Screening For Highly Regenerative Genotypes By Barley Mature Embryo Culture And Transforming Taznf Gene Into Barley Variety By Agrobacterium-mediated Transformation

Barley mature embryos culture owns advantages of without time limit and the numberof guarantees, and is a good test system for biotechnology research. The genotypedifference is very large and most of the varieties are difficult to regenerate. Therefore,screening for high-regenerative genotypes in barley mature embryo culture andtransforming TaZnF gene with salt and drought tolerances into barley variety byAgrobacterium-mediated will be able to increase the efficiency of mature embryo cultureand genetic transformation in barley.1In order to screen barley varieties for genetic transformation,mature embryo cultureefficiency of30barley varieties was compared using the modified culture medium in thispaper.The results showed the callus induction rate of mature embryo was significantdifference in30barley varieties.Morex callus induction rate reached94.95%,short awnpurple barley callus induction rate was less than6%.The differentiation rate was alsosignificant difference in30barley varieties,Zhexiu22differentiation rate reached97.6%,Zhudamai5differentiation rate is lower,only11.85%.In the regenerationrate,different varieties have significant difference,Golden promise regeneration rate ishigher,44.7%,Kenpimai9regeneration rate is relatively low,only1.4%.6varieties ofmaterials, including Golden promise, Zhepi8, Zhexiu12, Metcalfe, Frankin, Zheda8,were suitable for genetic transformation and had higher callus inductionfrequency(Mix=94.95%) and green plant regeneration rate(Mix=44.7%).2, Based on the established regeneration system of barley mature embryo culture,the TaZnF gene super-expression vector were transformated into the embryonic callus ofbarley varieties named Golden promise by Agrobacterium-mediated. The rate of resistantcallus was81.16%, green plant regeneration rate was10.25%.From132transgenic plantsusing PCR,6plants were amplified out of the Bar gene specific bands from theregeneration,the conversion rate was4.55%;13plants out of the bands of the primer1ofTaZnF gene,the conversion rate was9.85%;43plants out of the bands of the primer2of TaZnF gene, the conversion rate was32.57%.From those62plants,2plants wereamplified out of specific bands of both of the Bar gene primers and primer1of TaZnFgene;2plants out of spcefic bands of both of the Bar gene and primer2of TaZnF gene;5plants out specific bands of both of the primer1and2of TaZnF gene,1plant with threekinds of primer bands.These results indicated that the foreign gene had been integrated intothe barley genom.