摘要：为了防治烟草疫霉对烟草引起的病害，从不同烟田健康烟株根际土壤中筛选分离42株根际细菌，经平板对峙初筛、细菌发酵液115 ℃处理后复筛得到10株对烟草黑胫病病原菌（Phytophthora parasitica）具有较好抑制效果的芽孢杆菌。经分子生物学鉴定可将其分为解淀粉芽孢杆菌（Bacillus amyloliquefaciens）、枯草芽孢杆菌（B. subtilis）和贝莱斯芽孢杆菌（B. velezensis）3类。选取枯草芽孢杆菌GJ1、解淀粉芽孢杆菌GJ7和貝莱斯芽孢杆菌GJ11对各类芽孢杆菌进行纤维素酶和蛋白酶分泌特性检测、烟草根际土壤、根和茎定殖能力检测以及田间试验，得出这3种芽孢杆菌均可分泌纤维素酶和蛋白酶，均能在烟草根际土壤、根和茎中定殖，在根际土壤中定殖能力为GJ1>GJ7>GJ11，在根和茎中定殖能力均为GJ7>GJ1>GJ11。GJ1、GJ7、GJ11对烟草黑胫病田间相对防治效果分别为33.33%、80.00%、28.60%，初步确定拮抗芽孢杆菌田间防治效果取决于其在烟草内定殖能力。
关键词：烟草黑胫病病原菌（Phytophthora parasitica）；拮抗细菌；定殖能力；防治效果
中图分类号：S476；S435.72 文献标识码：A
文章编号：0439-8114（2018）18-0060-06
DOI：10.14088/j.cnki.issn0439-8114.2018.18.014 开放科学（资源服务）标识码（OSID）：
Selecting of Antagonistic Bacillus against Tobacco Black Shank and Detecting of Control Efficacy
SHI He-li，TAN Jun，DING Cai-fu，ZHAO Xiu-yun，XIANG Bi-kun，LUO Fang
（Enshi Tobacco Company of Hubei Province，Enshi 445000，Hubei，China）
Abstract： In order to control the tobacco black shank caused by Phytophthora parasitica var. nicotianae，42 bacterial strains were isolated from different health status tobacco rhizosphere soil，10 of them had an antagonistic effect on P. nicotianae by using the plate confronting method and treatment with fermentation filtrate after heated at 115 ℃. Three groups of Bacillus could be identified under the 16S rDNA sequence analysis：Bacillus amyloliquefaciens， B. subtilis and B. velezensis. The characteristics of cellulase and protease activity，colonization of Bacillus in tobacco rhizosphere soil，root and stem，and field experiments were carried out on B. subtilis GJ1，B. amyloliquefaciens GJ7 and B. velezensis GJ11. The results showed that GJ1，GJ7 and GJ11 strains could secrete cellulase and protease，and colonize in the rhizosphere soil，root and stem of tobacco. The ability of colonization in the rhizosphere soil was GJ1>GJ7>GJ11，in root and stem was GJ7>GJ1>GJ11. However，the control efficiency against tobacco black shank of GJ1，GJ7 and GJ11 were 33.33%，80.00% and 28.60%，respectively，suggesting that the control efficacy in field of antagonistic Bacillus may depend on its colonization ability in tobacco.
Key words： Phytophthora parasitica； antagonistic bacteria； colonization； control efficacy
烟草黑胫病是一种严重影响烟草生长的土传真菌病害，由烟草黑胫病病原菌疫霉菌（Phytophthora parasitica var. nicotianae）引起。主要发病期在烟草的成株期，病原侵染茎基部，以根茎部发病为主，向髓部扩展，病株叶片自下而上依次变黄，直至整株烟草枯死，给烟草种植带来巨大的经济损失[1]。近年来有关防治措施主要有化学防治、轮作、抗病品种选育等，但这些措施都存在缺陷。长期使用化学药剂可导致病原菌耐药性增强、烟叶农药残留、污染环境[2]，使用抗病品种会使烟叶质量下降，给卷烟的安全性和烟叶出口带来严重的不利影响[3]。生物防治具有无污染、无毒害、无残留、可持续、环境友好等特点，其已受到烟草病害防治的重视。