The invention discloses a construction method of an
agrobacterium tumefaciens mediated
masson pine genetic transformation
system, and belongs to the technical field of
genetic engineering. According to the construction method of the
agrobacterium tumefaciens-mediated
masson pine genetic transformation
system, immature
masson pine cones are collected, embryogenic calluses are induced and cultured, the embryogenic calluses of the masson pine in a PEM III stage are used as a material, suspension pre-culture is performed to obtain an
acceptor material, and the
acceptor material is subjected to genetic transformation to obtain the
agrobacterium tumefaciens-mediated masson pine genetic transformation
system. The method comprises the following steps: firstly, carrying out
tissue culture on a transgenic somatic
embryo, infecting by adopting an agrobacterium infection solution with an OD600 value of 0.2-0.6, carrying out co-culture,
recovery culture and screening culture, carrying out
histochemical staining by adopting GUS, carrying out
molecular identification to obtain a transgenic embryogenic
callus, and carrying out maturation treatment to finally obtain the transgenic somatic
embryo. The masson pine genetic transformation system established by the invention is stable in transformation performance and high in transformation positive rate, and when an
acceptor material is masson pine embryonic
callus with the
genotype of 1-3-5, the transformation positive rate is as high as 100%.