Novel method of low-gluten wheat germplasm creation
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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
Inactive Publication Date: 2012-10-17
INST OF CEREAL & OIL CROPS HEBEI ACAD OF AGRI & FORESTRY SCI
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After more than ten years of work, my country has bred a batch of high-quality weak-gluten wheat, but there are still some problems in breeding and production: lack of excellent parent resources; poor quality stability; processing quality still needs to be improved; comprehensive traits need to be further improved
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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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Abstract
The invention discloses a method for breeding low-gluten wheat. The low-gluten wheat breeding method provided by the invention comprises a step that: a Glu-1Ex gene is transferred into a target wheat, such that a transgenic wheat with a gluten strength lower than that of the target wheat is obtained. The method also comprises the steps that: the transgenic wheat is hybridized with a wheat with a gluten strength requiring lowering, such that a hybridized wheat with a gluten strength lower than that of the wheat with the gluten strength requiring lowering is obtained; the hybridized wheat is subjected to backcrossing with the wheat with the gluten strength requiring lowering, such that a backcrossing progeny is obtained; and the backcrossing progeny is subjected to selfing. As a result of experiments, with the transferring of the Glu-1Ex gene, a stably inherited HMW-GS expression co-suppression wheat material can be obtained. When the material is adopted as a breeding genetic material, and is used in backcrossing of other wheat materials with good comprehensive characteristic performances, wheat novel germplasm with reduced gluten strength can be obtained. Therefore, the creation of high-quality low-gluten wheat suitable for cookie and pastry processing can be promoted.
Description
technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for cultivating low-gluten wheat, in particular to a method for transforming Glu-1E b X gene method to obtain stable genetically co-suppressed HMW-GS expressing low-gluten wheat varieties. Background technique [0002] Wheat is one of the three important food crops (wheat, rice and corn) in the world. Wheat seed storage proteins account for about 50% of the total seed protein, and their structure and characteristics play an important role in the quality of flour processing. Wheat gluten is mainly composed of gliadin and glutenin, which determine the extensibility and elasticity of dough respectively. Glutenin is a multimer, which is formed by cross-linking multiple subunits through disulfide bonds, and its molecular mass is as high as several million Daltons (Da). After separation by isoelectric focusing and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAG...
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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00A01H1/02
Inventor 李辉胡梦芸张颖君刘茜
Owner INST OF CEREAL & OIL CROPS HEBEI ACAD OF AGRI & FORESTRY SCI
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