Construction Of Plant Expression Vector And Genetic Transformation Of α-Farnesene Synthase Gene From Peel Tissue Of Apple

Superficial scald, a physiological storage disorder of apples and pears, can reduce edible quality (especially appearance quality), market value, and the resistance to disease; hence lead substantial loss in economy.α-farnesene has been associated with the occurrence of scald. The accumulation ofα-farnesene in the skin of apple fruit during storage appears to be predominantly through classical mevalonate pathway .3-Hydroxy-3-methylglutaryl CoA reductase (HMGR), farnesyl diphosphate synthase (FPPS) andα-farnesene synthase (AFS) is the rate-limiting enzyme in the mevalonate pathway. AFS is the final, rate-limiting enzyme that converts farnesyl diphosphate (FPP) toα-farnesene in the mevalonic acid (MVA) pathway.In order to study the expression ofα-farnesene, we designed specific primers according to the full sequence ofα-farnesene synthase gene, constructed both sense and RNAi vector,and transformed both tobacco and apple (white pearman).To futher improve the regeneration system of apple (white pearman), factors related to the regeneration of apple (white pearman) were studied. We also transformed apple(white pea rman) Via Agrobacterim Tumeficience infection, in order to screen for new apple cultivars.1. Specific primers for PCR designed from AFS-1 had been registered in GenBank (No.AY563622)2. A recombinant plant expression vector pBI-AFS was constructed and it was transformed to Tobacco.3. A recombinant prokaryotic expression vector pET-AFS was constructed and it was transformed to E.Coli. Western blot analysis indicated that AFS gene expressed in tobacco.4. A recombinant plant expression vector RNAi was constructed and do genetic transformation.5. Basal medium, hormone concentration and composition and sucrose concentration, et.were investigated for their effects on apple adventitious shoot regeneration.6. On the basis of regeneration system from in vitro leaves of apple, factors influencing the frequency of gene transfer were examined and an efficient gene transformation system for apple was established.