Artificial enlarge method (of) Rhodobryum giganteum gametophyte

A technology of moss and gametophyte, applied in the field of bioengineering, can solve the problems of affecting the yield of resources and the consistency and stability of resource traits, scattered distribution and destruction of wild plant resources, etc. Chemical and high-density industrial production, with obvious economic benefits

Inactive Publication Date: 2011-05-25
SHANGHAI NORMAL UNIVERSITY
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  • Summary
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  • Application Information

AI Technical Summary

Problems solved by technology

Large-scale pharmaceutical production using wild warmland moss will face many problems: First, the natural growth process is often affected by changes in the natural environment, and there will be various mutated individuals or groups, which will affect the consistency and consistency of resource yield and resource traits. stability; secondly, different harvest periods and different origins will affect the stability of the quality of medicinal materials; more importantly, the distribution of wild plant resources in nature is scattered, and there is almost no phenomenon of large-scale flakes. Wild plants have become an important resource for development. On the one hand, it is difficult to meet the needs of the market by relying on wild resources alone. On the other hand, under the pressure of development and utilization, they are easily destroyed and pose a direct threat to the survival of species.
However, in the prior art, there is no culture technique for a large number of gametophytes in test tubes of Physcomitrella plants, and there is no relevant technical report on culturing the Physcomitrella plant tissue into gametophytes.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Disinfect the surface of the gametophytes of Physcomitrella warmosa without unfolding leaves growing in the natural environment, the disinfectant is 70-75% alcohol solution by volume and 0.1-0.2% mercury solution by mass fraction; after disinfection, cut the length Shoot tips of 0.5-1.0 cm were inoculated on the culture medium for inducing gametophytes to produce protonema. The culture conditions were: temperature 23±2°C, light intensity 2500±250lx, light time 10 hours / day, and culture for 50 days.

[0028] Prepare the culture medium that induces gametophytes to produce protonema according to conventional methods, and the formula consists of the following five types:

[0029] a) Improved Knop+5~10g / l sucrose;

[0030] b) Improved Knop+0.1mg / l 6-benzylaminopurine+5~10g / l sucrose;

[0031] c) Improved Knop+2.0mg / l 2,4-dichlorophenoxyacetic acid+5-10g / l sucrose;

[0032] d) Improved Knop+1.0mg / l 2,4-dichlorophenoxyacetic acid+0.05mg / l 6-benzylaminopurine+5-10g / l sucrose;...

Embodiment 2

[0043] Disinfect the surface of the gametophytes of Physcomitrella warmosa without unfolding leaves growing in the natural environment, the disinfectant is 70-75% alcohol solution by volume and 0.1-0.2% mercury solution by mass fraction; after disinfection, cut the length Inoculate the shoot tip of 0.5-1.0 cm on the medium for inducing gametophyte to produce protonema, the medium formula for inducing gametophyte to produce protonema is: improved Knop+2.0mg / l 2,4-dichlorophenoxy Acetic acid + 0.1mg / l 6-benzylaminopurine + 5-10g / l sucrose, wherein the formula of the modified Knop medium is: KCl 76.04mg / l, MgSO 4 ·7H 2 O 828.2mg / l, KH 2 PO 4 250.4mg / l, Ca(NO 3 ) 2 4H 2 O 1001.3mg / l, CuSO 4 ·5H2 O 0.027mg / l, KI0.014mg / l, ZnSO 4 ·7H 2 O0.027mg / l, H 3 BO 3 0.309mg / l, Na 2 MoO 4 2H 2 O 0.012mg / l, MnCl 2 4H 2 O0.198mg / l, CoCl 2 ·6H 2 O 0.027mg / l, FeSO 4 ·7H 2 O 27.8mg / l, Na 2 - EDTA 37.3mg / l, ammonium tartrate 0.1151mg / l, prepared as a medium for inducing gametop...

Embodiment 3

[0057] The green protonema obtained in Example 1 was transferred to the protonema propagation medium, and the culture conditions were: temperature 23±2°C, light intensity 3000±250lx, light time 12 hours / day, and cultivated for 55 days.

[0058] Prepare the protocelium propagation medium according to conventional methods, and the formula consists of the following four types:

[0059] a) Improved Knop medium + 1.0mg / l 2,4-dichlorophenoxyacetic acid + 5-10g / l sucrose;

[0060] b) Improved Knop medium + 2.0mg / l 2,4-dichlorophenoxyacetic acid + 5-10g / l sucrose;

[0061] c) Improved Knop medium + 1.0mg / l 6-furfurylaminopurine + 5-10g / l sucrose;

[0062] d) Modified Knop medium + 2.0 mg / l 6-furfurylaminopurine + 5-10 g / l sucrose.

[0063] The formula of the improved Knop medium is: KCl 76.04mg / l, MgSO 4 ·7H 2 O 828.2mg / l, KH 2 PO 4 250.4mg / l, Ca(NO 3 ) 2 4H 2 O 1001.3mg / l, CuSO 4 ·5H 2 O 0.027mg / l, KI0.014mg / l, ZnSO 4 ·7H 2 O 0.027mg / l, H 3 BO 3 0.309mg / l, Na 2 MoO ...

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Abstract

The invention discloses a method of manual extensive-reproduction of rhodobryum giganteum gametocyte, which includes steps in the following order: (1) sterilizing the surface of the rhodobryum giganteum gametocyte which grows in a natural environment with unexpanded leaf blades; cutting the stem tip and inoculating to the culture medium of inducing the gametocyte to generate the protonema and culturing for 40-60 days; (2) transplanting the protonema obtained in step (1) to the extensive-reproduction protonema culture medium and culturing for 40-60 days; (3) transplanting the extensive reproduced protonema in step (2) to the culture medium of inducing the protonema to generate the gametocyte and culturing for 40-60 days. The invention initiates the development of the method of manual extensive-reproduction of rhodobryum giganteum gametocyte and bridges the gap of the prior art. The manual extensive reproduction not only greatly facilitates the production of high intensification and high density industrialization, but also facilitates automatization of production control; which achieves the mass production of manual extensive-reproduction of rhodobryum giganteum in a short time and provides a market guarantee for the pharmacy field with significant economic benefits.

Description

technical field [0001] The invention relates to a method for artificially amplifying gametophytes of Physcomitrella euphratica, belonging to the technical field of bioengineering. Background technique [0002] Rhodobryum giganteum belongs to the genus Rhodobryum giganteum in the family Mossaceae, and it is collectively referred to as Rhodobryum roseum together with the same plant. Huixincao is a traditional local medicine, and it is also a national medicine used by the Yi, Dai, and Hani peoples. In recent years, the researches on the pharmacological effects, modern clinical researches and preparation application researches of Physcomitrella plants (mainly using Physcomitrella spp. efficacy in disease. This kind of medicinal plant preparation currently includes Huixinkang tablets produced by Yunnan Yingmao Biopharmaceutical Co., Ltd. and Huixincao tablets and injections produced by Shaanxi Changwu Pharmaceutical Factory. [0003] At present, the grass moss - Moss argentina...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00C12N5/04
Inventor 娄玉霞陈圆圆曹同郭水良
Owner SHANGHAI NORMAL UNIVERSITY
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