Dekkers L C, Mulders I H, Phoelich C C, Chin-A-Woeng T F, Wijfjes A H, Lugtenberg B J
Leiden University, Institute of Molecular Plant Sciences, Clusius Laboratory, The Netherlands.
Mol Plant Microbe Interact. 2000 Nov;13(11):1177-83. doi: 10.1094/MPMI.2000.13.11.1177.
We show that the disease tomato foot and root rot caused by the pathogenic fungus Fusarium oxysporum f. sp. radicis-lycopersici can be controlled by inoculation of seeds with cells of the efficient root colonizer Pseudomonas fluorescens WCS365, indicating that strain WCS365 is a biocontrol strain. The mechanism for disease suppression most likely is induced systemic resistance. P. fluorescens strain WCS365 and P. chlororaphis strain PCL1391, which acts through the production of the antibiotic phenazine-1-carboxamide, were differentially labeled using genes encoding autofluorescent proteins. Inoculation of seeds with a 1:1 mixture of these strains showed that, at the upper part of the root, the two cell types were present as microcolonies of either one or both cell types. Microcolonies at the lower root part were predominantly of one cell type. Mixed inoculation tended to improve biocontrol in comparison with single inoculations. In contrast to what was observed previously for strain PCL1391, mutations in various colonization genes, including sss, did not consistently decrease the biocontrol ability of strain WCS365. Multiple copies of the sss colonization gene in WCS365 improved neither colonization nor biocontrol by this strain. However, introduction of the sss-containing DNA fragment into the poor colonizer P. fluorescens WCS307 and into the good colonizer P. fluorescens F113 increased the competitive tomato root tip colonization ability of the latter strains 16- to 40-fold and 8- to 16-fold, respectively. These results show that improvement of the colonization ability of wild-type Pseudomonas strains by genetic engineering is a realistic goal.
我们发现,由致病真菌尖孢镰刀菌番茄根腐专化型引起的番茄根腐病,可通过用高效根系定殖菌荧光假单胞菌WCS365的细胞接种种子来控制,这表明菌株WCS365是一种生防菌株。病害抑制机制很可能是诱导系统抗性。荧光假单胞菌菌株WCS365和通过产生抗生素吩嗪-1-甲酰胺发挥作用的绿针假单胞菌菌株PCL1391,利用编码自发荧光蛋白的基因进行了差异标记。用这两种菌株的1:1混合物接种种子表明,在根的上部,两种细胞类型以一种或两种细胞类型的微菌落形式存在。根下部的微菌落主要是一种细胞类型。与单接种相比,混合接种往往能改善生防效果。与之前对菌株PCL1391的观察结果相反,包括sss在内的各种定殖基因的突变并没有持续降低菌株WCS365的生防能力。WCS365中sss定殖基因的多个拷贝既没有改善该菌株的定殖能力,也没有提高其生防效果。然而,将含有sss的DNA片段导入定殖能力差的荧光假单胞菌WCS307和定殖能力强的荧光假单胞菌F113中,分别使后两种菌株在番茄根尖的竞争定殖能力提高了16至40倍和8至16倍。这些结果表明通过基因工程提高野生型假单胞菌菌株的定殖能力是一个现实的目标。