CN102144545A - In vitro rapid propagating method of brachythecium procumbens gametophyte - Google Patents
In vitro rapid propagating method of brachythecium procumbens gametophyte Download PDFInfo
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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
The present invention relates to the blue or green moss culture technique of a kind of flagellum, the blue or green moss gametophyte of specifically a kind of flagellum rapid propagation in vitro method.
Background technology
Blue or green moss (Brachythecium procumbens (mitt.) jaeg.) bryophyte of flagellum, the blue or green Rhodobryum of blue or green moss section (Brachytheciaceae) (Brachythecium); Monoecism, the plant yellow green, the bodily form is tight, and is glossy.Stem is crawled, bending, irregular plagiotropic growth, the single or branch again of branch.Ye Misheng, imbricate is arranged, the upright stretching, extension; Cauline leaf is wealthy avette, indent, tool gauffer; Blade tip tool undercoat point, middle rib reaches the 2/3-3/4 of leaf length.Sporophyte is born in stem; Perichaetial bract is long and narrow, and is upright, the tip warp.The seta distortion, level and smooth, be about 2mm.
Bryophyte is simple in structure, and absorption affinity is strong to environmental pollutants.Prior art shows that bryophyte is phanerogamous 10 times to the reaction sensitivity of the factors such as heavy metal, air and environmental contaminants, and the element of bryophyte accumulation is considerably beyond the physiological requirements level.Even noxious material exists concentration very low in environment, bryophyte also can be adsorbed it, accumulate and concentrate in the plant corpus.Since to advantage bryophytes such as air pollutant sensitivities by internationally recognized for the environmental pollution indicator organism.
Utilizing the sedimentation pollution of bryophyte monitoring and research environment heavy metal is the important technological problems of international environmental science research.China mainly is by analyzing the constituent content in the liver moss sample tissue, at point pollution source quantitative analysis is carried out in atmospheric pollution, long-term Quantitative Monitoring being carried out in the atmospheric pollution in area on a large scale such as city.
Blue or green moss (Brachythecium procumbens (mitt.) jaeg.) the plant volume of flagellum is big, widely distributed, is easy to gather, and utilizes its indicative for environments to pollute and has clear superiority.By method for tissue culture, set up the fast numerous system of the blue or green moss of flagellum, can provide desirable test material for utilizing migration, distribution and the enrichment mechanism of the blue or green moss research atmosphere heavy metal element of flagellum in the bryophyte body.Domestic and international by retrieval patent documentation is extensively consulted domestic and international publication, and not seeing has the blue or green moss of flagellum to expand numerous gametophytic culture technique in vitro in a large number, does not see and utilizes the blue or green moss gametophyte of flagellum cultured in vitro to obtain gametophytic correlative study report.
Summary of the invention
The object of the present invention is to provide the gametophytic method of the blue or green moss of a kind of rapid propagation in vitro flagellum, realize manually a large amount of blue or green moss of numerous flagellum of expanding fast, for physiological responses and migration, distribution and the enrichment mechanism of pollutant in the bryophyte body of studying the bryophyte environmental pollution provides desirable test material.
The present invention seeks to realize like this:
The blue or green moss gametophyte of a kind of flagellum rapid propagation in vitro method, step is as follows:
(1) accurately take by weighing following material, insert in the container and mix, stir, the preparation medium:
KNO
3250mg/L, MgSO
47H
2O 250mg/L, KH
2PO
4250mg/L, Ca (NO
3)
24H
2O 1000mg/L, tartaric acid ammonia 0.115mg/L, NaFe-EDTA 36.7mg/L, MnSO
44H
2O 11.15mg/L, H
3BO
33.1mg/L, KI 0.415mg/L, ZnSO
47H
2O4.3mg/L, CuSO
45H
2O 0.0125mg/L, NaMoO
42H
2O 0.125mg/L, CoCL
26H
2O0.0125mg/L;
(2) cultivate aseptic gametophyte:
Take from the newborn branch surface sterilization of the blue or green moss gametophyte of the flagellum of growing in the right environment, be inoculated on the medium of step (1) preparation condition of culture: 25 ± 2 ℃ of temperature; Intensity of illumination 4000lx; Light application time 12 hours/day; Cultivated 10 days, screening obtains aseptic gametophyte;
(3) cultivate newborn gametophyte branch:
A prepares newborn gametophyte medium:
Medium+0.1～2.0mg/L 6-benzyl aminopurine+0.01～0.2mg/L methyl+10g/L the sucrose of step (1) preparation is inserted in the container and is mixed, stirs, and makes;
The aseptic gametophyte that B cultivates step (2) is transferred to newborn gametophyte medium, condition of culture: temperature 20-35 ± 2 ℃; Intensity of illumination 4000lx; Light application time 12 hours/day; Cultivated 30 days, and obtained newborn gametophyte branch;
(4) gametophyte propagation expands numerous:
A prepares gametophyte propagation and expands breeding culture medium:
Medium+0～0.5mg/L 6-benzyl aminopurine+0～0.05mg/L methyl+10g/L the sucrose of step (1) preparation is inserted in the container and is mixed, stirs, and makes;
B is inoculated into the segment of the newborn gametophyte branch cut growth 0.3cm-0.5cm of step (3) cultivation gametophyte propagation and expands on the breeding culture medium condition of culture: 25 ± 2 ℃ of temperature; Intensity of illumination 4000lx; Light application time 12 hours/day; Cultivated the gametophyte branch that each gametophyte segment can newborn One's name is legion 30 days;
30 seconds of alcoholic solution that the newborn branch surface sterilization of the blue or green moss gametophyte of flagellum is 75% volume fraction in the step (2) and the mercuric chloride solution of 0.1% volume fraction 3 minutes.
Select young tender branch cutting newborn on the blue or green moss gametophyte of flagellum among step (4) B for use.
Main points of the present invention are:
Take from the newborn branch of the blue or green moss gametophyte of the flagellum of growing in the right environment, be inoculated on the medium, cultivated 10 days, screening obtains aseptic gametophyte; Cultivate newborn gametophyte branch then: carry out gametophyte propagation again and expand numerous.
The medium of the present invention's preparation is the improvement Knop ' s medium of no any hormone; It is to increase necessary component on the improvement Knop ' of no any hormone s medium base that the newborn gametophyte medium that the present invention uses, gametophyte propagation expand breeding culture medium.
Because the blue or green moss plant of flagellum volume is big, widely distributed, be easy to gather, utilize its absorption environmental contaminants, indicative for environments pollution situation to have clear superiority.So the inventive method is implemented the back to heavy metal and environmental contaminants in the blue or green moss absorption of flagellum, the accumulation environment, has great importance for the protection environment.
The present invention has set up the method for the blue or green moss gametophyte of flagellum rapid propagation in vitro in the world first, and having filled up prior art does not have the blue or green moss gametophyte of flagellum rapid propagation in vitro method blank.Use the quick expanding propagation method that exsomatizes of the present invention, help highly intensification and the production of producing of high density worker; Helping automation control produces; Can realize artificial a large amount of blue or green moss gametophyte of numerous flagellum that expands at short notice, for physiological responses and migration, distribution and the enrichment mechanism of pollutant in the bryophyte body of studying the bryophyte environmental pollution provides desirable test material, have important scientific research, the value of environmental protection.
Advantage of the present invention is:
1, method is simple, and the blue or green moss reproduction speed of flagellum is fast.
2, be convenient to intensification and the production of producing of worker.
3, help protecting human environment.
4, provide desirable test material for Research of Environmental Sciences.
Embodiment
Below in conjunction with embodiment the present invention is done further detailed, complete explanation:
Embodiment 1:
The new branch of the blue or green moss gametophyte of flagellum that clip grows in the natural environment carries out surface sterilization, and disinfectant is the alcoholic solution of 75% volume fraction and the mercuric chloride solution of 0.1% volume fraction.After the sterilization, be inoculated on the improvement Knop ' s medium of no any hormone and cultivate cutting branch tip that length is about 0.5cm, to obtain aseptic explant.Condition of culture is: 25 ± 2 ℃ of temperature, and intensity of illumination 4000lx, light application time 12 hours/day was cultivated 10 days.
Prepare the alcoholic solution of 75% volume fraction and the mercuric chloride solution of 0.1% quality volume fraction according to a conventional method, be used for the gametophytic surface sterilizing of the blue or green moss of flagellum, bactericidal agent combination and sterilization time are following 3 kinds:
A) mercuric chloride solution of 30 seconds of alcoholic solution of 75% volume fraction and 0.1% quality volume fraction is 3 minutes;
B) mercuric chloride solution of 30 seconds of alcoholic solution of 75% volume fraction and 0.1% quality volume fraction is 5 minutes;
C) mercuric chloride solution of 30 seconds of alcoholic solution of 75% volume fraction and 0.1% quality volume fraction is 10 minutes;
Improvement Knop ' s the medium of the no any hormone of preparation is used for the screening and culturing of aseptic explant according to a conventional method, and culture medium prescription consists of: improve Knop ' s minimal medium+10g/L sucrose.
The result is as follows in the gametophyte sterilization:
A) after 3 minutes sterilization treatment of the mercuric chloride solution of 30 seconds of alcoholic solution of 75% volume fraction and 0.1% quality volume fraction, aseptic survival rate 38.8% is polluted survival rate 52.4%, lethality 9.8%;
B) after 5 minutes sterilization treatment of the mercuric chloride solution of 30 seconds of alcoholic solution of 75% volume fraction and 0.1% quality volume fraction, aseptic survival rate 12.2% is polluted survival rate 22.6%, lethality 65.2%;
C) after 10 minutes sterilization treatment of the mercuric chloride solution of 30 seconds of alcoholic solution of 75% volume fraction and 0.1% quality volume fraction, lethality 100%;
More above-mentioned gametophyte sterilization result is as can be seen: the blue or green moss gametophyte of flagellum to the antitoxin harmful ability of 0.1% quality volume fraction mercuric chloride solution a little less than, sterilization treatment can cause the phenomena of mortality to take place in 3 minutes, the sterilization treatment time, the whole death of gametophyte totally when reaching 10 minutes.
Embodiment 2:
The aseptic explant that example 1 is obtained is transferred on the medium of inducing newborn gametophyte generation, cultivates 30 days.Condition of culture is: 25 ℃ of temperature, intensity of illumination 4000Lx, light application time 12 hours/day;
The medium that newborn gametophyte produces is induced in preparation according to a conventional method, and culture medium prescription consists of following five kinds:
Improvement Knop ' s minimal medium prescription is: KNO
3250mg/L, MgSO
47H
2O250mg/L, KH
2PO
4250mg/L, Ca (NO
3)
24H
2O 1000mg/L, tartaric acid ammonia 0.115mg/L, NaFe-EDTA 36.7mg/L, MnSO
44H
2O 11.15mg/L, H
3BO
33.1mg/L, KI 0.415mg/L, ZnSO
47H
2O 4.3mg/L, CuSO
45H
2O 0.0125mg/L, NaMoO
42H
2O 0.125mg/L, CoCL
26H
2O 0.0125mg/L;
A) improvement Knop ' s minimal medium+0.1mg/L 6-benzylaminopurine+0.01mg/L methyl+10g/L sucrose;
B) improvement Knop ' s minimal medium+0.1mg/L 6-benzylaminopurine+0.05mg/L methyl+10g/L sucrose;
C) improvement Knop ' s minimal medium+0.5mg/L 6-benzylaminopurine+0.05mg/L methyl+10g/L sucrose;
D) improvement Knop ' s minimal medium+1.0mg/L 6-benzylaminopurine+0.1mg/L methyl+10g/L sucrose;
E) improvement Knop ' s minimal medium+2.0mg/L 6-furfuryl group aminopurine+0.2mg/L methyl+10g/L sucrose.
Cultivation results is as follows:
A) in improvement Knop ' s minimal medium+0.1mg/L 6-benzylaminopurine+0.01mg/L methyl+10g/L sucrose, aseptic branch tip elongation growth speed is slower, and newborn branch is less.When cultivating 30 days, aseptic branch average length is 1.7cm, and the length that produces on each branch reaches 3 of the above newborn branch amount average out to of 0.5cm;
B) in improvement Knop ' s minimal medium+0.1mg/L 6-benzylaminopurine+0.05mg/L methyl+10g/L sucrose, aseptic branch tip elongation growth speed is very fast, and newborn branch is more.When cultivating 30 days, aseptic branch average length is 3.2cm, and the length that produces on each branch reaches 5 of the above newborn branch amount average out to of 0.5cm;
C) in improvement Knop ' s minimal medium+0.5mg/L 6-benzylaminopurine+0.05mg/L methyl+10g/L sucrose, aseptic branch tip elongation growth speed is general, and newborn branch is more and lack.When cultivating 30 days, aseptic branch average length is 2.2cm, does not have length to reach the above newborn branch of 0.5cm on each branch;
D) in improvement Knop ' s minimal medium+1.0mg/L 6-benzylaminopurine+0.1mg/L methyl+10g/L sucrose, aseptic branch tip expansion chap and elongation growth speed are very slow, and newborn branch is many and short.When cultivating 30 days, be 1.1cm, do not have length to surpass the newborn branch of 0.5cm on each branch through measuring aseptic branch average length;
E) in improvement Knop ' s minimal medium+2.0mg/L 6-benzylaminopurine+0.2mg/L methyl+10g/L sucrose, the obvious chap swelling of aseptic branch tip, it is dark green that color becomes, and elongation speed is extremely slow, forms the green shot-like particle of many intensive new lives on the branch.When cultivating 30 days, aseptic branch average length is 0.8cm only, does not see newborn branch.
More above-mentioned cultivation results is improveing on the Knop ' s minimal medium basis as can be seen, and it is different to add two kinds of concentration of hormone combinations of 6-benzylaminopurine and methyl, and the growth of aseptic gametophyte tip and the formation of inducing of newborn branch are had a significant impact.0.1mg/L 6-benzylaminopurine+0.05mg/L methyl had not only helped the growth of aseptic gametophyte tip but also had helped the formation of newborn branch.
Embodiment 3:
The aseptic newborn branch that example 2 is obtained cuts into the segment that is about 0.3cm-0.5cm, is transferred on the optimal medium of inducing newborn gametophyte generation, cultivates 30 days.The described prescription of the optimal medium of newborn gametophyte generation of inducing is: improvement Knop ' s minimal medium+0.1mg/L 6-benzylaminopurine+0.05mg/L methyl+10g/L sucrose.Wherein improveing Knop ' s minimal medium prescription is: KNO
3250mg/L, MgSO
47H
2O 250mg/L, KH
2PO
4250mg/L, Ca (NO
3)
24H
2O 1000mg/L, tartaric acid ammonia 0.115mg/L, NaFe-EDTA 36.7mg/L, MnSO
44H
2O 11.15mg/L, H
3BO
33.1mg/L, KI 0.415mg/L, ZnSO
47H
2O4.3mg/L, CuSO
45H
2O 0.0125mg/L, NaMoO
42H
2O 0.125mg/L, CoCL
26H
2O 0.0125mg/L;
The medium that the conventional method preparation induces newborn gametophyte to produce, cultivate by following four kinds of condition of culture:
A) temperature is 20 ± 2 ℃, intensity of illumination 4000lx, and light application time 12 hours/day was cultivated 30 days;
B) temperature is 25 ± 2 ℃, intensity of illumination 4000lx, and light application time 12 hours/day was cultivated 30 days;
C) temperature is 30 ± 2 ℃, intensity of illumination 4000lx, and light application time 12 hours/day was cultivated 30 days;
D) temperature is 35 ± 2 ℃, intensity of illumination 4000lx, and light application time 12 hours/day was cultivated 30 days.
Cultivation results is as follows:
A) temperature is 20 ± 2 ℃, intensity of illumination 4000Lx, and light application time 12 hours/day, when cultivating 30 days, aseptic gametophyte segment color is bud green, produces 3 of the newborn branch amount average out to that length surpasses 0.5cm in each segment;
B) temperature is 25 ± 2 ℃, intensity of illumination 4000Lx, and light application time 12 hours/day, when cultivating 30 days, aseptic gametophyte segment color is bud green, produces 5 of the newborn branch amount average out to that length surpasses 0.5cm in each segment;
C) temperature is 30 ± 2 ℃, intensity of illumination 4000Lx, and light application time 12 hours/day, when cultivating 30 days, aseptic gametophyte segment color pale green, length surpasses 6 of the newborn branch amount average out to of 0.5cm in each segment, but it is thin and delicate to grow, and color is yellowish green;
D) temperature is 35 ± 2 ℃, intensity of illumination 4000Lx, and light application time 12 hours/day, when cultivating 30 days, aseptic gametophyte segment color is yellowish green, and the thin and delicate jaundice of newborn branch in each segment does not have length to surpass the newborn branch of 0.5cm.
More above-mentioned cultivation results as can be seen, cultivation temperature is formed with appreciable impact for blue or green gametophytic growth conditions of moss of flagellum and newborn branch, 25 ± 2 ℃ is comparison suitable culture temperature, the formation that all is unfavorable for gametophytic growth and newborn branch too high or too low for temperature.
Embodiment 4:
With b among the embodiment 3) the newborn gametophyte cutting that obtains of inducing culture shreds and is inoculated into gametophyte propagation and expands on numerous medium, cultivated 30 days.Condition of culture is: 25 ± 2 ℃ of temperature, intensity of illumination 4000lx, light application time 12 hours/day.
Prepare gametophyte propagation according to a conventional method and expand numerous medium, prescription consists of following four kinds:
A) improvement Knop ' s minimal medium+10g/L sucrose;
B) improvement Knop ' s minimal medium+0.1mg/L 6-benzylaminopurine+0.05mg/L methyl+10g/L sucrose;
C) improvement Knop ' s minimal medium+0.2mg/L 6-benzylaminopurine+0.05mg/L methyl+10g/L sucrose;
D) improvement Knop ' s minimal medium+0.5mg/L 6-benzylaminopurine+0.05mg/L methyl+10g/L sucrose;
Cultivation results is as follows:
A) in improvement Knop ' s minimal medium+10g/L sucrose, newborn branch is less on the branch in small, broken bits, and when cultivating 30 days, newborn branch average length is 1.2cm, 1 of the newborn branch amount average out to that produces on each branch in small, broken bits;
B) in improvement Knop ' s minimal medium+0.1mg/L 6-benzylaminopurine+0.05mg/L methyl+10g/L sucrose, newborn branch is less on the branch in small, broken bits, when cultivating 30 days, aseptic branch average length is 1.6cm, 2 of the newborn branch amount average out to that produces on each branch in small, broken bits;
C) in improvement Knop ' s minimal medium+0.2mg/L 6-benzylaminopurine+0.05mg/L methyl+10g/L sucrose, newborn branch is more on the branch in small, broken bits, when cultivating 30 days, be 1.4cm through measuring aseptic branch average length, 4 of the newborn branch amount average out to that produces on each branch in small, broken bits;
D) in improvement Knop ' s minimal medium+0.5mg/L 6-benzylaminopurine+0.05mg/L methyl+10g/L sucrose, newborn branch is less on the branch in small, broken bits, when cultivating 30 days, aseptic branch average length is 0.6cm, 2 of the newborn branch amount average out to that produces on each branch in small, broken bits;
More above-mentioned cultivation results is improveing on the Knop ' s minimal medium basis as can be seen, and it is different to add two kinds of concentration of hormone combinations of 6-benzylaminopurine and methyl, and gametophytic growth and propagation are had a significant impact.0.2mg/L 6-benzylaminopurine+0.05mg/L methyl had not only helped the formation of gametophyte branch but also had helped the growth of newborn branch.
The above is the preferred embodiments of the present invention only, is not limited to the present invention, and for a person skilled in the art, the present invention can have various changes and variation.Within the spirit and principles in the present invention all, any modification of being done, be equal to replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (3)
1. the blue or green moss gametophyte of flagellum rapid propagation in vitro method, step is as follows:
(1) accurately take by weighing following material, insert in the container and mix, stir, the preparation medium:
KNO
3250mg/L, MgSO
47H
2O 250mg/L, KH
2PO
4250mg/L, Ca (NO
3)
24H
2O 1000mg/L, tartaric acid ammonia 0.115mg/L, NaFe-EDTA 36.7mg/L, MnSO
44H
2O 11.15mg/L, H
3BO
33.1mg/L, KI 0.415mg/L, ZnSO
47H
2O4.3mg/L, CuSO
45H
2O 0.0125mg/L, NaMoO
42H
2O 0.125mg/L, CoCL
26H
2O0.0125mg/L;
(2) cultivate aseptic gametophyte:
Take from the newborn branch surface sterilization of the blue or green moss gametophyte of the flagellum of growing in the right environment, be inoculated on the medium of step (1) preparation condition of culture: 25 ± 2 ℃ of temperature; Intensity of illumination 4000lx; Light application time 12 hours/day; Cultivated 10 days, screening obtains aseptic gametophyte;
(3) cultivate newborn gametophyte branch:
A prepares newborn gametophyte medium:
Medium+0.1～2.0mg/L 6-benzyl aminopurine+0.01～0.2mg/L methyl+10g/L the sucrose of step (1) preparation is inserted in the container and is mixed, stirs, and makes;
The aseptic gametophyte that B cultivates step (2) is transferred to newborn gametophyte medium, condition of culture: temperature 20-35 ± 2 ℃; Intensity of illumination 4000lx; Light application time 12 hours/day; Cultivated 30 days, and obtained newborn gametophyte branch;
(4) gametophyte propagation expands numerous:
A prepares gametophyte propagation and expands breeding culture medium:
Medium+0～0.5mg/L 6-benzyl aminopurine+0～0.05mg/L methyl+10g/L the sucrose of step (1) preparation is inserted in the container and is mixed, stirs, and makes;
B is inoculated into the segment of the newborn gametophyte branch cut growth 0.3cm-0.5cm of step (3) cultivation gametophyte propagation and expands on the breeding culture medium condition of culture: 25 ± 2 ℃ of temperature; Intensity of illumination 4000lx; Light application time 12 hours/day; Cultivated the gametophyte branch that each gametophyte segment can newborn One's name is legion 30 days.
2. the blue or green moss gametophyte of flagellum according to claim 1 rapid propagation in vitro method is characterized in that:
30 seconds of alcoholic solution that the newborn branch surface sterilization of the blue or green moss gametophyte of flagellum is 75% volume fraction in the step (2) and the mercuric chloride solution of 0.1% volume fraction 3 minutes.
3. the blue or green moss gametophyte of flagellum according to claim 1 rapid propagation in vitro method is characterized in that: select young tender branch cutting newborn on the blue or green moss gametophyte of flagellum among step (4) B for use.
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| CN102511393A (en) * | 2011-12-12 | 2012-06-27 | 齐齐哈尔大学 | Method for establishing racomitrium japonicum gametophyte regeneration system |
| CN103651140A (en) * | 2013-12-10 | 2014-03-26 | 上海师范大学 | In-vitro rapid propagation method of Brachymenium nepalense gametophytes and culture medium thereof |
| CN103999772A (en) * | 2014-05-20 | 2014-08-27 | 上海师范大学 | In-vitro rapid propagation method of macromitrium cavaleriei card and ther |
| CN106386483A (en) * | 2016-08-30 | 2017-02-15 | 中国科学院昆明植物研究所 | Rapid propagation method for Brachythecium kuroishicum |
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2011
- 2011-01-11 CN CN2011100040429A patent/CN102144545B/en not_active Expired - Fee Related
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102511393A (en) * | 2011-12-12 | 2012-06-27 | 齐齐哈尔大学 | Method for establishing racomitrium japonicum gametophyte regeneration system |
| CN103651140A (en) * | 2013-12-10 | 2014-03-26 | 上海师范大学 | In-vitro rapid propagation method of Brachymenium nepalense gametophytes and culture medium thereof |
| CN103651140B (en) * | 2013-12-10 | 2015-10-14 | 上海师范大学 | A kind of gametophytic method of rapid propagation in vitro short moon moss and substratum thereof |
| CN103999772A (en) * | 2014-05-20 | 2014-08-27 | 上海师范大学 | In-vitro rapid propagation method of macromitrium cavaleriei card and ther |
| CN106386483A (en) * | 2016-08-30 | 2017-02-15 | 中国科学院昆明植物研究所 | Rapid propagation method for Brachythecium kuroishicum |
| CN106386483B (en) * | 2016-08-30 | 2018-06-22 | 中国科学院昆明植物研究所 | A kind of rapid propagation method for the leaf blueness moss that wrinkles |
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