Dracaena cochinchinensis loose callus induction method

Abstract

Provided is a dracaena cochinchinensis loose callus induction method. The method comprises the following steps: firstly, dracaena cochinchinensis seeds or buds are taken as explants, and disinfection is carried out; secondly, the explants after disinfection are placed in an MS basal medium, and buds or clustered buds are generated through induction; thirdly, the aseptic buds are placed in MS, B5, N6 media with 6-benzyladenine 6-BA and naphthylacetic acid NAA, and cultured, and callus is obtained; fourthly, the callus is placed in induction media with different hormones and cultured, and loose callus is obtained. The callus obtained through the method is peak green or milk white and loose and has a strong propagation capability. A lot of well-grown loose callus can be induced in short time, and materials are provided for production of dracaena cochinchinensis total alkaloids through cell suspension culture.

Description

technical field

[0001] The invention relates to a method for inducing plant loose callus, in particular to a method for inducing loose callus of Dracaena glabrata. Background technique

[0002] Dracaena cochinchinensis (Lour) S.C. Chen, also known as Millennium Wood, Blood Dracaena, and Cochin Dracaena, is a endangered species and a national second-class protected plant. Dracaena fragrans has high medicinal value, and the blood jelly extracted from its resin is called kylin jelly. It is a traditional Chinese medicinal material and has been used in my country for more than 1,500 years. According to the "Compendium of Materia Medica", dragon's blood is warm, flat, sweet, salty, non-toxic, enters the blood system, and returns to the lung, spleen, and kidney meridians. It has remarkable effects such as promoting muscle growth and suppressing sores, and has the reputation of "the holy medicine for promoting blood circulation". It is mainly distributed in Yunnan, Guangxi, Hainan ...

Examples

Embodiment 1

[0017] An example of the method for the induction of the loose callus of Dracaena glabrata of the present invention, comprises the steps:

[0018] (1) Disinfection of explants: first clean the dirt on the surface of the explants with an aqueous solution of detergent, rinse them with running water for 5 to 8 minutes in a beaker, then move them to an ultra-clean workbench, and decontaminate the seeds with 75v / v% alcohol. Soak with buds for 30s, rinse with sterile water once, and then use 0.1v / v% HgC1 2 Soak for 8, 12, and 15 minutes respectively, and soak in sterile water for 3 times. Blot dry the surface moisture of the explants with sterile filter paper and inoculate them, wherein the sterile water is distilled water sterilized by high temperature and high pressure; the explants are plump seeds or buds germinated from seeds sown in river sand;

[0019] (2) Acquisition of sterile seedlings: Inoculate the explants obtained in step (1) into MS induction medium respectively, at a...

Embodiment 2

[0023] (1) Disinfection of explants: first clean the dirt on the surface of the explants with an aqueous solution of detergent, rinse them with running water for 5 to 8 minutes in a beaker, then move them to an ultra-clean workbench, and decontaminate the seeds with 75v / v% alcohol. Soak with buds for 30s, rinse with sterile water once, and then use 0.1v / v% HgC1 2 Soak for 8, 12, and 15 minutes respectively, and soak in sterile water for 3 times. Blot dry the surface moisture of the explants with sterile filter paper and inoculate them, wherein the sterile water is distilled water sterilized by high temperature and high pressure; the explants are plump seeds or buds germinated from seeds sown in river sand;

[0024] (2) Acquisition of sterile seedlings: Inoculate the explants obtained in step (1) into MS induction medium respectively, at a culture temperature of 25±2°C, with a light intensity of 20-30 μmol / m 2 s, cultivated under the condition of light time of 12h / d, and sprou...

Embodiment 3

[0028] (1) Disinfection of explants: first clean the dirt on the surface of the explants with an aqueous solution of detergent, rinse them with running water for 5 to 8 minutes in a beaker, then move them to an ultra-clean workbench, and decontaminate the seeds with 75v / v% alcohol. Soak with buds for 30s, rinse with sterile water once, and then use 0.1v / v% HgC1 2 Soak for 8, 12, and 15 minutes respectively, and soak in sterile water for 3 times. Blot dry the surface moisture of the explants with sterile filter paper and inoculate them, wherein the sterile water is distilled water sterilized by high temperature and high pressure; the explants are plump seeds or buds germinated from seeds sown in river sand;

[0029] (2) Acquisition of sterile seedlings: Inoculate the explants obtained in step (1) into MS induction medium respectively, at a culture temperature of 25±2°C, with a light intensity of 20-30 μmol / m 2 s, cultivated under the condition of light time of 12h / d, and sprou...

technical field

[0001] The invention relates to a method for inducing plant loose callus, in particular to a method for inducing loose callus of Dracaena glabrata. Background technique

[0002] Dracaena cochinchinensis (Lour) S.C. Chen, also known as Millennium Wood, Blood Dracaena, and Cochin Dracaena, is a endangered species and a national second-class protected plant. Dracaena fragrans has high medicinal value, and the blood jelly extracted from its resin is called kylin jelly. It is a traditional Chinese medicinal material and has been used in my country for more than 1,500 years. According to the "Compendium of Materia Medica", dragon's blood is warm, flat, sweet, salty, non-toxic, enters the blood system, and returns to the lung, spleen, and kidney meridians. It has remarkable effects such as promoting muscle growth and suppressing sores, and has the reputation of "the holy medicine for promoting blood circulation". It is mainly distributed in Yunnan, Guangxi, Hainan ...

Examples

Embodiment 1

[0017] An example of the method for the induction of the loose callus of Dracaena glabrata of the present invention, comprises the steps:

[0018] (1) Disinfection of explants: first clean the dirt on the surface of the explants with an aqueous solution of detergent, rinse them with running water for 5 to 8 minutes in a beaker, then move them to an ultra-clean workbench, and decontaminate the seeds with 75v / v% alcohol. Soak with buds for 30s, rinse with sterile water once, and then use 0.1v / v% HgC1 2 Soak for 8, 12, and 15 minutes respectively, and soak in sterile water for 3 times. Blot dry the surface moisture of the explants with sterile filter paper and inoculate them, wherein the sterile water is distilled water sterilized by high temperature and high pressure; the explants are plump seeds or buds germinated from seeds sown in river sand;

[0019] (2) Acquisition of sterile seedlings: Inoculate the explants obtained in step (1) into MS induction medium respectively, at a...

Embodiment 2

[0023] (1) Disinfection of explants: first clean the dirt on the surface of the explants with an aqueous solution of detergent, rinse them with running water for 5 to 8 minutes in a beaker, then move them to an ultra-clean workbench, and decontaminate the seeds with 75v / v% alcohol. Soak with buds for 30s, rinse with sterile water once, and then use 0.1v / v% HgC1 2 Soak for 8, 12, and 15 minutes respectively, and soak in sterile water for 3 times. Blot dry the surface moisture of the explants with sterile filter paper and inoculate them, wherein the sterile water is distilled water sterilized by high temperature and high pressure; the explants are plump seeds or buds germinated from seeds sown in river sand;

[0024] (2) Acquisition of sterile seedlings: Inoculate the explants obtained in step (1) into MS induction medium respectively, at a culture temperature of 25±2°C, with a light intensity of 20-30 μmol / m 2 s, cultivated under the condition of light time of 12h / d, and sprou...

Embodiment 3

[0028] (1) Disinfection of explants: first clean the dirt on the surface of the explants with an aqueous solution of detergent, rinse them with running water for 5 to 8 minutes in a beaker, then move them to an ultra-clean workbench, and decontaminate the seeds with 75v / v% alcohol. Soak with buds for 30s, rinse with sterile water once, and then use 0.1v / v% HgC1 2 Soak for 8, 12, and 15 minutes respectively, and soak in sterile water for 3 times. Blot dry the surface moisture of the explants with sterile filter paper and inoculate them, wherein the sterile water is distilled water sterilized by high temperature and high pressure; the explants are plump seeds or buds germinated from seeds sown in river sand;

[0029] (2) Acquisition of sterile seedlings: Inoculate the explants obtained in step (1) into MS induction medium respectively, at a culture temperature of 25±2°C, with a light intensity of 20-30 μmol / m 2 s, cultivated under the condition of light time of 12h / d, and sprou...