The Function Ananlysis Of BpMYB41 Gene In Betula Platyphylla

MYB family is one of the largest members of transcription factors.Which is a structure combined by containing MYB domain of genes.And most belong to the R2R3 MYB proteins-the class.In plant cell differentiation,organ formation and morphology and so on has certain adjustment.First of all,This study get 11 MYB transcription factor gene,which have complete ORF.Then using Bioedit,clustalx and MEAG6 analysis software for bioinformatics analysis,and by using the RT-PCR technology which genes expressed in different under the condition of quantity are analyzed.These conditions mainly including root,stem and leaf of different groups;Methyl jasmonic acid and ethylene,abscisic acid and kinetin different hormone;NaCl and Mannitol stress under different processing time.The results showed:(1)Of all the MYB gene,The expression of BpMYB41 and BpMYB8 is highest in leaves,most of the MYB gene expression in root and stem volume were significantly higher than that of leaf of expression,This description MYB gene expression has tissue specificity;(2)Most MYB genes under four different hormone treatment the overall trend of the expression pattern of basic consistent:Gene expression of basic are expressed in the cut,which under jasmonic acid,ethylene and abscisic acid treatment.And When using kinetin(KT)treatment,the gene expression of all were higher expressed.But there are two more specific genes,one is BpMYB2 which is up-regulated under the four kinds of hormone treatment,the other is BpMYB11,it is the opposite of BpMYB2,which is down-regulated under the four kinds of hormone treatment.(3)Between the different genes,it expression has great difference,but the expression of the overall trend is basically the same.When treated with 200 mm NaCl and 300 mm Mannitol stress under different time.The BpMYB5?BpMYB9 and BpMYB41 introduced into onion epidermal cells by particle bombardment,the results showed that BpMYB5?BpMYB9 and BpMYB41 is a nuclear protein.Comprehensive bioinformatics analysis and the quantitative results,this paper selected the BpMYB41 genes for further studies.BpMYB41 gene transient expression is analyzed in birch.Instantaneous expression is also analyzed under different stress treatment research.The results showed:Overexpression of pROK ?-BpMYB41 restructuring carrier can make BpMYB41 gene expression in plant of excess.And suppression expression of RNAi-BpMYB41 recombinant expression vector can inhibit BpMYB41 the expression of genes in plants.And under different stress treatment,overexpression and suppress expression of BpMYB41 expression appeared significant change,This shows that BpMYB41 gene has certain function of resistance.overexpression and suppress expression of BpMYB41,empty expression vector plants and wild type birch using NBT,DAB,and Evans Blue for histochemical staining,The staining results showed that BpMYB41 gene can reduce the content of hydrogen peroxide and O2-in birch,which can reduce the cell damage level.These results demonstrated that BpMYB41 can enhance resistance of birch to stress.In order to study the influence of BpMYB41 gene for the expression of resistance and material related genes.By using the RT-PCR technology makes quantitative analysis.Quantitative PCR results of the material related genes show:under the influence of the pROK II-BpMYB41 almost all of the related gene expression quantity are up-regulated.But under the influence of the RNAi-BpMYB41 almost all of the related gene expression quantity are down-regulated.It indicate that BpMYB41 gene can regulate the expression of deposition of secondary cell wall.It prove BpMYB41 with these genes control the development of the xylem.Quantitative PCR results of the resistance related genes(SOD?POD?P5CDH?P5CS)show:Under NaCl stress treatment almost all of the related gene expression quantity are up-regulated.This shows that resistance genes can be subjected to the NaCl stress-induced.Which can be induced in response of Birch to salt stress.under the influence of the pROK ?-BpMYB41 almost all of the related gene expression quantity are up-regulated.This suggests that the pROK ?-BpMYB41 can induce the expression of resistance genes.Therefore proved BpMYB41 gene which has the function of a certain resistance.But under the influence of the RNAi-BpMYB41 almost all of the related gene expression quantity are down-regulated.It indicate that RNAi-BpMYB41 can inhibit the expression of resistance genes,It proved that BpMYB41 gene has certain function of resistance.The overexpression of BpMYB41 and wild type Arabidopsis were treated under NaCl,and mannitolstress.The results showed that the germination rate of the transgenic plants is higher than that of the wild type plants.It proved that BpMYB41 gene has certain function of resistance.Using yeast two-hybrid system for BpMYB41 gene transcription activation detection.Results indicate that BpMYB41 has no transcriptional activation activity.And the technology was used to study the interaction relationship between BpMYB41 with BpMYB3,BpMYB6 and BpMYB8.Results indicate that BpMYB41 gene and the other three BpMYB proteins have no interactions.