Transformation of transcription factor DREB1C gene into the fast-growing black locust mediated with Agrobacterium tumefaciens

Robinia pseudocacia,introduced from Hungary,is a fast-growing variety used in ornamental,soil and water conservation,fodder and nectar sources.Transformation of this woody plant using abiotic-stress-tolerant gene could meet the requirements for reclamation of arid or alkalid lands and vegetation restoration.In this paper,three aspects of R.pseudocacia were studied: shoot regeneration on MS medium with different combinations of 6-BA and NAA,optimization of β-glucuronidase(GUS) gene(carried by vector pCAMBIA 1301,and mediated by Agrobacterium tumefaciens) transformation,and gene transformation using the transcription factor AtDREB1C gene (abiotic-stress-tolerant gene carried by the same vector).Results showed that: 1) The highest shoot-regeneration rate 77.5% was obtained from the rachis of compound leaves cultured on MS base medium combimed with 6-BA 1.0 mg/L and NAA 0.5 mg/L;2) The best transformation efficiency was proved by the rachis of compound leaves proculturing for 0—2 days on the MS medium,immerging into the solution containing Agrobacterium and acetosyingone(AS) for 15-20 min,subsequencely co-culturing for two days on the same medium with AS 20 mg/L and hygromycin(Hyg) 7 mg/L;3) Following the method optimized,the AtDREB1C gene was introduced into R.pseudocacia,and transgenic plants were obtained and verified by PCR,PCR-Southern blotting and Southern blotting analysis.