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Stable transformation of Rhizoctonia solani with a modified hygromycin resistance gene

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Abstract

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 ⧎g/mL hygromycin. However, as the hygromycin concentration increased, the difference between the transformants and the control reduced, until at 100 ⧎ g/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.

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Authors and Affiliations

  1. School of Biological Sciences and Biotechnology, Murdoch University, 6150, Murdoch, WA, Australia

    Jiang Wu & Philip A. O’Brien

  2. Agritechnology, 2800, Orange, NSW, Australia

    Jiang Wu

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  1. Jiang Wu
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  2. Philip A. O’Brien
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Correspondence to Philip A. O’Brien.

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Wu, J., O’Brien, P.A. Stable transformation of Rhizoctonia solani with a modified hygromycin resistance gene. Australasian Plant Pathology 38, 79–84 (2009). https://doi.org/10.1071/AP08081

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  • DOI: https://doi.org/10.1071/AP08081

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