engleski

Proteome analysis for phylogenetic clarification in the Orobanchaceae

The most damaging weedy root parasites belong to the Orobanchaceae, and include the Orobanche and Phelipanche genera, some of which infect important crops and causing extensive damage. In this study we used a proteomic approach to contribute to phylogenetic clarification of twelve broomrape species, based on the seed protein profile, complementing previous studies that used other approaches. Proteins were detected by Sypro Ruby staining, and the gel images were digitalized and analyzed. The 2-DE maps contained from 30 to 140 protein spots in/for the analysed species. A virtual master gel was built with spots present in all species, revealing the presence of 260 protein spots, of which only one was monomorphic while 143 were unique to a single species. Proteins were scored for the presence or absence of homologous spots, to create a binary matrix. Phylogenetic analysis was carried out using the maximum parsimony (MP) and distance methods. The trees obtained were rooted using Cistanche phelypaea as out-group. Both analyses showed Orobanche as a monophyletic group. No conclusions can be taken on Phelipanche due to the limited number of species included. AMOVA analysis revealed phenotypic diversity among species within genera, as well as a considerable level of variation between genera. This work represents a first attempt of using protein markers as a tool in molecular systematics of the Orobanchaceae.

Orobanche; Phelipanche; Proteomics; two-dimensional electrophoresis; phylogenetics; AMOVA

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