Transgenic Arabidopsis Lines Expressing Gene ZmGnT- Ⅲ Affect The Chlorophyll Concentration Of Seedling

N-acetylglucosamine transferase III (GnT-III) is a special glycosyltransferase which transfers the N-acetylglucosamine (GlcNAc) to the core β Man of N-glycan and generates bisecting GlcNAc chain. Once the reaction completes, the other glycosyltransferases of N-glycan oligosaccharide chain formation will be inhibited. Resulting in the changes of N-glycan construction and the fuction of glycoprotein.The prophase of this project got a lot of insertional mutagenesis M1 and M2 groups by crossing the active Mu transposon material and maize B73. Amplified the Mu transposon inserted fragments which is different with genomic DNA by optimized thermal asymmetric PCR. Analysed the flanking sequences of target sites and screened the genes related to maize important agronomic traits. The results showed that albino seedling phenotypic was closely related to GnT-III gene mutation.To furtherly investigate the relationship between GnT III and chlorophyll content in plants, GnT-III gene of maize B73 was cloned. Then, GnT-III gene was connected to plant expression vector pMDC45/46/47/83/84/85 by Gateway technique and transformed Agrobacterium tumefaciens strain GV3101. Transfered the recombinant expression vectors into wild-type Arabidopsis plants by the dip method. Screened the transgenic plants by hygromycin resistance and ultimately obtained T3 transgenic lines 46,47,83,84. Identified that GnT-III transferred into wild-type Arabidopsis genome at the molecular level. Western blot experiment of transgenic lines showed that GnT-III expressed in the transgenic lines 46,83. Then, the green fluorescences of leaves and roots of transgenic lines 46,83 were observed. The result indicated that the fusion protein may be located in the cell membrane.The growth traits of transgenic lines 46,83 were analyzed. Comparing with wild-type Arabidopsis, the transgenic lines 46,83 has more leaves, larger blades, more lateral roots, and longer roots; Leaves of 46,83 transgenic plants, especially on the main stem, show oval and more trichomes which is different with wild-type under the same growing conditions. In addition, comparing with wild type, chlorophyll a, chlorophyll b and total chlorophyll content of transgenic lines 46 shows markedly significant difference, and the transgenic lines 83 only has significant difference in chlorophyll b and total chlorophyll content. These results indicate that overexpression of maize GnT III genes have an impact on Arabidopsis seedling growth and a substantial increase in chlorophyll content of seedlings.