Transformation of Xanthomonas campestris pathovar campestris with Plasmid DNA

Abstract

Procedures for the introduction of plasmid DNA into Gram-negative bacteria have been adapted and optimized to permit transformation of the plant pathogen Xanthomonas campestris pathovar campestris with the cloning vector pKT230 and other broad-host-range plasmids. The technique involves CaC1,-induced competence and heat shock and is similar to that routinely used for Escherichia coli. Wild-type X . c. campestris strains appear to restrict incoming unmodified DNA, so that plasmid DNA for transformation must be prepared from X . c. campestris (into which it has previously been introduced by conjugation). To overcome this disadvantage a restriction-deficient mutant has been isolated.

Xanthomonas

“…The properties of commonly used mutagenic agents and the types of mutations induced by them have been discussed (Carlton and Brown, 1981) and summarized (Chatterjee and Vidaver, 1986). Physicochemical agents have been used for mutagenesis of xanthomonads (Whitfield et al, 1981;Tsuchiya et al, 1982;Daniels et al, 1984a;Atkins et al, 1987;Harding et al, 1987;Thorne et al, 1987;Mellano and Cooksey, 1988;. Physicochemical agents have been used for mutagenesis of xanthomonads (Whitfield et al, 1981;Tsuchiya et al, 1982;Daniels et al, 1984a;Atkins et al, 1987;Harding et al, 1987;Thorne et al, 1987;Mellano and Cooksey, 1988;.…”

“…(Corey and Starr, 1957a,b;Yamasaki et al, 1966;Gabriel, 1984;Atkins et al, 1987;Murooka et al, 1987). Many xanthomonads do not respond to chemical treatments that induce competence necessary for transformation in other bacteria.…”

Resistance and systemic dispersal of Xanthomonas fragariae in strawberry germplasm (Fragaria L.)

“…4), it can be confirmed that the pathogen invades through the stomata, colonizes the intercellular space for multiplication and spreads systemically throughout the whole plant via the leaf veins. Electroporation with plasmid DNA on other Xanthomo-nas species has been carried out in previous investigations for the purpose of specific detection of the pathogen (Atkins et al, 1987;Yang et al, 1991). In a previous study with GFP-tagged X. fragariae by Kaste- 2013), a slightly lower virulence of the tagged strain was reported, but this was not found in the current investigation.…”

Plasmid and pathogenicity in Xanthomonas oryzae pathovar oryzae, the bacterial blight pathogen of Oryza sativa

“…The transformation of plasmid pMA36 into the cured strain was performed with some modification of the method described by Atkins et al (1987). Plasmid-cured log phase cells (5 ml) of Xoo were added to 50 ml of prewarmed Luria broth in a 250 ml Erlenmeyer flask and kept in an orbital shaker (30"C, 200 rev min-').…”