Method for induction and subculture of taxus chinensis leaf callus and culture medium used by method
A technology for inducing culture medium and callus, applied in the directions of botanical equipment and methods, horticultural methods, applications, etc., can solve the problem that taxus yew resources cannot meet the demand for paclitaxel production, and achieves low pollution rate and induction rate. High, fast growth effect
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Embodiment 1
[0028] A method for inducing a yew leaf callus, specifically comprising the steps of:
[0029] (1) Disinfection of explants
[0030] Take the young leaves of Taxus chinensis in the same year, wash them with running water for 4 hours, disinfect them with 70% alcohol for 30 seconds, disinfect them with 1% mercury chloride for 8 minutes, and rinse them with sterile water;
[0031] (2), callus induction culture
[0032] Scratch the leaves of the sterilized yew explants obtained in step (1) with a blade and inoculate them on the induction medium, and culture them in the dark at a controlled temperature of 20-22°C for 20 days to obtain callus after induction ;
[0033] The induction medium, calculated per liter, consists of 400mg NH 4 N0 3 , 556mg Ca(NO 3 ) 2 .4H 2 O, 990 mg K 2 SO 4 , 96 mg CaCl 2 .2H 2 O, 170 mg KH 2 PO 4 , 0.25 mg Na 2 MoO 4 .2H 2 O, 370 mg MgSO 4 .7H 2 O, 22.4mg MnSO 4 .H 2 O, 8.6mg ZnSO 4 .7H 2 O, 0.25mg CuSO 4 .5H 2 O, 27.8 mg FeSO 4 ....
Embodiment 2
[0038] A method for inducing a yew leaf callus, specifically comprising the steps of:
[0039] (1) Disinfection of explants
[0040] Take the young leaves of Taxus chinensis in the same year, rinse them with running water for 4 hours, disinfect them with 70% alcohol for 30 seconds, disinfect them with 1% mercury chloride for 8 minutes, and rinse them with sterile water;
[0041] (2), callus induction culture
[0042] Scratch the leaves of the sterilized yew explants obtained in step (1) with a blade and inoculate them on the induction medium, and culture them in the dark at a controlled temperature of 20-22°C for 20 days to obtain callus after induction ;
[0043] The induction medium, calculated per liter, consists of 400mg NH 4 N0 3 , 556mg Ca(NO 3 ) 2 .4H 2 O, 990 mg K 2 SO 4 , 96 mg CaCl 2 .2H 2 O, 170 mg KH 2 PO 4 , 0.25 mg Na 2 MoO 4 .2H 2 O, 370 mg MgSO 4 .7H 2 O, 22.4mg MnSO 4 .H 2 O, 8.6mg ZnSO 4 .7H 2 O, 0.25mg CuSO 4 .5H 2 O, 27.8 mg FeSO 4 ...
Embodiment 3
[0048] A method for inducing a yew leaf callus, specifically comprising the steps of:
[0049] (1) Disinfection of explants
[0050] Take the young leaves of Taxus chinensis in the same year, wash them with running water for 4 hours, disinfect them with 70% alcohol for 30 seconds, disinfect them with 1% mercury chloride for 8 minutes, and rinse them with sterile water;
[0051] (2), callus induction culture
[0052] Scratch the leaves of the sterilized yew explants obtained in step (1) with a blade and inoculate them on the induction medium, and culture them in the dark at a controlled temperature of 20-22°C for 20 days to obtain callus after induction ;
[0053] The induction medium is calculated per liter, consisting of 1650mg NH 4 NO 3 , 1900mg KNO 3 , 440mg CaCl 2 2H 2 O, 370mg MgSO 4 ·7H 2 O, 170mg KH 2 PO 4 , 0.83mg KI, 6.2 mgH 3 BO 3 , 22.3mg, MnSO 4 4H 2 O, 8.6mg ZnSO 4 ·7H 2 O, 0.25mg Na 2 MnO 4 2H 2 O, 0.025mg CuSO 4 ·5H 2 O, 0.025mg CoCl 2 ·6H...
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