Molecular characterization of beta-tubulin from Phakopsora pachyrhizi, the causal agent of Asian soybean rust

β-tubulins are structural components of microtubules and the targets of benzimidazole fungicides used to control many diseases of agricultural importance. Intron polymorphisms in the intron-rich genes of these proteins have been used in phylogeographic investigations of phytopathogenic fungi. In this work, we sequenced 2764 nucleotides of the β-tubulin gene (Pp tubB) in samples of Phakopsora pachyrhizi collected from seven soybean fields in Brazil. Pp tubB contained an open reading frame of 1341 nucleotides, including nine exons and eight introns. Exon length varied from 14 to 880 nucleotides, whereas intron length varied from 76 to 102 nucleotides. The presence of only four polymorphic sites limited the usefulness of Pp tubB for phylogeographic studies in P. pachyrhizi. The gene structures of Pp tubB and orthologous β-tubulin genes of Melampsora lini and Uromyces viciae-fabae were highly conserved. The amino acid substitutions in β-tubulin proteins associated with the onset of benzimidazole resistance in model organisms, especially at His 6 , Glu 198 and Phe 200 , were absent from the predicted sequence of the P. pachyrhizi β-tubulin protein

RAPD and SCAR markers linked to resistance to frogeye leaf spot in soybean

The soybean (Glycine max (L.) Merrill) frogeye leaf spot is caused by the fungus Cercospora sojina Hara and is a widespread disease in Brazil and other countries, causing severe losses in grain yield and also affecting seed quality. The availability of DNA markers linked to genes for resistance to this disease would accelerate breeding programs, particularly when other traits are also being evaluated. Bulked segregant analysis was applied to 3 F2 populations derived from crosses between the resistant cultivars Parana, Cristalina and Uberaba, and the susceptible cultivar Bossier. In the cross 'Parana' x 'Bossier', 2 RAPD markers were identified, CSOPA1(800C) and CSOPA2(1,250C), located at 4.4 ± 1.8 centiMorgans (cM) and 3.4 ± 1.7 cM respectively from the resistance locus. DNA fragments of similar molecular weight were observed in the population derived from the cross 'Cristalina' x 'Bossier' at 2.3 ± 1.2 and 4.7 ± 1.5 cM from the resistance locus, respectively. In the offspring of the cross 'Uberaba' x 'Bossier', a DNA fragment corresponding to marker CSOPA1(800C) was detected at 5.6 ± 2.1 cM from the resistance locus. Although marker CSOPA2(1,250C) was not observed in this population, an additional marker was detected (CSOUB1(1,100C)) at 6.7 ± 2.2 cM from the resistance locus. The 1,250 bp fragment of CSOPA2(1,250C) was cloned and converted into a SCAR marker, which amplified a single fragment whose size corresponded to the cloned segment of the crosses involving cultivars Cristalina and Parana. Markers CSOPA1(800C), CSOPA2(1,250C) and CSOUB1(1,100C) were mapped to soybean linkage group J with the aid of known SSRs linked to the Rcs3 locus, indicating that the RAPD and SCAR markers identified in our research also tag this resistance gene

Genetics of resistance to powdery mildew (Microsphaera diffusa) in Brazilian soybean populations

Crosses between resistant and susceptible soybean cultivars were performed and the F2 populations were obtained to study the inheritance of soybean resistance to powdery mildew and to estimate the number and action of genes related to resistance. The reaction to powdery mildew was studied in a greenhouse and pots carrying plants with symptoms were distributed among the pots carrying the genotypes to be tested as a source of inoculum. Individual plants were scored according to the method of Yorinori (1997), with modifications, and classified as resistant or susceptible. The results showed that adult soybeans plants can present resistance to powdery mildew, which is controlled by one major gene with a dominant effect