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In vitro rapid propagating method of brachythecium procumbens gametophyte

A technology of stolonite moss and gametophytes, which is applied in the field of in vitro rapid propagation of stolonite moss gametophytes, can solve problems such as no stolonite moss large-scale propagation gametophyte culture technology, no gametophyte research reports, etc., to achieve It is convenient for intensive and industrialized production, conducive to automatic control of production, and the effect of simple method

Inactive Publication Date: 2011-08-10
SHANGHAI NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After retrieving domestic and foreign patent documents and extensively reviewing domestic and foreign publications, there is no culture technique for the large-scale expansion of gametophytes in test tubes, and there is no relevant research report on the use of in vitro culture of gametophytes of Phenophytum moss

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The new branches of the gametocytes of the stolonite moss grown in the natural environment were clipped for surface disinfection, and the disinfectants were 75% alcohol solution and 0.1% mercuric chloride solution. After disinfection, the apex of the cut branch with a length of about 0.5 cm was inoculated on the modified Knop's medium without any hormone for culture to obtain sterile explants. The culture conditions are: temperature 25±2°C, light intensity 4000 lx, light time 12 hours / day, and culture for 10 days.

[0036] Prepare the alcohol solution of 75% volume fraction and the mercury chloride solution of 0.1% mass volume fraction according to conventional method, be used for the surface sterilization of stolonifer gametophyte, sterilant combination and sterilization time are the following 3 kinds:

[0037] a) 75% volume fraction of alcohol solution for 30 seconds and 0.1% mass volume fraction of mercury solution for 3 minutes;

[0038] b) 75% volume fraction of a...

Embodiment 2

[0047]The sterile explants obtained in Example 1 were transferred to the medium for inducing the production of new gametophytes and cultured for 30 days. The culture conditions are: temperature 25°C, light intensity 4000Lx, light time 12 hours / day;

[0048] Prepare the culture medium that induces newborn gametophytes to produce according to conventional methods, and the culture medium formula consists of the following five types:

[0049] The formula of improved Knop's basic medium is: KNO 3 250mg / L, MgSO 4 ·7H 2 O250mg / L, KH 2 PO 4 250mg / L, Ca(NO 3 ) 2 4H 2 O 1000mg / L, ammonium tartrate 0.115mg / L, NaFe-EDTA 36.7mg / L, MnSO 4 4H 2 O 11.15mg / L, H 3 BO 3 3.1mg / L, KI 0.415mg / L, ZnSO 4 ·7H 2 O 4.3mg / L, CuSO 4 ·5H 2 O 0.0125mg / L, NaMoO 4 2H 2 O 0.125mg / L, CoCL 2 ·6H 2 O 0.0125mg / L;

[0050] a) Improved Knop's basic medium+0.1mg / L 6-benzylaminopurine+0.01mg / L naphthaleneacetic acid+10g / L sucrose;

[0051] b) Improved Knop's basic medium+0.1mg / L 6-benzylaminopur...

Embodiment 3

[0063] The sterile new branches obtained in Example 2 were cut into sections of about 0.3cm-0.5cm in length, transferred to the best medium for inducing newborn gametophyte production, and cultivated for 30 days. The optimal medium formulation for inducing newborn gametophytes is: improved Knop's basic medium + 0.1mg / L 6-benzylaminopurine + 0.05mg / L naphthaleneacetic acid + 10g / L sucrose. The formula of the improved Knop's basic medium is: KNO 3 250mg / L, MgSO 4 ·7H 2 O 250mg / L, KH 2 PO 4 250mg / L, Ca(NO 3 ) 2 4H 2 O 1000mg / L, ammonium tartrate 0.115mg / L, NaFe-EDTA 36.7mg / L, MnSO 4 4H 2 O 11.15mg / L, H 3 BO 3 3.1mg / L, KI 0.415mg / L, ZnSO 4 ·7H 2 O4.3mg / L, CuSO 4 ·5H 2 O 0.0125mg / L, NaMoO 4 2H 2 O 0.125mg / L, CoCL 2 ·6H 2 O 0.0125mg / L;

[0064] Conventional methods are used to prepare the culture medium for inducing newborn gametocytes to be cultured according to the following four culture conditions:

[0065] a) The temperature is 20±2°C, the light intensity ...

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Abstract

The invention discloses an in vitro rapid propagating method of brachythecium procumbens gametophyte, comprising the following steps of: disinfecting the surface of the new branches of the brachytecium procumbens gametophyte growing in a natural environment, and inoculating to a culture medium without any hormone for cultivating for 10 days to obtain the sterile gametophyte; transplanting the sterile gametophyte to the new gametophyte culture medium for cultivating for 30 days to obtain new gametophyte branches; and cutting the new gametophyte branches into 0.3-0.5cm sections, and transplanting the sections to a gametophyte propagation culture enlarging culture medium for cultivating for 30 days, wherein each gametophyte section can newly generate many gametophyte branches. The method is simple and has the advantages of fast breeding speed of the brachytecium procumbens gametophyte, is convenient for intensive and industrial production, and beneficial to human environment protection. Furthermore, the method supplies an ideal testing material for the environmental science research.

Description

technical field [0001] The invention relates to a culture technology of stolonite moss, in particular to a method for in vitro rapid propagation of stolonite moss gametophytes. Background technique [0002] Bryophyte (Brachythecium procumbens (mitt.) jaeg.) Bryophyte, Brachytheciaceae (Brachytheciaceae) genus (Brachythecium); monoecious, yellow-green plant, compact and shiny. Main stem prostrate, curved, irregular oblique branches, single or rebranched. The leaves are dense, imbricate, erect and stretching; the stems and leaves are broadly ovate, concave, and wrinkled; the tips of the leaves are short-haired, and the middle rib reaches 2 / 3-3 / 4 of the length of the leaf. The sporophyte is born on the main stem; the female bracts are long and narrow, erect, and the apex recurses. Stem twisted, smooth, about 2mm long. [0003] Bryophytes have a simple structure and strong adsorption capacity for environmental pollutants. Existing technologies have shown that bryophytes are ...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 娄玉霞曹同郭水良
Owner SHANGHAI NORMAL UNIVERSITY
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