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Inheritance of restriction polymorphisms of S. scitamineum subtelomeric region in fungal sexual crosses in planta Sugarcane is a major source of food and renewable energy in the world. Among the biotic factors that reduce its productivity we highlight the sugarcane smut disease, caused by the biotrophic fungus Sporisorium scitamineum, whose damages to the crop can overpass 80%. However, few genetic studies have targeted this pathogen. Thus, the present work studied the dynamics of RFLP marks associated to the subtelomeric region (tel-RFLP) and GFP marks of three populations of the pathogen (a, b and c) obtained from consecutive disease cycles. Population a was initially formed with inoculation using teliospores F1 from the whip 39 obtained in previous work (REIS, 2012). Populations b and c were initially produced by controlled crosses of sexual compatible haploid sporidia with known tel-RFLP profiles as parents. In population b the parent sporidia (18A and 39B) were isolated from distinct teliospores, had more different tel-RFLP profiles (7 polymorphic marks) and simulated outcrossing. In population c, the parental sporidia 39Agfp and 39Bgfp had more similar tel-RFLP profiles. These mutants were obtained by heterologous insertion of gfp gene in sporidia obtained from single teliospore, simulating selfing (intra-tetrad mating type). The comparison between wild-type and GFP mutant sporidia tel-RFLP profiles had detected the gfp gene insertion in the subtelomeric region of the individual 39Agfp (parent). For each disease cycle wild-type and GFP mutant teliospores were obtained and two sporidia were isolated to perform controlled crosses that lead to the next disease cycle. The use of controlled crosses resulted in the production of genetically identical teliospores, which allowed inferring about same progeny meiotic occurring events. The sexual phenotyping of the sporidia that had their tel-RFLP analyzed reduced the bias resulted from deviation of expected 1:1 ratio segregation among the mating types of same meiotic division progeny. Besides, it helped in the recombinants detection, due to mating type loci segregation in meiosis I. The analysis of wild-type and GFP mutant progenies telRFLP marks, obtained for 2 and 1 disease cycles, has pointed out events of homologous chromosomes recombination and crossing-over events. Despite of the high amount of marks obtained, the tel-RFLP marks allowed the tracking of individuals from the same population, which can benefit future sugarcane smut disease epidemiologic evaluations. Moreover, the gfp gene expression of mutant probasidium cells led to the detection of crossing-over involving the GFP mark and the first observation of linear four-cells tetrads to this species. A high potential use of gfp gene in genetic studies applied to this pathosystem was observed. The recombination suggested for the subtelomeric region showed a recombination frequency haven’t described for this species that needs to be further studied.

Herança dos polimorfismos de restrição associados à região subtelomérica de Sporisorium scitamineum em análise de cruzamentos sexuados do fungo in planta