| Northern Corn Leaf Blight is the main disease on the leaves of corn.The pathogen is Setosphaeria turcica and it is a heterothallism fungus.There are StMAT1-1 and StMAT1-2 mating type genes in the genome.The strain with StMA1-1 gene in the genome is A mating type strain,the strain with only StMAT1-2 gene is a mating type strain,and the strain with double mating type genes is Aa mating type.Previous studies found that the strains lacking the StMAT1-1 gene or the strains lacking the StMAT1-2 gene lost the sexual reproductive ability and the asexual reproduction ability decreased,indicating that the SiMAT1-1 gene and the StMA T1-2 gene are play an important role in reproduction.Based on the previous research,we systematically analyzed the composition of the mating type of Setosphaeria turcica in different corn production areas in China from 2016 to 2018,and have a preliminary understanding of the mating type composition of each province in different years The wild-type strain 01-23,?Stmat1-1 mutant and ?Stmat1-2 mutant were sequenced by high-throughput sequencing and analysisedusing bioinformatics analysis methods for-GO and KEGG.The expression and selection of differentially expressed genes related to sexual reproduction provide a preliminary basis for the study of the mechanism of mating genes in the sexual reproduction process of S.turcica.The main results are as follows:1.In this study,the pathogens of Setosphaeria turcica collected from different provinces in different years in the field were isolated and the mating type was identified.Among the 278 corn leaf spot pathogens from 2016 to 2018,22 isolates were A mating type,with the isolation frequency of 7.91%;234 isolates were a mating type,with the isolation frequency of 84.18%;22 isolates were Aa mating type,with the isolation frequency of 7.91%.2.Collected the mycelium of wild-type strains 01-23,mutant strains ?Stmat1-1 and?Stmat1-2 cultured at 25? for 5 days for RNA-Seq sequencing.A total of 13,896 expressed genes were detected,including 11253 known genes,2643 new genes;21809 expressed transcripts,of which 10936 known transcripts and 10873 new transcripts.3.Perform gene annotation analysis on the detected genes with six major databases(NR,Swiss-prot,Pfam,COG,GO and KEGG).8122 genes were annotated to GO database,3749 genes were annotated to KEGG database,9442 genes were annotated to COG database,11,179 genes were annotated to NR database,6695 genes were annotated to Swiss-Prot database,7929 genes were annotated to the Pfam database.4.Perform gene differential expression analysis between different sample groups.A total of 3099 differentially expressed genes were obtained in the WT_vs_StMAT1 sample group,of which 1393 were up-regulated genes and 1706 were down-regulated genes;in the WT_vs_StMAT2 sample group,there were 3666 differentially expressed genes,1523 genes were up-regulated and 2143 genes were down-regulated;in the StMAT1_vs_StMAT2 sample group,there were 1 949 differentially expressed genes,758 up-regulated genes and 1191 down-regulated genes.5.Enrich the differentially expressed genes of different sample groups separately.GO enrichment shows that it is related to metabolic activity.Enzyme activity and membrane composition are significantly enriched.KEGG enrichment analysis found that Pathway enrichment related to the amino acid metabolism signaling pathway was significant.6.Six genes related to pheromone and 25 genes related to meiosis were screened out from the differentially expressed genes.The homologous protein search of the differentially expressed genes related to sexual reproduction showed that the deletion of the hybrid gene of Setosphaeria turcica affected meiosis,cell process and signal transduction in the process of sexual reproduction?... |
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