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A kind of method and culture medium thereof for in vitro rapid propagation of Physcomitrella brevis gametophytes

A technology of gametophyte and tyrae moss, applied in the field of bioengineering, to achieve the effect of facilitating automatic control production, highly intensive and high-density factory production, and enriching provenance

Inactive Publication Date: 2015-10-14
SHANGHAI NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in the prior art, there is no culture technique for a large number of gametocytes in a culture bottle of Physcomitrella brevis, and there is no relevant research report on obtaining gametophytes by in vitro culture of Physcomitrella brevis spore capsules

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Cut the spore capsules of Lueres brevis growing in the natural environment for surface disinfection. The disinfectant is 75% alcohol solution and 0.1% mercury solution by volume; after disinfection, the spores in the sterile capsules are inoculated Cultivate in modified Knop's medium without any hormones to obtain sterile protonema. The culture conditions are: temperature 23±2°C, light intensity 2000 lx, light time 14 hours / day, and observe the culture results after 10 days of culture.

[0024] Prepare a 75% volume fraction alcohol solution and a 0.1% mass volume fraction mercuric chloride solution according to the conventional method, and use them for surface sterilization of the gametophytes of Physcomitrella brevis.

[0025] a) 75% by volume alcohol solution for 30 seconds and 0.1% by volume mercuric solution for 3 minutes;

[0026] b) 75% by volume alcohol solution for 30 seconds and 0.1% by volume mercuric solution for 5 minutes;

[0027] c) 75% volume fraction o...

Embodiment 2

[0036] The sterile protonema obtained in Example 1 was transferred to a medium for inducing the differentiation of the protonema to form gametophytes, and cultured for 30 days. The culture conditions are: temperature 23°C, light intensity 2000Lx, light time 14 hours / day;

[0037] Prepare the culture medium for inducing differentiation gametocytes to produce according to the conventional method, and the culture medium formula consists of the following 6 types:

[0038] a) Improved Knop's basal medium-125mg / L KNO 3 +10g / L sucrose +6g / L agar powder;

[0039] b) Improved Knop's basic medium + 125mg / L KNO 3 +10g / L sucrose +6g / L agar powder;

[0040] c) Improved Knop's minimal medium-500mg / L Ca(NO 3 ) 2 4H 2 O+10g / L sucrose+6g / L agar powder;

[0041] d) Improved Knop's basic medium + 500mg / L Ca(NO 3 ) 2 4H 2 O+10g / L sucrose+6g / L agar powder;

[0042] e) Improved Knop's minimal medium-125mg / L KH 2 PO 4 +10g / L sucrose +6g / L agar powder;

[0043] f) Improved Knop's basic ...

Embodiment 3

[0054] Inoculate the gametophytes obtained by the induction culture in f) of Example 2 onto the culture medium for gametophyte multiplication and culture for 30 days. The culture conditions are: temperature 23±2°C, light intensity 2000 lx, light time 14 hours / day.

[0055] Prepare the medium for gametophyte proliferation and expansion according to conventional methods, and the formula consists of the following four types:

[0056] a) MS basic medium + 10g / L sucrose + 6g / L agar powder;

[0057] b) 1 / 2 MS basic medium (the amount of macroelement compound is 1 / 2 of the amount specified in the formula) + 10g / L sucrose + 6g / L agar powder;

[0058] c) 1 / 3 MS basic medium (the amount of macroelement compound is 1 / 3 of the amount specified in the formula) + 10g / L sucrose + 6g / L agar powder;

[0059] d) 1 / 4MS basic medium (the amount of macroelement compound is 1 / 4 of the amount specified in the formula) + 10g / L sucrose + 6g / L agar powder;

[0060] The formula of the MS basic medium...

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Abstract

The invention discloses an in-vitro rapid propagation method of Brachymenium nepalense gametophytes, comprising the following steps of: (1) performing surface sterilization on capsulea of Brachymenium nepalense growing in a natural environment, and then, inoculating the capsulea to an improved Knop culture medium without any hormone to be cultured for 10 days, after spores germinate into protonemae, culturing 30 days again to obtain more protonemae after the growth of the primary protonemae; (2) transferring the protonemae obtained in (1) to a culture medium used for inducing the generation of the gametophytes, culturing for 30 days to obtain gametophyte branches; and (3) inoculating individual gametophyte branches obtained by induction culture in (2) to a culture medium for subculture multiplication of the gametophytes, and culturing for 30 days, wherein each gametophyte can be used to breed numerous new gametophytes. According to the in-vitro rapid propagation method of the Brachymenium nepalense gametophytes, the artificial propagation expanding of a large amount of Brachymenium nepalense gametophytes can be realized within a short time, a large amount of ideal materials are provided for indicating and monitoring atmospheric pollution by using bryophytes, and rich seedlings are provided for application of the bryophytes to garden landscaping.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method for rapidly propagating gametophytes of Physcomitrella brevis in vitro by using plant tissue culture technology. Background technique [0002] Brachymenium nepalense Hook. is a bryophyte belonging to the genus Brachymenium Schwaegr. belonging to the family Bryaceae. The plants are strong, medium-sized, clustered longitudinally, with no or slightly shiny stems and leaves, yellow-green to dark green. The stem is upright, with many new branches, up to 2cm high (about 1cm in northern my country, which varies greatly due to different environments). The base has reddish-brown rhizoids, and the leaves are clustered at the top of the branches in a rosette shape. The lower leaves are sparse and small. Shrinked when dry, tightly spiraled on the stem. The leaves are oblong tongue-shaped, oblong spoon-shaped or egg-shaped oblong, acuminate, except the upper ed...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04A01H4/00
CPCY02A40/22
Inventor 娄玉霞丁雪郭水良胡治祥
Owner SHANGHAI NORMAL UNIVERSITY
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