CN108617297A - The cuttage transplantation method of sea-buckthorn tissue-cultured seedling - Google Patents
The cuttage transplantation method of sea-buckthorn tissue-cultured seedling Download PDFInfo
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- CN108617297A CN108617297A CN201810403338.XA CN201810403338A CN108617297A CN 108617297 A CN108617297 A CN 108617297A CN 201810403338 A CN201810403338 A CN 201810403338A CN 108617297 A CN108617297 A CN 108617297A
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- cuttage
- cultured seedling
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G2/00—Vegetative propagation
- A01G2/10—Vegetative propagation by means of cuttings
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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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- Developmental Biology & Embryology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
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- Cultivation Of Plants (AREA)
Abstract
The present invention provides the cuttage transplantation method of sea-buckthorn tissue-cultured seedling, includes the following steps：(1) tissue-cultured seedling is prepared；(2) cutting medium is prepared；(3) root-growing agent is prepared：6‑BA 50mg/L+IAA 30mg/L；(4) cuttage and culture of sea-buckthorn tissue-cultured seedling：Tissue-cultured seedling is immersed in 3min in root-growing agent, it inserts it into cutting medium later, after cuttage, the same day is drenched matrix with water, with covered rearing with plastic film on seedlings nursing plate, it is moved rapidly into the shade that shade density is 75% 85%, water drenching, weeding, fertilising and the prevention and control of plant diseases, pest control is carried out during cultivating cuttage seeding；(5) it is out of the garden.The present invention merges taking root for tissue-cultured seedling in one step with transplanting; it need not again be transplanted in culture in glassware to taking root; compared to traditional tissue culture asexual reproduction method; further shorten seedling raise period; simplify process simultaneously; cost is reduced, is conducive to realize large-scale cultivation sea-buckthorn tissue-cultured seedling in a short time.
Description
Technical field
The invention belongs to the plant regeneration technical fields by tissue culture technique, and in particular to the cuttage of sea-buckthorn tissue-cultured seedling
Transplantation method.
Background technology
Sea-buckthorn (Classification system：Hippophae rhamnoides Linn.) it is perennial deciduous fruit tree, shrub or little Qiao
Wood.Its fruit, pericarp, blade, bark and its fabricated product contain 280 several physiological active substances, have strong anti-inflammatory, kill
Bacterium, analgesic and the special efficacy for promoting regeneration.Many of which ingredient is killing and is inhibiting tumour cell, radioresistance, anti-freezing
Blood, blood pressure lowering prevent blood vessel embolism, anti-aging, antifatigue, enhancing body energy and immunity etc. from all showing mystery
Therapeutic effect.
Sea-buckthorn branches and leaves are luxuriant, lateral root is flourishing, root turion is extremely strong, and the root that can go here and there rapidly forms intensive group from tiller, and can be with
The symbiosis such as actinomyces, bacterium, mycobacteria form a large amount of root nodules, and having nitrogen fixing capacity more stronger than soybean, (per hectare can fixed nitrogen
180kg).In addition, rapid with growth, drought-resistant, saline-alkali tolerant is barren, drives in freezing winter and in sultry summer, have conserve water and soil, check winds and fix drifting sand,
The obvious effect for improveing soil and the adaptable equal promotion ecological balance, is not only the pioneer tree of arid windy and sandy area afforestation
Kind, also it is important firewood forests and fodder forest seeds.So in current conceding the land to forestry, agricultural industry restructuring, plantation
Sea-buckthorn has important nutritive value, the ecological value and economic value.
Sea-buckthorn dioecism is difficult to keep the characteristic of improved seeds by seminal propagation in production；Root turion breeds line of breeding
Number is relatively low.The main method of sea-buckthorn breeding at present has：Two kinds of generative propagation and vegetative propagation, wherein it is three that vegetative propagation, which is divided into,
Kind：Cuttage, semi-lignified cuttage and tissue cultures.Tissue cultures are not by time restriction, in the few situation the following group of material
It is the best approach to knit culture, while being also to realize nursery stock large-scale production, carry out gene genetic conversion breeding, preserve germplasm money
One of the mechanism study such as source, plant cold resistance salt tolerant etc. essential approach.But traditional tissue culture propagation is in squamous subculture
Afterwards, it the step of passing through culture of rootage, allows in culture medium of the tissue culture subculture bud in bottle and takes root, then transplant again into matrix,
Complex procedures, culture of rootage requires stringent sterile working just to can guarantee Quality of Seedlings in bottle, and the cost is relatively high, in addition
Transplanting culture, more needs to consume more manpower and materials.
Invention content
In order to solve the problems in the existing technology, the present invention provides the cuttage transplantation methods of sea-buckthorn tissue-cultured seedling.
The present invention provides the cuttage transplantation method of sea-buckthorn tissue-cultured seedling, includes the following steps：
(1) tissue-cultured seedling is prepared；
(2) cutting medium is prepared；
(3) root-growing agent is prepared：Root-growing agent is：6-BA 50mg/L+IAA 30mg/L；
(4) cuttage and culture of sea-buckthorn tissue-cultured seedling：Tissue-cultured seedling is immersed in 3min in root-growing agent, later, is inserted it into
In cutting medium, after cuttage, the same day is drenched matrix with water, and with covered rearing with plastic film on seedlings nursing plate, it is quickly moved
Enter in the shade that shade density is 75%-85%, cultivates conventionally water drenching, weeding, fertilising and disease pest during cuttage seeding
Evil prevention；
(5) it is out of the garden.
Preferably, in step (1), the method for preparing tissue-cultured seedling is：The stem apex for choosing sea-buckthorn current-year branch is made
For explant, disinfection is seeded to aseptic culture medium and carries out bud inducement cultivation, and after obtaining sterile bud, sterile bud is transferred to after being commissioned to train
It supports and carries out shoot proliferation culture on base, choose proliferation 25-30d, the long good tufted seedling of 3-5cm upgrowth situations is as cuttage material
Material：The culture medium of root is cleaned with clear water, tufted seedling is cut into single plant, only retains the 2-3 piece blades of stem top.
Preferably, in step (2), the cutting medium is by perlite, vermiculite and turfy soil according to 1:2:4 volume
Than mixture prepared.
Preferably, the preparation sequence of step (1)-(3) can be interchanged.
Preferably, in step (4), it is 20-30 DEG C to keep temperature in shade, relative humidity 80%-90%.
Preferably, in step (4), the cuttage of sea-buckthorn tissue-cultured seedling is carried out in the annual 4-7 months.
Preferably, in step (4), the 1/5-1/4 that part accounts for entire tissue-cultured seedling length is impregnated.
Preferably, in step (5), when height of seedling 45cm or more, is out of the garden.
The present invention merges taking root for tissue-cultured seedling in one step with transplanting, need not again be moved in culture in glassware to taking root
It plants, compared to traditional tissue culture asexual reproduction method, further shortens seedling raise period, while simplifying process, reduce into
This, is conducive to realize large-scale cultivation sea-buckthorn tissue-cultured seedling in a short time.
Specific implementation mode
Embodiment below facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments
Method is unless otherwise specified conventional method.Test material as used in the following examples is unless otherwise specified city
It sells.
The cuttage transplantation method of the sea-buckthorn tissue-cultured seedling of the present invention includes the following steps：
(1) tissue-cultured seedling is prepared：The stem apex of sea-buckthorn current-year branch is chosen as explant, disinfection is seeded to aseptic culture medium
Bud inducement cultivation is carried out, after obtaining sterile bud, sterile bud is transferred to progress shoot proliferation culture on subculture medium, chooses and increases
25-30d is grown, is about the good tufted seedling of 3-5cm upgrowth situations as cuttage material：The culture medium of root is cleaned with clear water,
Tufted seedling is cut into single plant, only retains the 2-3 piece blades of stem top；
(2) cutting medium is prepared：By perlite, vermiculite and turfy soil according to 1:2:4 volume ratio is mixed with to obtain skewer
Matrix is inserted, after disinfection liquid disinfectant, cutting medium is fitted into seedlings nursing plate.Thimerosal can be in the prior art any one
Kind, for example mass concentration is 3% ferrous sulfate aqueous solution, such as the liquor potassic permanganate etc. of 3g/L.
There are many cutting mediums, and the present invention is to a variety of host materials such as perlite, peat, red heart soil, vermiculite and bentonite
It is combined, then screening obtains optimization formula：The volume ratio of perlite, vermiculite and turfy soil is 1:2:4, in the formula,
With following root-growing agent compatibilities, the rooting rate highest of sea-buckthorn.
(3) root-growing agent is prepared：Root-growing agent is：6- benzyls aminoadenine (6-BA) 50mg/L+ heteroauxins (IAA) 30mg/
L。
The sequence of step (1)-(3) can be interchanged.
(4) cuttage and culture of sea-buckthorn tissue-cultured seedling：The cuttage of sea-buckthorn tissue-cultured seedling is carried out in the annual 4-7 months.By tissue-cultured seedling
It is immersed in 3min in root-growing agent, the 1/5-1/4 that immersion part accounts for entire tissue-cultured seedling length inserts it into cutting medium later
In, after cuttage, the same day is moved rapidly into shade density with water matrix of drenching with covered rearing with plastic film on seedlings nursing plate
It is 20-30 DEG C, relative humidity 80%-90% in the shade of 75%-85%, to keep temperature in shade, cultivates cuttage
Conventionally water drenching, weeding, fertilising and the prevention and control of plant diseases, pest control between seedling stage.
(5) it is out of the garden：Height of seedling 45cm or more is out of the garden.
The selection of the soaking time of root-growing agent and sea-buckthorn tissue-cultured seedling in root-growing agent：
Select three hormon, hormone concentration and soaking time factors as variable.Select respectively hormone for 6-BA,
IAA, 6-BA+IAA, hormone concentration are set as 10mg/L, 30mg/L, 50mg/L, 70mg/L, soaking time be selected as 1min,
3min、5min、7min.Per 50 plants of tissue-cultured seedling of group selection, the cuttage transplanting of sea-buckthorn tissue-cultured seedling is carried out according to the method described above.As a result it sends out
Existing, under the conditions of above-mentioned each, the average rooting rate range of each group is between 30-98%, wherein when root-growing agent is：6-BA (6- benzyls
Aminoadenine) 50mg/L+IAA (heteroauxin) 30mg/L, when soaking time is 3min, seedling goes out root after cuttage 6d, average raw
Root rate is up to 98%, and root system is long after growth, and root system is white, and aerial part color is green.
Embodiment 1
The cuttage transplantation method of the sea-buckthorn tissue-cultured seedling of the present invention includes the following steps：
(1) tissue-cultured seedling is prepared：The stem apex of sea-buckthorn current-year branch is chosen as explant, disinfection is seeded to aseptic culture medium
Bud inducement cultivation is carried out, after obtaining sterile bud, sterile bud is transferred to progress shoot proliferation culture on subculture medium, chooses and increases
28d is grown, is about the good tufted seedling of 4cm upgrowth situations as cuttage material：The culture medium of root is cleaned with clear water, will be grown thickly
Seedling is cut into single plant, only retains 2 blades of stem top；
(2) cutting medium is prepared：By perlite, vermiculite and turfy soil according to 1:2:4 volume ratio is mixed with to obtain skewer
Matrix is inserted, after disinfection liquid disinfectant, cutting medium is fitted into seedlings nursing plate.Thimerosal is the ferrous sulfate that mass concentration is 3%
Aqueous solution.
(3) root-growing agent is prepared：Root-growing agent is：6-BA (6- benzyls aminoadenine) 50mg/L+IAA (heteroauxin) 30mg/
L。
(4) cuttage of sea-buckthorn tissue-cultured seedling：The cuttage of sea-buckthorn tissue-cultured seedling is carried out in annual May.Tissue-cultured seedling is immersed in life
3min in root agent impregnates part and accounts for the 1/5 of entire tissue-cultured seedling length, later, inserts it into cutting medium, cuttage terminates
Afterwards, the same day is drenched matrix with water, with covered rearing with plastic film on seedlings nursing plate, is moved rapidly into the shading that shade density is 80%
In canopy, keep shade in temperature be 25 DEG C, relative humidity 85%, cultivate cuttage seeding during conventionally water drenching, remove
Grass, fertilising and the prevention and control of plant diseases, pest control.
(5) it is out of the garden：Height of seedling 45cm or more is out of the garden.
Embodiment 2
The cuttage transplantation method of the sea-buckthorn tissue-cultured seedling of the present invention includes the following steps：
(1) tissue-cultured seedling is prepared：The stem apex of sea-buckthorn current-year branch is chosen as explant, disinfection is seeded to aseptic culture medium
Bud inducement cultivation is carried out, after obtaining sterile bud, sterile bud is transferred to progress shoot proliferation culture on subculture medium, chooses and increases
25d is grown, is about the good tufted seedling of 3cm upgrowth situations as cuttage material：The culture medium of root is cleaned with clear water, will be grown thickly
Seedling is cut into single plant, only retains 3 blades of stem top；
(2) cutting medium is prepared：By perlite, vermiculite and turfy soil according to 1:2:4 volume ratio is mixed with to obtain skewer
Matrix is inserted, after disinfection liquid disinfectant, cutting medium is fitted into seedlings nursing plate.Thimerosal is the liquor potassic permanganate of 3g/L.
(3) root-growing agent is prepared：Root-growing agent is：6-BA (6- benzyls aminoadenine) 50mg/L+IAA (heteroauxin) 30mg/
L。
(4) cuttage of sea-buckthorn tissue-cultured seedling：The cuttage of sea-buckthorn tissue-cultured seedling is carried out in annual April.Tissue-cultured seedling is immersed in life
3min in root agent impregnates part and accounts for the 1/5 of entire tissue-cultured seedling length, later, inserts it into cutting medium, cuttage terminates
Afterwards, the same day is drenched matrix with water, with covered rearing with plastic film on seedlings nursing plate, is moved rapidly into the shading that shade density is 75%
In canopy, keep shade in temperature be 20 DEG C, relative humidity 80%, cultivate cuttage seeding during conventionally water drenching, remove
Grass, fertilising and the prevention and control of plant diseases, pest control.
(5) it is out of the garden：Height of seedling 45cm or more is out of the garden.
Embodiment 3
The cuttage transplantation method of the sea-buckthorn tissue-cultured seedling of the present invention includes the following steps：
(1) tissue-cultured seedling is prepared：The stem apex of sea-buckthorn current-year branch is chosen as explant, disinfection is seeded to aseptic culture medium
Bud inducement cultivation is carried out, after obtaining sterile bud, sterile bud is transferred to progress shoot proliferation culture on subculture medium, chooses and increases
30d is grown, is about the good tufted seedling of 5cm upgrowth situations as cuttage material：The culture medium of root is cleaned with clear water, will be grown thickly
Seedling is cut into single plant, only retains 2 blades of stem top；
(2) cutting medium is prepared：By perlite, vermiculite and turfy soil according to 1:2:4 volume ratio is mixed with to obtain skewer
Matrix is inserted, after disinfection liquid disinfectant, cutting medium is fitted into seedlings nursing plate.Thimerosal is the ferrous sulfate that mass concentration is 3%
Aqueous solution.
(3) root-growing agent is prepared：Root-growing agent is：6-BA (6- benzyls aminoadenine) 50mg/L+IAA (heteroauxin) 30mg/
L。
(4) cuttage of sea-buckthorn tissue-cultured seedling：The cuttage of sea-buckthorn tissue-cultured seedling is carried out in annual July.Tissue-cultured seedling is immersed in life
3min in root agent impregnates part and accounts for the 1/4 of entire tissue-cultured seedling length, later, inserts it into cutting medium, cuttage terminates
Afterwards, the same day is drenched matrix with water, with covered rearing with plastic film on seedlings nursing plate, is moved rapidly into the shading that shade density is 85%
In canopy, keep shade in temperature be 30 DEG C, relative humidity 90%, cultivate cuttage seeding during conventionally water drenching, remove
Grass, fertilising and the prevention and control of plant diseases, pest control.
(5) it is out of the garden：Height of seedling 45cm or more is out of the garden.
Finally it should be noted that：The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention,
Although the present invention is described in detail referring to the foregoing embodiments, for those skilled in the art, still may be used
With technical scheme described in the above embodiments is modified or equivalent replacement of some of the technical features.
All within the spirits and principles of the present invention, any modification, equivalent replacement, improvement and so on should be included in the present invention's
Within protection domain.
Claims (8)
1. the cuttage transplantation method of sea-buckthorn tissue-cultured seedling, it is characterised in that：Include the following steps：
(1) tissue-cultured seedling is prepared；
(2) cutting medium is prepared；
(3) root-growing agent is prepared：Root-growing agent is：6-BA 50mg/L+IAA 30mg/L；
(4) cuttage and culture of sea-buckthorn tissue-cultured seedling：Tissue-cultured seedling is immersed in 3min in root-growing agent, later, inserts it into cuttage
In matrix, after cuttage, the same day is moved rapidly into screening with water matrix of drenching with covered rearing with plastic film on seedlings nursing plate
Cloudy degree is in the shade of 75%-85%, and conventionally water drenching, weeding, fertilising and pest and disease damage are anti-during cultivating cuttage seeding
It controls；
(5) it is out of the garden.
2. according to the method described in claim 1, it is characterized in that：In step (1), the method for preparing tissue-cultured seedling is：Choosing
Take the stem apex of sea-buckthorn current-year branch as explant, disinfection is seeded to aseptic culture medium and carries out bud inducement cultivation, obtains sterile
After bud, sterile bud is transferred to progress shoot proliferation culture on subculture medium, proliferation 25-30d, long 3-5cm is chosen and grows shape
The good tufted seedling of condition is as cuttage material：The culture medium of root is cleaned with clear water, tufted seedling is cut into single plant, only retains stem
The 2-3 piece blades on top.
3. according to the method described in claim 1, it is characterized in that：In step (2), the cutting medium is by perlite, leech
Stone and turfy soil are according to 1:2:4 volume ratio is mixture prepared.
4. according to the method described in claim 1, it is characterized in that：The preparation sequence of step (1)-(3) can be interchanged.
5. according to the method described in claim 1, it is characterized in that：In step (4), it is 20-30 DEG C to keep temperature in shade,
Relative humidity is 80%-90%.
6. according to the method described in claim 1, it is characterized in that：In step (4), sea-buckthorn tissue culture is carried out in the annual 4-7 months
The cuttage of seedling.
7. according to the method described in claim 1, it is characterized in that：In step (4), impregnates part and account for entire tissue-cultured seedling length
1/5-1/4。
8. according to the method described in claim 1, it is characterized in that：In step (5), when height of seedling 45cm or more, is out of the garden.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110122084A (en) * | 2019-05-23 | 2019-08-16 | 宁德师范学院 | Cuttage breeding method in a kind of Morinda officinalis tissue-cultured seedling room |
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| CN107173226A (en) * | 2017-06-13 | 2017-09-19 | 新疆阿勒泰地区园林场 | A kind of sea-buckthorn tissue culture outside sprout-cultivating-bottle radication method |
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| CN100998315A (en) * | 2006-01-09 | 2007-07-18 | 尹伟伦 | Direct sometic embryogenesis of sea-buckthorn, and regeneration of sea-buckthorn plants |
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| CN110122084A (en) * | 2019-05-23 | 2019-08-16 | 宁德师范学院 | Cuttage breeding method in a kind of Morinda officinalis tissue-cultured seedling room |
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Application publication date: 20181009 |