Induction method of callus of sterile seedling prepared from root bud of Amomum villosum Lour.

Abstract

The invention provides an induction method of callus of a sterile seedling prepared from the root bud of Amomum villosum Lour. The induction method comprises the following steps: taking the sterile seedling of the root bud of the Amomum villosum Lour. as an explant, sterilizing, inoculating and performing induction culture to obtain the callus, wherein an induction medium for induction culture isMS+1.0 to 2.5 mg / L 6-BA+0.5 to 1.5 mg / L 2,4-D+0.5 to 1.5 mg / L NAA; the illumination time of a culture environment is 10 h, the light intensity is 2000 Lx, and the culture temperature is 25 DEG C. Thestem and the root tip of the sterile seedling of the rhizome of the Amomum villosum Lour. Are used as the explant, and the asexual reproduction can retain the complete medicinal properties of a parent, and variation is not easy to generate, thereby making the medicine property stable and ensuring the effect of the drug treatment, and the explant material is easy to obtain and can be taken at any time for breeding culture.

Description

technical field

[0001] The invention relates to a callus induction method, which belongs to the technical field of tissue culture. Background technique

[0002] Amomum villosum Lour. is a perennial evergreen herb of the genus Amomum Roxb. in the ginger family. It is used as medicine with its dried and ripe fruit. It is also known as "Amomum villosum Lour." , has a history of more than 1300 years of application. Spring amomum is warm in nature and pungent in taste. It has the effects of reducing dampness and appetizing, warming spleen and stopping diarrhea, regulating qi and preventing miscarriage. Yangchun sand is mainly distributed in Guangdong, Guangxi, Yunnan and other places, among which the amomum produced in Guangdong Yangchun has the best medicinal effect. Amomum has a large demand, and the annual demand is estimated to exceed 2.2×10 6 kg, due to the high fruit drop rate and low yield of Amomum edulis, the output of Amomum amomum is far from meeting the demand of t...

Examples

Embodiment 1

[0035] A method for inducing callus of aseptic seedlings prepared from the rhizome buds of P. chinensis, comprising the following steps: cutting the stem of the aseptic seedling of P. chinensis into 2cm segments, and aseptically operating and sterilizing the stem segments Finally, vertically place the inoculation in the prepared and sterilized medium for cultivation, inoculate 5 bottles, 1 block per bottle, observe and record every 3 days. The well-growing callus was selected by culture, and the successfully induced callus was subcultured once every 30 days.

[0036] The environmental conditions for cultivation are as follows: temperature: 22°C; relative air humidity: 50%; light control is 10 hours of light, and light intensity is 1500Lx.

[0037] The medium formula is: MS+1.0mg / L 6-BA+0.5mg / L 2,4-D+0.5mg / L NAA

Embodiment 2

[0039] The steps of the method for inducing the callus of the aseptic seedlings prepared from the rhizome buds of Yangchunsha in this example are the same as those in Example 1.

[0040] The cultivation environment conditions in this embodiment are as follows: temperature: 25°C; relative air humidity: 80%; light control is 15 hours of light, and the light intensity is 2000Lx.

[0041] The medium formula is: MS+2.5mg / L 6-BA+1.5mg / L 2,4-D+1.5mg / L NAA

Embodiment 3

[0043] The steps of the method for inducing the callus of the aseptic seedlings prepared from the rhizome buds of Yangchunsha in this example are the same as those in Example 1.

[0044] The cultivation environment conditions in this embodiment are as follows: temperature: 25°C; relative air humidity: 80%; light control is 10 hours of light, and light intensity is 2000Lx.

[0045] The medium formula is: MS+1.5mg / L 6-BA+1.0mg / L 2,4-D+0.5mg / L NAA

technical field

[0001] The invention relates to a callus induction method, which belongs to the technical field of tissue culture. Background technique

[0002] Amomum villosum Lour. is a perennial evergreen herb of the genus Amomum Roxb. in the ginger family. It is used as medicine with its dried and ripe fruit. It is also known as "Amomum villosum Lour." , has a history of more than 1300 years of application. Spring amomum is warm in nature and pungent in taste. It has the effects of reducing dampness and appetizing, warming spleen and stopping diarrhea, regulating qi and preventing miscarriage. Yangchun sand is mainly distributed in Guangdong, Guangxi, Yunnan and other places, among which the amomum produced in Guangdong Yangchun has the best medicinal effect. Amomum has a large demand, and the annual demand is estimated to exceed 2.2×10 6 kg, due to the high fruit drop rate and low yield of Amomum edulis, the output of Amomum amomum is far from meeting the demand of t...

Examples

Embodiment 1

[0035] A method for inducing callus of aseptic seedlings prepared from the rhizome buds of P. chinensis, comprising the following steps: cutting the stem of the aseptic seedling of P. chinensis into 2cm segments, and aseptically operating and sterilizing the stem segments Finally, vertically place the inoculation in the prepared and sterilized medium for cultivation, inoculate 5 bottles, 1 block per bottle, observe and record every 3 days. The well-growing callus was selected by culture, and the successfully induced callus was subcultured once every 30 days.

[0036] The environmental conditions for cultivation are as follows: temperature: 22°C; relative air humidity: 50%; light control is 10 hours of light, and light intensity is 1500Lx.

[0037] The medium formula is: MS+1.0mg / L 6-BA+0.5mg / L 2,4-D+0.5mg / L NAA

Embodiment 2

[0039] The steps of the method for inducing the callus of the aseptic seedlings prepared from the rhizome buds of Yangchunsha in this example are the same as those in Example 1.

[0040] The cultivation environment conditions in this embodiment are as follows: temperature: 25°C; relative air humidity: 80%; light control is 15 hours of light, and the light intensity is 2000Lx.

[0041] The medium formula is: MS+2.5mg / L 6-BA+1.5mg / L 2,4-D+1.5mg / L NAA

Embodiment 3

[0043] The steps of the method for inducing the callus of the aseptic seedlings prepared from the rhizome buds of Yangchunsha in this example are the same as those in Example 1.

[0044] The cultivation environment conditions in this embodiment are as follows: temperature: 25°C; relative air humidity: 80%; light control is 10 hours of light, and light intensity is 2000Lx.

[0045] The medium formula is: MS+1.5mg / L 6-BA+1.0mg / L 2,4-D+0.5mg / L NAA

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