A kind of preparation method of sagittarius protoplast
A technology of protoplasts and sagittarius, applied in the field of preparation of sagittarius protoplasts, can solve problems such as difficulties in conventional breeding of sagittarius, single genetic basis of cultivated varieties, and difficulties in breeding of sagittarius
Active Publication Date: 2017-12-29
扬州食研室食品科技有限公司
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Problems solved by technology
The conventional breeding of Sagittarius is difficult, and it is easy to cause a single genetic basis of cultivated varieties, which hinders the improvement of Sagittarius varieties
[0003] Before the present invention was made, the conventional breeding of Sagittarius was very difficult, and it was easy to cause a single genetic basis of cultivars, which hindered the improvement of Sagittarius varieties.
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[0026] 1. Arrowhead shoot tip culture: take 0.5-1cm length of Arrowhead shoot tip and inoculate it on the shoot tip medium (MS+0.5mg / L 6-BA+0.4mg / L NAA), and place it in a tissue culture room (temperature 25°C , light intensity 2500lx, light time 16h / d) for 30 days.
[0027] 2. Hydroponic pretreatment of test-tube plantlets: move the test-tube plantlets in step (1) into a jar filled with sterile water, and cultivate them for 15 days under the same culture conditions as in step (1).
[0028] 3, the enzymatic hydrolysis of protoplast: the blade that the test-tube seedling of clipping step (2) grows newly is placed in the aseptic petri dish that is covered with filter paper, blade is cut into 1-2mm wide strip with scalpel blade, Add enzyme solution, seal with parafilm, and enzymatically hydrolyze at 28°C for 12 hours in the dark. The composition of the enzyme solution is: MS+0.25% cellulase+0.25% isolated enzyme+0.25% hemicellulase+0.0055%NaH 2 PO 4 +0.06% 2-morpholineethanesu...
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The invention relates to a preparation method of sagittarius protoplast. According to the invention, the sagittarius protoplast is obtained by cultivating the shoot tip of the sagittarius, hydroponic pretreatment of test-tube seedlings, enzymatic hydrolysis of the protoplast, purification of the protoplast, and suspension of the protoplast. The invention overcomes the difficulties in conventional breeding of Sagittarius, which easily causes single genetic basis of cultivars and hinders the improvement of Sagittarius varieties. The present invention has utilized the stem tip tissue culture technique to obtain aseptic strong seedlings of Sagittarius sagittarius, which provides the best explants for the separation of protoplasts, and then adopts enzymatic hydrolysis to isolate high-quality sagittarius protoplasts, and its yield is between 2 More than 106 cells / g, the vitality is more than 80%, most of them are spherical, the cell membrane is complete and rich in chloroplast, with high vitality, which can meet the needs of various follow-up research.
Description
technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for preparing sagittarius protoplasts. Background technique [0002] Sagittarius is a perennial aquatic herbaceous plant, which is often cultivated as an annual crop in production and is native to China. It is widely cultivated in South China and the Yangtze River Basin, and the main production areas are Taihu Lake, Lixiahe and Pearl River Delta in Jiangsu. The conventional breeding of Sagittarius is difficult, and it is easy to cause a single genetic basis of cultivated varieties, which hinders the improvement of Sagittarius varieties. [0003] Before the present invention was made, conventional breeding of Sagittarius was very difficult, which easily resulted in a single genetic basis of cultivars and hindered the improvement of Sagittarius varieties. Contents of the invention [0004] The purpose of the present invention is just to overcome above-mentioned defect, dev...
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IPC IPC(8): C12N5/04
Inventor 徐小勇钱荣陈学好李良俊程立宝徐强高红胜齐晓花
Owner 扬州食研室食品科技有限公司