Studies On The Molecular Characteristics And Resistance Adversity Of Wheat Kinase Genes TaMAPKK1 And TaMAPKK3

During the growth process of plants,they will encounter various stresses of adversity.For better survival,plants have evolved various response mechanisms to respond to adverse environments.In recent years,studies have found that the mitogen-activated protein kinase(MAPK)cascade pathway plays an important role in regulating plant growth and development,hormone signal transduction,response to biological or abiotic stress,and so on.In this study,wheat stem tips and model plant tobacco leaves were used as explants to obtain transgenic plants.The function and molecular mechanisin of wheat kinase genes TaMAPKK1 and TaMAPKK3 in response to low phosphorus stress were investigated.Below are key research findings.1.Obtain the test genes TaMAPKK1,TaMAPKK3,the nucleotide sequence,the encoded amino acid sequence,andthrough NCBI,and analyze the evolution characteristics of amino acid level.Studies have shown that the proteins encoded by TaMAPKK1 and TaMAPKK3 have the typical functions of MAPKKs protein.2.Analyze the expression characteristics of the tested genes under different adversities.The results showed that,compared with normal culture,TaMAPKKI expression increased under wheat nitrogen deficiency,but the difference was not obvious under low phosphorus,simulated drought and salt stress;TaMAPKK3 expression increased during wheat phosphorus deficiency,at low nitrogen,Simulated drought and salinity,the expression difference is small.3.Using DNA recombination technology,construct pCAMBIA3301 binary expression vectors fused with TaMAPKK1,TaMAPKK3 sense and antisense sequences respectively,and use Agrobacterium-mediated genetic transformation methods to establish tobacco and wheat transformation strains to obtain overexpression and silent expression for Genetically modified tobacco and wheat plants tested.Using transgenic and wild-type tobacco strains as materials,the functions of the tested genes under different adversities were identified.The results showed that when the nitrogen supply was inadequate,compared with the wild type,tobacco plants overexpressing TaMAPKK1 had better growth,strong roots,and increased dry matter accumulation.On the contrary,the overall growth of silent expression plants was poor.Under low phosphorus stress,compared with wild type,transgenic tobacco lines over-expressing TaMAPKK3 grew vigorously,accumulated more dry matter,increased the volume of roots,and the growth of silent expression lines was poorer.Less accumulation.4.Using DNA recombination technology,fuse TaMAPKK3 amplification product upstream of pCAMBIA3300 containing GFP,construct TaMAPKK3 subcellular localization vector,transform tobacco epidermal cells by injection infection method,and use fusion protein detection method of test gene and reporter gene to identify the subcellular localization characteristics of TaMAPKK3.The results showed that TaMAPKK3 was localized in the plasma membrane after endoplasmic reticulum sorting5.Identify cytoprotective enzyme activity of wild-type and TaMAPKK3 transgenic tobacco lines under low phosphorus stress and related gene expression characteristics of cytoprotective enzyme family The results showed that under low phosphorus stress,compared with wild-type tobacco,the activities of SOD,POD,and CAT in TaMAPKK3 overexpressing plants were significantly increased,the MDA content was decreased,and the expression levels of NtSOD2,NtMnSOD2,NtCAT1,NtPOD1;2 and other increased,indicating that TaMAPKK3 may be improve the antioxidant capacity of plants by regulating the expression of related antioxidant enzyme genes6.Analyze the expression characteristics of the PIN family gene related to the PT family gene that mediates phosphorus uptake by wild-type tobacco and the root formation under Ta stress.The results showed that the transcriptional abundance of PT overexpressing plants NtPt1,NtPt2 and NtPt3 increased significantly,indicating that TaMAPKK3 may improve the transport and utilization efficiency of phosphorus by regulating the expression of the above-mentioned phosphorus transporter family genes;TaMAPKK3 overexpressing plants NtPiN1b,The increased transcriptional abundance of NtP1N2 indicates that TaMAPKK3 may improve the establishment of the root system and the whole plant by regulating the expression of related PIN genes.