23 Jul Corn transgenic method using ultrasound-assisted agrobacterium mediation of pollen
Abstract
The invention discloses a non-tissue culture transgenic method using ultrasound-assisted agrobacterium mediation of pollen and belongs to the technical field of genetic engineering. The method comprises (1) carrying out liquid culture on a target gene-containing agrobacterium strain and diluting the culture to 0.6-0.8 OD with a nutrient solution, (2) dissolving pollen in a sucrose solution with a concentration of 10-20%, carrying out ultrasonic treatment, carrying out standing and removing the supernatant and (3) mixing the agrobacterium suspension and a pollen receptor, adding 50 to 200 micromoles per liter of acetosyringone, standing the mixture for 5 to 15 minutes, transferring the precipitated treated pollen to plant stigma through a fine brush, sleeving the plant stigma with a bag, carrying out sign hanging labeling, carrying out treatment once every two days, repeating the process 2-3 times, harvesting the plant seeds after the transgenic plant seeds mature, and carrying out sowing and screening a transgenic plant through a screening label gene. The method has a clear transformation mechanism, is easy to operate, improves a conversion rate and a seed setting rate, has good stability and has a great practical value.
Year: 2017
Country: CN
Doc No: 107475283