Transferring Pyruvate Orthophosphate Dikinase (PPDK) Gene Form Echinochloa Crusgalli Var Frumentacea Into Wheat(Triticum Aestivum L.) Mediatede By Agrobacterium Tumefaciens

The dependence of genotype sensitivity of receptor restrained the scope of the agrobacterium-mediated wheat receptor.The single access to obtain explant limits the number of receptor materials, while the wheat transformation system which we had was not versatility result in the low efficiency of agrobacterium-mediated transformation,and the low efficiency was one of the main reasens why the gene transformation of wheat was fall behind compared with other field crops.In order to reform the C₄ gene of wheat genome,and improve photosynthetic efficiency,we must increase the efficiency of gene transformation.Various factors were found to restrain the wheat transformation system:such as receptor gene,vector type,infection factor,differentiation factor and random factor.This paper began to expand the numbers of the explant and identify the wheat species which are sensitivity patterns,by the method of transformation expression of gus gene,aimed to integrate the limiting factor mentioned above,improved transformation efficiency,and gain the transformation plant,to build a high efficiency conversion system.The results showed that:1)The recover ratios of mature embryo of jimai21,shandong01-35,handan3475,shiluan02-1 and jin57 were above 60%,the number of explants can be increased by increasing the number of inducing mature embryo,and which can be used for optimization system materials.The recover ratios of 14 immature embryos were above 85%,and they can be used for the main transformation materials.2)Identified the agrobacterium-infected toxicity of strain C58C1,the results showed that the toxicity levels were fit to infect.Infected 34 different materials of mature embryos and 14 calli of immature embryos,detected Yangmai10,yangmai158, shiluan02-1,shijiazhuang5,zhoumai18,shandong01-35,taishan008,Y9-63,handan3475 by the method of PCR,The Pyruvate Orthophosphate Dikinase Gene and hpt can be expressed,so they were all sensitivity receptor wheats;The genes of the immature embryos of yangmai10,yangmai158 and the mature embryos of shiluan02-1 can be expressed,and the best gene expression ratios were that the regeneration recovery which had been trained for 14 days,so they were optimization system materials. 3)Yangmai10,yangrnai158,shiluan02-1as optimize materials,detected their recover ratios and genetic expressions,the best infection effectiveness of yangmai158 and yangmai10 were that the callis had been preincubated for 60 days,and the bacilli concentrations were OD600 0.6-0.8 and OD₆₀₀0.5-0.7,the best infection time were 40-50min and 30-40min respectively;while the best infection effectiveness of shiluan02-1 was that the calli had been preincubated for 180 days,the bacilli concentrations and infection time were OD₆₀₀0.4-0.6 and 20-30min.Sterile filter paper can be used as buffer preparatory for improving Gus gene expression,The differentiation of Greenpoint was induced in the culture medium which had any excitatory autacoid,and for promoting the bud points to shape, the concentration of NAA and 6-BA were adjusted to 1mg/L,0.2 mg/L.In order to promote bud points differentiated and rooted,the bud points were provided low nutrition and growth hormone.