Molecular Cloning And Expression Analyses Of Genes Related To Potassium Channel And Transporter From Mulberry

Potassium is an essential nutrient for plants to survive,and can participate in a series of important physiological functions in plant.Due to the constant change of potassium content in soil and the existence of various forms of stress in nature,plants need to be able to transfer potassium flexibly.The aim of this study is to verify the cloning and related functions of potassium channels and transporters in mulberry trees,delve deeply into the molecular mechanism of potassium being used efficiently in plants,which created a credible molecular basis for the development of potassium innovation varieties,and improved the living environment of human beings and the sustainable development of silkworm mulberry industry.The main research contents are as follows:1.Physiological and biochemical indicators,preliminary analysis of transcriptome sequencing of low potassium stress of mulberry and identification of the different expression genes.Under the condition of low potassium stress,using the kits to determine the expression levels of POD,SOD,MDA,chlorophyll,BCA and PRO were different in different degree.The mulberry samples with normal growth and low potassium stress was selected for transcriptome sequencing,and 12.91Gb Clean Data was obtained,all samples Clean Data reached 6.44Gb and Q30 base percentage is over 87.33%,the efficiency was different from 51.62%to 53.31%.A series of bioinformatics analysis were conducted according to the comparison results,according to the expression of gene,the expression analysis,function enrichment and annotation were performed,and the expression status of 12 different expressed genes were analyzed.2.Cloning,sequence analysis and expression analysis of KAT2 gene of mulberry's internal rectification K~+channel.The MaKAT2 gene,which was obtained from the clonal population of Lu sang yu71-1,and its length was 2528bp.The length of the open reading frame was 2385bp,encoding 794 amino acid residues,theoretically,its protein molecular weight and isoelectric point are 90.073kD,7.11 respectively,belonging to the ANK-2 super family.Using bioinformatics to explore its relationship with different plant species to understand its evolutionary state.The quantitative PCR results showed that,the expression of MaKAT2 gene in drought,high salt,low temperature and low potassium these four kinds of stress conditions had different responses.3.Cloning,sequence analysis and expression analysis of AKT2 gene of K~+channel in weak inward rectified in mulberry.The MaAKT2 gene,which was obtained from the clonal population of Lu sang yu71-1,and its length was 2703bp.The length of the open reading frame was 2556bp,encoding 851 amino acid residues,theoretically,its protein molecular weight and isoelectric point are 96.901kD,6.3 respectively,belonging to the ANK-2 super family.Using bioinformatics to explore its relationship with different plant species to understand its evolutionary state.The quantitative PCR results showed that,the expression of MaAKT2 gene in drought,high salt,low temperature and low potassium these four kinds of stress conditions had different responses.4.Cloning,sequence analysis and expression analysis of KCO5 gene from mulberry KCO channel.The MaKCO5 gene,which was obtained from the clonal population of Lu sang yu71-1,and its length was 1282bp.The length of the open reading frame was 1137bp,encoding 378 amino acid residues,theoretically,its protein molecular weight and isoelectric point are 44.435kD,6.35 respectively,belonging to the Ion-trans-2 super family.Using bioinformatics to explore its relationship with different plant species to understand its evolutionary state.Gene cloning technology was used for prokaryotic expression,and the target gene MaKCO5 and pET-28a were construct an expression vector and transfer to Escherichia coli.The quantitative PCR results showed that,the expression of MaKCO5 gene in drought,high salt,low temperature and low potassium these four kinds of stress conditions had different responses.5.Cloning,sequence analysis and expression analysis of HAK5 gene of high affinity potassium transporter in mulberry.The MaHAK5 gene,which was obtained from the clonal population of Lu sang yu71-1,and its length was 2625bp.The length of the open reading frame was 2478bp,encoding 825 amino acid residues,theoretically,its protein molecular weight and isoelectric point are 92.165kD,8.37 respectively,belonging to the K-trans super family.Using bioinformatics to explore its relationship with different plant species to understand its evolutionary state.The quantitative PCR results showed that,the expression of MaHAK5 gene in drought,high salt,low temperature and low potassium these four kinds of stress conditions had different responses.