Application of corn ZmWx gene in increase of corn yield and improvement of grain characteristics
A corn and gene technology, applied in the direction of plant genetic improvement, application, genetic engineering, etc., can solve the problem of few reports on the content of amylose in grains
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Embodiment 1
[0031] Example 1: Overexpression of GBSSI improves maize grain yield and amylose content Wx gene cloning and recombination
[0032]Take about 0.1 g of corn kernels in the mid-filling stage, grind them into powder with liquid nitrogen, transfer them to a 1.5 mL centrifuge tube filled with 800 μL extract solution (100 mM Tris, pH 9.0; 200 mM NaCl; 1% sodium laurate; 20 mM EDTA; 5 mDTT), vortex Vortex for 1 minute. Centrifuge at 12,000 rpm for 10 minutes at 4°C, then take 600 μL of the supernatant, add 1 / 10 volume of NaAc (2M, pH 4.0), mix well, add 300 μL of chloroform isoamyl alcohol (24:1) and 300 μL of water-saturated phenol, Vortex for 2 minutes. Then centrifuge at 12000 rpm at 4°C for 10 minutes, take 500 μL of supernatant, add 500 μL of chloroform-isoamyl alcohol (24:1), and vortex extract for 2 minutes. Then centrifuge at 12,000 rpm for 10 minutes at 4°C, take 360 μL of supernatant, add 120 μL of 8MLiCl, precipitate at 4°C for 3 hours, and centrifuge at 12,000 rpm for...
Embodiment 2
[0053] Example 2: Polymerization of transgenic ZmWx and mutant AGPase genes improves corn grain yield and starch content
[0054] The ability of GBSSI to control amylose synthesis is not only affected by the protein content of GBSSI, but also restricted by factors such as the substrate ADPG. In the case of sufficient supply of the substrate ADPG, the content of amylose synthesized by GBSSI may be further increased. In the process of starch synthesis, AGPase catalyzes G1P and ATP to generate ADPG and inorganic phosphate PPi, which provide precursor substance ADPG for starch synthesis. It is generally believed that the synthesis of ADPG is an important regulatory point of starch synthesis, and AGPase is an important rate-limiting enzyme in the starch synthesis pathway. Therefore, the obtained mutant AGPase-overexpressed transgenic maize was cross-polymerized with the GBSSI-overexpressed transgenic maize, and the grain traits, starch content and composition were detected.
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