Cloning And Stress Resistance Of MhIMT1 Gene From Mirabilis Himalaica Edgew

The dry root of Mirabilis himalaica is one of the “Five Medicine”,the Tibetan transliteration: wazhi,the plant is a strong growth,strong drought resistance of plants.According to previous laboratory studies,Mirabilis in the Himalayas under drought stress will enhance its tolerance and adaptability to drought stress by means of osmotic regulators to maintain cell turgor and enzyme activity to protect cell membrane from damage.The main drought-resistant morphological characteristics of the root system: the root system expanded in volume and had high vertical depth;It is conducive to water absorption and drought resistance,and some branches and leaves on the ground are compact and horizontal,which is conducive to water conservation.,leaves of the plant surface local villi and keratinization degree is high,reduce the evaporation and loss of water.To sum up,Mirabilis himalaica is a plant of great drought resistance research value.In recent years,scholars at home and abroad have carried out a series of studies on Mirabilis in tissue culture,artificial cultivation,chemical constituents of its roots,etc.,but no reports have been made on its drought-resistant genes or related mechanisms.This study started by Mirabilis himalaica drought and ck ratio on the transcriptome data were analyzed,and the excavated Inositol O-methyltransferase,Inositol methyltransferase(IMT1)gene in Mirabilis himalaica drought have increased expression in the transcriptome data capacity,thus to clone,through bioinformatics analysis,plant expression vector of build,inflorescence infection method into arabidopsis and resistance to its research,the research results are as follows:1.In this study,the MhIMT1 gene was cloned from Mirabilis himalaica by PCR.Through homology comparison,the similarity between the MhIMT1 gene and the flower of succulent plants reached86%.2.This is the first systematic screening of the reference gene Mirabilis himalaica,eventually passed in seven internal candidate gene Ge Norm,Norm Finder,Best Keeper three internal stability analysis software has elected in Mirabilis himalaica stable internal genetic GAPDH and TUBa,Mirabilis himalaica gene expression spectrum analysis for the future provide a reference basis.3.Through to the Mirabilis himalaica MhIMT1 protein sequence of bioinformatics analysis,results show that the open reading frame of 1110 bp,the molecular weight of 40649.93 Da,theory of isoelectric point of5.91.MhIMT1 protein is a stable protein with a stability coefficient of30.09,containing 12% acidic amino acids and 10% alkaline amino acids.MhIMT1 protein is a hydrophobic protein with no peptide conductivity,no signal peptide and no transmembrane transport domain.And through the homologous modeling to predict its MhIMT1 tertiary structure.4.The target gene was linked to the p HB vector by double digestion,,and the plant expression vector was constructed.which was transformed into agrobacterium GV3101,and transformed into arabidopsis thaliana by inflocence infection.After 3 generations screening,the transgenic arabidopsis thaliana pure line was obtained.5.Observe the morphological differences between wild type transgenic plants and MhIMT1 transgenic plants under normal growth conditions,and find that MhIMT1 transgenic plants have more branches than wild type plants.The chlorophyll content of MhIMT1 transgenic plants was also significantly higher than that of wild type.6.The dry weight of MhIMT1 transgenic plants was higher than that of the wild type,which indicated that the overexpression of MhIMT1 enhanced the plant’s tolerance to dehydration.