000363
neg_320	新生霉素 Novobiocin	0	0	0.000288
neg_617	多球壳菌素 Myriocin	0	0	0.00028
neg_513	四环素 Tetracycline	0.000129	0.000133	1.25E-06
neg_667	依诺沙星 Enoxacin	7.17E-06	5.7E-06	0.000149

绛红褐链霉菌CC2-6抑制大白菜根肿病的效果及机制分析

Biocontrol Efficacy and Mechanisms of Streptomyces purpeofuscus CC2-6 Against Chinese Cabbage Clubroot Caused by Plasmodiophora brassicae

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