Browsing by Author "Zhong, Shaobin"
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Item Comparative Genome Structure, Secondary Metabolite, and Effector Coding Capacity across Cochliobolus Pathogens(PLoS Genetics, 2013-01-24) Steffenson, Brian; Condon, Bradford J; Leng, Yueqiang; Wu, Dongliang; Bushley, Kathryn E; Ohm, Robin A; Otillar, Robert; Martin, Joel; Schackwitz, Wendy; Grimwood, Jane; MohdAzinudin, NurAinizzati; Xue, Chunsheng; Wang, Rui; Manning, Viola A; Dhillon, Braham; Tu, Zheng Jin; Salamov, Asaf; Sun, Hui; Lowry, Steve; LaButti, Kurt; Han, James; Copeland, Alex; Lindquist, Erika; Barry, Kerrie; Schmutz, Jeremy; Baker, Scott E; Ciuffetti, Lynda M; Grigoriev, Igor V; Zhong, Shaobin; Turgeon, B.GillianThe genomes of five Cochliobolus heterostrophus strains, two Cochliobolus sativus strains, three additional Cochliobolus species (Cochliobolus victoriae, Cochliobolus carbonum, Cochliobolus miyabeanus), and closely related Setosphaeria turcica were sequenced at the Joint Genome Institute (JGI). The datasets were used to identify SNPs between strains and species, unique genomic regions, core secondary metabolism genes, and small secreted protein (SSP) candidate effector encoding genes with a view towards pinpointing structural elements and gene content associated with specificity of these closely related fungi to different cereal hosts. Whole-genome alignment shows that three to five percent of each genome differs between strains of the same species, while a quarter of each genome differs between species. On average, SNP counts among field isolates of the same C. heterostrophus species are more than 25× higher than those between inbred lines and 50× lower than SNPs between Cochliobolus species. The suites of nonribosomal peptide synthetase (NRPS), polyketide synthase (PKS), and SSP–encoding genes are astoundingly diverse among species but remarkably conserved among isolates of the same species, whether inbred or field strains, except for defining examples that map to unique genomic regions. Functional analysis of several strain-unique PKSs and NRPSs reveal a strong correlation with a role in virulence.Item Genetic and molecular characterization of mating type genes in Cochliobolus sativus(Mycologia, 2001-09-01) Steffenson, Brian; Zhong, ShaobinGenetic and molecular approaches were used to characterize the mating type (MAT) genes in Cochliobolus sativus. One hundred and four ascospore progeny derived from a cross of C. sativus isolates ND93-1 (MAT-1) X ND9OPr (MAT-2) were backcrossed with their parents to determine mating type, but only five progeny produced pseudothecia with asci and/or ascospores. When degenerate primers from the conserved high mobility group (HMG) protein domain encoded by the MAT-2 gene in Cochliobolus species were used in polymerase chain reaction (PCR) with genomic DNA of C. sativus as templates, an amplicon of predicted size was amplified only from MAT-2 isolates. The presence of a MAT-2 homolog in these MAT-2 isolates was confirmed by Southern hybridization with the HMG box as a probe. Additionally, the presence or absence of the HMG homolog in the progeny segregated in a 1:1 ratio, as expected for the single gene control of mating type. Using primers based on the conserved regions at the 5' and 3' flanks of the idiomorphs in the MAT genes of other Cochliobolus species, the full-length MAT-1 and MAT-2 idiomorphs were cloned by PCR from C. sativus isolates ND93-1 and ND9OPr, respectively. DNA sequence analysis indicated that these two idiomorphs are organized in a manner similar to their respective counterparts in other Cochliobolus species. DNA hybridization and PCR amplification analysis of 54 field isolates of C. sativus collected worldwide showed that both mating types exist in populations round the world. The low frequency of successful backcrosses of progeny to parents in the ND93-1 X ND9OPr cross, combined with the fact that many crosses between isolates of opposite mating type are unsuccessful, suggests that genetic factors other than MAT genes affect the fertility of the fungus.