Novel method of low-gluten wheat germplasm creation
A technology for wheat and hybrid wheat, which is applied to low-gluten wheat varieties that co-suppress HMW-GS expression and cultivates low-gluten wheat, can solve the problems of improved processing quality, further improvement of comprehensive traits, poor quality stability and the like
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Embodiment 1
[0036] Embodiment 1, transfer Glu-1E b Obtaining x-gene Wheat and Identification of Its HMW-GS
[0037] 1. Recombinant expression vector pCAMBIA3301-Glu-1E b construction of x
[0038] Glu-1E derived from Thinopyrum bessarabicum b The x gene (GenBank: AY525782, sequence 1) was used as a template, and the upstream and downstream primers were used as degenerate primers to amplify Glu-1E with restriction sites Xba I and Sac I at both ends. b x gene fragment.
[0039] Upstream primer: 5′-GAC TCTAGAA TGGCTAAGCGGC / TTA / GG-3' (the underlined part is the recognition site of Xba I, and the following sequence is the 1st-25th position of sequence 1)
[0040] Downstream primer: 5′-CG GAGCTC GCTATCACTGGCTG / AGCC-3' (the underlined part is the recognition site of Sac I, and the 10th-27th position is the reverse complementary sequence of the 1794-1818th position of sequence 1)
[0041] After the PCR product was double-digested with Xba I and Sac I, it was ligated to the backbone fragm...
Embodiment 2
[0063] Embodiment 2, obtain low-gluten wheat variety by hybridization
[0064] 1. Obtaining hybrid wheat and backcross progeny and identification of HMW-GS expression
[0065] 1. Acquisition of new germplasm of wheat with complete deletion of HMW-GS and identification of HMW-GS expression
[0066] T obtained in Example 1 1 Generation Lg-1-1 was used as the male parent, and the recipient wild-type wheat variety Gaocheng 8901, wheat variety Xiaoyan 54 and Jinmai No. 1 The generation Lg-1-1 was used as the female parent, and the wild-type wheat variety Gaocheng 8901 was used as the male parent for backcrossing. On the one hand, SDS-PAGE electrophoresis was performed on the seeds of the above four combined hybrid progenies, and on the other hand, Western hybridization was performed to analyze the expression of HMW-GS. For the experimental operation method, refer to the relevant content of Example 1. Experiments were repeated three times.
[0067] SDS-PAGE results such as Fi...
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