Genetic Transformation Of Fertility Pear With α-Farnesene Synthase Gene

α-Farnesene, an acyclic sesquiterpene hydrocarbon that existed in organisms, has been proved to play an important role in attraction of codling moth chilling injury and development of scald on cold-storage apples. As one of aroma related volatiles in apple fruit,α-farnesene appears to be predominantly through classical mevalonate pathway.α-farnesene synthase (AFS) is the final, rate-limiting enzyme that converts farnesyl diphosphate (FPP) toα- farnesene in the mevalonic acid (MVA) pathway of terpene biosynthesis.Specific primers according to the full sequence ofα-farnesene synthase gene were designed. Transgenic plant was got by Real time RT-PCR and Western Blot analysis,and physiological characteristic of transgenic plant was studied .The main resμLts were as follow:(1) A recombinant plant expression vector pBI-AFS was constructed and was used to transform to'Fertility'pear.(2) Adventitious bud regeneration from the leaflet of fertility pear and efficient gene transformation system were optimized. The resμLts showed that the optimum medium for adventitious bud regeneration was MS+BA1.0mg/L+IBA0.2mg/L+sucrose30.0g/L+agar 6.0g/L, and 1/2MS+ IBA 0.3mg/L+sucrose30.0g/L+agar 6.0g/L for root regeneration.(3) RT-PCR and Real-time PCR analysis showed that AFS was highly expressed in wild type plants, much lower in transgenic pear.(4) A recombinant prokaryotic expression vector pEASY-E1-AFS was constructed by cloning the fragment of AFS and was transformed to E.Coli. Western blot analysis indicated that AFS gene expressed in the pear.(5) The transgenic plants have lower adaptability to high temperature than wild type plants. It coμLd not keep higher photosynthesis rate than the wild one in high temperature stress.