Method for enlarging population quantity of Carpinus putoensis Cheng
A Putuo hornbeam content technology, applied in the field of botany, can solve problems such as no ecological effect, failure to meet basic requirements, limited quantity, etc.
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Embodiment 1
[0107] Embodiment 1, the preparation of basic culture medium
[0108] The basic medium (Putoensis basal medium, referred to as PT medium) was prepared as follows:
[0109] Prepare basic medium according to Table 1, pH 5.8, and sterilize at 121°C for 15-25 minutes. Among them, the preparation method of the banana extract is as follows: take 100 g of peeled fresh bananas, mash them, add 500 ml of distilled water to boil, filter with gauze after cooling, collect the filtrate, dilute the filtrate to 1000 ml with distilled water, and store it at 4°C for later use. This medium was used for both shoot induction and elongation.
[0110] Table 1. Components of PT medium
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Embodiment 2
[0113] Embodiment 2, Putuo hornbeam in vitro culture method 1
[0114] (1) Cover the branches of Putuo hornbeam with a nylon mesh bag with a hole diameter of 0.1mm×0.1mm for 1-3 months (the most suitable time is April-July every year), and after the new branches grow, take strong branches for future use .
[0115] (2) above-mentioned strong branch is cut into the cutting section of about 10cm, cleans with running water, blots the surface water droplet, rinses the leaf axil of branch with the ethanol of 70% in syringe for 3 times, then rinses branch with sterile water in syringe. Leaf axils were removed for 5 times, most of the leaves were excised, and the materials were arranged into initial explants ( figure 1 A).
[0116] Sterilize with 70% ethanol for 80 seconds on the ultra-clean workbench, soak in 0.1% mercuric chloride for 5 minutes, rinse with sterile water 8 times, dry the water droplets on the surface of the sterile material with sterile filter paper, and cut the in...
Embodiment 3
[0125] Embodiment 3, Putuo hornbeam in vitro culture method 2
[0126] (1) Take the clean and strong branches of Putuo hornbeam for later use.
[0127] (2) above-mentioned strong branch is cut into the cutting section of about 9cm, cleans with running water, blots the surface water droplet, rinses the leaf axil of branch with 70% ethanol in syringe for 5 times, then rinses branch with sterile water in syringe. Leaf axils were cut 3 times, most of the leaves were excised, and the materials were arranged into initial explants.
[0128] Sterilize with 70% ethanol for 40 seconds on the ultra-clean workbench, soak in 0.1% mercury liter for 8 minutes, rinse with sterile water for 5 times, dry the water droplets on the surface of the sterile material with sterile filter paper, and cut the initial explants into pieces Cut into 1-3cm sections for later use.
[0129](3) After inoculating the above cut section in PT medium supplemented with 1mg / l BA and 0.4mg / l NAA and culturing for 5 ...
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