CN106561453B - The rapid propagation method of Campanumoea javanica - Google Patents
The rapid propagation method of Campanumoea javanica Download PDFInfo
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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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- 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
- A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
- A01G9/02—Receptacles, e.g. flower-pots or boxes; Glasses for cultivating flowers
- A01G9/029—Receptacles for seedlings
- A01G9/0299—Handling or transporting of soil blocks or seedlings
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
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Abstract
The invention discloses a kind of rapid propagation methods of Campanumoea javanica, using Campanumoea javanica ripe berry as explant, berry is aseptically cut after sterilized, sterilizing and takes out Campanumoea javanica seed, and the sprouting seedling obtained through sprouting culture carries out Initial culture, squamous subculture, culture of rootage and acclimatization and transplants successively.The present invention greatly reduces the possibility of variation by the way that its seed using Campanumoea javanica ripe berry as explant, is taken to induce Multiple Buds after sprouting and cultivating and without callus phase, is conducive to keep the merit of seedling.
Description
Technical field
The present invention relates to the tissue culture technology fields of Campanumoea javanica.It is more particularly related to a kind of great Hua money
The rapid propagation method of leopard.
Background technology
Campanumoea javanica is Campanulaceae leopard platymiscium Campanumoea javanica also known as climing campanulaceae, milk starches root,
Bavin Radix Codonopsis (Guizhou), osmanthus Radix Codonopsis (Guangdong), tetragonous ginseng, cowtail (Yunnan).Herbaceous stem winds liana, has milk, has carrot shape
Root.Stem is hairless, multi-branched.Leaf to life, few alternate, have long handle, blade is heart-shaped or heart shaped is avette, and there are shallow sawtooth, pole in edge
Few full edge, it is 3-11 centimetres long, it is 2-9 centimetres wide, it is hairless or back side sparsely grow becomes mildewed sometimes.Single leave armpit is spent, each portion is hairless, flower
Calyx is separated with ovary, and 5 split to nearly base portion, sliver ovum shape lanceolar or lanceolar, 1-1.8 centimetres long；Corolla is big, and corolla is upper,
White or yellow green, inner face purple, mitriform are split to middle part；5 pieces of stamen；Column cap 4-5 is split, ovary and capsule Room 5.The black purple of berry
Color, aubergine are spherical.Seed is irregular, is often short cylinder, there is netted decorative pattern on surface.
Campanumoea javanica is born in the shrubbery below 2400 meters of height above sea level and in sparse woods.Main product is in Yunnan, Guizhou, Guangxi and wide
Most of counties and cities in east.Fruit is sweet, edible.Root is used as medicine, sweet slight bitter temperature, can strengthening the spleen and stomach, tonifying lung gas, expelling phlegm and arresting coughing.Cure mainly void
Labor internal injury, cough due to deficiency of the lung, deficiency syndrome of the lung diarrhea, milk are few, infantile malnutrition.Root also can cooked food, have the benefits of heat-clearing, calm, control nerve
It is weak to wait diseases.
Campanumoea javanica is mostly use among the people as medicinal material, is that wild resource is used as medicine at present, with the increasingly increasing of dosage
Add, wild resource is deficient, and Campanumoea javanica have effects that it is similar to Radix Codonopsis, with good exploitation prospect.Cause
This, it is quite necessary to it is bred and artificial cultivation conducts a research.However, through consulting literatures data, great Hua is not yet found at present
Research in terms of leopard artificial cultivation and breeding is reported.
The content of the invention
It is an object of the invention to solve at least the above, and provide the advantages of at least will be described later.
It is a still further object of the present invention to provide a kind of rapid propagation method of Campanumoea javanica, by with Campanumoea javanica
Ripe berry is explant, its seed is taken to induce Multiple Buds without callus phase after sprouting and cultivating, is subtracted significantly
Lack the possibility of variation, be conducive to keep the merit of seedling.
A further object of the invention is by being carried out with suitable proliferated culture medium and prescription of rooting medium after being commissioned to train
It supports and culture of rootage, is quickly obtained a large amount of, test tube seedling of high quality, convenient for large-scale industrial production application, and production cost
It is cheap.
In order to realize these purposes and further advantage according to the present invention, a kind of quick breeding of Campanumoea javanica is provided
Method using Campanumoea javanica ripe berry as explant, aseptically cuts berry and takes out great Hua gold after sterilized, sterilizing
Money leopard seed, the sprouting seedling obtained through sprouting culture carry out Initial culture, squamous subculture, culture of rootage and acclimatization and transplants successively.
Preferably, the rapid propagation method of the Campanumoea javanica, Campanumoea javanica seed are seeded to sprouting culture
It carries out sprouting on base cultivating 7-10 days and must sprout seedling, the formula of the germination medium is：MS+1.5mg/L 6-BA+0.5mg/
LGA3+ 300mg/L PVP, the condition for sprouting culture are：Temperature is 24-28 DEG C, intensity of illumination 2000Lx, light application time are
10h/ days.
Preferably, the rapid propagation method of the Campanumoea javanica, Campanumoea javanica ripe berry is as explant
Disinfection, sterilizing methods be：Campanumoea javanica ripe berry is taken, the alcohol that volume fraction is 75% is being used on superclean bench
After impregnating 5~10S, with the HgCl that mass fraction is 0.1%2Then solution sterilization 15min sways washing 4 times, nothing with sterile water
Bacterium blotting paper blots moisture.
Preferably, the rapid propagation method of the Campanumoea javanica, Initial culture concretely comprise the following steps：It is trained sprouting
It supports obtained sprouting seedling and cuts to be seeded on inducing culture behind root and cultivate 15-20 days, obtain Multiple Buds, wherein, just it is commissioned to train
Foster condition is：Temperature is 24-28 DEG C, intensity of illumination 2000Lx, light application time are 10h/ days；The formula of inducing culture
For：MS+2.5mg/L 6-BA+0.5mg/L KT+0.5mg/L TDZ+0.2mg/L NAA+300mg/L PVP+200mg/L are hydrolyzed
Casein.
Preferably, the rapid propagation method of the Campanumoea javanica, squamous subculture concretely comprise the following steps：It will just be commissioned to train
It supports obtained Multiple Buds and aseptically cuts the bud clump containing 2-3 bud and be forwarded in proliferated culture medium progress after being commissioned to train
It supports, cultivation cycle is 15-20 days, and the condition of squamous subculture is：24-28 DEG C of temperature, intensity of illumination 2000Lx, light application time are
10h/ days；The formula of proliferated culture medium is：MS+2.0mg/L 6-BA+0.5mg/L KT+0.2mg/L TDZ+0.2mg/L NAA.
Preferably, the rapid propagation method of the Campanumoea javanica, culture of rootage concretely comprise the following steps：When after being commissioned to train
When supporting obtained sprout length to more than 2cm, single bud is cut into from base portion, is forwarded to cultivate 20 days in root media and be taken root
Seedling, the condition of culture of rootage are：24-28 DEG C of temperature, intensity of illumination 2000Lx, light application time are 10h/ days；Root media
Formula be：1/2MS+1.5mg/L IBA+0.5mg/L ABT1。
Preferably, the rapid propagation method of the Campanumoea javanica, acclimatization and transplants concretely comprise the following steps：To take root training
It supports obtained rooted seedling to uncap hardening 4 days, then take out rooted seedling, clear water washes away the root media of its root, and transplanting is extremely
Transplanted seedling is obtained on the seedbed of fine sand river, relative humidity is 95% in 10 days inner retaining ring borders after transplanting, and it is 23- to control day temperature
25 DEG C, nocturnal temperature is 15-18 DEG C, upper cup plantation behind 15 days after transplanting, and field production is carried out behind 30 days after transplanting.
Preferably, the rapid propagation method of the Campanumoea javanica, the rooted seedling are cleaned after root, and transplanting is extremely
Before, operations described below is also carried out：The root system of rooted seedling is placed in strengthening root liquid and impregnates 30min, contains 3mg/L in the strengthening root liquid
Moringa leaf juice and 1mg/L far-infared ceramic powder, the solvent of the strengthening root liquid is magnetized water, and the Moringa leaf juice is leaf of Moringa
Crushed after 100 mesh sieves with magnetized water according to mass ratio is 1:It grinds after 5 mixing and uniformly obtains；
In addition, after transplantation of seedlings of taking root, 1g far infrareds pottery is uniformly spread fertilizer over the fields with every square metre of seedbed on the seedbed of the fine sand river
Porcelain powder.
The present invention includes at least following advantageous effect：
1) overall procedure quickly bred of Campanumoea javanica of the invention is easy to operate, and production cost is low, growth rate
Soon, rooting and transplant survival rate is high, convenient for large-scale industrial production application, can largely, quick, persistently acquisition high quality test tube
Seedling, it is highly practical；
2) present invention Multiple Buds after sprouting and cultivating are induced with Campanumoea javanica seed and without callus phase,
The possibility of variation is greatly reduced, is conducive to keep the merit of seedling；
3) proliferated culture medium of the invention can realize the multiplication of Multiple Buds, and proliferation times are up to 13.46/20d；The present invention
Root media can realize taking root for sprout, for rooting rate up to 100%, transplanting survival rate is up to 92.04%；
4) present invention is greatly reinforced by spreading fertilizer over the fields far-infared ceramic powder using the immersion of strengthening root liquid and seedbed to rooted seedling
The vigor of rooted seedling root system so that transplanting survival rate is up to 99.4%.
Part is illustrated to embody by further advantage, target and the feature of the present invention by following, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Specific embodiment
With reference to specific embodiment, the present invention is described in further detail, those skilled in the art's reference to be made to say
Bright book word can be implemented according to this.
It should be noted that experimental method described in following embodiments, is conventional method unless otherwise specified, institute
Reagent and material are stated, unless otherwise specified, is commercially obtained.
6-BA：6- benzyl aminoadenines；GA3：Gibberellin；PVP：Polyvinylpyrrolidone；KT：Kinetin；TDZ：It is new
Plant growth regulator Thidiazuron；NAA：Methyl α-naphthyl acetate；IBA：Indolebutyric acid；ABT1：Plant with difficult rooting cuttage and seedling culture is special to take root
Powder；
Embodiment 1：
A kind of rapid propagation method of Campanumoea javanica, using Campanumoea javanica ripe berry as explant, sterilized, sterilizing
Berry is aseptically cut afterwards and takes out Campanumoea javanica seed, and the sprouting seedling obtained through sprouting culture carries out just being commissioned to train successively
Foster, squamous subculture, culture of rootage and acclimatization and transplants.
Wherein, Campanumoea javanica seed, which is seeded on germination medium sprout cultivating 7-10 days, must sprout seedling, described
The formula of germination medium is：MS+1.5mg/L 6-BA+0.5mg/LGA3+ 300mg/L PVP, the condition for sprouting culture are：Temperature
Spend for 24-28 DEG C, intensity of illumination 2000Lx, light application time be 10h/ days.
Wherein, Campanumoea javanica ripe berry is as the disinfection of explant, sterilizing methods：Take the ripe slurry of Campanumoea javanica
Fruit is 0.1% with mass fraction after the alcohol that volume fraction is 75% is used to impregnate 5~10S on superclean bench
HgCl2Then solution sterilization 15min sways washing 4 times with sterile water, sterile blotting paper blots moisture.
Wherein, Initial culture concretely comprises the following steps：Induction is seeded to after the sprouting seedling that sprouting culture obtains is cut root
It is cultivated 15-20 days on culture medium, obtains Multiple Buds, wherein, the condition of Initial culture is：Temperature is 24-28 DEG C, intensity of illumination is
2000Lx, light application time are 10h/ days；The formula of inducing culture is：MS+2.5mg/L 6-BA+0.5mg/L KT+0.5mg/L
TDZ+0.2mg/L NAA+300mg/L PVP+200mg/L caseinhydrolysates.
Wherein, squamous subculture concretely comprises the following steps：The Multiple Buds that Initial culture is obtained aseptically cut containing
The bud clump of 2-3 bud, which is forwarded in proliferated culture medium, carries out squamous subculture, and cultivation cycle is 15-20 days, the condition of squamous subculture
For：24-28 DEG C of temperature, intensity of illumination 2000Lx, light application time are 10h/ days；The formula of proliferated culture medium is：MS+2.0mg/
L 6-BA+0.5mg/L KT+0.2mg/L TDZ+0.2mg/L NAA。
Wherein, culture of rootage concretely comprises the following steps：When the sprout length that squamous subculture obtains is to more than 2cm, cut from base portion
It into single bud, is forwarded to cultivate 20 days in root media and obtains rooted seedling, the condition of culture of rootage is：24-28 DEG C of temperature, light
According to intensity be 2000Lx, light application time is 10h/ days；The formula of root media is：1/2MS+1.5mg/L IBA+0.5mg/L
ABT1。
Wherein, acclimatization and transplants concretely comprise the following steps：The rooted seedling that culture of rootage is obtained is uncapped hardening 4 days, then will be raw
Offspring takes out, and clear water washes away the root media of its root, transplants to the seedbed of fine sand river to obtain transplanted seedling, in 10 days after transplanting
It is 95% to keep relative humidity in environment, and controlling day temperature, nocturnal temperature is 15-18 DEG C, 15 days after transplanting for 23-25 DEG C
Upper cup is planted afterwards, and field production is carried out behind 30 days after transplanting.
The method of the present embodiment 1 can realize the multiplication of Multiple Buds, proliferation times 13.46/20d, and rooting rate 100% moves
Plant survival rate 92.04%.
Embodiment 2：
A kind of rapid propagation method of Campanumoea javanica, on the basis of embodiment 1, using tender shoots or stem with bud as explant
Explant is directly inoculated into inducing culture and carries out Initial culture by body, and in addition to explant, other are same as Example 1.
The method of the present embodiment 2 cannot induce Multiple Buds.
Embodiment 3：
A kind of rapid propagation method of Campanumoea javanica, on the basis of embodiment 1, the formula of the proliferated culture medium replaces
It is changed to：MS、6-BA 1.5mg/L、KT 0.75mg/L、NAA 1.0mg/L.
In addition to the formula of proliferated culture medium, other are same as Example 1.
The method of the present embodiment 3 can realize the multiplication of Multiple Buds, and proliferation times are 3.46 times, and transplanting survival rate reaches
90.3%.
Embodiment 4：
A kind of rapid propagation method of Campanumoea javanica, on the basis of embodiment 1, the formula of the root media replaces
It is changed to 1/2MS+1.5mg/L IBA.
In addition to the formula of root media, other are same as Example 1.
The method of the present embodiment 4 can realize taking root for test tube seedling, rooting rate 82.4%, and transplanting survival rate is up to 91.5%.
Embodiment 5：
On the basis of embodiment 1, the rapid propagation method of the Campanumoea javanica, rooted seedling cleaning root it
Afterwards, transplant to before, also carry out operations described below：The root system of rooted seedling is placed in strengthening root liquid and impregnates 30min, in the strengthening root liquid
The far-infared ceramic powder of Moringa leaf juice and 1mg/L containing 3mg/L, the solvent of the strengthening root liquid is magnetized water；The leaf of Moringa
It is 1 that juice, which is that leaf of Moringa crushed after 100 mesh sieves with magnetized water according to mass ratio,:It grinds after 5 mixing and uniformly obtains, wherein magnetized water
For 5 parts, leaf of Moringa powder is 1 part；
In addition, after transplantation of seedlings of taking root, 1g far infrareds pottery is uniformly spread fertilizer over the fields with every square metre of seedbed on the seedbed of the fine sand river
Porcelain powder.
The method of the present embodiment 5 can realize rooting rate 100%, while transplanting survival rate is up to 99.4%.Contain in strengthening root liquid
Moringa leaf juice and far-infared ceramic powder, can promote that root system is sturdy, prevents root diseases, improves its transplanting survival rate；And in seedling
A certain amount of far-infared ceramic powder is spread fertilizer over the fields on bed, it is possible to reduce the disease rate after transplanting improves survival rate.
Although the embodiments of the present invention have been disclosed as above, but its be not restricted in specification and embodiment it is listed
With it can be fully applied to various fields suitable for the present invention, for those skilled in the art, can be easily
Realize other modification, therefore without departing from the general concept defined in the claims and the equivalent scope, it is of the invention and unlimited
In specific details and shown here as the embodiment with description.
Claims (3)
1. a kind of rapid propagation method of Campanumoea javanica, which is characterized in that using Campanumoea javanica ripe berry as explant, warp
Berry is aseptically cut after disinfection, sterilizing and takes out Campanumoea javanica seed, through sprouting the sprouting seedling cultivated and obtained successively
Carry out Initial culture, squamous subculture, culture of rootage and acclimatization and transplants；
Campanumoea javanica seed, which is seeded on germination medium sprout cultivating 7-10 days, must sprout seedling, the germination medium
Formula be：MS+1.5mg/L 6-BA+0.5mg/LGA3+ 300mg/L PVP, the condition for sprouting culture are：Temperature is 24-28
DEG C, intensity of illumination 2000Lx, light application time be 10h/ days；
Initial culture concretely comprises the following steps：It the obtained sprouting seedling of culture will be sprouted cuts to be seeded on inducing culture behind root and train
It supports 15-20 days, obtains Multiple Buds, wherein, the condition of Initial culture is：Temperature is 24-28 DEG C, intensity of illumination 2000Lx, light
It it is 10h/ days according to the time；The formula of inducing culture is：MS+2.5mg/L 6-BA+0.5mg/L KT+0.5mg/L TDZ+
0.2mg/L NAA+300mg/L PVP+200mg/L caseinhydrolysates；
Squamous subculture concretely comprises the following steps：The Multiple Buds that Initial culture is obtained aseptically are cut containing 2-3 bud
Bud clump, which is forwarded in proliferated culture medium, carries out squamous subculture, and cultivation cycle is 15-20 days, and the condition of squamous subculture is：Temperature 24-
28 DEG C, intensity of illumination 2000Lx, light application time be 10h/ days；The formula of proliferated culture medium is：MS+2.0mg/L 6-BA+
0.5mg/L KT+0.2mg/L TDZ+0.2mg/L NAA；
Culture of rootage concretely comprises the following steps：When the sprout length that squamous subculture obtains is to more than 2cm, single bud is cut into from base portion,
It is forwarded to cultivate 20 days in root media and obtains rooted seedling, the condition of culture of rootage is：24-28 DEG C of temperature, intensity of illumination are
2000Lx, light application time are 10h/ days；The formula of root media is：1/2MS+1.5mg/L IBA+0.5mg/L ABT1；
Acclimatization and transplants concretely comprise the following steps：The rooted seedling that culture of rootage is obtained is uncapped hardening 4 days, then takes out rooted seedling,
Clear water washes away the root media of its root, transplants to thin river sand seedbed to obtain transplanted seedling, 10 days inner retaining ring borders after transplanting
Middle relative humidity is 95%, and controlling day temperature, nocturnal temperature is 15-18 DEG C for 23-25 DEG C, upper cup kind behind 15 days after transplanting
It plants, field production is carried out behind 30 days after transplanting.
2. the rapid propagation method of Campanumoea javanica as described in claim 1, which is characterized in that Campanumoea javanica ripe berry
Disinfection, sterilizing methods as explant are：Campanumoea javanica ripe berry is taken, is being with volume fraction on superclean bench
After 75% alcohol impregnates 5~10S, with the HgCl that mass fraction is 0.1%2Solution sterilization 15min, is then swayed with sterile water
Washing 4 times, sterile blotting paper blots moisture.
3. the rapid propagation method of Campanumoea javanica as described in claim 1, which is characterized in that the rooted seedling cleans root
Afterwards, transplant to before, also carry out operations described below：The root system of rooted seedling is placed in strengthening root liquid and impregnates 30min, the strengthening root liquid
In the far-infared ceramic powder of the Moringa leaf juice containing 3mg/L and 1mg/L, the solvent of the strengthening root liquid is magnetized water, the Moringa
It is 1 that leaf juice, which is that leaf of Moringa crushed after 100 mesh sieves with magnetized water according to mass ratio,:It grinds after 5 mixing and uniformly obtains；
In addition, after transplantation of seedlings of taking root, 1g far-infared ceramic powders are uniformly spread fertilizer over the fields with every square metre of seedbed on the thin river sand seedbed.
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