CN107018902A - A kind of method for tissue culture of dendrobium candidum - Google Patents
A kind of method for tissue culture of dendrobium candidum Download PDFInfo
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- CN107018902A CN107018902A CN201710332651.4A CN201710332651A CN107018902A CN 107018902 A CN107018902 A CN 107018902A CN 201710332651 A CN201710332651 A CN 201710332651A CN 107018902 A CN107018902 A CN 107018902A
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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
Abstract
The invention discloses a kind of method for tissue culture of dendrobium candidum, dendrobium candidum protocorm is taken, is cultivated in root media, until growing the dendrobium candidum seedling that 12 true leaves are highly 1.5 2.5cm；It is 1.5 2.5cm, the dendrobium candidum seedling containing 12 true leaves to take height, is inoculated in seedling culture medium, cultivates 50 60 days, produces.When the inventive method carries out tissue cultures to dendrobium candidum, proliferation rate is up to 358.5%, and seedling growth is good.
Description
Technical field
The present invention relates to a kind of method for tissue culture of dendrobium candidum, the group of the high dendrobium candidum of particularly a kind of proliferation rate
Knit cultural 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, have 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 the effects such as, it is antitumor, improve human body
In terms of immunity, treatment enterogastric diseases, anti-aging, anti-oxidant, platelet aggregation-against, reduction blood glucose, treatment cataract
There is good efficacy.
Modern pharmacology research shows that dendrobium candidum has humidification to non-specific and specific immunity, anti-swollen
In terms of the treatment of knurl, anti-oxidant, reduction blood glucose and cataract, 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 that Dendrobium Sw is peculiar, and wherein dendrobine is earliest discovery, and research is most
Many alkaloids；HERBA DENDROBII alkali is similar to non-steroid anti-inflammatory drug pharmacological action, with immunological regulation, anti-inflammatory, expansion blood vessel
Deng effect.
At present, certain achievement has been obtained in preserving seed and quick proliferation to dendrobium candidum using tissue culture technology, but
Improved seeds are through subculture for several times or because of factors such as Corticosteroids are improper, culture environments, proliferation rate and low.
The content of the invention
The purpose of the present invention, is to provide a kind of method for tissue culture of dendrobium candidum, and the inventive method is entered to dendrobium candidum
During row tissue cultures, proliferation rate is up to 358.5%, and seedling growth is good.
What the present invention was realized in.
A kind of method for tissue culture of dendrobium candidum, takes dendrobium candidum protocorm, is cultivated in root media, until long
Go out the dendrobium candidum seedling that 1-2 pieces true leaf is highly 1.5-2.5cm；It is 1.5-2.5cm, the iron sheet of the true leaf of piece containing 1-2 to take height
Stem of noble dendrobium seedling, is inoculated in seedling culture medium, cultivates 50-60 days, produces.
The method for tissue culture of foregoing dendrobium candidum, the root media calculates main by coptis root 5-15 by weight
Part, 10-20 parts of the root of herbaceous peony, 5-15 parts of dragon's blood, 10-20 parts of pseudo-ginseng, 5-15 parts of ginger, 5-15 parts of Poria cocos, 1-10 parts of peptone, potato
5-15 parts of 1-10 parts, 1-5 parts of potato and soya bean are made.
The method for tissue culture of foregoing dendrobium candidum, the root media is calculated by weight, mainly by coptis root 10
Part, 15 parts of the root of herbaceous peony, 10 parts of dragon's blood, 15 parts of pseudo-ginseng, 10 parts of ginger, 10 parts of Poria cocos, 5 parts of peptone, 5 parts of potato, 3 parts of potato and
10 parts of soya bean is made.
The method for tissue culture of foregoing dendrobium candidum, the root media is prepared in the steps below：
(1) coptis root, the root of herbaceous peony, dragon's blood, pseudo-ginseng, ginger and Poria cocos are taken, plus 16-18 times is measured decocting and boiled after 5-6h, filtering, filter residue
Discard, collect filtrate, obtain A product；
(2) fetch earth beans and potato, add 5-6 times and measure water, break into slurry, obtain B product；
(3) soya bean is taken, powder is ground into, C product are obtained；
(4) A product, the mixing of B product, are heated to 50-60 DEG C, add after C product and peptone, stirring 5-10min, are cooled to 5-10
DEG C, produce.
The method for tissue culture of foregoing dendrobium candidum, the seedling culture medium calculates main by cooked maize mud by weight
20-30 parts of 10-20 parts, 10-20 parts of Chinese yam mud, 10-20 parts of taro paste, 1-5 parts of agar and Chinese medicine slag are mixed, and are produced.
The method for tissue culture of foregoing dendrobium candidum, the seedling culture medium is calculated by weight, mainly by cooked maize
15 parts of mud, 15 parts of Chinese yam mud, 15 parts of taro paste, the 25 parts of mixings of 3 parts of agar and Chinese medicine slag, are produced.
The method for tissue culture of foregoing dendrobium candidum, the main root of kudzu vine 50-60 by calculating by weight of the Chinese medicine slag
20-30 parts of part, 30-40 parts of the root of Chinese clematis, 20-30 parts of selfheal, 10-20 parts of cortex albiziae, 1-10 parts of Herba Andrographitis and rhizoma atractylodis are by following
It is prepared by method：Take the root of kudzu vine, the root of Chinese clematis, selfheal, cortex albiziae, Herba Andrographitis and rhizoma atractylodis plus 5-6 times is measured water, decoct 2-3h, filter, filter
Slag dry, pulverize into coarse granule, produce.
The method for tissue culture of foregoing dendrobium candidum, the Chinese medicine slag it is main by calculate by weight 55 parts of the root of kudzu vine,
25 parts of 35 parts of the root of Chinese clematis, 25 parts of selfheal, 15 parts of cortex albiziae, 5 parts of Herba Andrographitis and rhizoma atractylodis are prepared as follows：Take the root of kudzu vine, prestige
Medicine xiaolingxian, selfheal, cortex albiziae, Herba Andrographitis and rhizoma atractylodis add 5.5 times of amount water, decoct 2.5h, and filtering, filter residue and drying is ground into thick
Grain, is produced.
Applicant has carried out substantial amounts of experimental study to the present invention, and part Experiment is as follows：
Experimental example
1 method
2 treatment groups, i.e., of the present invention group and contrast groups are set.
Of the present invention group：Tissue cultures are carried out by embodiment 1；
Contrast groups：Coptis root, the root of herbaceous peony, dragon's blood, three are free of in method for tissue culture be the same as Example 1, but the root media
7th, Chinese medicine slag is free of in ginger and Poria cocos, the seedling culture medium.
Observed after 60d, count the growing state of tissue-cultured seedling.
Statistical method：
Average plant height during average plant height-subculture during growth value (cm)=statistics；
Proliferation rate (%)=(seedling sum during one subculture of seedling sum during statistics)/initial seedling sum × 100%.
2 results
The influence of 2.1 pairs of candidum tissue culturing seedling growths
Growing state after being cultivated through 50d is shown in Table 1.
The influence that table 1 grows to tissue-cultured seedling
| Group | Growth value/cm | Proliferation rate/% |
| Contrast groups | 1.43 | 253.6 |
| Of the present invention group | 1.85 | 358.5 |
As seen from table, present invention group growth value and appreciation rate are above contrast groups.
Compared with prior art, when carrying out tissue cultures with present invention side, tissue-cultured seedling grows, growth value and proliferation rate
Hormone is free of in height, and culture medium.
Compared with prior art, when the inventive method carries out tissue cultures to dendrobium candidum, proliferation rate is up to 358.5%,
Seedling growth is good.
Embodiment
Embodiment 1.
A kind of method for tissue culture of dendrobium candidum, takes dendrobium candidum protocorm, is cultivated in root media, until long
Go out the dendrobium candidum seedling that 1-2 pieces true leaf is highly 1.5-2.5cm；It is 1.5-2.5cm, the iron sheet of the true leaf of piece containing 1-2 to take height
Stem of noble dendrobium seedling, is inoculated in seedling culture medium, cultivates 50-60 days, produces.
The prescription of rooting medium：Coptis root 10kg, root of herbaceous peony 15kg, dragon's blood 10kg, pseudo-ginseng 15kg, ginger 10kg, Poria cocos
10kg, peptone 5kg, potato 5kg, potato 3kg and soya bean 10kg.
The root media is prepared in the steps below：
(1) take coptis root, the root of herbaceous peony, dragon's blood, pseudo-ginseng, ginger and Poria cocos, plus 17 times of amount decoctings to boil after 5.5h, filter, filter residue is abandoned
Go, collect filtrate, obtain A product；
(2) fetch earth beans and potato, add 5.5 times of amount water, break into slurry, obtain B product；
(3) soya bean is taken, powder is ground into, C product are obtained；
(4) A product, the mixing of B product, are heated to 50-60 DEG C, add after C product and peptone, stirring 8min, are cooled to 5-10 DEG C,
Produce.
The Chinese medicine slag is by root of kudzu vine 55kg, root of Chinese clematis 35kg, selfheal 25kg, cortex albiziae 15kg, Herba Andrographitis 5kg and rhizoma atractylodis
25kg is prepared as follows：The root of kudzu vine, the root of Chinese clematis, selfheal, cortex albiziae, Herba Andrographitis and rhizoma atractylodis plus 4 times of amount water are taken, are decocted
2.5h, filtering, filter residue and drying is ground into coarse granule, produced.
Embodiment 2.
A kind of method for tissue culture of dendrobium candidum, takes dendrobium candidum protocorm, is cultivated in root media, until long
Go out the dendrobium candidum seedling that 1-2 pieces true leaf is highly 1.5-2.5cm；It is 1.5-2.5cm, the iron sheet of the true leaf of piece containing 1-2 to take height
Stem of noble dendrobium seedling, is inoculated in seedling culture medium, cultivates 60 days, produces.
The prescription of rooting medium：Coptis root 15kg, root of herbaceous peony 20kg, dragon's blood 15kg, pseudo-ginseng 20kg, ginger 15kg, Poria cocos
15kg, peptone 10kg, potato 10kg, potato 5kg and soya bean 15kg.
The root media is prepared in the steps below：
(1) take coptis root, the root of herbaceous peony, dragon's blood, pseudo-ginseng, ginger and Poria cocos, plus 18 times of amount decoctings to boil after 6h, filter, filter residue is discarded,
Filtrate is collected, A product are obtained；
(2) fetch earth beans and potato, add 6 times of amount water, break into slurry, obtain B product；
(3) soya bean is taken, powder is ground into, C product are obtained；
(4) A product, the mixing of B product, are heated to 50-60 DEG C, add after C product and peptone, stirring 10min, are cooled to 5-10
DEG C, produce.
The Chinese medicine slag is by root of kudzu vine 60kg, root of Chinese clematis 40kg, selfheal 30kg, cortex albiziae 20kg, Herba Andrographitis 10kg and grey
Art 30kg is prepared as follows：The root of kudzu vine, the root of Chinese clematis, selfheal, cortex albiziae, Herba Andrographitis and rhizoma atractylodis plus 6 times of amount water are taken, are decocted
3h, filtering, filter residue and drying is ground into coarse granule, produced.
Embodiment 3.
A kind of method for tissue culture of dendrobium candidum, takes dendrobium candidum protocorm, is cultivated in root media, until long
Go out the dendrobium candidum seedling that 1-2 pieces true leaf is highly 1.5-2.5cm；It is 1.5-2.5cm, the iron sheet of the true leaf of piece containing 1-2 to take height
Stem of noble dendrobium seedling, is inoculated in seedling culture medium, cultivates 50 days, produces.
The prescription of rooting medium：Coptis root 5kg, root of herbaceous peony 10kg, dragon's blood 5kg, pseudo-ginseng 10kg, ginger 5kg, Poria cocos 5kg,
Peptone 1kg, potato 1kg, potato 1kg and soya bean 5kg.
The root media is prepared in the steps below：
(1) take coptis root, the root of herbaceous peony, dragon's blood, pseudo-ginseng, ginger and Poria cocos, plus 16 times of amount decoctings to boil after 5h, filter, filter residue is discarded,
Filtrate is collected, A product are obtained；
(2) fetch earth beans and potato, add 5 times of amount water, break into slurry, obtain B product；
(3) soya bean is taken, powder is ground into, C product are obtained；
(4) A product, the mixing of B product, are heated to 50-60 DEG C, add after C product and peptone, stirring 5min, are cooled to 5-10 DEG C,
Produce.
The Chinese medicine slag is by root of kudzu vine 50kg, root of Chinese clematis 30kg, selfheal 20kg, cortex albiziae 10kg, Herba Andrographitis 1kg and rhizoma atractylodis
20kg is prepared as follows：The root of kudzu vine, the root of Chinese clematis, selfheal, cortex albiziae, Herba Andrographitis and rhizoma atractylodis plus 5 times of amount water are taken, 2h is decocted,
Filtering, filter residue and drying is ground into coarse granule, produced.
Claims (8)
1. a kind of method for tissue culture of dendrobium candidum, it is characterised in that：Dendrobium candidum protocorm is taken, is trained in root media
Support, until growing the dendrobium candidum seedling that 1-2 pieces true leaf is highly 1.5-2.5cm；Take height true for 1.5-2.5cm, piece containing 1-2
The dendrobium candidum seedling of leaf, is inoculated in seedling culture medium, cultivates 50-60 days, produces.
2. the method for tissue culture of dendrobium candidum as claimed in claim 1, it is characterised in that：The root media is by weight
Part calculates main by 5-15 parts of coptis root, 10-20 parts of the root of herbaceous peony, 5-15 parts of dragon's blood, 10-20 parts of pseudo-ginseng, 5-15 parts of ginger, Poria cocos 5-15
5-15 parts of part, 1-10 parts of peptone, 1-10 parts of potato, 1-5 parts of potato and soya bean are made.
3. the method for tissue culture of dendrobium candidum as claimed in claim 2, it is characterised in that：The root media is by weight
Part calculate, mainly by 10 parts of coptis root, 15 parts of the root of herbaceous peony, 10 parts of dragon's blood, 15 parts of pseudo-ginseng, 10 parts of ginger, 10 parts of Poria cocos, 5 parts of peptone,
10 parts of 5 parts of potato, 3 parts of potato and soya bean are made.
4. the method for tissue culture of dendrobium candidum as claimed in claim 2 or claim 3, it is characterised in that：The root media is pressed
It is prepared by following step：
（1）Coptis root, the root of herbaceous peony, dragon's blood, pseudo-ginseng, ginger and Poria cocos are taken, plus 16-18 times is measured decocting and boiled after 5-6h, filtering, filter residue is abandoned
Go, collect filtrate, obtain A product；
（2）Fetch earth beans and potato, add 5-6 times and measure water, break into slurry, obtain B product；
（3）Soya bean is taken, powder is ground into, C product are obtained；
（4）A product, the mixing of B product, are heated to 50-60 DEG C, add after C product and peptone, stirring 5-10min, are cooled to 5-10 DEG C,
Produce.
5. the method for tissue culture of dendrobium candidum as claimed in claim 1, it is characterised in that：The seedling culture medium is by weight
Part calculates main by 10-20 parts of cooked maize mud, 10-20 parts of Chinese yam mud, 10-20 parts of taro paste, 1-5 parts of agar and Chinese medicine slag 20-
30 parts of mixings, are produced.
6. the method for tissue culture of dendrobium candidum as claimed in claim 6, it is characterised in that：The seedling culture medium is by weight
Part calculates, and mainly by 15 parts of cooked maize mud, 15 parts of Chinese yam mud, 15 parts of taro paste, the 25 parts of mixings of 3 parts of agar and Chinese medicine slag, produces.
7. the method for tissue culture of dendrobium candidum as claimed in claims 6 or 7, it is characterised in that：The Chinese medicine slag it is main by
50-60 parts of the root of kudzu vine calculated by weight, 30-40 parts of the root of Chinese clematis, 20-30 parts of selfheal, 10-20 parts of cortex albiziae, Herba Andrographitis 1-
10 parts prepare as follows with 20-30 parts of rhizoma atractylodis：Take the root of kudzu vine, the root of Chinese clematis, selfheal, cortex albiziae, Herba Andrographitis and rhizoma atractylodis plus 5-6
Amount water, decocts 2-3h again, and filtering, filter residue and drying is ground into coarse granule, produced.
8. the method for tissue culture of dendrobium candidum as claimed in claim 8, it is characterised in that：The Chinese medicine slag is main by by weight
25 parts of 55 parts of the root of kudzu vine, 35 parts of the root of Chinese clematis, 25 parts of selfheal, 15 parts of cortex albiziae, 5 parts of Herba Andrographitis and the rhizoma atractylodis of part calculating are measured by following
It is prepared by method：The root of kudzu vine, the root of Chinese clematis, selfheal, cortex albiziae, Herba Andrographitis and rhizoma atractylodis plus 5.5 times of amount water are taken, 2.5h is decocted, filters, filter
Slag dry, pulverize into coarse granule, produce.
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Cited By (8)
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| CN107548992A (en) * | 2017-10-23 | 2018-01-09 | 贵州济生农业科技有限公司 | A kind of imitating wild planting process of the good dendrobium candidum of growing way |
| CN107616085A (en) * | 2017-10-23 | 2018-01-23 | 贵州济生农业科技有限公司 | A kind of pseudo-wild cultivating method of the high dendrobium candidum of polyoses content |
| CN107926713A (en) * | 2017-12-28 | 2018-04-20 | 贵州济生农业科技有限公司 | A kind of candidum tissue culturing for improving rooting rate supports method |
| CN107926703A (en) * | 2017-12-04 | 2018-04-20 | 浙江东来天然生物制品有限公司 | A kind of high selenium-enriched officinal dendrobium stem seeding cultivating method of transplanting survival rate |
| CN108207626A (en) * | 2017-12-29 | 2018-06-29 | 贵州济生农业科技有限公司 | A kind of tissue of the good dendrobium candidum of seedling growth supports method |
| CN108782203A (en) * | 2018-06-25 | 2018-11-13 | 湖南省南岳树木园 | A method of imitating the wild plantation stem of noble dendrobium |
| CN109156360A (en) * | 2018-10-28 | 2019-01-08 | 江西兼济堂农业开发有限公司 | A kind of method for tissue culture of Paris polyphylla |
| CN113598049A (en) * | 2021-08-12 | 2021-11-05 | 福建省中科生物股份有限公司 | Chinese herbal medicine pre-cooking liquid for inhibiting tissue culture plant endophyte pollution and application method |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107548992A (en) * | 2017-10-23 | 2018-01-09 | 贵州济生农业科技有限公司 | A kind of imitating wild planting process of the good dendrobium candidum of growing way |
| CN107616085A (en) * | 2017-10-23 | 2018-01-23 | 贵州济生农业科技有限公司 | A kind of pseudo-wild cultivating method of the high dendrobium candidum of polyoses content |
| CN107926703A (en) * | 2017-12-04 | 2018-04-20 | 浙江东来天然生物制品有限公司 | A kind of high selenium-enriched officinal dendrobium stem seeding cultivating method of transplanting survival rate |
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| CN108207626A (en) * | 2017-12-29 | 2018-06-29 | 贵州济生农业科技有限公司 | A kind of tissue of the good dendrobium candidum of seedling growth supports method |
| CN108782203A (en) * | 2018-06-25 | 2018-11-13 | 湖南省南岳树木园 | A method of imitating the wild plantation stem of noble dendrobium |
| CN109156360A (en) * | 2018-10-28 | 2019-01-08 | 江西兼济堂农业开发有限公司 | A kind of method for tissue culture of Paris polyphylla |
| CN113598049A (en) * | 2021-08-12 | 2021-11-05 | 福建省中科生物股份有限公司 | Chinese herbal medicine pre-cooking liquid for inhibiting tissue culture plant endophyte pollution and application method |
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Application publication date: 20170808 |