Method for gene expression by transient transformation of wild rice seed using agrobacterium

A method for gene expression by transient transformation of a wild rice seed using Agrobacterium, comprising the following steps:

• (1) inoculating a sterilized wild rice seed into an induction medium to allow induction culture of a callus, conducting subculture, and selecting a resulting callus with a dense structure to allow pre-culture to obtain a pre-cultured callus;
• (2) transforming a 1305Ubi-Ubi-GFP-H plasmid carrying a green fluorescent protein (GFP) gene into an Agrobacterium strain EHA105 or LBA4404 to allow culture at 28° C. for 2 d, selecting a resulting single clone to allow PCR and electrophoresis detection to obtain a positive Agrobacterium strain;
• transferring the positive Agrobacterium strain into a yeast extract broth (YEB) liquid medium to allow culture, subjecting an obtained bacterial solution to centrifugation to collect a bacterial cell, resuspending the bacterial cell in an infection buffer until a resulting bacterial suspension has an optical density (OD) value of 0.02 to 0.5 at a wavelength of 600 nm to obtain an infection bacterial solution;
• and
• (3) infecting the pre-cultured callus with the infection bacterial solution, inoculating a resulting infected callus into a co-culture medium to allow co-culture in the dark at 22° C.±2° C. for 2 d to 4 d;
• transferring a resulting co-cultured callus into a recovery medium to allow recovery culture in the dark at 28° C.±2° C. for 3 d;
• observing an infection status of a recovered callus under a stereo fluorescence microscope, wherein the GFP gene serving as an exogenous target gene is successfully expressed in the callus of the wild rice seed if the recovered callus appears green fluorescence under ultraviolet light;
• and calculating a transient expression rate.