Method for separating Eucheuma seaweed protoplast and culturing regenerated plant
A technology for protoplast and plant regeneration, applied in the fields of botanical equipment and methods, plant regeneration, horticultural methods, etc., can solve the problems of occupying a large area of water, unable to improve varieties, unstable seed breeding and production, etc., and achieve breeding efficiency. The effect of high and shortened seedling cycle
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[0019] The present invention will be further described through specific embodiments below in conjunction with the accompanying drawings.
[0020] The experimental material of this embodiment is Eucheuma ( Eucheuma denticulatum ).
[0021] 1. Protoplast isolation method
[0022] Take 1-2 grams of Eucheuma, grind or cut it into small tissue pieces or small branches, use marine shellfish (balone) digestive glands to extract and purify, and make seaweed according to the method of biochemically purifying proteins (enzymes) For tool enzymes, add 2 M glucose as an osmotic agent, and immerse small tissue pieces or small branches in the seaweed tool enzyme solution at 20-25°C for 2-3 hours, stirring with a glass rod during this period. Use a 300-mesh sieve to filter the cell mixture to remove undigested cells, cell clusters, and debris; take the supernatant and centrifuge at 800 rpm for 10 minutes to make the single cells (protoplasts) sink, and the cell debris remains in the super...
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