栽培墨兰根腐病病原鉴定与生防菌筛选

doi:10.11924/j.issn.1000-6850.casb2021-0239

中国农学通报 ›› 2021, Vol. 37 ›› Issue (19): 128-133.doi: 10.11924/j.issn.1000-6850.casb2021-0239

所属专题：植物保护

栽培墨兰根腐病病原鉴定与生防菌筛选

唐婕¹(), 陈健鑫¹, 尼玛此姆¹, 魏玉倩¹, 落追¹, 韩雨庭¹, 吕则佳¹, 马焕成², 伍建榕¹^,²()

¹西南林业大学/云南省高校森林灾害预警控制重点实验室,昆明 650224
²西南林业大学西南地区生物多样性保育国家林业局重点实验室,昆明 650224

收稿日期:2021-03-10 修回日期:2021-05-14 出版日期:2021-07-05 发布日期:2021-07-29
通讯作者: 伍建榕
作者简介:唐婕,女,1994年出生,硕士,研究方向：森林病理学。E-mail： 519779006@qq.com。
基金资助:
国家自然科学基金项目“墨兰菌根真菌多样性与根腐病发生的抑病机理研究”(31860208);国家自然科学基金项目“地生兰-菌根真菌-松三联共生关系研究”(31360198);西南林业大学木棉纤维人工林产业化培育省创新团队培育项目(2018CH014);云南省“云岭教学名师”培育项目

Pathogen Identification and Biocontrol Screening of Root Rot of Cymbidium sinense

Tang Jie¹(), Chen Jianxin¹, Nima Cimu¹, Wei Yuqian¹, Luo Zhui¹, Han Yuting¹, Lv Zejia¹, Ma Huancheng², Wu Jianrong¹^,²()

¹Key Laboratory of Forest Disaster Warning and Control in Yunnan Higher Education Institutions/ Southwest Forestry University, Kunming 650224
²Key Laboratory of Biodiversity Conservation in Southwest China of State Forest Administration, Southwest Forestry University, Kunming 650224

Received:2021-03-10 Revised:2021-05-14 Online:2021-07-05 Published:2021-07-29
Contact: Wu Jianrong

摘要/Abstract

摘要：

为明确墨兰根腐病的病原,提出绿色生防措施,以发生根腐病的墨兰为材料,采用组织分离法分离病原菌,并对其进行形态学鉴定及分子生物学鉴定,通过柯赫氏法则验证致病性。同时,采集野外健康墨兰根部和土壤,采用稀释涂布法分离获得20株细菌,通过平板对峙培养,筛选出1株细菌MJ5-1能同时拮抗2株病原真菌。通过16S rRNA基因序列分析,该拮抗菌为短小芽孢杆菌(Bacillus pumilus)。结果表明,导致云南墨江栽培墨兰根腐病的病原是腐皮镰刀菌(Fusarium solani),短小芽孢杆菌(Bacillus pumilus)MJ5-1能抑制腐皮镰刀菌菌丝及孢子的生长。研究结果可为产业化栽培墨兰的根腐病诊断及无公害防治提供理论依据。

关键词: 墨兰根腐病, 组织分离法, 病原菌, 鉴定, 拮抗菌, 筛选

Abstract:

The paper aims to identify the pathogen of root rot and put forward green biological control measures. The pathogen of root rot of Cymbidium sinense was isolated by tissue isolation method, and identified by morphology and molecular biology. The pathogenicity was verified by Koch’s law. At the same time, 20 strains of bacteria were isolated from the roots and soil of wild healthy C. sinense by dilution coating method. Through plate confrontation culture, one strain of bacteria MJ5-1 was screened, which could antagonize two pathogens. The 16S rRNA gene sequence analysis showed that the antagonistic strain was Bacillus pumilus. The results showed that Fusarium solani was the pathogen of root rot of cultivated C. sinense in Mojiang. B. pumilus MJ5-1 could inhibit the growth of mycelium and spore of F. solani. This study could provide a theoretical basis for the diagnosis and pollution-free control of root rot of cultivated C. sinense.

Key words: root rot of Cymbidium sinense, tissue isolation method, pathogen, identification, antagonistic bacteria, screening

中图分类号:

S432.1

唐婕, 陈健鑫, 尼玛此姆, 魏玉倩, 落追, 韩雨庭, 吕则佳, 马焕成, 伍建榕. 栽培墨兰根腐病病原鉴定与生防菌筛选[J]. 中国农学通报, 2021, 37(19): 128-133.

Tang Jie, Chen Jianxin, Nima Cimu, Wei Yuqian, Luo Zhui, Han Yuting, Lv Zejia, Ma Huancheng, Wu Jianrong. Pathogen Identification and Biocontrol Screening of Root Rot of Cymbidium sinense[J]. Chinese Agricultural Science Bulletin, 2021, 37(19): 128-133.

图/表 8

图1. 墨兰人工种植多年根腐病及地上部分枯萎症状

图2. 回接病原腐皮镰刀菌(Fusarium solani)ML6-1的发病症状

图3. 喷洒ML6-1菌丝悬浮液后的墨兰发病初期

图4. 墨兰典型根腐病症状分离菌株ML6-1的形态学特征

图5. 采用PAUP*4.0b10MP法建立的墨兰根腐病病原菌ITS序列的分子系统进化树

图6. 菌株MJ1-1、MJ7-2、MJ5-1的16S rRNA电泳图谱 M为100~2000 bp DNA Marker,CK为ddH2O对照

供试菌株F. Solani	抑菌直径/mm	无菌滤液抑菌率/%
MJ1-1	30.6±1.2b	96.68±0.214bc
MJ5-1	43.6±1.2a	100.68±1.13ab
MJ7-2	29.6±1.2b	97.68±1.2bc

表1. 短小芽孢杆菌对墨兰根腐病病原真菌的抑菌直径和抑菌率

图7. 拮抗菌短小芽孢杆菌MJ5-1与菌株ML6-1平板对峙结果左为菌株ML6-1对照,右为短小芽孢杆菌MJ5-1与菌株ML6-1对峙

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栽培墨兰根腐病病原鉴定与生防菌筛选

Pathogen Identification and Biocontrol Screening of Root Rot of Cymbidium sinense

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