CN105409775B - A kind of dendrobium candidum strengthening seedling and rooting culture medium and preparation method and cultural method - Google Patents
A kind of dendrobium candidum strengthening seedling and rooting culture medium and preparation method and cultural method Download PDFInfo
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- CN105409775B CN105409775B CN201510874078.0A CN201510874078A CN105409775B CN 105409775 B CN105409775 B CN 105409775B CN 201510874078 A CN201510874078 A CN 201510874078A CN 105409775 B CN105409775 B CN 105409775B
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- 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
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Abstract
The additive that dendrobium candidum strengthening seedling and rooting culture medium disclosed by the invention is made an addition in basal medium by basal medium and additionally is constituted, and the pH value of the culture medium is 6.0 ~ 6.4, and addition of the additive in basal medium is respectively：Coconut milk 1 ~ 3%, peeled potatoes 1 ~ 3%, peeling banana 3 ~ 5%, dry soybean 3 ~ 5%, powdered activated carbon 0.05 ~ 0.10%, sucrose 2 ~ 3%, agar 0.55 ~ 0.75%；Culture medium of the present invention replaces classical hormonal using natural additive, effectively prevent conventional medium and adds the problem of seedling stability that hormone debug time length, workload are big, turn out is poor, aberration rate is high；The preparation method of culture medium of the present invention is easy, easy to operate, and raw material sources are wide, cost is low；Short using the strengthening seedling and rooting time of medium culture of the present invention, typically at 11 ~ 12 weeks, and seedling body is sturdy, dimensionally stable, best in quality, well developed root system, transplanting survival rate is up to more than 99.2%, and no variation situation occurs, and can greatly improve efficiency, survival rate and the quality of dendrobium candidum plantation.
Description
Technical field
The invention belongs to plant biotechnology field, and in particular to a kind of dendrobium candidum strengthening seedling and rooting culture medium and system
Preparation Method and cultural method.
Background technology
Dendrobium candidum（Dendrobium officinale）For orchid family Dendrobium herbaceos perennial,《Taoist Scriptures》In
Be listed in first of " Chinese nine big celestial grass ", with strong cloudy strengthening the essence, nourishing the stomach to improve the production of body fluid, three high drop, improve body immunity, anti-aging,
Antitumor the effects such as.Because it has effects that uniqueness, scholar of the dendrobium candidum since ancient times always by vast health favors.It is wild
Dendrobium candidum is mainly distributed on the ground such as Zhejiang, Anhui, Guizhou, Jiangxi, Yunnan, and Seeds of Dendrobium Candidum is because of its unique construction, nothing
Method is sprouted alone, nature sprouting is only possible in the case where encountering suitable fungal component, people are to wild resource in addition
Mad excavation, present dendrobium officinale resource is endangered.With the development of modern biotechnology tissue culture technology, current iron
The artificial breeding technology of skin stem of noble dendrobium seedling has been gradually formed.
In traditional dendrobium candidum tissue culture, strengthening seedling and rooting is usually sharp by addition on the basis of basal medium
Element such as NAA, 6-BA promote the growth and increment of seedlings of Dendrobium officinale.But the reasonable species of hormone addition, addition are difficult the palm
Hold, generally require to spend many time and efforts to carry out substantial amounts of basic research.Improper or excessive addition can cause group
Situations such as seedling is slow-growing, small and weak, yellowing leaf is mottled, overall chlorosis, aberration rate are raised is trained, characterizing part often has table in addition
Existing hysteresis quality, these unfavorable factors have a strong impact on efficient production and the survival rate of field production of seedling.
The content of the invention
First technical problem to be solved by this invention is to provide one kind and added without hormone, without harmful chemical component, soon
Speed promotes the dendrobium candidum strengthening seedling and rooting culture medium of strengthening seedling and rooting.
Second technical problem to be solved by this invention is to provide a kind of workable dendrobium candidum strengthening seedling and rooting
The preparation method of culture medium.
3rd technical problem to be solved by this invention is to provide a kind of strong using above-mentioned medium culture dendrobium candidum
The method of seedling.
The present invention solve technical scheme that above-mentioned first technical problem used for：A kind of dendrobium candidum strengthening seedling and rooting training
Base is supported, the additive that the culture medium is made an addition in basal medium by basal medium and additionally is constituted, the pH value of the culture medium
For 6.0 ~ 6.4, in terms of the weight of described basal medium, the addition of described additive in described basal medium
Respectively：The % of coconut milk 1 ~ 3, the % of peeled potatoes 1 ~ 3, peeling banana 3 ~ 5 %, the % of dry soybean 3 ~ 5, powdered activated carbon 0.05 ~
0.10 %, the % of sucrose 2 ~ 3, the % of agar 0.55 ~ 0.75.
Described basal medium is 1/3 ~ 2/3MS basal mediums.MS basal mediums are at present using most common
Culture medium, its composition includes a great number of elements, trace element, molysite, organic principle, inositol etc., can make or select city by oneself
Sell product.1/3MS basal mediums are that a great number of elements is changed into 1/3 original culture medium, and similarly 2/3MS basal mediums are
A great number of elements is changed into 2/3 original culture medium.The application selects 1/3 ~ 2/3MS basal mediums, can advantageously promote iron sheet
The growth of stem of noble dendrobium seedling.
The present invention solve technical scheme that above-mentioned second technical problem used for：A kind of dendrobium candidum strengthening seedling and rooting training
The preparation method of base is supported, is comprised the following steps：
1）The pretreatment of additive：Fresh coconut is squeezed the juice and obtains coconut milk, potato and banana are removed the peel, by dry soybean
Sent out 8 ~ 12 hours in 20 ~ 28 DEG C of bubbly waters, and get out powdered activated carbon, sucrose, agar, it is standby；
2）The preparation of basal medium：Prepare basal medium；
3）The addition of additive：In terms of the weight of basal medium, each additive is weighed, each additive is in basal medium
In addition be respectively：The % of coconut milk 1 ~ 3, the % of peeled potatoes 1 ~ 3, peeling banana 3 ~ 5 %, the % of dry soybean 3 ~ 5, powdery are lived
Property charcoal 0.05 ~ 0.10 %, the % of sucrose 2 ~ 3, the % of agar 0.55 ~ 0.75, by coconut milk, peeled potatoes, peeling banana, soybean,
Powdered activated carbon, sucrose and agar are added in basal medium successively, are obtained dendrobium candidum strengthening seedling and rooting culture medium, are used NaOH
The pH value of dendrobium candidum strengthening seedling and rooting culture medium is adjusted to 6.0 ~ 6.4 by solution；
4）Packing and sterilizing：Divide after dendrobium candidum strengthening seedling and rooting culture medium is heated into 85 ~ 90 DEG C, 5 ~ 8 min of insulation
Dress, then sterilize 20 ~ 25 min in 121 DEG C of high steam, finally that the culture medium cooling for bacterium of having gone out is standby.
The present invention solve technical scheme that above-mentioned 3rd technical problem used for：One kind utilizes above-mentioned medium culture
The method of dendrobium candidum strong sprout, incubation is：The female seedling of dendrobium candidum that inoculation height is 1 ~ 1.5 cm in the medium, 24
Daily illumination 11 ~ 13 hours at ~ 28 DEG C, intensity of illumination is 2500 ~ 3000 Lux, until strengthening seedling and rooting.
Compared with prior art, the advantage of the invention is that：Dendrobium candidum strengthening seedling and rooting culture medium of the present invention, using natural
Additive replaces classical hormonal, no harmful chemical component addition, effectively prevent conventional medium addition hormone debug time length,
The problem of workload is big, turn out seedling stability is poor, aberration rate is high；The system of dendrobium candidum strengthening seedling and rooting culture medium of the present invention
Preparation Method is easy, easy to operate, and raw material sources are wide, cost is low；Using medium culture of the present invention dendrobium candidum strengthening seedling and rooting when
Between it is short, typically at 11 ~ 12 weeks, and culture seedling seedling body it is sturdy, dimensionally stable is best in quality, well developed root system, transplanting survival rate
Up to 99.2 more than %, no variation situation occurs, thus culture medium of the present invention can greatly improve efficiency that dendrobium candidum plants, into
Motility rate and quality.
Embodiment
The present invention is described in further detail with reference to embodiments.
The dendrobium candidum strengthening seedling and rooting culture medium of embodiment one, makes an addition to by 1/3MS basal mediums and additionally basic training
The additive composition in base is supported, the pH value of the culture medium is 6.0, and in terms of the weight of 1/3MS basal mediums, additive is 1/
Addition in 3MS basal mediums is respectively：The % of coconut milk 1, the % of peeled potatoes 1, peeling banana 3 %, the % of dry soybean 3,
The % of powdered activated carbon 0.05, the % of sucrose 2, the % of agar 0.55, the preparation method of the dendrobium candidum strengthening seedling and rooting culture medium include with
Lower step：
1）The pretreatment of additive：Fresh coconut is squeezed the juice and obtains coconut milk, potato and banana are removed the peel, by dry soybean
Sent out 10 hours in 25 DEG C of bubbly waters, and get out powdered activated carbon, sucrose, agar, it is standby；
2）The preparation of basal medium：Prepare 1/3MS basal mediums；
3）The addition of additive：In terms of the weight of 1/3MS basal mediums, each additive is weighed and successively added to 1/
In 3MS basal mediums, dendrobium candidum strengthening seedling and rooting culture medium is obtained, with NaOH solution by dendrobium candidum strengthening seedling and rooting culture medium
PH value be adjusted to 6.0；
4）Packing and sterilizing：Dispensed after dendrobium candidum strengthening seedling and rooting culture medium is heated into 85 DEG C, 5 min of insulation, then
Sterilize 20 min in 121 DEG C of high steam, finally cools down the culture medium for bacterium of having gone out, that is, the culture of embodiment one is made
Base.
Using the method for the medium culture dendrobium candidum strong sprout of embodiment one, incubation is：It is inoculated with the medium
It is highly 1 ~ 1.5 cm female seedling of dendrobium candidum, daily illumination 13 hours at 24 DEG C, intensity of illumination is 2500 Lux, is measured
Finished product seedling is obtained after 84 days, the seedling seedling body is sturdy, and well developed root system, transplanting survival rate is 99.2%, and aberration rate is zero.
The dendrobium candidum strengthening seedling and rooting culture medium of embodiment two, makes an addition to by 1/2MS basal mediums and additionally basic training
The additive composition in base is supported, the pH value of the culture medium is 6.2, and in terms of the weight of 1/2MS basal mediums, additive is 1/
Addition in 2MS basal mediums is respectively：The % of coconut milk 2, the % of peeled potatoes 2, peeling banana 4 %, the % of dry soybean 4,
The % of powdered activated carbon 0.075, the % of sucrose 2.5, the % of agar 0.62, the preparation method of the dendrobium candidum strengthening seedling and rooting culture medium include
Following steps：
1）The pretreatment of additive：Fresh coconut is squeezed the juice and obtains coconut milk, potato and banana are removed the peel, by dry soybean
Sent out 9 hours in 28 DEG C of bubbly waters, and get out powdered activated carbon, sucrose, agar, it is standby；
2）The preparation of basal medium：Prepare 1/2MS basal mediums；
3）The addition of additive：In terms of the weight of 1/2MS basal mediums, each additive is weighed and successively added to 1/
In 2MS basal mediums, dendrobium candidum strengthening seedling and rooting culture medium is obtained, with NaOH solution by dendrobium candidum strengthening seedling and rooting culture medium
PH value be adjusted to 6.2；
4）Packing and sterilizing：Dispensed after dendrobium candidum strengthening seedling and rooting culture medium is heated into 88 DEG C, 6.5 min of insulation, so
Sterilize 22 min in 121 DEG C of high steam afterwards, finally cools down the culture medium for bacterium of having gone out, that is, the training of embodiment two is made
Support base.
Using the method for the medium culture dendrobium candidum strong sprout of embodiment two, incubation is：It is inoculated with the medium
It is highly 1 ~ 1.5 cm female seedling of dendrobium candidum, daily illumination 12 hours at 26 DEG C, intensity of illumination is 3000 Lux, is measured
Finished product seedling is obtained after 80 days, the seedling seedling body is sturdy, well developed root system, transplanting survival rate is 99.5 %, and aberration rate is zero.
The dendrobium candidum strengthening seedling and rooting culture medium of embodiment three, makes an addition to by 2/3MS basal mediums and additionally basic training
The additive composition in base is supported, the pH value of the culture medium is 6.4, and in terms of the weight of 2/3MS basal mediums, additive is 2/
Addition in 3MS basal mediums is respectively：The % of coconut milk 3, the % of peeled potatoes 3, peeling banana 5 %, the % of dry soybean 5,
The % of powdered activated carbon 0.10, the % of sucrose 3, the % of agar 0.75, the preparation method of the dendrobium candidum strengthening seedling and rooting culture medium include with
Lower step：
1）The pretreatment of additive：Fresh coconut is squeezed the juice and obtains coconut milk, potato and banana are removed the peel, by dry soybean
Sent out 11 hours in 22 DEG C of bubbly waters, and get out powdered activated carbon, sucrose, agar, it is standby；
2）The preparation of basal medium：Prepare 2/3MS basal mediums；
3）The addition of additive：In terms of the weight of 2/3MS basal mediums, each additive is weighed and successively added to 2/
In 3MS basal mediums, dendrobium candidum strengthening seedling and rooting culture medium is obtained, with NaOH solution by dendrobium candidum strengthening seedling and rooting culture medium
PH value be adjusted to 6.4；
4）Packing and sterilizing：Dispensed after dendrobium candidum strengthening seedling and rooting culture medium is heated into 90 DEG C, 8 min of insulation, then
Sterilize 25 min in 121 DEG C of high steam, finally cools down the culture medium for bacterium of having gone out, that is, the culture of embodiment three is made
Base.
Using the method for the medium culture dendrobium candidum strong sprout of embodiment three, incubation is：It is inoculated with the medium
It is highly 1 ~ 1.5 cm female seedling of dendrobium candidum, daily illumination 11 hours at 28 DEG C, intensity of illumination is 3000 Lux, is measured
Finished product seedling is obtained after 78 days, the seedling seedling body is sturdy, and well developed root system, transplanting survival rate is 99.6%, and aberration rate is zero.
Claims (3)
1. a kind of dendrobium candidum strengthening seedling and rooting culture medium, it is characterised in that the culture medium is by basal medium and additionally makes an addition to
Additive composition in basal medium, the pH value of the culture medium is 6.0~6.4, and described basal medium is 1/3~2/
3MS basal mediums, in terms of the weight of described basal medium, described additive adding in described basal medium
Dosage is respectively：It is coconut milk 1~3%, peeled potatoes 1~3%, peeling banana 3~5%, dry soybean 3~5%, powdered activated
Charcoal 0.05~0.10%, sucrose 2~3%, agar 0.55~0.75%.
2. the preparation method of the dendrobium candidum strengthening seedling and rooting culture medium described in a kind of claim 1, it is characterised in that including following
Step：
1) pretreatment of additive：Fresh coconut is squeezed the juice and obtains coconut milk, potato and banana are removed the peel, by dry soybean in 20
~28 DEG C of bubbly waters are sent out 8~12 hours, and get out powdered activated carbon, sucrose, agar, standby；
2) preparation of basal medium：Prepare basal medium；
3) addition of additive：In terms of the weight of basal medium, each additive is weighed, each additive is in basal medium
Addition is respectively：Coconut milk 1~3%, peeled potatoes 1~3%, peeling banana 3~5%, dry soybean 3~5%, powdery are lived
Property charcoal 0.05~0.10%, sucrose 2~3%, agar 0.55~0.75%, by coconut milk, peeled potatoes, peeling banana, big
Beans, powdered activated carbon, sucrose and agar are added in basal medium successively, obtain dendrobium candidum strengthening seedling and rooting culture medium, are used
The pH value of dendrobium candidum strengthening seedling and rooting culture medium is adjusted to 6.0~6.4 by NaOH solution；
4) packing and sterilizing：Dispensed after dendrobium candidum strengthening seedling and rooting culture medium is heated into 85~90 DEG C, 5~8min of insulation, so
Sterilize 20~25min in 121 DEG C of high steam afterwards, finally that the culture medium cooling for bacterium of having gone out is standby.
3. a kind of method of the medium culture dendrobium candidum strong sprout described in utilization claim 1, it is characterised in that incubation
For：Inoculation height is the 1~1.5cm female seedling of dendrobium candidum in the medium, daily illumination 11~13 hours at 24~28 DEG C,
Intensity of illumination is 2500~3000Lux, until strengthening seedling and rooting.
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| CN107094627A (en) * | 2017-05-12 | 2017-08-29 | 江苏绿洲园艺绿化有限公司 | A kind of dendrobium candidum rooting of vitro seedling culture medium and preparation method thereof |
| CN106962165B (en) * | 2017-05-22 | 2020-09-15 | 东莞市睿绅生物技术有限公司 | Preparation method of movable dendrobium officinale culture medium and integrated seedling culture method |
| CN109496781B (en) * | 2018-10-16 | 2021-04-20 | 皖西学院 | Dendrobium huoshanense culture medium |
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| CN101564008B (en) * | 2008-04-24 | 2011-03-23 | 中国医学科学院药用植物研究所 | Hormone-free cultivation and rapid propagation method of dendrobium candidum axenic seedlings |
| CN102613086B (en) * | 2012-03-31 | 2013-12-11 | 南京农业大学 | Hormone-free tissue culture method for dendrobium candidum |
| CN104067940A (en) * | 2014-06-30 | 2014-10-01 | 广州花都先锋园艺有限公司 | Hormone-free tissue culture method for dendrobium officinale |
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