摘要
The expression vector pBPC30, which carries the high molecular weight glutenin subunit (HMW-GS) 1Dx5 and 1Dy10 genes, was transferred into hexaploid winter wheat cv. Jinghua No. 1, Jing411 and Jingdong No. 6 ex-plants of immature embryos and immature inflorescence by particle bombardment. A large number of resistant trans-genic plants were obtained under the selection of herbicide bialaphos or phosphinothricin (PPT). Confirmed transgenic plants of T0 generation showed successful integration of HMW-GS genes and bar gene into the wheat genome. T1 generation of transgenic plants can resist 20150 mg/L PPT. Protein analysis of T2 seed by SDS-PAGE showed that HMW-GS 1Dx5 and 1Dy10 genes were well expressed in offspring seed of transgenic lines by co-expression with or substitution of endogenous 1Dx2 or 1Dy10. In one transgenic line, TG3-74, a new protein band between endogenous pro-tein subunits 7 and 8 (marked as 8*) of glutenin appeared, but endogenous subunit 8 (encoded by 1By8 gene) was ab-sent. Analysis of gluten rheological quality on seed proteins of 102 T3 plants showed that the sedimentation value of 5 transgenic lines (44.249.0 mL) was remarkably improved, 59.6%64.3% higher than that of wild type Jinghua No. 1 and Jingdong No. 6, similar to bread wheat Cheyenne (48.0 mL). Analysis of dough rheological properties of transgenic lines showed that the dough stable time of 5 transgenic lines range from 16 to 30 min, whereas the dough stable time of wild type was only between 37 min. Our research suggests that introducing novel HMW-GS genes into wheat is an effi-cient way to improve its bread-making quality.
The expression vector pBPC30, which carries the high molecular weight glutenin subunit (HMW-GS) 1Dx5 and 1Dy10 genes, was transferred into hexaploid winter wheat cv. Jinghua No. 1, Jing411 and Jingdong No. 6 ex-plants of immature embryos and immature inflorescence by particle bombardment. A large number of resistant trans-genic plants were obtained under the selection of herbicide bialaphos or phosphinothricin (PPT). Confirmed transgenic plants of T0 generation showed successful integration of HMW-GS genes and bar gene into the wheat genome. T1 generation of transgenic plants can resist 20150 mg/L PPT. Protein analysis of T2 seed by SDS-PAGE showed that HMW-GS 1Dx5 and 1Dy10 genes were well expressed in offspring seed of transgenic lines by co-expression with or substitution of endogenous 1Dx2 or 1Dy10. In one transgenic line, TG3-74, a new protein band between endogenous pro-tein subunits 7 and 8 (marked as 8*) of glutenin appeared, but endogenous subunit 8 (encoded by 1By8 gene) was ab-sent. Analysis of gluten rheological quality on seed proteins of 102 T3 plants showed that the sedimentation value of 5 transgenic lines (44.249.0 mL) was remarkably improved, 59.6%64.3% higher than that of wild type Jinghua No. 1 and Jingdong No. 6, similar to bread wheat Cheyenne (48.0 mL). Analysis of dough rheological properties of transgenic lines showed that the dough stable time of 5 transgenic lines range from 16 to 30 min, whereas the dough stable time of wild type was only between 37 min. Our research suggests that introducing novel HMW-GS genes into wheat is an effi-cient way to improve its bread-making quality.
基金
This work was supported by the National Special Program of Transgenic Plants Research and Development(Grant No.J2000-B-017).
关键词
转基因技术
麦谷蛋白
冬小麦
基因表达
wheat (Triticum aestivum L.), HMW-GS, transgenic, expression, transgene inheritance, bread-making quality.
