Cloning And Expression Analysis Of Mating-type Genes(STE2,HMG,Alpha-box)of Oidium Heveae

Powdery mildew in rubber tree,caused by obligate parasitic fungus Oidium heveae B A.Steinmann,greatly reduced rubber production while serious harm.However,the study of Oidium heveae focused on observation to pathogen biolo-gy and cell biology,very little research have been done on O.heveae about the molecule.For exploring the molecular regulated mechanism and gene functions in infection of Oidium heveae,we conducted transcriptome sequencing of rubber powdery mildew interactions during infection,find out 3 genes of MAT(mating-type),analyzed the genes expression of pathogenicity-related by RT-qPCR,It provides the theoretical foundation for exploring the molecular regulated mechanism and MAT gene functions in infection of Oidium heveae,further verify its specific function.The main results as follows:1 Based on the genome of Oidium heveae,we used BLAST on NCBI and ho mologous cloning,and get 3 MAT gene,named OH-MAT2,OH-MATT-2,OH-M AT1-1.Sequence analysis indicated that the DNA length of OH-MAT2 was 964 bp,cDNA was 921 bp,the homologous rate of MAT STE2 protein between Blumeria graminis f.sp.Hordei and Oidium heveae was 93%;the DNA length of OH-MAT1-2 was 1071 bp,cDNA was 858 bp,the homologous rate of M AT alpha 1 protein between Erysiphe necator and Oidium heveae was 99%;th e DNA length of OH-MAT1-1 was 930 bp,cDNA was 930 bp,the homologo us rate of MAT alpha 1 protein between Erysiphe necator and Oidium heveae was 80%;OH-MAT2,OH-MAT1-2,OH-MAT1-1 should be MAT gene.2 Germination and appressorium formation of 12d post inoculation fresh conidium was better than the 30 d,and leaf age 7 to 10 d rubber leaves suitable conidium in detail.Total RNA was extracted from Oidium heveae at four infection stages,4 h,8 h,12 h,24 h,3 d,5 d.Based on Sequence analysis and Relative fluorescence quantitative on this 3 genes of Oidium heveae,we found that:this genes expressed at different time points,and the difference was significant,At 4h they are up-regulated,and then OH-MAT2 gene is also up-regulated,OH-MAT1-2 gene and OH-MAT1-1 gene are down-regulated.3 Extract DNA from Oidium heveae collected from 7 sites in Hainan and 6 sites in Yunnan,clone MAT gene and analysis Sequences.The results show that:the OH-MAT2 gene OH-MAT1-2 gene and OH-MAT1-1 gene are all existing in Oidium heveae from 13 sites in Hainan and Yunnan.Phylogenetic analysis show There is no obvious difference on these MAT genes among these 14 Oidium heveae.