A kind of method for improving wheat processing quality
A wheat and genetically modified wheat technology, applied in chemical instruments and methods, botanical equipment and methods, biochemical equipment and methods, etc., can solve the problems of poor baking quality, easy rheology of dough, and difficult processing.
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Embodiment 1
[0088] Embodiment 1, the acquisition of wheat mutant gene 1Dx2m2
[0089] 1. PCR amplification of the full-length sequence of the wheat 1Dx2 gene coding region
[0090] Genomic DNA of common wheat cultivar Konong 199 was used as a template to perform PCR amplification using specific primer combination Dx2F and Dx2R, high-fidelity PrimeSTAR HS DNA polymerase and 2×PrimeSTAR GC Buffer (Takara, Dalian). The obtained PCR amplified fragment 1Dx2 ( figure 1 ) is the full-length sequence of the coding region of the 1Dx2 gene. The nucleotide sequences of the primer combinations Dx2F and Dx2R are respectively Sequence 1 and Sequence 2 in the sequence listing.
[0091] The PCR program for amplifying the full-length sequence of the 1Dx2 coding region was as follows: pre-denaturation at 98°C for 3 min; denaturation at 98°C for 30 sec, renaturation and extension at 72°C for 3 min, 38 cycles; and finally extension at 72°C for 10 min.
Embodiment 2、1Dx2
[0101] Embodiment 2, the acquisition of 1Dx2m2 transgenic wheat
[0102] 1. Creation of transgenic receptor materials
[0103] The transgenic acceptor material used in the present invention is the high-molecular-weight glutenin subunit 1Dx2 deletion mutant of common wheat variety Kenong 199 (KN199), denoted as kn2 - . kn2 - Mutants were obtained by EMS mutagenesis method. The specific process is as follows: (1) choose 1000 plump wild-type Ke Nong 199 seeds and use 0.4% EMS (Sigma, M0880) to treat them in the dark for 24 hours and then wash them with running water for 24 hours. These seeds are denoted as M 0 ;(2) M 0 After 4 weeks of normal germination and vernalization (4°C), the seeds were planted in the Beijing Changping Experimental Base of the Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, and managed in the normal field. The harvested seeds were recorded as M 1 ; using SDS-PAGE method from M 1 The 1Dx2 subunit deletion mutant was scree...
Embodiment 3、1
[0116] Example 3, Determination of processing quality traits of 1Dx2m2 transgenic lines and comparative analysis with recipients, wild-type Kenong 199 and 1Dx2 transgenic lines
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