Technique for preparing Sudan grass protoplast by using vacuum enzymolysis method
A technology of protoplast and enzymatic hydrolysis, which is applied in the direction of plant cells, etc., to achieve the effect of good cell integrity, less cell death, and simple method
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[0017] Embodiment 1: Preparation of Sudan Grass S722 protoplasts
[0018] 1. Preparation of sudan grass seedlings: select plump sudan grass S722 seeds, plant them in a nutrient bowl, and cultivate them at 25°C for 8 days.
[0019] 2. Treatment of young leaves and stems: cut 1 g of young leaves and stems of sudan grass S722, and cut the leaves into about 1 mm length with a blade.
[0020] 3. Vacuum enzymatic hydrolysis: put the cut stems and leaves into the triangular flask with 10mL of enzymatic hydrolysis solution, and put it in a vacuum environment (-0.1MPa) for 1 hour. After taking it out, cover the mouth of the triangular flask with tin foil ℃ shaker, rotating speed 40rpm, enzymatic hydrolysis for 3 hours. (enzymolysis solution formula: 0.645M mannitol, 10mM potassium chloride, 1mM calcium chloride, 8mM 2-morpholineethanesulfonic acid, pH5.5, 1.5% cellulase and 0.3% isolated enzyme R-10).
[0021] 4. Dissociation of protoplasts: Pour off the enzymatic solution, add 10ml ...
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