Transformation of ACC Synthase Gene in Tree Peony, PsACS1, into Nicotiana tabacum

摘要

Ethylene plays important roles in the regulation of a number of physiological and developmental processes, and is involved in flower opening and senescence of many species.Further researches have been demonstrating that 1-aminocyclopropane-1-carboxylate synthase (ACC synthase, ACS) is a key enzyme regulating the ethylene biosynthetic pathway.To clarify the function of PsACS1 in tree peony, we constructed the sense expression vector of PsACS1 and transferred it into Nicotiana tabacum.Total RNA from the petals of cut tree peony (Paeonia suffruticosa 'Luoyang Hong') was obtained using the CTAB method.The full length eDNA of PsACS1 was cloned by PCR with primers added BamHI and SalI site sequence and transformed into pGM-T vector.After digestion with BamHI and SalI,PsACS1 was ligated with plant expression vector pBI121N in the sense way and the sense plant expression vetor pBIN-35S::PsACS1 was successfully constructed.Moreover, pBIN-35S::PsACS1 was transferred into Agrobacterium tumefaciens GV3101 and then transformed into Nicotiana tabacum 'NC89' through leaf discs.The adventitious shoots and plants were obtained under the selection pressure of 100 mg/L kanamycin.PCR analysis confirmed that PsACS1 was successfully transferred into the genome of Nicotiana tabacum.