CN103999776A - Orchid rapid-propagation culture medium formula - Google Patents
Orchid rapid-propagation culture medium formula Download PDFInfo
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- CN103999776A CN103999776A CN201410264595.1A CN201410264595A CN103999776A CN 103999776 A CN103999776 A CN 103999776A CN 201410264595 A CN201410264595 A CN 201410264595A CN 103999776 A CN103999776 A CN 103999776A
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Abstract
The invention relates to an orchid rapid-propagation culture medium formula. Culture mediums for fast propagation of orchids include starting culture mediums, proliferation culture mediums and rooting and seedling strengthening culture mediums which are used in three culture stages. According to the culture medium formula, no plant hormones are added, and the characteristic of wild imitation cultivation is achieved. Orchid tissue culture seedlings obtained through the formula are good in growth vigor, regular and high in transplantation survival rate.
Description
Technical field
The present invention relates to field of plant tissue culture technique, especially relate to a kind of culture medium prescription of orchid tissue-culturing quick-propagation.
Background technology
Orchid is extended familys in angiosperm, be in higher plant except composite family maximum section, the known orchid in the whole world approximately has nearly 25000 kinds of 800 genus according to statistics, be distributed widely in the various terrestrial ecosystem except two-stage and extreme drought desert area, wherein abundant with torrid areas kind.The orchid that China is traditional, mainly refers to epidendrum, is referred to as China blue, and orchid not only has vast fan as integrating to view and admire with medicinal plant, and becomes a part of traditional Chinese culture.Yet traditional modes of reproduction of orchid is division propagation, reproduction coefficient is low, speed is slow, be subject to the restriction in geographical environment and season, be difficult to reach fast, the object of efficient breeding, though and cymbidium seed is many under field conditions (factors), but because it does not have full grown embryo, be difficult to sprout.By tissue, cultivating this technological means and method is the effective measures that solve at present orchid breeding, but the medium overwhelming majority who uses adds chemosynthesis plant hormone to promote micropropagation of plants and growth, and often resistance is poor, transplanting lethality is higher for group training seedling.In a kind of orchid tissue-culturing quick-propagation culture medium prescription provided by the invention, do not add any chemosynthesis plant hormone, not only high, the fast growth of growth coefficient, and group training seedling is healthy, adaptive capacity to environment is strong, transplanting survival rate is high.
Summary of the invention
Technical problem to be solved by this invention is a kind of orchid tissue-culturing quick-propagation culture medium prescription providing for above-mentioned prior art present situation, to reach orchid Fast-propagation and the strong object of group training seedling adaptive capacity to environment.
In order to achieve the above object, the present invention is achieved by the following technical solutions:
The present invention is a kind of orchid fast propagating culture medium formula, and the medium of the Fast-propagation of described orchid comprises following three medium that cultivation stage is used, and the content of the medium in each stage is:
A. start medium: spend precious No. 1 0.5 ~ 2.5g/L, spend precious No. 2 0.2 ~ 2.0g/L, peptone 0.1 ~ 1.0g/L, potato 20 ~ 45g/L, Coconut Juice 50 ~ 200ml/L, sucrose 10 ~ 30g/L;
B. proliferated culture medium: 1/2MS, potato 15 ~ 40g/L, banana 30 ~ 70g/L, sucrose 15 ~ 40g/L, peptone 0.5 ~ 1.5g/L, active carbon 0.5 ~ 1.5g/L;
C. Rooting and hardening-off culture base: 1/2MS, potato 15 ~ 45g/L, banana 30 ~ 80g/L, sucrose 10 ~ 30g/L, peptone 0.5 ~ 1.5g/L, apple 20 ~ 50g/L, active carbon 0.5 ~ 1.5g/L.
Preferred: the medium of the Fast-propagation of orchid comprises following three medium that cultivation stage is used, and the content of the medium in each stage is:
A. start medium: spend precious No. 1 1.0 ~ 2.0g/L, spend precious No. 2 0.5 ~ 1.5g/L, peptone 0.3 ~ 0.8g/L, potato 25 ~ 40g/L, Coconut Juice 80 ~ 120ml/L, sucrose 10 ~ 20g/L;
B. proliferated culture medium: 1/2MS, potato 25 ~ 35g/L, banana 40 ~ 60g/L, sucrose 20 ~ 30g/L, peptone 0.7 ~ 1.2g/L, active carbon 0.8 ~ 1.3g/L;
C. Rooting and hardening-off culture base: 1/2MS, potato 20 ~ 40g/L, banana 40 ~ 60g/L, sucrose 15 ~ 25g/L, peptone 0.8 ~ 1.2g/L, apple 30 ~ 40g/L, active carbon 0.8 ~ 1.2g/L.
Preferred: the medium of the Fast-propagation of orchid comprises following three medium that cultivation stage is used, and the content of the medium in each stage is:
A. start medium: spend precious No. 1 1.5g/L, spend precious No. 2 1.0g/L, peptone 0.5g/L, potato 30g/L, Coconut Juice 100ml/L, sucrose 15g/L;
B. proliferated culture medium: 1/2MS, potato 30g/L, banana 50g/L, sucrose 25g/L, peptone 1.0g/L, active carbon 1.0g/L;
C. Rooting and hardening-off culture base: 1/2MS, potato 30g/L, banana 50g/L, sucrose 20g/L, peptone 1.0g/L, apple 35g/L, active carbon 1.0g/L.
Preferred: the medium of the Fast-propagation of orchid comprises following three medium that cultivation stage is used, and the content of the medium in each stage is:
A. start medium: spend precious No. 1 1.5g/L, spend precious No. 2 1.0g/L, peptone 0.5g/L, potato 25g/L, Coconut Juice 80ml/L, sucrose 20g/L;
B. proliferated culture medium: 1/2MS, potato 35g/L, banana 45g/L, sucrose 25g/L, peptone 1.0g/L, active carbon 1.0g/L;
C. Rooting and hardening-off culture base: 1/2MS, potato 30g/L, banana 55g/L, sucrose 25g/L, peptone 1.0g/L, apple 40g/L, active carbon 1.0g/L.
Preferred: the medium of the Fast-propagation of orchid comprises following three medium that cultivation stage is used, and the content of the medium in each stage is:
A. start medium: spend precious No. 1 1.2g/L, spend precious No. 2 1.5g/L, peptone 0.5g/L, potato 35g/L, Coconut Juice 120ml/L, sucrose 20g/L;
B. proliferated culture medium: 1/2MS, potato 30g/L, banana 55g/L, sucrose 25g/L, peptone 1.0g/L, active carbon 1.0g/L;
C. Rooting and hardening-off culture base: 1/2MS, potato 35g/L, banana 60g/L, sucrose 20g/L, peptone 1.0g/L, apple 35g/L, active carbon 1.0g/L.
Preferred: the PH of the medium of three cultivation stages is 5.8 ~ 6.2.
Preferred: the seed that the explant that startup cultivation is used is orchid.
Preferred: three kinds of medium are take the solid culture medium that agar is supporter.
Beneficial effect of the present invention: do not add any chemosynthesis plant hormone in culture medium prescription of the present invention, with this medium culture orchid high, fast growth of growth coefficient not only, and group training seedling is healthy, adaptive capacity to environment is strong, transplanting survival rate is high.
Accompanying drawing explanation
Fig. 1 utilizes culture medium prescription of the present invention to cultivate the candidum tissue culturing seedling of 30d
Fig. 2 utilizes culture medium prescription of the present invention to cultivate the candidum tissue culturing seedling of 90d.
Embodiment
In order to deepen the understanding of the present invention, below in conjunction with drawings and Examples, the present invention is described in further detail, this embodiment, only for explaining the present invention, does not form and limits protection scope of the present invention.
As shown in Figure 1-2, the present invention is a kind of orchid fast propagating culture medium formula, and the medium of the Fast-propagation of described orchid comprises following three medium that cultivation stage is used, and the content of the medium in each stage is:
A. start medium: spend precious No. 1 0.5 ~ 2.5g/L, spend precious No. 2 0.2 ~ 2.0g/L, peptone 0.1 ~ 1.0g/L, potato 20 ~ 45g/L, Coconut Juice 50 ~ 200ml/L, sucrose 10 ~ 30g/L;
B. proliferated culture medium: 1/2MS, potato 15 ~ 40g/L, banana 30 ~ 70g/L, sucrose 15 ~ 40g/L, peptone 0.5 ~ 1.5g/L, active carbon 0.5 ~ 1.5g/L;
C. Rooting and hardening-off culture base: 1/2MS, potato 15 ~ 45g/L, banana 30 ~ 80g/L, sucrose 10 ~ 30g/L, peptone 0.5 ~ 1.5g/L, apple 20 ~ 50g/L, active carbon 0.5 ~ 1.5g/L;
The seed that the explant that startup cultivation is used is orchid, three kinds of medium are take the solid culture medium that agar is supporter, start medium, proliferated culture medium and Rooting and hardening-off culture base and all take the solid culture medium that agar is supporter, the PH of the medium of three cultivation stages is 5.8 ~ 6.2.
Preferred: the medium of the Fast-propagation of described orchid comprises following three medium that cultivation stage is used, and the content of the medium in each stage is:
A. start medium: spend precious No. 1 1.0 ~ 2.0g/L, spend precious No. 2 0.5 ~ 1.5g/L, peptone 0.3 ~ 0.8g/L, potato 25 ~ 40g/L, Coconut Juice 80 ~ 120ml/L, sucrose 10 ~ 20g/L;
B. proliferated culture medium: 1/2MS, potato 25 ~ 35g/L, banana 40 ~ 60g/L, sucrose 20 ~ 30g/L, peptone 0.7 ~ 1.2g/L, active carbon 0.8 ~ 1.3g/L;
C. Rooting and hardening-off culture base: 1/2MS, potato 20 ~ 40g/L, banana 40 ~ 60g/L, sucrose 15 ~ 25g/L, peptone 0.8 ~ 1.2g/L, apple 30 ~ 40g/L, active carbon 0.8 ~ 1.2g/L.
The seed that the explant that startup cultivation is used is orchid, three kinds of medium are take the solid culture medium that agar is supporter, start medium, proliferated culture medium and Rooting and hardening-off culture base and all take the solid culture medium that agar is supporter, the PH of the medium of three cultivation stages is 5.8 ~ 6.2.
Embodiment 1
Dendrobium candidum tissue is cultivated Fast-propagation
(1) explant: dendrobium candidum fruit pod, the complete free from flaw of fruit pod;
(2) explant sterilization: fruit pod is cleaned with clear water, put into 75% alcohol 20s, sterile water wash 2 times, then fruit pod is put into 0.1% mercuric chloride 12min, sterile water wash 5 ~ 7 times;
(3) inoculation and startup are cultivated: fruit pod one side under gnotobasis, sterilization being completed is opened an osculum, seed is sowed in aseptic culture medium, start in medium, the formula that starts medium is: spend precious No. 1 1.5g/L+ to spend precious No. 2 1.0g/L+ peptone 0.5g/L+ potato 30g/L+ Coconut Juice 100ml/L+ sucrose 15g/L+ agar 5.5g/L, PH 6.0;
(4) propagation is cultivated: start and cultivate after 25 ~ 35d, dendrobium candidum seedling or protocorm are transferred in proliferated culture medium, proliferation culture medium formula is: 1/2MS+ potato 30g/L+ banana 50g/L+ sucrose 25g/L+ peptone 1.0g/L+ active carbon 1.0g/L+ agar 5.5g/L, and PH 6.2;
(5) Rooting and hardening-off culture: propagation is cultivated 30 ~ 45d, group training seedling is transferred in Rooting and hardening-off culture base, Rooting and hardening-off culture based formulas is: 1/2MS+ potato 30g/L+ banana 50g/L+ sucrose 20g/L+ peptone 1.0g/L+ apple 35g/L+ active carbon 1.0g/L+ agar 5.5g/L, PH 5.8.Cultivate 40 ~ 60d and complete Rooting and hardening-off culture;
(6) culture environment condition: first secretly cultivate 10d except starting to cultivate, then carry out outside illumination cultivation, other cultivation stage all needs illumination cultivation, intensity of illumination and cycle: 2000 ~ 3000LX, 14h/d; 23 ± 2 ℃ of temperature; Humidity 40%, is first secretly cultivated 10d that is when starting cultivation, then carries out illumination cultivation, and all carries out illumination cultivation in propagation cultivation and Rooting and hardening-off culture stage.
Embodiment 2
OncidiumLuridum tissue-culturing quick-propagation
(1) explant: oncidiumLuridum fruit pod, the complete free from flaw of fruit pod;
(2) explant sterilization: fruit pod is cleaned with clear water, put into 75% alcohol 20s, sterile water wash 2 times, then fruit pod is put into 0.1% mercuric chloride 12min, sterile water wash 5 ~ 7 times;
(3) inoculation and startup are cultivated: fruit pod one side under gnotobasis, sterilization being completed is opened an osculum, seed is sowed in aseptic culture medium, being about to seed sows in aseptic startup medium, the formula of medium is: spend precious No. 1 1.5g/L+ to spend precious No. 2 1.0g/L+ peptone 0.5g/L+ potato 25g/L+ Coconut Juice 80ml/L+ sucrose 20g/L+ agar 5.0g/L, PH 5.8;
(4) propagation is cultivated: start and cultivate after 30d, oncidiumLuridum seedling or protocorm are transferred in proliferated culture medium, proliferation culture medium formula is: 1/2MS+ potato 35g/L+ banana 45g/L+ sucrose 25g/L+ peptone 1.0g/L+ active carbon 1.0g/L+ agar 5.0g/L, and PH 6.0;
(5) Rooting and hardening-off culture: propagation is cultivated 40 ~ 60d, group training seedling is transferred in Rooting and hardening-off culture base, culture medium prescription is: 1/2MS+ potato 30g/L+ banana 55g/L+ sucrose 25g/L+ peptone 1.0g/L+ apple 40g/L+ active carbon 1.0g/L+ agar 5.0g/L, PH 6.0.Cultivate 60d left and right and complete Rooting and hardening-off culture;
(6) culture environment condition: first secretly cultivate 7d except starting to cultivate, then carry out outside illumination cultivation, other cultivation stage all needs illumination cultivation; Intensity of illumination and cycle: 1800 ~ 2500LX, 14h/d; 24 ± 2 ℃ of temperature; Humidity 35% is first secretly cultivated 7d when starting cultivation, then carries out illumination cultivation, and all carries out illumination cultivation in propagation cultivation and Rooting and hardening-off culture stage.
Embodiment 3
Bletilla striata tissue-culturing quick-propagation
(1) explant: bletilla striata fruit pod, the complete free from flaw of fruit pod;
(2) explant sterilization: fruit pod is cleaned with clear water, put into 75% alcohol 20s, sterile water wash 2 times, then fruit pod is put into 0.1% mercuric chloride 12min, sterile water wash 5 ~ 7 times;
(3) inoculation and startup are cultivated: fruit pod one side under gnotobasis, sterilization being completed is opened an osculum, seed is sowed in aseptic culture medium, the formula of medium is: spend precious No. 1 1.2g/L+ to spend precious No. 2 1.5g/L+ peptone 0.5g/L+ potato 35g/L+ Coconut Juice 120ml/L+ sucrose 20g/L+ agar 5.0g/L, PH 6.0;
(4) propagation is cultivated: start and cultivate after 25d, oncidiumLuridum seedling or protocorm are transferred in proliferated culture medium, culture medium prescription is: 1/2MS+ potato 30g/L+ banana 55g/L+ sucrose 25g/L+ peptone 1.0g/L+ active carbon 1.0g/L+ agar 5.0g/L, and PH 6.0;
(5) Rooting and hardening-off culture: propagation is cultivated after 45d, group training seedling is transferred in Rooting and hardening-off culture base, culture medium prescription is: 1/2MS+ potato 35g/L+ banana 60g/L+ sucrose 20g/L+ peptone 1.0g/L+ apple 35g/L+ active carbon 1.0g/L+ agar 5.0g/L, PH 6.0.Cultivate 60d left and right and complete Rooting and hardening-off culture.
(6) culture environment condition: first secretly cultivate 10d except starting to cultivate, then carry out outside illumination cultivation, other cultivation stage all needs illumination cultivation; Intensity of illumination and cycle: 1800 ~ 3000LX, 14h/d; 24 ± 2 ℃ of temperature; Humidity 40%.
In culture medium prescription of the present invention, do not add any chemosynthesis plant hormone, with this medium culture orchid high, fast growth of growth coefficient not only, and group training seedling is healthy, adaptive capacity to environment is strong, transplanting survival rate is high, in this culture medium prescription, do not add any plant hormone, the feature with imitative wild cultivation, the orchid group training seedling growing way of using formula of the present invention to obtain is good and neat, and transplanting survival rate is high.
Claims (8)
1. an orchid fast propagating culture medium is filled a prescription, and it is characterized in that: the medium of the Fast-propagation of described orchid comprises following three medium that cultivation stage is used, and the content of the medium in each stage is:
A. start medium: spend precious No. 1 0.5 ~ 2.5g/L, spend precious No. 2 0.2 ~ 2.0g/L, peptone 0.1 ~ 1.0g/L, potato 20 ~ 45g/L, Coconut Juice 50 ~ 200ml/L, sucrose 10 ~ 30g/L;
B. proliferated culture medium: 1/2MS, potato 15 ~ 40g/L, banana 30 ~ 70g/L, sucrose 15 ~ 40g/L, peptone 0.5 ~ 1.5g/L, active carbon 0.5 ~ 1.5g/L;
C. Rooting and hardening-off culture base: 1/2MS, potato 15 ~ 45g/L, banana 30 ~ 80g/L, sucrose 10 ~ 30g/L, peptone 0.5 ~ 1.5g/L, apple 20 ~ 50g/L, active carbon 0.5 ~ 1.5g/L.
2. a kind of orchid fast propagating culture medium is filled a prescription according to claim 1, it is characterized in that: the medium of the Fast-propagation of described orchid comprises following three medium that cultivation stage is used, and the content of the medium in each stage is:
A. start medium: spend precious No. 1 1.0 ~ 2.0g/L, spend precious No. 2 0.5 ~ 1.5g/L, peptone 0.3 ~ 0.8g/L, potato 25 ~ 40g/L, Coconut Juice 80 ~ 120ml/L, sucrose 10 ~ 20g/L;
B. proliferated culture medium: 1/2MS, potato 25 ~ 35g/L, banana 40 ~ 60g/L, sucrose 20 ~ 30g/L, peptone 0.7 ~ 1.2g/L, active carbon 0.8 ~ 1.3g/L;
C. Rooting and hardening-off culture base: 1/2MS, potato 20 ~ 40g/L, banana 40 ~ 60g/L, sucrose 15 ~ 25g/L, peptone 0.8 ~ 1.2g/L, apple 30 ~ 40g/L, active carbon 0.8 ~ 1.2g/L.
3. a kind of orchid fast propagating culture medium is filled a prescription according to claim 1, it is characterized in that: the medium of the Fast-propagation of described orchid comprises following three medium that cultivation stage is used, and the content of the medium in each stage is:
A. start medium: spend precious No. 1 1.5g/L, spend precious No. 2 1.0g/L, peptone 0.5g/L, potato 30g/L, Coconut Juice 100ml/L, sucrose 15g/L;
B. proliferated culture medium: 1/2MS, potato 30g/L, banana 50g/L, sucrose 25g/L, peptone 1.0g/L, active carbon 1.0g/L;
C. Rooting and hardening-off culture base: 1/2MS, potato 30g/L, banana 50g/L, sucrose 20g/L, peptone 1.0g/L, apple 35g/L, active carbon 1.0g/L.
4. a kind of orchid fast propagating culture medium is filled a prescription according to claim 1, it is characterized in that: the medium of the Fast-propagation of described orchid comprises following three medium that cultivation stage is used, and the content of the medium in each stage is:
A. start medium: spend precious No. 1 1.5g/L, spend precious No. 2 1.0g/L, peptone 0.5g/L, potato 25g/L, Coconut Juice 80ml/L, sucrose 20g/L;
B. proliferated culture medium: 1/2MS, potato 35g/L, banana 45g/L, sucrose 25g/L, peptone 1.0g/L, active carbon 1.0g/L;
C. Rooting and hardening-off culture base: 1/2MS, potato 30g/L, banana 55g/L, sucrose 25g/L, peptone 1.0g/L, apple 40g/L, active carbon 1.0g/L.
5. a kind of orchid fast propagating culture medium is filled a prescription according to claim 1, it is characterized in that: the medium of the Fast-propagation of described orchid comprises following three medium that cultivation stage is used, and the content of the medium in each stage is:
A. start medium: spend precious No. 1 1.2g/L, spend precious No. 2 1.5g/L, peptone 0.5g/L, potato 35g/L, Coconut Juice 120ml/L, sucrose 20g/L;
B. proliferated culture medium: 1/2MS, potato 30g/L, banana 55g/L, sucrose 25g/L, peptone 1.0g/L, active carbon 1.0g/L;
C. Rooting and hardening-off culture base: 1/2MS, potato 35g/L, banana 60g/L, sucrose 20g/L, peptone 1.0g/L, apple 35g/L, active carbon 1.0g/L.
6. according to a kind of orchid fast propagating culture medium formula described in any one of claim 1-5, it is characterized in that: the PH of the medium of described three cultivation stages is 5.8 ~ 6.2.
7. according to a kind of orchid fast propagating culture medium formula described in any one of claim 1-5, it is characterized in that: the seed that the explant that described startup cultivation is used is orchid.
8. according to a kind of orchid fast propagating culture medium formula described in any one of claim 1-5, it is characterized in that: three kinds of described medium are take the solid culture medium that agar is supporter.
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| CN104996301A (en) * | 2015-08-06 | 2015-10-28 | 云南省农业科学院花卉研究所 | Rapid propagation method for holcoglossum flavescens |
| CN105248284A (en) * | 2015-11-09 | 2016-01-20 | 广西壮族自治区药用植物园 | Tissue culture rapid propagation method of bulbophyllum delitescens hance |
| CN106718933A (en) * | 2017-01-23 | 2017-05-31 | 中国林业科学研究院林业研究所 | Spend more the culture medium of class pocket orchid aseptic seeding and sprouting and rooting |
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| CN109287488A (en) * | 2018-11-09 | 2019-02-01 | 翁源县天下泽雨农业科技有限公司 | A kind of 18 sound of laughing stem tip tissue culture method of hybrid orchid |
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| CN110521604A (en) * | 2019-09-25 | 2019-12-03 | 盐城工学院 | A kind of clavus dendrobium nobile tissue culture medium (TCM) and cultural method |
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| CN109122325A (en) * | 2018-11-09 | 2019-01-04 | 翁源县天下泽雨农业科技有限公司 | A kind of aseptic seeding quick-breeding method of sword-leaved cymbidium seed |
| CN109287488A (en) * | 2018-11-09 | 2019-02-01 | 翁源县天下泽雨农业科技有限公司 | A kind of 18 sound of laughing stem tip tissue culture method of hybrid orchid |
| CN109964815A (en) * | 2019-03-26 | 2019-07-05 | 成都大学 | A kind of bletilla striata aseptic seedling rapid induction Multiple Buds and fast numerous method of taking root |
| CN110521604A (en) * | 2019-09-25 | 2019-12-03 | 盐城工学院 | A kind of clavus dendrobium nobile tissue culture medium (TCM) and cultural method |
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