Cloning And Functional Study Of A Regulation Gene Related To Phyllotaxy Develope In Maize (Zea Mays L.)

Maize with the opposite phyllotaxy and also initiating ears in opposite pairs is an aberrant mutant.The opposite phyllotaxy character is a new type of phyllotaxy location of maize and it is also a precious material to maize breeding and plant evolution. Methoding the gene chip, we studied the difference of between Maize with the opposite and alternate phyllotaxy at3-level:H4D/d, BeiD/d,5003D/d. The gene, treatment called Zmedp, may control the develop of Maize with the opposite phyllotaxy in3-level of Maize with the opposite and alternate phyllotaxy.The research of ZMedp gene and Osedp gene is by constructing the molecular regulation of overexpression vector and RNAi expression vector. And we selected inbreed lines included wild maize and transgenic maize, compared the endogenesis hormone by ELISA method. Results are follows:1. After ZMedp gene was obtained by RT-PCR, the overexpression vector and RNAi expression vector of ZMedp gene were constructed and transformed into maize mediated by Agrobacterium. Finally transgenic maize for ZMedp overexpression and RNAi were picked out by PCR.2. We studied the tissue-specific expression in ZMedp by RT-PCR method. The result shows that the ZMedp gene is espressed in roots, stems, and lesves and shoot tip. The strongest expression is in the shoot tip, followed by the root, again is in the stem, and expression in the leaves could hardly see.3. To determine the subcellular localization of the ZMedp protein, we generated p1302-ZMedp-mGFP fusion construct under the control of CaMV35S promoter. Then fusion constructs was bombarded into the onion epidermal cells. The results of this experiment revealed that the fusion protein is expressed in the nuclei of the onion epidermal cells. This data thus indicate that the ZMedp protein is nuclear protein.4. The study coppare the contents of endogenous Cytokinin and auxin in wild maize and transgen maize by the ELISA method. It suggests that the contents of Cytokinin were decresed while that the contents of auxin was improved. This shows that ZMedp gene work for cytokinin and auxin synthesis, yet this gene weren’t reported in the study of corn. It is a new regulation of endogenous hormone gene.5. Find the homologous gene of ZMedp in rice, and select300bp as RNAi fragment. The fragment was obtained by PCR, the RNAi expression vector of Osedp gene was constructed and transformed into rice mediated by Agrobacterium. Finally transgenic maize for Osedp RNAi were picked out by PCR.