Study On Transformation System Of Anti-aphid Gene In Maize

The corn is one of the most widely distributed food crops in the world. Itsplant Acreage is the third in the world after wheat and rice door. The corn has always playeda very important position in China’s agricultural production and the national economy. Butin recent years, with the corn acreage has expanded each year, corn aphids and other pestsand diseases occured generally, then the corn yield and quality were badly affected.In response to this situation, the experimental maize immature embryo cultureregeneration system based on the use of Agrobacterium-mediated genetic transformation ofaphid resistance gene, designed to aphid resistance gene extracted from mint TransformationSystem EβF turninto the receptor cells and regenerated plants, and create a new maizegermplasm resistance to aphids, and lay the foundation for breeding maize varieties forresistance to aphids.Literature review a general introduction of the transgenic technology, and then focus onmaize genetic transformation of methods, the conversion of receptor conversion vectors andtransforming marker gene, etc. describes the maize genetic transformation of the researchprogress, pointed out that the situation of domestic transgenic corn is about to enter thecommercialization ofto learn from the experience of transgenic cotton industry, strictvarious aspects of genetically modified corn so that the health of genetically modified cornindustrialization step, orderly, and rapid development path, and ultimately the purpose ofgenetically modified corn industry.In this study, from the corn receptor material system optimization, optimization ofexpression vector, Agrobacterium transformation system optimization, optimization ofseveral aspects of antibiotic selection system exploration On a suitable corn transfer of genesand tissue culture optimization system.In the corn receptor material system from callus callus phenotype and the ability toreproduce the differences, and come to the Chang7-2×Hi II A good callus culture material.Therefore, the following test will be Chang7-2×Hi II A callus receptor conversionefficiency of transformation.The optimization of the expression vector system to compare the conversion of two expression vectors difference to pNCX carrier and to pBJ40for the carrier, whether it is thestate of the callus from the screening, screening resistant callus derived from the number ofblocks, orfinal number of regenerated plants have the effect of conversion to pBJ40as thecarrier as the carrier pNCX.Determine the transformation efficiency of the optimization ofAgrobacterium-mediated transformation system of Agrobacterium infection time, the callusof maize materials Zheng22×87-1and Chang7-2×Hi II A dip time must not exceed15min, to10min more suitable for the callus of different materials due to their different stateof a difference.In the optimization of antibiotic selection system, compare the differences in theantibiotic G418and Kan callus screening, an indication of the15mg/L of G418concentration screening time has been effectively inhibited the growth of maize callus, andeven to set thethe highest concentration of20mg/L, Kan, after screening twice still havefailed to wither and die in callus, corn callus is sensitive to the antibiotic G418and right Kanis not sensitive enough, prone to false positive plants.Experiments complete optimization of the system at the same time elaborated intomaize callus by Agrobacterium-mediated aphid resistance gene EβF the process, includingcallus induction, Agrobacterium infection, the resistant calliscreening, callus, seedlingemergence and root, the last transplant survival. Molecular detection, and ultimately thesurvival of plants has been a positive plants.