Validov S, Kamilova F, Qi S, Stephan D, Wang J J, Makarova N, Lugtenberg B
Clusius Laboratory, Institute of Biology, Leiden University, Leiden, Netherlands.
J Appl Microbiol. 2007 Feb;102(2):461-71. doi: 10.1111/j.1365-2672.2006.03083.x.
Tomato foot and root rot (TFRR), caused by Fusariumoxysporum f. sp. radicis-lycopersici (Forl), is an economically important disease of tomato. The aim of this study was to develop an efficient protocol for the isolation of bacteria, which controls TFRR based on selection of enhanced competitive root-colonizing bacteria from total rhizosphere soil samples.
A total of 216 potentially enhanced bacterial strains were isolated from 17 rhizosphere soil samples after applying a procedure to enrich for enhanced root tip colonizers. Amplified ribosomal DNA restriction analysis, in combination with determination of phenotypic traits, was introduced to evaluate the presence of siblings. One hundred sixteen strains were discarded as siblings. Thirty-eight strains were discarded as potential pathogens based on the sequence of their 16S rDNA. Of the remaining strains, 24 performed equally well or better than the good root colonizer Pseudomonas fluorescens WCS365 in a competitive tomato root tip colonization assay. Finally, these enhanced colonizers were tested for their ability to control TFRR in stonewool, which resulted in seven new biocontrol strains.
The new biocontrol strains, six Gram-negative and one Gram-positive bacteria, were identified as three Pseudomonas putida strains and one strain each of Delftia tsuruhatensis, Pseudomonas chlororaphis, Pseudomonas rhodesiae and Paenibacillus amylolyticus.
We describe a fast method for the isolation of bacteria able to suppress TFRR in stonewool, an industrial plant growth substrate. The procedure minimizes the laborious screens that are a common feature in the isolation of biocontrol strains.
由尖孢镰刀菌番茄根腐专化型(Fusarium oxysporum f. sp. radicis-lycopersici,Forl)引起的番茄根腐病是番茄一种具有重要经济影响的病害。本研究的目的是开发一种高效的细菌分离方案,该方案基于从根际土壤总样本中筛选具有更强竞争定殖能力的细菌来控制番茄根腐病。
在采用一种富集根尖定殖菌的程序后,从17个根际土壤样本中总共分离出216株潜在的优势细菌菌株。引入核糖体DNA扩增片段限制性分析,并结合表型特征测定来评估是否存在亲缘菌株。116株菌株因是亲缘菌株而被舍弃。基于16S rDNA序列,38株菌株被作为潜在病原体舍弃。在剩余的菌株中,24株在竞争性番茄根尖定殖试验中表现与优良的根定殖菌荧光假单胞菌WCS365相当或更优。最后,对这些增强定殖菌在岩棉中控制番茄根腐病的能力进行了测试,得到了7株新的生防菌株。
新的生防菌株包括6株革兰氏阴性菌和1株革兰氏阳性菌,分别鉴定为3株恶臭假单胞菌、1株鹤见德尔夫特菌、1株绿针假单胞菌、1株罗兹假单胞菌和1株解淀粉芽孢杆菌。
我们描述了一种快速分离能够抑制岩棉(一种工业植物生长基质)中番茄根腐病的细菌的方法。该程序最大限度地减少了生防菌株分离过程中常见的繁琐筛选工作。