UBC Undergraduate Research

Protocol for identification of Cronartium ribicola x Cronartium comandrae hybrids in populations of Pinus monticola Chalmers, Eric


Cronartium ribicola is a rust fungi affecting soft pines throughout British Columbia. It is believed that C. ribicola is capable hybridizing with C. comandrae and is doing so in areas where the two rusts are in close proximity with each other. Establishing a means of identifying hybrid individuals is an important step in studying the hybrid individuals. Through the use of digestion enzymes and gel electrophoresis, hybrid aeciospores DNA can be differentiated from non-hybrid spore DNA. The restriction endonuclease BtsCI selectively cleaves C. ribicola DNA segments between the ITS1f and ITS4 binding sites while leaving C. comandrae segments intact. The result of the selectivity of this enzyme in the presence of C. comandrae, C. ribicola, and hybrid DNA is one, two, and three bands, respectively, when observed through gel electrophoresis. This relatively easy and cost effective means of identifying these hybrids will facilitate further study of these and other inter-specific hybrids, as well as the relationships of other hybrid fungi.

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