Cloning Of APETALA2 Homologs From Apple And Strawberry And Genetic Transformation Of Royal Gala

In this study, MAP2 and SAP2, two homologs ofAP2 in Arabidopsis and HAP2 in Hyacinthus orientalis L., were isolated from the flower buds of apple (Mains pumila Mill.) and strawberry (Fragaria L) respectively. The sequence of both genes and the expression pattern ofMAP2 were also analyzed. The results are as follows:(1) Two API homologs, MAP2 and SAP2 were isolated from flower buds of apple and strawberry by using RT-PCR. Their accession numbers in GenBank are AF332215 and AF365367, respectively. The size ofMAP2 is 2212 bp, and it encodes a putative polypeptide of 549 amino acids. SAP2 is 1820 bp in full length, and encode a putative polypeptide of 436 amino acids. The sequencing and blast search results demonstrated that the residues of MAP2 and SAP2 are 48.8% and 40.0% identical with AP2 respectively. Both MAP2 and SAP2 have two typical AP2 domains and a putative nuclear location signal, KKSR. Taken together, MAP2 and SAP2 belong to an enlarging family: AP2 transcription factor. They may have some functions in the development of flower and seed.(2) Southern blot analysis Q?MAP2 and SAP2 showed that there are more than one copy of these genes in the genome of apple and strawberry.(3) Because the expression signal of MAP2A was hard to be detected by the northern hybridization, we combined RT-PCR and Southern hybridization to determine its expression. We found that MAP2A gene is expressed in both non-floral and floral organs, which is the same as the pattern ofAP2 and HAP2.(4) We constructed A?P2-pQE30 expression vector. The SDS-PAGE shows MAP2 gene can produce fusion protein in the E. coll..(5) Sense and anti-sense MAP2A gene expression vector under control of CaMV35S promoter were constructed. Using the leaves of 'Royal Gala' as the explants, both vectors were transformed via Agrobacterium-mediated transgene system. Some transgenic plants have been obtained now.