Functional Characterization Of Arabidopsis Amino Acid Permease1(AtAAP1) In Transgenic Maize

Amino acid permease(AAP)is a transporter related to amino acid,which is located on the cell membrane.Recent studies have shown that many plant amino acid transporters have been identified in model plant Arabidopsis thaliana.In this study,we use turn endosperm-specific promoter maize and transfer molding promoter maize as materials to study the function of AtAAP1 gene in maize.In this study,the expression levels of AtAAP1 gene in the seeds of turn endosperm-specific promoter maize are analyzed by qRT-PCR,the physiological and biochemical indexes of roots,leaves and seeds of turn endosperm-specific promoter maize positive and negative plants were also determined: free amino acid content,soluble protein content,total nitrogen content,20 kinds of amino acids and composition,alpha and beta gliadin content,proline content and lysine content.In order to understand the function of AtAAP1 gene and the mechanism of nitrogen utilization in maize,and to make the AAP genes using in industrialization.The main results were as follows: 1: Determination the content of free amino acid,soluble protein,proline and lysine in two kinds of transgenic seeds of different growth stage,founded that the content of physiological index in turn endosperm-specific promoter maize seeds are the highest.In the silking stage,the content of free amino acid and soluble protein are highest in the two kinds of transgenic maize.2: The expression of AtAAP1 gene in seeds of turn endosperm-specific promoter maize are highest compared with the negative seeds.The expression of AtAAP1 gene was highest in mature stage.3: Determination the content of free amino acids and soluble protein of turn endosperm-specific promoter maize's positive and negative plants,at different growth of roots,leaves and seeds,founded that the contents of free amino acid and soluble protein were the highest in the roots and leaves of transgenic maize at the time of silking stage,and soluble protein was the highest in transgenic seeds at maturity.Positive maize plants of roots and seeds of free amino acids and soluble protein content were higher than negative maize plants,while the content of free amino acids and the soluble protein in the leaves is opposite to that in the roots.The free amino acid content in roots of positive maize plants was 1.44 times of that in negative maize plants.The soluble protein content was 1.69 times of that in negative maize plants,free amino acids content in positive maize seeds was 1.36 times of that in negative maize seeds,the content of soluble protein was 1.69 times of that in negative maize seeds.4: Analysis the content of total nitrogen,20 kinds of amino acids in roots,leaves and seeds of turn endosperm-specific promoter maize's positive and negative plants,founded that positive maize plant's reproduction organs are higher than negative maize plants.The content of neutral amino acid and glutamate in positive maize seeds increased obviously.5: Use the BandScan to analysis the gliadin content of turn endosperm-specific promoter maize seeds,the results showed that the gliadin content of positive maize seeds are higher than it in negative maize seeds.