| One of the important aims of wheat quality breeding is improve the breadmaking quality. It is very important to breadmaking quality of wheat that if the HMW-Glu(5+10)is exist and how much it is. In order to get wheat lines with over-expression of high molecular weight glutenin subunit 5 and subunit 10, Yumai 18, a widely used wheat cultivar in Henan Province ever was transformed with a high-efficient expression vector with the target genes by means of biolistic bombardment and Agrobacterium tumefaciens. To improve the transgenic efficiency, this paper also investigated some parameters of biolistic bombardment and Agrobacterium-mediated transformation at the same time. Below is the result:A high-efficient expression vector was constructed in order to enchance expression of target genes and overcome gene silencing. The method was mainly to attach KDEL gene after target genes, and insert Mar of wheat into vector. Thus the over-expression vector with over-expression of high molecular weight glutenin subunit 5 and subunit 10 was constructed. It was tested that the vector was constructed successfully via enzyme cleavage.Calli derived from 9565 wheat immature embryos were transformed by biolistic bombardment, and 169 regenerated plantlets were gotten. PCR identification showed that among all transgenic plants there were 137 showing sub 5, The PCR positive ratio was 81.1%;There were 142 showing sub 10, The PCR positive ratio was 84%. and 116 sub 5+10 positive, The PCR positive ratio was 68.6%. Most of the regeneration plantlets had seeds, but several had no seeds.The several key parameters were also improved. The results showed that the immature embryos with 3 d pre-cultured were better than that with 10 d pre-cultured. The average green spot ratio of Au dosage of 250 μg per shot and target distance of 12 cm were the highest. 20 mg· L-1 Hygromycin was best as resistant selective reagent...
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