23 Jul Method for genetic transformation of Bt gene of No.23 Sichuan sugarcane by mediating agrobacterium tumefaciens
Abstract
The invention discloses a method for genetic transformation of a Bt gene of No.23 Sichuan sugarcane by mediating agrobacterium tumefaciens. The method comprises the following steps: getting an embryonic callus of receptor required by genetic transformation of the No.23 Sichuan sugarcane; pre-culturing and pre-processing the embryonic callus; preparing agrobacterium engineering bacteria liquid containing a plant expression vector and a target gene; carrying out dip dyeing to the embryonic callus through the engineering bacteria liquid; co-culturing the embryonic callus and the agrobacterium; screening and culturing kanamycin-resistant callus tissue, resistant bud and resistant seedlings; and carrying out PCR (Polymerase Chain Reaction) detection to a resistant plant. According to the method, the genetic transformation of the No.23 Sichuan sugarcane is successfully realized within a shorter dip dyeing time (1.5 to 2 minutes) through agrobacterium liquid (OD 600 of about 0.1) with lower concentration by utilizing various processing methods.
Year: 2013
Country: CN
Doc No: 103103212