23 Jul Soybean embryonic tip growth point transformation method
Abstract
The invention relates to a soybean embryonic tip growth point transformation method which comprises the steps of obtaining an explant of soybean with one cotyledon and exposing the growth point; performing vacuum penetration-assisted infection by use of agrobacteria; transforming to obtain transgenic soybean. The explant of soybean is an embryonic tip exposing the growth point and obtained by removing seed coats, one cotyledon and all prophyll after sterile water culture and germination in a sterile environment. The method comprises the steps of piercing the embryonic tip with a needle point dipped with agrobacteria, and putting into agrobacteria liquid; performing vacuum penetration-assisted infection for 5-8 minutes under a pressure of 0.05MPa; absorbing the bacterial liquid on the surface; drying, culturing in sterile water for 3 days in total under a dark condition, and culturing for 7 days in total under an illumination condition; transferring into a greenhouse till inoculation, and detecting the T1 generation. Without a tissue culture stage or requiring strict sterile environment, the method provided by the invention can obtain a great quantity of regenerated plants within relatively short time, is applied to a basic operating system of soybean genetic engineering, and is of great theoretical and practical significance to the soybean quality improvement.
Year: 2014
Country: CN
Doc No: 103695463