Hylogenetic analysis was according to 45 PTI1 protein sequences. Species abbreviations are as follows. At: A. thaliana; Os: O. sativa; Zm: Z. mays; Sl: S. lycopersicum; Gm: G. max; Nt: N. tabacum. Several sequence alignments of PTI1 amino-acid sequences were performed using ClustalX, and the phylogenetic was constructed working with MEGA7 by the maximum likelihood method and 1000 bootstrap replicates. The tree was divided into six phylogenetic subgroups, designated I-VI. Letters outside from the tree indicate the defined groupsV.IIZmI1 -Si PT IDQ101-I PT Si.ZmZm1-At PTI1 -At PTI1 -70 AYDQ80388 .Huangfu et al. BMC Plant Biology(2021) 21:Page 5 ofFig. 3 Phylogenetic relationships, gene structure and architecture of conserved protein motifs in PTI1 genes from mTORC1 Activator site foxtail millet. The phylogenetic tree was constructed determined by the full-length sequences of foxtail millet PTI1 proteins using MEGA7 application (A). Exon-intron structure of foxtail millet PTI1 genes. Green boxes indicate untranslated 5- and 3-regions; yellow boxes indicate exons; black lines indicate introns (B). The motif composition of foxtail millet PTI1 proteins. The motifs, numbers ten, are displayed in various colored boxes (C). The sequence info for each and every motif is provided in Extra file two. The length of protein is often estimated utilizing the scale at the bottomand SiPTI1 had eight introns. The rest of SiPTI1 genes had six introns. Exon-intron structural analysis indicated that members of some PTI1 subfamilies have related exon-intron structures. Comparable benefits have been also discovered in maize [31] and other studies. The motif patterns among SiPTI1s were investigated (Fig. three C and Extra file 3). A total of 10 motifs have been found and 5 of them had been located to become very conserved. Also, all of SiPTI1s contained motifs 1, 2, three, four and five. Except for SiPTI10, all of other SiPTI1s include motifs 6 and 9. Moreover, motif eight was located in three of your SiPTI1s members (SiPTI1, SiPTI11 and SiPTI12), though motif 10 was only presented in two members (SiPTI1, SiPTI11). Interestingly, the motif distribution of SiPTI1 was distinctive from other members on the loved ones, in that motifs three, five, 9 seem twice each. Regardless of the distinction of motif types in between groups, members within precisely the same group for instance SiPTI1 and SiPTI1, SiPTI1 and SiPTI1, SiPTI11 and SiPTI1 are likely to exhibit equivalent motif patterns (Fig. 3 A and C), which indicate functional similarity in between them. Amino acid sequence analyses showed that the SiPTI1s contain the representative kinase domains, which include STKC_IRAK, Pkinase_Tyr, STYKc, and SPS1 (information not shown). As recognized that the catalytic domain of serine/threonine NPY Y2 receptor Agonist Biological Activity kinases contains 11 subdomains [31, 32], the pileup evaluation also showed that the 12 SiPTI1 kinases also contained the conserved 11 subdomains like recognized PTI1 gene of SlPTI1 in tomato (Supplementary Fig. two). Also, when compared the SiPTI1s sequence of foxtail millet with all the PTI1 sequences of maize and rice, we located that thecatalytic domain of serine/threonine kinases also contains 11 subdomains, which were constant together with the results of SiPTI1s and SlPTI1 sequence evaluation (Supplementary Fig. 3).Cis-acting components and subcellular localization of PTI1 genes in foxtail milletCis-elements analysis showed that all SiPTI1 genes promoter contained MYB, MYC and ABA-responsive (ABRE) components. In addition, excepted for SiPTI12, each CGTCA-motif and TGACG-motif cis-elements have been present in foxtail millet PTI1 genes loved ones (F.