Verification Of Biological Function Of The Gene ZmLTP3 In Maize And The Analysis Of The Promoter Cloing

Studies have shown that plants under stress conditions,the expression of ZmLTP3 gene will be greatly improved.But in no stress the gene expression level has been rarely reported.In this study,the sense and antisense chain of ZmLTP3 gene are each connected the expression vector and then transformed into maize cultivar 2416 by pollen-tube pathway to verify its biological function.At the same time,The upstream promoter sequence(1302bp)of the ZmLTP3 gene was cloned and connected expression vector,than transform the vector into Arabidopsis thaliananto,the promoter activity was verified by GUS staining,the mechanism of upstream regulation was confirmed:1? Under drought stress the over-expression of ZmLTP3 gene of maize lines in physiological and biochemical reactions are better than CK,with the prolongation of treatment time,transgenic plant height,stem diameter,leaf area,root number,root length,fresh weight,dry weight,three protective enzymes(SOD,POD,CAT)expression.Chlorophyll content is significantly higher than CK.MDA content and electric conductivity are significantly lower than the CK,after water,these differences are not reduced under high salt stress;over-expression of ZmLTP3 gene of maize strains in physiological and biochemical reactions are better than the CK,with high salt treatment time,transgenic plant height,stem diameter,Leaf area,root number,root length,fresh weight,dry weight,three protective enzymes(SOD,POD,CAT)activity,chlorophyll content is significantly higher than that of the CK,MDA content and electric conductivity was significantly lower than CK.The antisense expression under drought and salt stress of transgenic lines and CK differences were not significant.2? The promoter sequence was cloning by PCR,connected to T vector for sequencing.The sequencing results showed that Sequence length from the upstream primer to ATG is only 1302 bp,but not 2492 bp in the NCBI database,the sequencing results with Plantcare online analysis found it contains CAAT-box in the clone sequence,TATA-box promoter role of essential elements,in addition to it also containing many response to biotic stress,abiotic stress components.3? The promoter sequence was linked to the GUS gene to construct the plant expression vector and transformed into Arabidopsis thaliana.The results of transgenic Arabidopsis thaliana histochemical staining GUS showed that the sequence had promoter activity.