Xanthomonas axonopodis pv. glycines ( Xag ) causes bacterial pustule disease which can significantly reduce the production of soybean. A collection of 26 isolates of Xag from different soybean-production areas of Thailand was shown to differ with regard to aggressiveness on soybean. They also differed in their ability to induce a hypersensitive response (HR) on four cultivars of tobacco and on other plant species including pepper, tomato, cucumber, pea and sesame. Tomato was most sensitive to HR induction by Xag . Isolate KU-P-34017 caused an HR on all the plant species tested. The minimal concentration of KU-P-34017 needed to induce HR on tobacco was approximately 5 × 10 8 CFU mL − 1 . A bacterium-plant interaction period of at least 2·5 h was necessary for HR, and different temperatures, relative humidity and light periods did not affect HR development. Inhibitors of eukaryotic metabolism, including cobalt chloride, lanthanum chloride and sodium orthovanadate (completely), and cycloheximide (partially) blocked the HR on tobacco, indicating the association of an active plant response. In contrast, the HR on tomato was inhibited only by cobalt chloride.
Agrobacterium vitis, the causal agent of grape crown gall, can have severe economic effects on grape production. The bacterium survives systemically in vines and, therefore, is disseminated in propagation material. We developed an assay for use in indexing programs that is efficient and sensitive for detecting A. vitis in grape tissue. Initially, real-time polymerase chain reaction (PCR) primers specific for diverse tumorigenic strains of A. vitis were developed using the virD2 gene sequence. To overcome the effects of PCR inhibitors present in plant tissue, DNA extraction methods that included magnetic capture hybridization (MCH), immunomagnetic separation (IMS), and extraction with the Mo Bio Powerfood kit were compared. The assays incorporating MCH or IMS followed by real-time PCR were 10,000-fold more sensitive than direct real-time PCR when tested using boiled bacterial cell suspensions, with detection thresholds of 10(1) CFU/ml compared with 10(5) CFU/ml. DNA extraction with the Powerfood DNA extraction kit was 10-fold more sensitive than direct real-time PCR, with a detection threshold of 10(4) CFU/ml. All three assays were able to detect A. vitis in healthy-appearing grapevine cuttings taken from infected vines.
Nontumorigenic Agrobacterium vitis strain F2/5 is able to prevent crown gall caused by tumorigenic A. vitis on grape but not on other plant species such as tobacco. Mutations in a quorum-sensing transcription factor, aviR, and in caseinolytic protease (clp) component genes clpA and clpP1 resulted in reduced or loss of biological control. All mutants were complemented; however, restoration of biological control by complemented clpA and clpP1 mutants was dependent on the copy number of vector that was used as well as timing of application of the complemented mutants to grape wounds in relation to inoculation with pathogen. Mutations in other quorum-sensing and clp genes and in a gene associated with polyketide synthesis did not affect biological control. It was determined that, although F2/5 inhibits transformation by tumorigenic A. vitis strains on grape, it does not affect growth of the pathogen in wounded grape tissue over time.
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