Herbicide Resistant Gene Genetic Transformation Of Alfalfa

Alfalfa(Medicago sativa L.),as the world's oldest cultivated legume with the longest cultivation history,the largest planting area and the highest utilization value,is an indispensable resource for the development of grass and animal husbandry all over the world.With the transformation of agricultural and animal husbandry production patterns,herbicides have been widely promoted and used.Alfalfa seedling length and slow plant growth,causing rapid weed growth and impeding the growth and development of alfalfa seedlings,seriously affecting their yields.With the advent of GM crops,herbicide-tolerant GM crops have also become a common planting requirement.Since the first successful transgene in 1986,the genetic transformation of alfalfa has made some progress.However,the conversion efficiency of the existing alfalfa genetic transformation system is very low,which limits the transformation of alfalfa by exogenous genes and reduces the breeding speed of transgenic alfalfa varieties.In this experiment,leaves of the alfalfa tissue culture were used as explants.Agrobacterium-mediated method was used to successfully transfer the CP4 EPSPS gene to the receptor.During the experiment,the factors affecting genetictransformation were optimized,including glufosinate screening concentration(2mg/L),co-cultivation time(3d),agrobacterium infestation concentration(OD600=0.8),agrobacterium species(LBA105),Glyphosate spraying concentration(1%).73 strains of resistant plants were obtained.24 transgenic plants were verified by test strips and PCR tests.6 strains of positive plants were determined by test strips and glyphosate spraying.