ERAF17 Gene Function In Differentiation Of Cucumber Female Flower

ERAF17 gene, a kind of plant MADS-box gene controlling flower development, is isolated from cucumber by differential display of RNA expression.ERAF17 gene expresses differently between male and female reproductive organs. Its expression is induced by ethylene or ethylene released compounds that can promote femaleness. No evidence directly shows the relationship between ERAF17 gene and femaleness, however. So this work focus on proving the positive correlation between the expression of ERAF17 gene induced by ethylene and development of female flowers,then search for the signal pathway of cucumber female reproductive organ development.In this work the cucumber ERAF17 gene fragment was isolated via plant total RNA extraction and RT-PCR. Using PCR,enzyme digestion,ligation and bacterium transformation the vector express ERAF17 and silence ERAF17(hairpin structure) were constructed into Agrobecterium (EHA105) which were used for cucumber transformation. After infection,regeneration and Kanamycin selection,two series of transformants were yielded. We found that the phenotype of them was changed correspondingly. Normal cucumber generates male flower at lower node and female flower at a higher node position. The transformants within hairpin DNA resulted silence as expected,generating male flowers on all the node positions. Male flowers were also only generated at all the node positions in transformants with overexpression of ERAF17 gene but no female flowers. This suggested that the main reason may be due to co-suppression of ERAF17. The proofs above indicated that the ERAF17 gene indeed played an important role in female flower differentiation.We also applied E. coli to express fusion ERAF17 protein to immunize mice and the polyclonal antibodies were generated, which could be used for transformant identification.