CN108207626A - A kind of tissue of the good dendrobium candidum of seedling growth supports method - Google Patents
A kind of tissue of the good dendrobium candidum of seedling growth supports method Download PDFInfo
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- CN108207626A CN108207626A CN201711474321.5A CN201711474321A CN108207626A CN 108207626 A CN108207626 A CN 108207626A CN 201711474321 A CN201711474321 A CN 201711474321A CN 108207626 A CN108207626 A CN 108207626A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
Tissue the invention discloses a kind of good dendrobium candidum of seedling growth supports method,Take Adult form dendrobium candidum stem section,After being rinsed well with flowing water,It is impregnated 39 minutes with 0.1% mercuric chloride solution,With aseptic water washing 45 times,It puts on superclean bench,Length is cut into as 1 2cm,Small stem section with 23 sections,It is impregnated 13 minutes with 0.1% mercuric chloride solution again,After aseptic water washing 45 times,It is inoculated in sterile inducing culture,In 24 26 DEG C of temperature,24 hour/day of daylight light irradiation,After being cultivated 40 days under 800 1200lx of intensity of illumination,Stem section goes out seedling in internode director,It is grown after seedling to 1 2cm,It is transferred in root media and takes root,It is 2000 3000Lx in illumination,Temperature is cultivated 50 60 days for 20 25 DEG C,To obtain the final product.When the method for the present invention carries out tissue cultures to dendrobium candidum, rooting rate is high, and seedling growth is good.
Description
Technical field
The present invention relates to a kind of method for tissue culture of dendrobium candidum, the good dendrobium candidum of particularly a kind of seedling growth
Tissue supports method.
Background technology
Dendrobium candidum nickname iron sheet orchid, bracketplant, Yunnan iron sheet, iron sheet bracketplant are orchid family Dendrobium herbaceos perennials,
Dendrobium candidum is the top grade of Shihu " medicinal materials, is known as the laudatory title of " gold in Chinese celestial grass-medicine ", is used as medicine with stem, contains polysaccharide, stone
The active ingredients such as dry measure used in former times alkali, alkaloid, with nourishing Yin and clearing heat, the beneficial stomach that promotes the production of body fluid, wet one's whistle improving eyesight and other effects, in antitumor, raising human body
Immunity, anti-aging, anti-oxidant, platelet aggregation-against, reduces blood glucose, treatment cataract etc. at treatment enterogastric diseases
There is good efficacy.
Modern pharmacology research shows that dendrobium candidum has humidification to non-specific and specific immunity, anti-swollen
Knurl, treatment that is anti-oxidant, reducing blood glucose and cataract etc., pharmacological action is notable.Its chemical composition is more complicated, it has been determined that
Type of compounds mainly have：Alkaloids, polysaccharide, luxuriant and rich with fragrance class, bibenzyl parent nucleus class, wherein alkaloids, polysaccharide pharmacology are lived
Property is more notable.Stem of noble dendrobium alkaline sesquiterpenoids alkaloid is peculiar for Dendrobium Sw, and wherein dendrobine is to find earliest, and research is most
More alkaloids；HERBA DENDROBII alkali is similar to non-steroid anti-inflammatory drug pharmacological action, has immunological regulation, anti-inflammatory, expansion blood vessel
The effects that.
At present, certain achievement has been obtained in preserving seed and quick proliferation to dendrobium candidum using tissue culture technology, but
Improved seeds cause seedling growth bad, rooting rate pole through subculture for several times or because of factors such as Corticosteroids are improper, culture environments
It is low.
Invention content
The purpose of the present invention, the tissue for being to provide a kind of good dendrobium candidum of seedling growth support method, the method for the present invention pair
When dendrobium candidum carries out tissue cultures, rooting rate is high, and seedling growth is good.
The invention is realized in this way：A kind of tissue of the good dendrobium candidum of seedling growth supports method, takes Adult form iron sheet
Stem of noble dendrobium stem section after being rinsed well with flowing water, is impregnated 3-9 minutes with 0.1% mercuric chloride solution, with aseptic water washing 4-5 times, put super
On net workbench, small stem section of the length for 1-2cm, with 2-3 section is cut into, then impregnated 1-3 minutes with 0.1% mercuric chloride solution,
After aseptic water washing 4-5 times, be inoculated in sterile inducing culture, 24-26 DEG C of temperature, 24 hour/day of daylight light irradiation,
After being cultivated 40 days under intensity of illumination 800-1200lx, stem section goes out seedling in internode director, is grown after seedling to 1-2cm, is transferred to
Take root in root media, illumination be 2000-3000Lx, temperature for 20-25 DEG C cultivate 50-60 days to get.
The tissue of the good dendrobium candidum of aforementioned seedling growth is supported in method, and the inducing culture is：It is trained to 1LMS
Support and NAA2.0-4.0mg, mashed potatoes 5-10g and sucrose 15-25g added in base, mixing to get.
The tissue of the good dendrobium candidum of aforementioned seedling growth is supported in method, and the inducing culture is：It is trained to 1LMS
Support and NAA 3.0mg, mashed potatoes 7g and sucrose 20g added in base, mixing to get.
The tissue of the good dendrobium candidum of aforementioned seedling growth is supported in method, and the root media is：It is trained to 1LMS
It supports and bananas juice 5-15mg, peptone 5-15mg, bletilla striata extract 1-6mg, Radix Salviae Miltiorrhizae juice 2-5mg, Astragalus Root P.E 3- is added in base
9mg, herba hedyotis diffusae extract 5-10mg, licorice 1-5mg and Fructus Jujubae extract 6-12mg, mixing to get.
The tissue of the good dendrobium candidum of aforementioned seedling growth is supported in method, and the root media is：It is trained to 1LMS
It supports and bananas juice 8-12mg, peptone 8-12mg, bletilla striata extract 2-4mg, Radix Salviae Miltiorrhizae juice 3-4mg, Astragalus Root P.E 5- is added in base
7mg, herba hedyotis diffusae extract 7-8mg, licorice 2-4mg and Fructus Jujubae extract 8-10mg, mixing to get.
The tissue of the good dendrobium candidum of aforementioned seedling growth is supported in method, and the root media is：It is trained to 1LMS
It supports and bananas juice 10mg, peptone 10mg, bletilla striata extract 3mg, Radix Salviae Miltiorrhizae juice 4mg, Astragalus Root P.E 6mg, long-noded pit viper is added in base
Diffusa extract 7mg, licorice 2mg and Fructus Jujubae extract 9mg, mixing to get.
The tissue of the good dendrobium candidum of aforementioned seedling growth is supported in method, and the bletilla striata extract is prepared：It takes
The bletilla striata after adding in the 8-10 times of water measured immersion 10-15min, decocts 1-2h, filtering, and filter residue adds 5-6 times and measures water extraction 2-3
Secondary, each 20-30min is filtered, and merging filtrate is spare after being condensed into concentrate；Filter residue adds 5-6 times and measures 50-60%'s
Ethanol solution, refluxing extraction 2-3 times, each 20-30min, filtering, merging filtrate recycle ethyl alcohol, obtain alcohol extract, alcohol extract
Merge with above-mentioned Aqueous extracts, when being concentrated into 50-60 DEG C relative density be 1.10-1.20 medicinal extract, dry, pulverize to get.
The tissue of the good dendrobium candidum of aforementioned seedling growth is supported in method, and the Astragalus Root P.E is prepared：It takes
Radix Astragali, adds in the 6-8 times of ethanol solution refluxing extraction for measuring 60-70% 2-3 times, each 20-30min, filtering, and merging filtrate returns
Ethyl alcohol is received, obtains alcohol extract, the dense medicinal extract to relative density at 50-60 DEG C for 1.10-1.20 is condensed into, dry, pulverize, i.e.,
.
The tissue of the good dendrobium candidum of aforementioned seedling growth is supported in method, and the licorice is prepared：It takes
Radix Glycyrrhizae is cleaned, after drying, is placed in demulcen container, sprays vinegar with the method for spray, is stirred in sprinkling until uniformly, connecing
After bored profit exhausts for 35 minutes by licorice medicinal materials completely to vinegar liquid, heating, fry to yellow color it is slightly dry when, after taking-up cools 6h, in
Dry 85 hours at 34-40 DEG C, coarse granule is ground into get turning within during which every 12 hours that medicine is primary, the weight ratio of Radix Glycyrrhizae and vinegar
It is 35:3；Above-mentioned licorice root particles are taken, the ethyl alcohol for adding in 9 times of amounts a concentration of 80% extracts 2 times, and 1.5 hours every time, extracting solution closed
And filter, filtrate recycling ethanol and relative density is the medicinal extract of 1.10-1.20 when being concentrated into 50-60 DEG C, extract dry crushes,
To obtain the final product.
The tissue of the good dendrobium candidum of aforementioned seedling growth is supported in method, and the herba hedyotis diffusae extract is made in this way
It is standby：Oldenlandia diffusa is taken, adds 8-10 times to measure water and decocts, after 2-3h, filtering, relative density is when filtrate is concentrated into 50-60 DEG C
The medicinal extract of 1.10-1.20 to get；The Fructus Jujubae extract is prepared：Jujube is taken, adds 8-10 times to measure water and decocts, after 2-3h,
Filtering, when filtrate is concentrated into 50-60 DEG C relative density be 1.10-1.20 medicinal extract to get；The Radix Salviae Miltiorrhizae juice is prepared：
Radix Salviae Miltiorrhizae adds 3-5 times to measure water, break into juice to get.
Applicant has carried out a large amount of experimental study to the present invention, and part Experiment is as follows：
Experimental example
1 method
3 processing groups, i.e., of the present invention group, contrast groups 1 and contrast groups 2 are set.
Of the present invention group：Tissue cultures are carried out by embodiment 1；
Contrast groups 1：Method for tissue culture is free of bletilla striata extract, Radix Salviae Miltiorrhizae with embodiment 1 in the root media
Juice, Astragalus Root P.E, herba hedyotis diffusae extract, licorice and Fructus Jujubae extract.
Contrast groups 2：Method for tissue culture is with embodiment 1, but in the root media：
The bletilla striata extract is prepared：The bletilla striata adds 8 times of amount water to decoct 3, and each 2h, decoction filtering is condensed into 50-60
DEG C when relative density be 1.10-1.20 medicinal extract, dry, pulverize to get.
The Astragalus Root P.E is prepared：Radix Astragali is taken, 8 times of amount water is added to decoct 3, each 2h, decoction filtering is condensed into
At 50-60 DEG C relative density be 1.10-1.20 medicinal extract, dry, pulverize to get.
The licorice is prepared：Extracting liquorice adds 8 times of amount water to decoct 3, and each 2h, decoction filtering is condensed into
At 50-60 DEG C relative density be 1.10-1.20 medicinal extract, dry, pulverize to get.
Plant height, the number of blade, newly-increased radical and the body weight increase rate of each group plant are measured after inoculation 60d.
Every group of result be subject to and organize in average value.
Body weight increase rate (%)=(weight before the tissue culture plant inoculation of weight-every bottle after every bottle of tissue-cultured seedling culture)/every bottle
Weight × 100% before tissue culture plant inoculation.
2 results
The influence of 2.1 pairs of candidum tissue culturing seedling growths
Growing state after 60d is cultivated is shown in Table 1.
The influence that table 1 grows tissue-cultured seedling
| Group | Plant height/cm | The number of blade/piece | Newly-increased radical | Body weight increase rate/% |
| Contrast groups 1 | 3.60±0.15 | 4.06±0.26 | 2.52±0.12 | 193.32±24.66 |
| Contrast groups 2 | 4.22±0.11 | 4.57±0.48 | 2.94±0.07 | 222.53±32.41 |
| Of the present invention group | 4.89±0.12 | 5.48±0.39 | 3.56±0.10 | 259.21±30.15 |
As seen from table, the plant height of present invention group plant, the number of blade, newly-increased radical and body weight increase rate are above contrast groups 1
With contrast groups 2.
Compared with prior art, when carrying out tissue cultures with present invention side, tissue-cultured seedling is grown, the plant height of plant, blade
Number, newly-increased radical and body weight increase rate are high, and hormone is free of in culture medium.
Specific embodiment
Embodiment 1.
A kind of tissue of the good dendrobium candidum of seedling growth supports method, takes Adult form dendrobium candidum stem section, is rinsed with flowing water
It after clean, is impregnated 6 minutes with 0.1% mercuric chloride solution, with aseptic water washing 5 times, puts on superclean bench, be cut into length as 1-
2cm, the small stem section with 2-3 section, then impregnated 2 minutes with 0.1% mercuric chloride solution, after aseptic water washing 4 times, it is inoculated in
In sterile inducing culture, 40 are cultivated under 24-26 DEG C of temperature, 24 hour/day of daylight light irradiation, intensity of illumination 800-1200lx
After it, stem section goes out seedling in internode director, is grown after seedling to 1-2cm, is transferred in root media and takes root, and is in illumination
2000-3000Lx, temperature for 20-25 DEG C cultivate 60 days to get.
The inducing culture is：NAA 3.0mg, mashed potatoes 7g and sucrose 20g are added in into 1LMS culture mediums, is mixed
It is even to get.
The root media is：Bananas juice 10mg, peptone 10mg, bletilla striata extract are added in into 1LMS culture mediums
3mg, Radix Salviae Miltiorrhizae juice 4mg, Astragalus Root P.E 6mg, herba hedyotis diffusae extract 7mg, licorice 2mg and Fructus Jujubae extract 9mg,
Mixing to get.
The bletilla striata extract is prepared：The bletilla striata is taken, after adding in the water immersion 12min of 9 times of amounts, decocts 1.5h, mistake
Filter, filter residue add 6 times of amount water and extract 2 times, each 25min, filtering, and merging filtrate is spare after being condensed into concentrate；Filter residue
The ethanol solution of 5 times of amounts 55% is added, refluxing extraction 2 times, each 25min filters, merging filtrate, recycles ethyl alcohol, obtains alcohol extracting
Liquid is taken, alcohol extract merges with above-mentioned Aqueous extracts, and relative density is the medicinal extract of 1.10-1.20 when being concentrated into 50-60 DEG C, dry, powder
It is broken to get.
The Radix Salviae Miltiorrhizae juice is prepared：Radix Salviae Miltiorrhizae add 4 times amount water, break into juice to get.
The Astragalus Root P.E is prepared：It takes Radix Astragali, adds in the ethanol solution refluxing extraction 2 times of 7 times of amounts 65%, often
Secondary 25min, filtering, merging filtrate, recycle ethyl alcohol, obtain alcohol extract, be condensed into it is dense to relative density at 50-60 DEG C be 1.10-
1.20 medicinal extract, dry, pulverize to get.
The herba hedyotis diffusae extract is prepared：Oldenlandia diffusa is taken, after 9 times of amount water is added to decoct 2.5h, mistake
Filter, when filtrate is concentrated into 50-60 DEG C relative density be 1.10-1.20 medicinal extract to get；
The licorice is prepared：Extracting liquorice is cleaned, after drying, is placed in demulcen container, with the side of spray
Method sprays vinegar, is stirred in sprinkling until uniformly, after then bored profit is exhausted for 35 minutes by licorice medicinal materials completely to vinegar liquid, adding
Heat, fry to yellow color it is slightly dry when, after taking-up cools 6h, dry 85 hours at 34-40 DEG C, be ground into coarse granule to get during which
Turn within every 12 hours that medicine is primary, the weight ratio of Radix Glycyrrhizae and vinegar is 35:3；Above-mentioned licorice root particles are taken, add in 9 times of amounts a concentration of 80%
Ethyl alcohol extract 2 times, 1.5 hours every time, extracting solution merged, and filtering, filtrate recycling ethanol is simultaneously concentrated into relatively close at 50-60 DEG C
Spend the medicinal extract for 1.10-1.20, extract dry, crush to get.
The Fructus Jujubae extract is prepared：Jujube is taken, after 9 times of amount water is added to decoct 2.5h, filtering, filtrate is concentrated into
At 50-60 DEG C relative density be 1.10-1.20 medicinal extract to get；
Embodiment 2.
A kind of tissue of the good dendrobium candidum of seedling growth supports method, takes Adult form dendrobium candidum stem section, is rinsed with flowing water
It after clean, is impregnated 9 minutes with 0.1% mercuric chloride solution, with aseptic water washing 5 times, puts on superclean bench, be cut into length as 1-
2cm, the small stem section with 2-3 section, then impregnated 3 minutes with 0.1% mercuric chloride solution, after aseptic water washing 5 times, it is inoculated in
In sterile inducing culture, 40 are cultivated under 24-26 DEG C of temperature, 24 hour/day of daylight light irradiation, intensity of illumination 800-1200lx
After it, stem section goes out seedling in internode director, is grown after seedling to 1-2cm, is transferred in root media and takes root, and is in illumination
2000-3000Lx, temperature for 20-25 DEG C cultivate 50 days to get.
The inducing culture is：NAA4.0mg, mashed potatoes 10g and sucrose 25g are added in into 1LMS culture mediums, is mixed
It is even to get.
The root media is：Bananas juice 10mg, peptone 10mg, bletilla striata extract are added in into 1LMS culture mediums
3mg, Radix Salviae Miltiorrhizae juice 4mg, Astragalus Root P.E 6mg, herba hedyotis diffusae extract 7mg, licorice 2mg and Fructus Jujubae extract 9mg,
Mixing to get.
The bletilla striata extract is prepared：The bletilla striata is taken, after adding in the water immersion 15min of 10 times of amounts, decocts 2h, mistake
Filter, filter residue add 6 times of amount water and extract 3 times, each 30min, filtering, and merging filtrate is spare after being condensed into concentrate；Filter residue
The ethanol solution of 6 times of amounts 60% is added, refluxing extraction 3 times, each 30min filters, merging filtrate, recycles ethyl alcohol, obtains alcohol extracting
Liquid is taken, alcohol extract merges with above-mentioned Aqueous extracts, and relative density is the medicinal extract of 1.10-1.20 when being concentrated into 50-60 DEG C, dry, powder
It is broken to get.
The Radix Salviae Miltiorrhizae juice is prepared：Radix Salviae Miltiorrhizae add 5 times amount water, break into juice to get.
The Astragalus Root P.E is prepared：It takes Radix Astragali, adds in the ethanol solution refluxing extraction 3 times of 8 times of amounts 70%, often
Secondary 30min, filtering, merging filtrate, recycle ethyl alcohol, obtain alcohol extract, be condensed into it is dense to relative density at 50-60 DEG C be 1.10-
1.20 medicinal extract, dry, pulverize to get.
The herba hedyotis diffusae extract is prepared：Oldenlandia diffusa is taken, 10 times of amount water is added to decoct, after 2-3h, mistake
Filter, when filtrate is concentrated into 50-60 DEG C relative density be 1.10-1.20 medicinal extract to get；
The licorice is prepared：Extracting liquorice is cleaned, after drying, is placed in demulcen container, with the side of spray
Method sprays vinegar, is stirred in sprinkling until uniformly, after then bored profit is exhausted for 35 minutes by licorice medicinal materials completely to vinegar liquid, adding
Heat, fry to yellow color it is slightly dry when, after taking-up cools 6h, dry 85 hours at 34-40 DEG C, be ground into coarse granule to get during which
Turn within every 12 hours that medicine is primary, the weight ratio of Radix Glycyrrhizae and vinegar is 35:3；Above-mentioned licorice root particles are taken, add in 9 times of amounts a concentration of 80%
Ethyl alcohol extract 2 times, 1.5 hours every time, extracting solution merged, and filtering, filtrate recycling ethanol is simultaneously concentrated into relatively close at 50-60 DEG C
Spend the medicinal extract for 1.10-1.20, extract dry, crush to get.
The Fructus Jujubae extract is prepared：Jujube is taken, after 10 times of amount water is added to decoct 3h, filtering, filtrate is concentrated into 50-
At 60 DEG C relative density be 1.10-1.20 medicinal extract to get；
Embodiment 3.
A kind of tissue of the good dendrobium candidum of seedling growth supports method, takes Adult form dendrobium candidum stem section, is rinsed with flowing water
It after clean, is impregnated 3 minutes with 0.1% mercuric chloride solution, with aseptic water washing 4 times, puts on superclean bench, be cut into length as 1-
2cm, the small stem section with 2-3 section, then impregnated 1 minute with 0.1% mercuric chloride solution, after aseptic water washing 4 times, it is inoculated in
In sterile inducing culture, 40 are cultivated under 24-26 DEG C of temperature, 24 hour/day of daylight light irradiation, intensity of illumination 800-1200lx
After it, stem section goes out seedling in internode director, is grown after seedling to 1-2cm, is transferred in root media and takes root, and is in illumination
2000-3000Lx, temperature for 20-25 DEG C cultivate 50 days to get.
The inducing culture is：Addition NAA2.0mg, mashed potatoes 5g and sucrose 15g into 1LMS culture mediums, mixing,
To obtain the final product.
The root media is：Bananas juice 8mg, peptone 8mg, bletilla striata extract are added in into 1LMS culture mediums
2mg, Radix Salviae Miltiorrhizae juice 3mg, Astragalus Root P.E 5mg, herba hedyotis diffusae extract 7mg, licorice 2mg and Fructus Jujubae extract 8mg,
Mixing to get.
The bletilla striata extract is prepared：The bletilla striata is taken, after adding in the water immersion 10min of 8 times of amounts, 1h is decocted, filters,
Filter residue adds 5 times of amount water and extracts 2 times, each 20min, filtering, and merging filtrate is spare after being condensed into concentrate；Filter residue adds again
Enter the ethanol solution of 5 times of amounts 50%, refluxing extraction 2 times, each 20min filters, merging filtrate, recycles ethyl alcohol, obtains alcohol extracting
Liquid, alcohol extract merge with above-mentioned Aqueous extracts, and relative density is the medicinal extract of 1.10-1.20 when being concentrated into 50-60 DEG C, be dry, pulverize,
To obtain the final product.
The Radix Salviae Miltiorrhizae juice is prepared：Radix Salviae Miltiorrhizae add 3 times amount water, break into juice to get.
The Astragalus Root P.E is prepared：It takes Radix Astragali, adds in the ethanol solution refluxing extraction 2 times of 6 times of amounts 60%, often
Secondary 20min, filtering, merging filtrate, recycle ethyl alcohol, obtain alcohol extract, be condensed into it is dense to relative density at 50-60 DEG C be 1.10-
1.20 medicinal extract, dry, pulverize to get.
The herba hedyotis diffusae extract is prepared：Oldenlandia diffusa is taken, after 8 times of amount water is added to decoct 2h, filtering,
When filtrate is concentrated into 50-60 DEG C relative density be 1.10-1.20 medicinal extract to get；
The licorice is prepared：Extracting liquorice is cleaned, after drying, is placed in demulcen container, with the side of spray
Method sprays vinegar, is stirred in sprinkling until uniformly, after then bored profit is exhausted for 35 minutes by licorice medicinal materials completely to vinegar liquid, adding
Heat, fry to yellow color it is slightly dry when, after taking-up cools 6h, dry 85 hours at 34-40 DEG C, be ground into coarse granule to get during which
Turn within every 12 hours that medicine is primary, the weight ratio of Radix Glycyrrhizae and vinegar is 35:3；Above-mentioned licorice root particles are taken, add in 9 times of amounts a concentration of 80%
Ethyl alcohol extract 2 times, 1.5 hours every time, extracting solution merged, and filtering, filtrate recycling ethanol is simultaneously concentrated into relatively close at 50-60 DEG C
Spend the medicinal extract for 1.10-1.20, extract dry, crush to get.
The Fructus Jujubae extract is prepared：Jujube is taken, after 8 times of amount water is added to decoct 2h, filtering, filtrate is concentrated into 50-
At 60 DEG C relative density be 1.10-1.20 medicinal extract to get.
Claims (10)
1. a kind of tissue of the good dendrobium candidum of seedling growth supports method, it is characterised in that：Adult form dendrobium candidum stem section is taken, is used
After flowing water is rinsed well, impregnated 3-9 minutes with 0.1% mercuric chloride solution, with aseptic water washing 4-5 times, put on superclean bench,
Small stem section of the length for 1-2cm, with 2-3 section is cut into, then is impregnated 1-3 minutes with 0.1% mercuric chloride solution, uses aseptic water washing
It after 4-5 times, is inoculated in sterile inducing culture, in 24-26 DEG C of temperature, 24 hour/day of daylight light irradiation, intensity of illumination 800-
After being cultivated 40 days under 1200lx, stem section goes out seedling in internode director, is grown after seedling to 1-2cm, is transferred in root media
Take root, illumination be 2000-3000Lx, temperature for 20-25 DEG C cultivate 50-60 days to get.
2. the tissue of the good dendrobium candidum of seedling growth as described in claim 1 supports method, it is characterised in that：The induction
Culture medium is：Add in NAA2.0-4.0mg, mashed potatoes 5-10g and sucrose 15-25g into 1LMS culture mediums, mixing to get.
3. the tissue of the good dendrobium candidum of seedling growth as claimed in claim 2 supports method, it is characterised in that：The induction
Culture medium is：Add in NAA 3.0mg, mashed potatoes 7g and sucrose 20g into 1LMS culture mediums, mixing to get.
4. the tissue of the good dendrobium candidum of seedling growth as described in claim 1 supports method, it is characterised in that：Described takes root
Culture medium is：Bananas juice 5-15mg, peptone 5-15mg, bletilla striata extract 1-6mg, Radix Salviae Miltiorrhizae juice 2- are added in into 1LMS culture mediums
5mg, Astragalus Root P.E 3-9mg, herba hedyotis diffusae extract 5-10mg, licorice 1-5mg and Fructus Jujubae extract 6-12mg,
Mixing to get.
5. the tissue of the good dendrobium candidum of seedling growth as claimed in claim 4 supports method, it is characterised in that：Described takes root
Culture medium is：Bananas juice 8-12mg, peptone 8-12mg, bletilla striata extract 2-4mg, Radix Salviae Miltiorrhizae juice 3- are added in into 1LMS culture mediums
4mg, Astragalus Root P.E 5-7mg, herba hedyotis diffusae extract 7-8mg, licorice 2-4mg and Fructus Jujubae extract 8-10mg,
Mixing to get.
6. the tissue of the good dendrobium candidum of seedling growth as claimed in claim 5 supports method, it is characterised in that：Described takes root
Culture medium is：Bananas juice 10mg, peptone 10mg, bletilla striata extract 3mg, Radix Salviae Miltiorrhizae juice 4mg, Radix Astragali are added in into 1LMS culture mediums
Extract 6mg, herba hedyotis diffusae extract 7mg, licorice 2mg and Fructus Jujubae extract 9mg, mixing to get.
7. the tissue of the good dendrobium candidum of seedling growth as described in any one of claim 4-6 supports method, it is characterised in that：
The bletilla striata extract is prepared：The bletilla striata is taken, after adding in the 8-10 times of water measured immersion 10-15min, 1-2h is decocted, filters,
Filter residue adds 5-6 times and measures water extraction 2-3 times, each 20-30min, filters, merging filtrate is spare after being condensed into concentrate；
Filter residue adds the 5-6 times of ethanol solution for measuring 50-60%, refluxing extraction 2-3 times, and each 20-30min is filtered, merging filtrate, is returned
Ethyl alcohol is received, obtains alcohol extract, alcohol extract merges with above-mentioned Aqueous extracts, and relative density is 1.10-1.20's when being concentrated into 50-60 DEG C
Medicinal extract, dry, pulverize to get.
8. the tissue of the good dendrobium candidum of seedling growth as described in any one of claim 4-6 supports method, it is characterised in that：
The Astragalus Root P.E is prepared：It takes Radix Astragali, adds in 6-8 times of ethanol solution refluxing extraction for measuring 60-70% 2-3 times, every time
20-30min, filtering, merging filtrate, recycle ethyl alcohol, obtain alcohol extract, be condensed into it is dense to relative density at 50-60 DEG C be 1.10-
1.20 medicinal extract, dry, pulverize to get.
9. the tissue of the good dendrobium candidum of seedling growth as described in any one of claim 4-6 supports method, it is characterised in that：
The licorice is prepared：Extracting liquorice is cleaned, after drying, is placed in demulcen container, sprays food with the method for spray
Vinegar is stirred in sprinkling until uniformly, after then bored profit is exhausted for 35 minutes by licorice medicinal materials completely to vinegar liquid, heating is fried to Huang
It is 85 hours dry at 34-40 DEG C after taking-up cools 6h when color is slightly dry, coarse granule is ground into get during which every 12 hours
Turn over that medicine is primary, the weight ratio of Radix Glycyrrhizae and vinegar is 35:3；Above-mentioned licorice root particles are taken, add in the ethyl alcohol extraction of 9 times of amounts a concentration of 80%
2 times, 1.5 hours every time, extracting solution merged, filtering, filtrate recycling ethanol and relative density is 1.10- when being concentrated into 50-60 DEG C
1.20 medicinal extract, extract dry, crush to get.
10. the tissue of the good dendrobium candidum of seedling growth as described in any one of claim 4-6 supports method, feature exists
In：The herba hedyotis diffusae extract is prepared：Oldenlandia diffusa is taken, adds 8-10 times to measure water and decocts, after 2-3h, filtering,
When filtrate is concentrated into 50-60 DEG C relative density be 1.10-1.20 medicinal extract to get；The Fructus Jujubae extract is prepared：It takes
Jujube adds 8-10 times to measure water and decocts, and after 2-3h, filtering, relative density is the leaching of 1.10-1.20 when filtrate is concentrated into 50-60 DEG C
Cream to get；The Radix Salviae Miltiorrhizae juice is prepared：Radix Salviae Miltiorrhizae adds 3-5 times to measure water, break into juice to get.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101213941A (en) * | 2008-01-18 | 2008-07-09 | 中国科学院昆明植物研究所 | Fast replication and in-vitro conservation method for dendrobium |
| CN107018902A (en) * | 2017-05-12 | 2017-08-08 | 贵州济生农业科技有限公司 | A kind of method for tissue culture of dendrobium candidum |
-
2017
- 2017-12-29 CN CN201711474321.5A patent/CN108207626A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101213941A (en) * | 2008-01-18 | 2008-07-09 | 中国科学院昆明植物研究所 | Fast replication and in-vitro conservation method for dendrobium |
| CN107018902A (en) * | 2017-05-12 | 2017-08-08 | 贵州济生农业科技有限公司 | A kind of method for tissue culture of dendrobium candidum |
Non-Patent Citations (1)
| Title |
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| 张书萍等: "铁皮石斛的组织培养与快速繁殖", 《辽宁农业科学》 * |
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