Saussurea medusa, Saussurea hypsipeta and Saussurea obvallata are typical alpine snowline plants growing in the Qinghai-Tibet plateau. They are characterized by a specialized morphology. S. medusa and S. hypsipeta have very dense trichomes on whole plant, whereas S. obvallata has transparent bracts covered inflorescences. The different forms reflect their adaptation to cold environments. To investigate the different mechanisms of adaptation of these species to cold temperatures, transcriptome sequencing was performed in three species of Saussurea DC. A total of 116394 137237 and 113879 Unigenes were identified from S. medusa, S. hypsipeta and S. obvallata, respectively. Of these, 55909 (48.03%), 65519 (47.74%) and 51889 (45.56%) Unigenes were matched in public databases. GO analysis identified that most of annotated Unigenes in the three species of plants were related to cellular, metabolic, and single−organism processes, and binding and catalytic activities. The differential expression of 37 genes related to environmental adaptation were discovered by pairwise comparisons. Of these, two candidate genes (Interaptin-like and CSLB3) related to trichome development were identified only in S. medusa and S. hypsipeta, which was consistent with their distinct morphology. Our data can provide a valuable resource for the further studies on the adaptive mechanisms of molecular and functional ecology in Saussurea DC.
Anthocyanins are ubiquitous in Compositae and MYB regulates the expression of DFR and plays an important role in anthocyanin synthesis. Here, the regulation pathway that MYB protein of subgroup 6 in Compositae promotes dihydroflavonol reductase (DFR) expression was analyzed and verified by yeast one-hybrid experiment in Saussurea medusa. The results of the branch model and site model analysis revealed that MYB gene underwent purification selection, and the motif of bHLH protein [DE]Lx(2)[RK]x(3)Lx(6)Lx(3))R and anthocyanin-related motif ANDV underwent strong purification selection during evolution. DFR promoter analysis showed that there are MYB binding site (GAGTTGAATGG) and bHLH binding site (CANNTG) at the sense strand of 84–116 nucleotide residues from the start codon, and the two motifs are separated by 9–10 nucleotide residues, and this rule exists in DFR promoters of many Compositae plants. Yeast one-hybrid experiment proved that SmMYB1 can activate the promoter of SmDFR. Our results provide a reference for further functional studyof DFR gene in Compositae.
Anthocyanins are ubiquitous in Compositae and MYB regulates the expression of DFR and plays an important role in anthocyanin synthesis. Here, the regulation pathway that MYB protein of subgroup 6 in Compositae promotes dihydroflavonol reductase (DFR) expression was analyzed and verified by yeast one-hybrid experiment in Saussurea medusa. The results of the branch model and site model analysis revealed that MYB gene underwent purification selection, and the motif of bHLH protein [DE]Lx(2)[RK]x(3)Lx(6)Lx(3))R and anthocyanin-related motif ANDV underwent strong purification selection during evolution. DFR promoter analysis showed that there are MYB binding site (GAGTTGAATGG) and bHLH binding site (CANNTG) at the sense strand of 84–116 nucleotide residues from the start codon, and the two motifs are separated by 9–10 nucleotide residues, and this rule exists in DFR promoters of many Compositae plants. Yeast one-hybrid experiment proved that SmMYB1 can activate the promoter of SmDFR. Our results provide a reference for further functional studyof DFR gene in Compositae.
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