Stable transformation of Rhizoctonia solani with a modified hygromycin resistance gene
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A vector for Agrobacterium tumefaciens-mediated transformation of basidiomycetes was constructed by insertion of a modified hygromycin resistance gene into the plant binary vector pBin19. The hygromycin coding region is flanked by a basidiomycete promoter and terminator. Isolates from different anastomosis groupings (AG) of the phytopathogenic fungus Rhizoctonia solani (Kuhn) were transformed with this vector using A. tumefaciens. Hygromycin-resistant transformants were isolated from a single AG6 isolate, but not from AG3 or AG4 isolates. Of six transformants isolated, five showed enhanced growth on agar containing either 25 or 50 μg/mL hygromycin. However, as the hygromycin concentration increased, the difference between the transformants and the control reduced, until at 100 μg/mL there was no difference. The resistance phenotype was retained through repeated subcultures on non-selective media. The presence of the gene in the transformants was confirmed by PCR analysis and Southern hybridisation.
|Publication Type:||Journal Article|
|Murdoch Affiliation:||School of Biological Sciences and Biotechnology|
|Copyright:||2009 Australasian Plant Pathology Society|
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