Analysis Of WRKY And SnRK2 Gene Family In Five Thinopyrum Species

Thinopyrum is a gennus of Trticeae,a perennial important wheat related species.The genus Brassica has many excellent disease resistance(powder disease,rust),drought resistance,and salt and alkali resistance.Isogenic genes provide the basia for the utilization of exogenous genes.The WRKY gene family and the SnRK2 gene family are ubiquitous in plants and play a role in both biotic and abiotic stresses.In this study,the transcriptome data of five species of genus Thinopyrum were used to exploit the gene members of the WRKY and SnRK2 gene families in five species by bioinformatics method,which laid a foundation for the excavation of some stress-resistant genes.The main resuts of this study are as follows:1.Sequencing and assembly transcriptome: using transcriptome second-generation sequencing technology for intermediates of Thinopyrum intermedium,Th.Elongatum,Th.Ponticum Pseudoroegneria strigosa and Th.bessrabicum were sequenced and assembled using the transcriptome sequencing.Through data filtering,quality detection and assembly and splicing,the number of transcripts was up to 70000 the above.The length of the N50 is above 1000 bp.2.Bioinformatics analysis of the WRKY gene family:(1)Sequence search and identification: The family members identified by WRKY were downloaded from the plant gene database,and the core sequence WRKYGQK and zinc finger strcture of the WRKY protein domain were ompared to finally identify,There are 21,29,49,33 and 33 family members in Pseudoroegneria strigosa,Thinopyrum intermedium,Th.Ponticum,Th.bessarabicum and Th.Elongatum(2)Physicochemical properties of the protein were analyzed: the most of the five species showed hydrophilic protein.Moet of the proteins were unstable proteins,and the fat solubility index was it is expressed as a fat-soluble protein.(3)In the analysis of protein secondary structure: ?-helix and random coil accounted for the highest proportion.The content of TbWRKY was higher than that of the other four species,the content of TpWRKY is higher than that of the other four species.(4)Construction of phylogenetic tree: The classification there are no separate branches.They are divided into three groups,indicating that they hace the sane evolution pattern as the five species.(5)In terms of conserved motif analysis: 42 WRKY protein sequences with higher quality were selected in the WRKY gene family and the conserved motifs between the subgroups were different and the motif models and numbers were also different.The exact sequences matched are also different.These results indicate that the five species in the WRKY gene family have different evlution patterns.Ten conserved motifs in the WRKY protein sequence of five species,indicating a higher conservation between the two gene family members.3.Biological analysis of the SnRK2 gene family:(1)Sequence search and identification: The protein conserved domain was detected by the highly conserved kinase domain DFGYKSSVLHSQPKIGOGPAYIAPE of SnRK2 protein,the final identification Pseudoroegneria strigosa,Thinop-yrum intermedium,Th.Ponticum,Th.bessarabicum and Th.Elongatum each of these five species has 10 family members.(2)Physicochemical properties of protein: The SnRK2 gene family has the highest number of the proteins are unstable proteins;the fat-soluble index is a fat-soluble protein.(3)Analysis of protein secondary steucture: ?-helix and random coil accounted for the highest proportion,the proportion of ?-helix in the TiSnRK2 was high,the ratio of irregular curl of TeSnRK2 was higher than of the other four species high.(4)Costruction of phylogenetic tree analysis: They are divided into three groups,indicating that they same as the five apecies.The evolutionary model also has some induction effects on some ABA induction,heat stress and low temperature treatment.(5)Conservative motif analysis: There are 10 types of conserved motifs in 50 protein sequences of five species.The conserved motifs between the subgroups are different,and the motif models and numbers are also different.The exact sequences are also different,indicating that the members of the SnRK2 gene family have different evolution patterns,indicating that the members of the SnRK2 gene family are highly conserved.4.SnRK2 gene sequence cloning verification: sequence cloning of SnRK2 gene members of five species,the sequence obtained by cloning is compared with the original sequence,which is consistent with the sequencing sequence obtained by the transcriptome,which provides functional verification for the next gene.