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Exploration of the Use of the “Bialaphos Genes” for Improving Bioherbicide Efficacy
- R. Charudattan, V. J. Prange, J. T. Devalerio
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- Journal:
- Weed Technology / Volume 10 / Issue 3 / September 1996
- Published online by Cambridge University Press:
- 12 June 2017, pp. 625-636
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We are studying the possibility of altering the virulence and host range of a phytopathogen by transferring and expressing certain genes from the soil-dwelling saprophyte, Streptomyces hygroscopicus, in a plant pathogen model, Xanthomonas campestris pv. campestris (XCC). The genes, referred to herein as the “bialaphos genes,” encode the production of bialaphos, a potent glutamine-synthetase-inhibiting herbicide. This cluster of genes was originally isolated from several biosynthetically blocked mutants of S. hygroscopicus and constructed into a plasmid vector, pBG9. We have transferred a fragment of the gene cluster into pLAFR3, a plasmid that functions in both Escherichia coli and XCC and contains a tetracycline resistance marker. The resulting plasmid, named pIL-1, was used to transform E. coli and was incorporated into XCC by conjugation. The transfer of the fragment was confirmed by Southern analysis. The genes were maintained in XCC for about 47 generations in the absence of selection for tetracycline, and no changes in cultural phenotypes were seen in the transformed XCC (XCC/pIL-1). The XCC/pIL-1 cells were pathogenic to their natural hosts cabbage and broccoli, but induced an altered hypersensitive response in the nonhosts bean, pepper, sunflower, and tobacco. The pathogenic host-reaction, induced by the parent XCC, XCC/pLAFR3, and XCC/pIL-1, was a typical black rot disease in inoculated leaves of the two hosts. The nonhost reaction on the nonhost leaves was necrotic hypersensitivity, induced by XCC and XCC/pLAFR3, or the inhibition of hypersensitivity accompanied by only chlorosis at sites inoculated with XCC/pIL-1. We hypothesize that the altered hypersensitivity phenotype may be due to the transformed XCC becoming more compatible with the nonhosts, a step toward acquiring nonhost-virulence, or due to disruption of the normal expression of the hypersensitivity and pathogenicity genes in the transformed XCC. More work is needed to confirm that the introduced genes are being expressed in XCC. With further understanding, this approach may provide a useful model to study host range, virulence, and strain improvement of plant pathogens for biological control of weeds.
Citrus canker: a new disease of Mexican lime (Citrus aurantifolia) and sour orange (C. aurantium) in Ethiopia
- Eshetu Derso, Kamaruzaman Sijam
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Introduction. Citrus canker, caused by the bacterium Xanthomonas axonopodis pv. citri, is a serious disease of most commercial citrus cultivars and some citrus relatives. Surveys were conducted in small-scale farms, commercial plantations, backyard orchards and nurseries in order to quantitatively study the occurrence and distribution of the disease and determine the intensity and field host range. Materials and methods. Seventeen locations in four regional states of Ethiopia were surveyed. Incidence on leaves was determined by counting total number of leaves and expressed as a proportion of leaves with at least one lesion. Incidence on fruits was determined on attached fruits, recorded as presence or absence of the symptoms. Severity was also measured on leaves and fruits. The identity of the pathogen was confirmed by morphological, biochemical and physiological characterization and detached leaf tests of representative isolates. Results. The surveys indicated the occurrence of citrus canker in Ethiopia. Overall incidence on leaves was 71.4% of leaves with at least one lesion and severity was 26.8% of leaf area infected. Incidence on fruits was 30% of fruits infected and severity was 21.25% fruit area infected. Morphological, biochemical and physiological characterization and detached leaf tests of isolates indicated characteristics similar to X. axonopodis pv. citri. The host range for citrus canker in Ethiopia was limited to Mexican lime (C. aurantifolia) and sour orange (C. aurantium). Based on the field host range and detached leaf tests, it appears that the X. axonopodis pv. citri variant that occurs in Ethiopia has similar host ranges to that of atypical Asiatic (Xac-A*) form. Canker severity significantly correlated with temperature but not with rainfall, elevation or tree age. Conclusion. This is the first confirmed report of the disease in Ethiopia.