| Pugionium cornutum(L.)Gaertn.is a biennial herb used for medicine and food.In recent years,it has been used to control soil erosion in many western regions.Due to the particularity of its growth environment,there must be a corresponding evolutionary mechanism within it.Therefore,studying the molecular mechanism of Pugionium cornutum(L.)Gaertn.seedlings under drought stress is constructive to discover stress resistance genes,and it has important guiding significance for the development and utilization of Pugionium cornutum plants and the cultivation of new germplasm.In this thesis,the seeds collected from the Mu Us sandy land were used for drought treatment,and combined analysis of transcriptomics and proteomics was used to screen different genes,and PcHSP26.5 and PcHSP17.8 were selected for full-length cloning and expression analysis.The specific results as follows:1.Through the analysis of the transcriptome and proteome sequencing results,84468 differentially expressed genes(DEGs)and 4588 differential proteins(DEPs)were found respectively,and then the two were jointly analyzed.Through data comparison between different treatments,a total of 6639 were found DEGs and 795 DEPs.Quantitative analysis showed that 109 genes/proteins were regulated at these two omics levels.KEGG analysis showed that the 109 genes/proteins were mainly involved in metabolic pathways including starch and Sucrose metabolism,mutual conversion of pentose and glucuronic acid esters,glutathione metabolism,biosynthesis of secondary metabolites,biosynthesis of phenylpropane,metabolism of amino sugars and nucleotide sugars,biosynthesis of amino acids.Further analysis of these 109 genes/proteins shows that the proportion of heat shock proteins is the most abundant,about one-fifth.2.Further,selecting PcHSP26.5 and PcHSP17.8 as representatives to gene cloning.The full-length c DNA of PcHSP26.5 and PcHSP17.8 are 699 bp and 462 bp,respectively,the number of encoded amino acids are 282 and 153,the molecular weights are 26.48 kDa and 17.49 kDa,and the theoretical isoelectric points(PI)are 6.36 and 5.99,neither the transmembrane structure and signal peptide site are both hydrophilic proteins.Subcellular location prediction and phylogenetic tree analysis showed that PcHSP26.5 is mainly located in the mitochondria and belongs to the MII subfamily;while PcHSP17.8is mainly located in the cytoplasm and belongs to the CI subfamily.3.Real-time fluorescent PCR results showed that PcHSP26.5 and PcHSP17.8 had a strong response to high temperature and Na Cl induction,followed by the response in drought,indicating that they all participated in the process of response of Pugionium cornutum(L.)Gaertn.to high temperature,Na Cl and drought stress,and especially sensitive to high temperature stress. |
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