ABSTRACT
The soft rot Enterobacteriaceae (SRE) Pectobacterium and Dickeya species (formerly classified as pectinolytic Erwinia spp.) cause important diseases on potato and other arable and horticultural crops. They may affect the growing potato plant causing blackleg and are responsible for tuber soft rot in storage thereby reducing yield and quality. Efficient and cost-effective detection and identification methods are essential to investigate the ecology and pathogenesis of the SRE as well as in seed certification programmes. The aim of this review was to collect all existing information on methods available for SRE detection. The review reports on the sampling and preparation of plant material for testing and on over thirty methods to detect, identify and differentiate the soft rot and blackleg causing bacteria to species and subspecies level. These include methods based on biochemical characters, serology, molecular techniques which rely on DNA sequence amplification as well as several less-investigated ones.
ABSTRACT
AIMS: Dickeya zeae is a pectinolytic bacterium responsible for soft rot disease in flower bulb crops. In this study, the possibility of controlling soft rot disease in hyacinth by using antagonistic bacteria isolated from hyacinth bulbs was explored. METHODS AND RESULTS: Bacterial isolates with potential for biocontrol were selected on the basis of antibiosis against D. zeae, siderophore production, and the N-acyl homoserine lactones (AHLs)-inactivation. In in vitro assays, 35 out of 565 hyacinth-associated bacterial isolates produced antimicrobial substances against D. zeae, whereas 20 degraded AHLs, and 35 produced siderophores. Isolates of interest were identified by 16S rDNA sequence analysis and reaction in BIOLOG tests. Twenty-six isolates that differed in characteristics were selected for pathogenicity testing on hyacinth cultivars, Pink Pearl and Carnegie. Two strains identified as Rahnella aquatilis and one as Erwinia persicinus significantly reduced tissue maceration caused by D. zeae 2019 on hyacinth bulbs, but not on leaves. CONCLUSIONS: Hyacinth bulbs harbour bacteria belonging to different taxonomic groups that are antagonistic to D. zeae, and some can attenuate decay of bulb tissue. SIGNIFICANCE AND IMPACT OF THE STUDY: Selected hyacinth-associated bacterial isolates have potential for control of soft rot disease caused by D. zeae in hyacinth bulb production.
Subject(s)
Anti-Bacterial Agents/pharmacology , Antibiosis , Eichhornia/microbiology , Pest Control, Biological/methods , Plant Diseases/microbiology , 4-Butyrolactone/analogs & derivatives , 4-Butyrolactone/analysis , Anti-Bacterial Agents/isolation & purification , Antibiosis/genetics , Bacteria/genetics , Bacteria/isolation & purification , Bacteria/metabolism , Carboxylic Ester Hydrolases , Enterobacteriaceae/isolation & purification , Plant Roots/genetics , Plant Roots/microbiology , RNA, Ribosomal, 16S/genetics , Siderophores/analysisABSTRACT
Quorum sensing plays a role in the regulation of soft rot diseases caused by the plant pathogenic bacterium Pectobacterium carotovorum subsp. carotovorum. The signal molecules involved in quorum sensing in P. carotovorum subsp. carotovorum belong to the group of N-acyl homoserine lactones (AHLs). In our study, we screened bacteria isolated from the potato rhizosphere for the ability to degrade AHLs produced by P. carotovorum subsp. carotovorum. Six isolates able to degrade AHLs were selected for further studies. According to 16S rDNA sequence analysis and fatty acid methyl ester profiling, the isolates belonged to the genera Ochrobactrum, Rhodococcus, Pseudomonas, Bacillus, and Delftia. For the genera Ochrobactrum and Delftia, for the first time AHL-degrading isolates were found. Data presented in this study revealed for the first time that Ochrobactrum sp. strain A44 showed the capacity to inactivate various synthetic AHL molecules; the substituted AHLs were inactivated with a lower efficiency than the unsubstituted AHLs. Compared with the other isolates, A44 was very effective in the degradation of AHLs produced by P. carotovorum subsp. carotovorum. It was verified by polymerase chain reaction, DNA-DNA hybridization, and a lactone ring reconstruction assay that Ochrobactrum sp. strain A44 did not possess AHL lactonase activity. AHL degradation in Ochrobactrum sp. strain A44 occurred intracellularly; it was not found in the culture supernatant. AHL-degrading activity of A44 was thermo sensitive. Experiments in planta revealed that Ochrobactrum sp. strain A44 significantly inhibited the maceration of potato tuber tissue. Since A44 did not produce antibiotics, the attenuation of the decay might be due to the quenching of quorum- sensing-regulated production of pectinolytic enzymes. The strain can potentially serve to control P. carotovorum subsp. carotovorum in potato.