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作物杂志,2024, 第2期: 242–248 doi: 10.16035/j.issn.1001-7283.2024.02.030

• 植物保护 • 上一篇    下一篇

马铃薯青枯病拮抗菌的筛选及温室防治效果研究

李盛(), 李翔, 朱美如, 王夏, 李昊阳, 谭欣如, 王海燕()   

  1. 四川大学生命科学学院/生物资源与生态环境教育部重点实验室，610065，四川成都
  • 收稿日期:2023-03-06 修回日期:2023-03-23 出版日期:2024-04-15 发布日期:2024-04-15
  • 通讯作者: 王海燕，研究方向为微生物遗传学，E-mail：hayawang@scu.edu.cn
  • 作者简介:李盛，研究方向为植物细菌病害，E-mail：1798861540@qq.com
  • 基金资助:
    国家自然科学基金(32060720)

Screening of Antagonistic Bacteria against Potato Bacterial Wilt and Study on Its Control Effect in Greenhouse

Li Sheng(), Li Xiang, Zhu Meiru, Wang Xia, Li Haoyang, Tan Xinru, Wang Haiyan()   

  1. College of Life Sciences, Sichuan University/Key Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, Chengdu 610065, Sichuan, China
  • Received:2023-03-06 Revised:2023-03-23 Online:2024-04-15 Published:2024-04-15

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摘要：

马铃薯青枯病是由青枯雷尔氏菌（Ralstonia solanacearum E.F. Smith）引起的毁灭性土传病害，筛选对马铃薯青枯病有良好防效的生防菌株具有重要意义。本研究从健康马铃薯、番茄、茄子和辣椒植株根际土壤中筛选鉴定出对马铃薯、番茄和烟草青枯病菌均具有较好抑制活性的6株拮抗菌，经分子生物学鉴定为赖氨酸芽胞杆菌（Lysinibacillus sp.）、假单胞菌（Pseudomonas sp.）和科萨克氏菌（Kosakonia sp.），分别具有产蛋白酶、铁载体和生物膜抑菌代谢物的能力。拮抗菌假单胞菌FC-17对“陇薯7号”和“大西洋”马铃薯青枯病的温室防治效果分别达到79.46%和70.83%，具有应用前景。

关键词: 马铃薯青枯病, 拮抗细菌, 抑菌代谢物, 防治效果

Abstract:

Bacterial wilt of potato is a destructive soil-borne disease caused by Ralstonia solanacearum. Screening antagonistic bacteria against pathogen is a vital work for the biocontrol of potato bacterial wilt. Six isolates with good inhibitory effect on three R.solanacearum strains of potato, tomato and tobacco were isolated from the rhizosphere soil of healthy potato, tomato, eggplant and pepper plants. They were identified as Lysinibacillus sp., Pseudomonas sp. and Kosakonia sp. through molecular detection, and it had the ability to produce protease, siderophor and biofilm, respectively. In greenhouse, strain FC-17 significantly reduced disease incidence by 79.46% and 70.83% on potato cultivars “Longshu 7” and “Atlantic”, respectively, indicating good potential for biocontrol of potato bacterial wilt.

Key words: Potato bacterial wilt, Antagonistic bacteria, Antimicrobial metabolites, Control effect

表1

盆栽防治效果试验设计

处理Treatment 添加物（盆）Supplement (pot)
R 20 mL无菌水+10 mL青枯菌菌液
FR 20 mL拮抗菌菌液+10 mL青枯菌菌液
F 20 mL拮抗菌菌液
CK 30 mL无菌水

图1

拮抗菌对3株青枯菌的抑制作用 (a)~(c) 培养基里依次包埋马铃薯、番茄和烟草青枯菌，样品孔里为不同拮抗菌的菌液50 μL（OD600nm=5.0）。

表2

6株拮抗菌对3种青枯菌的抑菌圈直径

菌株名称
Strain name		 青枯菌来源Source of R. solanacearum
马铃薯Potato 番茄Tomato 烟草Tobacco
XC-1 4.40±0.74c 3.91±0.30d 4.38±0.74d
XC-2 7.38±0.98b 9.41±0.45b 9.03±0.59b
FC-11 6.61±0.34b 5.54±0.82c 6.64±0.34c
FC-17 6.90±0.79b 8.26±0.62b 8.41±0.34b
HN-9 7.11±0.91b 8.83±0.80b 8.96±0.81b
NX-18 10.14±0.74a 11.98±0.80a 10.70±0.53a

图2

拮抗菌16S rRNA基因及基因间隔区PCR扩增产物电泳图 (a)为16S rRNA基因扩增产物电泳图，M为λ-Eco I T-14 digest；(b)为16S至23S基因间隔区扩增产物电泳图，M为1kb plus DNA Ladder。1~6分别为菌株XC-1、XC-2、FC-11、FC-17、HN-9和NX-18。

图3

拮抗菌菌落形态 菌株划线后在30 ℃静置培养48 h。

图4

拮抗菌抑菌物质的测定 (a) 蛋白酶；(b) 铁载体；(c) 生物膜；“+”：短小芽孢杆菌SCU11，“-”：大肠杆菌MG1655，CK：空白对照。

图5

拮抗菌FC-17对马铃薯青枯病的防治作用 (a)和(b)分别为“陇薯7号”和“大西洋”马铃薯，均拍摄于播种后45 d，最右侧为患病叶特写。

表3

盆栽试验FC-17对马铃薯青枯病的防治效果统计

马铃薯品种
Potato cultivar		 处理
Treatment		 病情指数
Disease index		 防治效果
Control effect (%)
陇薯7号
Longshu 7		 R 45.37±18.49 －
FR 9.52±16.49* 79.46
F － －
CK － －
大西洋
Atlantic		 R 52.68±16.04 －
FR 15.18±15.64* 70.83
F － －
CK － －
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