Thielaviopsis

About: Thielaviopsis is a research topic. Over the lifetime, 79 publications have been published within this topic receiving 3149 citations.

Cyanide production by Pseudomonas fluorescens helps suppress black root rot of tobacco under gnotobiotic conditions

Abstract: Pseudomonas fluorescens CHA0 suppresses black root rot of tobacco, a disease caused by the fungus Thielaviopsis basicola. Strain CHA0 excretes several metabolites with antifungal properties. The importance of one such metabolite, hydrogen cyanide, was tested in a gnotobiotic system containing an artificial, iron-rich soil. A cyanidenegative (hcn) mutant, CHA5, constructed by a gene replacement technique, protected the tobacco plant less effectively than did the wild-type CHA0. Complementation of strain CHA5 by the cloned wild-type hcn+ genes restored the strain's ability to suppress disease. An artificial transposon carrying the hcn+ genes of strain CHA0 (Tnhcn) was constructed and inserted into the genome of another P.fluorescens strain, P3, which naturally does not produce cyanide and gives poor plant protection. The P3::Tnhcn derivative synthesized cyanide and exhibited an improved ability to suppress disease. All bacterial strains colonized the roots similarly and did not influence significantly the survival of T.basicola in soil. We conclude that bacterial cyanide is an important but not the only factor involved in suppression of black root rot.

Suppression of Root Diseases by Pseudomonas fluorescens CHA0: Importance of the Bacterial Secondary Metabolite 2,4-Diacetylphloroglucinol

Abstract: Pseudomonas fluorescens strain CHAO suppresses Thielaviopsis basicola-induced black root rot of tobacco and Gaeumannomyces graminis var. tritici-induced take-all of wheat. Strain CHAO produces 2,4-diacetylphloroglucinol, a metabolite with antifungal, antibacterial, and phytotoxic activity. The role of this compound in disease suppression was tested under gnotobiotic conditions. A P. fluorescens mutant, obtained by Tn5 insertion, did not produce 2,4-diacetylphloroglucinol, showed diminished inhibition of T. basicola and of G. g. var. tritici in vitro, and had a reduced suppressive effect on tobacco black root rot and on take-all of wheat, compared with wild-type CHAO. Complementation of the mutant with an 11-kb DNA fragment from a genomic library of wild-type CHAO largely restored production of the metabolite, inhibition of the fungal pathogens in vitro and disease suppression. The Tn5 insertion was physically mapped using a 5.8-kb complementing fragment as a probe. 2,4- Diacetylphloroglucinol was shown to be produced in the rhizosphere of wheat by strain CHAO and by the complemented mutant, but not by the mutant defective in 2,4-diacetylphloroglucinol synthesis. These results support the importance of 2,4-diacetylphloroglucinol production by strain CHAO in the suppression of soilborne plant pathogens in the rhizosphere.

Global control in Pseudomonas fluorescens mediating antibiotic synthesis and suppression of black root rot of tobacco.

Abstract: Pseudomonas fluorescens CHA0 colonizes plant roots, produces several secondary metabolites in stationary growth phase, and suppresses a number of plant diseases, including Thielaviopsis basicola-induced black root rot of tobacco. We discovered that mutations in a P. fluorescens gene named gacA (for global antibiotic and cyanide control) pleiotropically block the production of the secondary metabolites 2,4-diacetylphloroglucinol (Phl), HCN, and pyoluteorin. The gacA mutants of strain CHA0 have a drastically reduced ability to suppress black root rot under gnotobiotic conditions, supporting the previous observations that the antibiotic Phl and HCN individually contribute to the suppression of black root rot. The gacA gene is directly followed by a uvrC gene. Double gacA-uvrC mutations render P. fluorescens sensitive to UV irradiation. The gacA-uvrC cluster is homologous to the orf-2 (= uvrY)-uvrC operon of Escherichia coli. The gacA gene specifies a trans-active 24-kDa protein. Sequence data indicate that the GacA protein is a response regulator in the FixJ/DegU family of two-component regulatory systems. Expression of the gacA gene itself was increased in stationary phase. We propose that GacA, perhaps activated by conditions of restricted growth, functions as a global regulator of secondary metabolism in P. fluorescens.

Comparison of rhizobacterial community composition in soil suppressive or conducive to tobacco black root rot disease.

Abstract: Work on soils suppressive to Thielaviopsis basicola-mediated tobacco black root rot has focused on antagonistic pseudomonads to date. The role of non-Pseudomonas rhizosphere populations has been neglected, and whether they differ in black root rot-suppressive versus -conducive soils is unknown. To assess this possibility, tobacco was grown in a suppressive and a conducive soil of similar physicochemical properties, and rhizobacterial community composition was compared using a 16S rRNA taxonomic microarray. The microarray contains 1033 probes and targets 19 bacterial phyla. Among them, 398 probes were designed for Proteobacteria, Firmicutes, Actinomycetes, Cyanobacteria and Bacteroidetes genera/species known to include strains relevant for plant protection or plant growth promotion. Hierarchical clustering as well as principal component analysis of microarray data discriminated clearly between black root rot-suppressive and -conducive soils. In contrast, T. basicola inoculation had no impact on rhizobacterial community composition. In addition to fluorescent Pseudomonas, the taxa Azospirillum, Gluconacetobacter, Burkholderia, Comamonas and Sphingomonadaceae, which are known to comprise strains with plant-beneficial properties, were more prevalent in the suppressive soil. Mycobacterium, Bradyrhizobium, Rhodobacteraceae, Rhodospirillum and others were more prevalent in the conducive soil. For selected taxa, microarray results were largely corroborated by quantitative PCR and cloning/sequencing. In conclusion, this work identified novel bacterial taxa that could serve as indicators of disease suppressiveness in soil-quality assessments, and it extends the range of bacterial taxa hypothesized to participate in black root rot suppression.

In vitro inhibition of soil microorganisms by 2-phenylethyl isothiocyanate

Abstract: Isothiocyanates (ITCs) are produced by the enzymatic hydrolysis of glucosinolates in cruciferous plants and can kill fungi, oomycetes and bacteria. The effect of 2-phenylethyl ITC (2-PE ITC), the main ITC liberated from the roots of canola, was tested in vitro on a range of fungi, oomycetes and bacteria. Bacteria were generally more tolerant than the eukaryotic pathogens to 2-PE ITC, although both groups showed considerable variability in response (ED90 ranging from 0·005 to 1·5 mm for eukaryotes, and from 0·33 to greater than 3·34 mm for complete inhibition of bacterial growth). While intraspecies variability was low, interspecies variability was high within some genera. Amongst the eukaryotes, Trichoderma spp. were the most tolerant to 2-PE ITC, while other genera, including Aphanomyces, Gaeumannomyces, Phytophthora and Thielaviopsis, were very sensitive. A range of responses was exhibited by Pythium spp. and the different anastomosis groups of Rhizoctonia solani. A simple laboratory screening may be an effective way to identify candidate soilborne pathogens for control in rotation cropping systems which include a Brassica crop phase.