Method for establishing adventitious root cultivation system of Psammosilene tuniceoides W. C. Wu et C. Y. Wu and expanding cultivation method of Psammosilene tuniceoides W. C. Wu et C. Y. Wu
A technology for expanding the cultivation and cultivation system, which is applied in the establishment and expansion of the cultivation system of the adventitious root of Aurantia chinensis, and can solve the problems of retention, growth stability, cultivation conditions and various adjustment parameters, such as the contradiction between cell growth and the accumulation of secondary metabolites , to achieve stable output and quality, rapid and convenient scale production, and reduce production costs
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Embodiment 1
[0059] (1) Select Saccharomyces auricosa as explants, and carry out disinfection treatment: soak in running water for 25 minutes with Tween 20, rinse with sterile water for 3 times; then use 0.1% HgCl 2 Treat for 8 minutes, rinse with sterile water for 3 times; then treat with 75% ethanol for 8 seconds, rinse with sterile water for 3 times, and finally dry the water with sterile filter paper;
[0060] (2) Callus induction: inoculate the explants in step (1) in callus induction medium: MS medium + 2,4-D 0.5 mg / L + NAA 0.5 mg / L + KT 0.1mg / L+cysteine 1.0mg / L, pH 6.0, cultivate in dark at 25±1°C. The leaves were inoculated in the induction medium for about 10 days and began to grow milky white callus; about 20 days, the callus gradually covered the explant, and at 25 days, the callus with loose texture and good growth was selected for subculture;
[0061] (3) Induced callus subculture: Take 0.5 g of callus obtained in step (2) and inoculate it into MS medium + sucrose 40g / L + ...
Embodiment 2
[0067] (1) Select the 1.0-1.5cm length of the golden iron lock stem section as the explant, and carry out disinfection treatment: after washing with running water, soak it in Tween 20 for 25 minutes, rinse it with sterile water for 3 times; then use 0.1% HgCl 2 Treat for 8 minutes, rinse with sterile water for 3 times; then treat with 75% ethanol for 8 seconds, rinse with sterile water for 3 times, and finally dry the water with sterile filter paper;
[0068] (2) Callus induction: inoculate the explants in step (1) in callus induction medium: MS medium + 2,4-D 0.5 mg / L + NAA 0.5 mg / L + KT 0.1mg / L+ cysteine 2.0mg / L, pH 6.0, cultivate in dark at 25±1°C. The leaves were inoculated in the induction medium for about 10 days and began to grow milky white callus; about 20 days, the callus gradually covered the explant, and at 25 days, the callus with loose texture and good growth was selected for subculture;
[0069] (3) Induced callus subculture: Take 0.5 g of callus obtained in...
Embodiment 3
[0076] (1) Select the 2.0-3.0cm length of the golden iron lock stem section as the explant, and carry out disinfection treatment: soak in Tween 20 for 25 minutes after washing with running water, and rinse with sterile water for 3 times; then use 0.1% HgCl 2 Treat for 8 minutes, rinse with sterile water for 3 times; then treat with 75% ethanol for 8 seconds, rinse with sterile water for 3 times, and finally dry the water with sterile filter paper;
[0077] (2) Callus induction: inoculate the explants in step (1) in callus induction medium: MS medium + 2,4-D 0.5 mg / L + NAA 0.5 mg / L + KT 0.1mg / L, pH 6.0, cultured at 25±1°C under natural light. The leaves were inoculated in the induction medium for about 10 days and began to grow milky white callus; about 20 days, the callus gradually covered the explant, and at 25 days, the callus with loose texture and good growth was selected for subculture;
[0078] (3) Induced callus subculture: Take 0.5 g of callus obtained in step (2) an...
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