CN107372105A - A kind of tissue culture and rapid propagation method of flower plants and nursery stock - Google Patents
A kind of tissue culture and rapid propagation method of flower plants and nursery stock Download PDFInfo
- Publication number
- CN107372105A CN107372105A CN201710519637.5A CN201710519637A CN107372105A CN 107372105 A CN107372105 A CN 107372105A CN 201710519637 A CN201710519637 A CN 201710519637A CN 107372105 A CN107372105 A CN 107372105A
- Authority
- CN
- China
- Prior art keywords
- parts
- nursery stock
- culture
- flower plants
- rapid propagation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
-
- 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
-
- 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
-
- 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
-
- 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
- A01N65/40—Liliopsida [monocotyledons]
- A01N65/42—Aloeaceae [Aloe family] or Liliaceae [Lily family], e.g. aloe, veratrum, onion, garlic or chives
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Developmental Biology & Embryology (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Environmental Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Agronomy & Crop Science (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The present invention relates to agricultural biological technical field, more particularly to a kind of tissue culture and rapid propagation method of flower plants and nursery stock, using stem apex as explant, sterilized by explant, bud inducement cultivation, Multiplying culture, Rooting and hardening-off culture, hardening and transplanting and other steps, wherein disinfectant used in explant sterilization are mainly made up of raw materials such as garlic, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry, garden burnet, windproof, gelatinized corn starch, natural gum, castor oil, Chinese juniper oil, spaonin powder and tea seed cake powders.The antibacterial efficiency high of tissue culture and rapid propagation method of the flower plants and nursery stock, it is pollution-free to explantation tissue's fanout free region, the bud induction rate, rooting rate and survival rate of flower plants and nursery stock are effectively improved, promote flower plants and nursery stock breeds production.
Description
Technical field
The present invention relates to agricultural biological technical field, and in particular to a kind of tissue culture and rapid propagation method of flower plants and nursery stock.
Background technology
Because by environment, season, soil and species, factors are limited in itself etc., therefore flower plants and nursery stock is bred also
It is restricted, wants to realize that its scale industrialization production is extremely difficult, and be difficult to breed out high-quality kind.City at present
Most of on field is all to carry out breeding for flower plants and nursery stock using asexual reproduction methods such as conventional cuttage, press strip or graftings, still
The speed that these methods are bred is slow, and the variety and quality bred out is bad, it is impossible to meets the growing market demand.
Plant Tissue Breeding is the theory for having totipotency according to plant cell, using the in vitro organ of plant (such as root,
Stem, leaf, stem apex etc.), tissue (such as forming layer, epidermis, cortex, marrow cell, endosperm) or cell (such as megaspore, microspore,
Body cell etc.) and protoplast, under the conditions of sterile and suitable synthetic medium and temperature etc. are artificial, callus can be induced
Tissue, adventitious bud, adventitious root, eventually form the subject of complete plant.Plant Tissue Breeding is widely used to agricultural production
In multiple fields, in seedling detoxification production, seedling quickly breed, the scale application of plant new resources, plant new product
Kind seed selection etc. generates far-reaching influence to modern agriculture, is that agricultural biotechnologies apply upper model in modern agriculture,
It is the important component of modern kind industry.At present, China's Plant tissue culture seedling is in flowers, fruit tree, shade plant, medicinal plant
Etc. the popularization and application for achieving large area.And plant tissue culture technique is by factors such as culture medium nutrient, bacterium, holders
Influence so that the production bred of flower plants and nursery stock is restricted, and at this moment needing to take appropriate measures improves these problems, example
Such as explant is carried out disinfection to reduce pollution of the bacterium to culture medium, carried out at present usually using alcohol, calcium hypochlorite, mercuric chloride
Explant sterilizes, although the bacterium on explant surface can be eliminated, can have certain injury to the tissue of explant, and mercuric chloride has
Poison, hardly possible cleaning, pollutes environment；Therefore, optimization plant tissue culture technique is the key point bred of flower plants and nursery stock.
The content of the invention
The goal of the invention of the present invention is：For above-mentioned problem, there is provided a kind of tissue-culturing rapid propagation side of flower plants and nursery stock
Method, antibacterial efficiency high is pollution-free to explantation tissue's fanout free region, is effectively improved bud induction rate, the rooting rate of flower plants and nursery stock
And survival rate, promote flower plants and nursery stock breeds production.
To achieve these goals, the technical solution adopted by the present invention is as follows：
A kind of tissue culture and rapid propagation method of flower plants and nursery stock, comprises the following steps：
(1) make explant from the disease-free stem apex sprouted then of robust growth, after scrubbing clean, use detergent solution
Explant surface is scrubbed with banister brush after immersion 5-7min, the dust on its surface and part thalline are removed, then use running water
Volumetric concentration is placed in after rinsing well to soak 5-10s in 70-75% alcohol, is equipped with being placed into after aseptic water washing 2-3 times
Immersion 50-60min in the container of disinfectant, immersion use aseptic water washing 2-3 times after terminating, finally dry surface with aseptic filter paper
The globule, it is standby；
The disinfectant is mainly made up of following raw material：Garlic, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry,
Garden burnet, windproof, gelatinized corn starch, natural gum, castor oil, Chinese juniper oil, spaonin powder and tea seed cake powder；
(2) first holder is added in blake bottle, add sterilizing after bud inducement cultivation base or proliferated culture medium or
Rooting and hardening-off culture base, then the explant after sterilization is inoculated into culture medium, successively carries out bud inducement cultivation, propagation successively
Culture and Rooting and hardening-off culture, finally obtain the nursery stock that takes root, standby；
(3) big Tanaka is transplanted to after selecting the nursery stock progress hardening of taking root of well-grown and stalwartness.
Preferably, in step (1), the disinfectant is mainly made up of following raw material：Garlic 20-30 parts, tealeaves
25-35 parts, madder 15-20 parts, folium artemisiae argyi 20-30 parts, aloe 17-20 parts, Sophora alopecuroide 17-20 parts, cogongrass cliff berry 15-17 parts, garden burnet
15-17 parts, windproof 17-20 parts, gelatinized corn starch 5-7 parts, gummy 4-6 parts, castor oil 2-3 parts, Chinese juniper oil 2-3 parts, spaonin powder 3-5 parts
And tea seed cake powder 3-5 parts.
Preferably, the disinfectant is prepared according to said ratio, specifically includes following steps：
(a) by fresh garlic, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry, garden burnet and it is windproof removal of impurities and it is clear
After washing, it is put into physiological saline and soaks 8-10min, then with rinsed with sterile water, it is standby；
(b) by the garlic handled well, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry, garden burnet and windproof chopping, then
According to 1:30-40 ratio mixes with sterilized water decocts 30-40min, is removed the gred with filtered through gauze, obtains filtered fluid, standby；
(c) gelatinized corn starch, natural gum, spaonin powder and tea seed cake powder are added in warm water dissolve it is well mixed after, pour into step (2)
Filtered fluid in, add castor oil and Chinese juniper oil, be uniformly mixed, obtain the disinfectant.
Preferably, holder employs aseptic filter paper bridge described in step (2) and solid carbon dioxide tongue is combined, i.e., by aseptic filter paper
Bridge and solid carbon dioxide tongue, which are soaked in deionized water, is made its expansion.
Preferably, the addition of holder described in step (2) is the 1/5-2/5 of blake bottle volume, wherein aseptic filter paper
The volume ratio of bridge and solid carbon dioxide tongue is 1：4-5.
Preferably, bud inducement cultivation base or proliferated culture medium or Rooting and hardening-off culture base are in MS bases described in step (2)
It is beautiful that peptone, caseinhydrolysate, glucose, bananas juice, potato juice, natrium nitrosum, azanol, dihydro are with the addition of in basal culture medium
Meter Su, isopentenyl gland purine, benzac, urea, BNOA, sodium phenate and activated carbon.
Preferably, Rooting and hardening-off culture base is also filled with carbon dioxide described in step (2).
Preferably, each cultivation stage provides illumination using cold-cathode fluorescence lamp in step (2), and temperature is 23-27 DEG C,
Relative humidity is 65-75%, and pH is 5.0-6.5, wherein the incubation of the bud inducement cultivation base and proliferated culture medium
Middle intensity of illumination is 1500-2000LUX, and intensity of illumination is 3000- in the incubation of the Rooting and hardening-off culture base
3500LUX。
In summary, by adopting the above-described technical solution, the beneficial effects of the invention are as follows：
1st, alcohol has stronger penetration power, is denatured bacterium protein, and bactericidal effect is good, at the same it also have it is stronger
Humidification, the air on material can be excluded, beneficial to the infiltration of other disinfectants.The tissue-culturing rapid propagation side of the flower plants and nursery stock of the present invention
Method kills part bacterium first with the alcohol-pickled explant several seconds, and after excluding the air on material, then be soaked in disinfectant and disappear
Poison, Disinfection Effect is improved, reduces the injury of explant.The disinfectant of the present invention is mainly by garlic, tealeaves, madder, folium artemisiae argyi, reed
The raw material systems such as luxuriant growth, Sophora alopecuroide, cogongrass cliff berry, garden burnet, windproof, gelatinized corn starch, natural gum, castor oil, Chinese juniper oil, spaonin powder and tea seed cake powder
Into；Wherein, sulfur-containing compound in garlic, containing materials such as alkaloid, catechins in tealeaves, rubican, folium artemisiae argyi are contained in madder
In contain aromatic oil, contain aloin in aloe, the materials such as organic acid, flavonoids and alkaloid, cogongrass cliff berry contained in Sophora alopecuroide
In contain flavones, contain a variety of tannin compositions in garden burnet, contain the materials such as volatile oil, mannitol, bitter taste glucoside, these plants in windproof
Contained material all there is certain antibiotic and sterilizing to act in thing, and the antibacterial range of every kind of material differs, each other
Cooperate, effectively increase the antibiotic and sterilizing scope of disinfectant；Gelatinized corn starch, natural gum, castor oil and Chinese juniper oil belong to stick together
Agent, it is mixed into plant extraction liquid, plant extraction liquid can be attached on to explant surface, performance, spaonin powder and tea is sticked together in raising
Seed cake powder belongs to wetting agent, effectively reduces the surface tension of plant extraction liquid, explant is soaked and is extended rapidly, beneficial to infiltration
Inside explant；Disinfectant made of these materials can quickly penetrate into explant surfaces externally and internally, and it is thin effectively to kill explant surface
Bacterium, antibacterial efficiency high, and do not injure explantation tissue, and non-environmental-pollution, be effectively improved flower plants and nursery stock bud induction rate,
Rooting rate and survival rate, promote flower plants and nursery stock breeds production.
2nd, the tissue culture and rapid propagation method of flower plants and nursery stock of the invention makees culture medium holder using filter paper bridge and solid carbon dioxide platform, reduces
Absorption of the holder to nutrient solution, gas permeability is improved, ensure that explant nutrient abundance, grown beneficial to explant.
3rd, each culture medium of the tissue culture and rapid propagation method of flower plants and nursery stock of the invention with the addition of albumen in MS minimal mediums
Peptone, caseinhydrolysate, glucose, bananas juice, potato juice, natrium nitrosum, azanol, dihydro zeatin, isopentenyl gland purine,
The nutrient such as benzac, urea, BNOA, sodium phenate and activated carbon, carbon source, the nitrogen source for providing abundance for explant,
Enable explant healthy growth, and carbon dioxide is also filled with Rooting and hardening-off culture base, there is provided sufficient illumination condition, explant
Physical efficiency has effectively facilitated the fast-growth of explant by photosynthesis supplementary carbon source.
Embodiment
Technical scheme will be clearly and completely described below, it is clear that described embodiment is only
Part of the embodiment of the present invention, rather than whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art
The every other embodiment obtained under the premise of creative work is not made, belongs to the scope of protection of the invention.
Unless otherwise defined, all of technologies and scientific terms used here by the article is with belonging to technical field of the invention
The implication that technical staff is generally understood that is identical.Term used in the description of the invention herein is intended merely to description tool
The purpose of the embodiment of body, it is not intended that in the limitation present invention.Term as used herein " and/or " include one or more phases
The arbitrary and all combination of the Listed Items of pass.
Embodiment 1
1st, the preparation of disinfectant
Disinfectant is mainly made up of following raw material：20 parts of garlic, 25 parts of tealeaves, 15 parts of madder, 20 parts of folium artemisiae argyi, reed
17 parts of luxuriant growth, 17 parts of Sophora alopecuroide, 15 parts of cogongrass cliff berry, 15 parts of garden burnet, windproof 17 parts, 5 parts of gelatinized corn starch, 4 parts of natural gum, 2 parts of castor oil, Chinese juniper
3 parts of 2 parts of pitch, 3 parts of spaonin powder and tea seed cake powder.
The expectation of said ratio is prepared into disinfectant using following methods, specifically includes following steps：
(a) by fresh garlic, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry, garden burnet and it is windproof removal of impurities and it is clear
After washing, it is put into physiological saline and soaks 8min, then with rinsed with sterile water, it is standby；
(b) by the garlic handled well, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry, garden burnet and windproof chopping, then
According to 1:30 ratio mixes with sterilized water decocts 30min, is removed the gred with filtered through gauze, obtains filtered fluid, standby；
(c) gelatinized corn starch, natural gum, spaonin powder and tea seed cake powder are added in warm water dissolve it is well mixed after, pour into step (2)
Filtered fluid in, add castor oil and Chinese juniper oil, be uniformly mixed, obtain the disinfectant.
2nd, prepared by culture medium
Bud inducement cultivation base or proliferated culture medium or Rooting and hardening-off culture base are to the addition of albumen in MS minimal mediums
Peptone, caseinhydrolysate, glucose, bananas juice, potato juice, natrium nitrosum, azanol, dihydro zeatin, isopentenyl gland purine,
Benzac, urea, BNOA, sodium phenate and activated carbon, after being uniformly mixed, pH is adjusted to 5, then in high temperature
Lower sterilizing, cools down and produces.
3rd, prepared by culture medium holder
Aseptic filter paper bridge and solid carbon dioxide tongue phase are employed in bud inducement cultivation base, proliferated culture medium and Rooting and hardening-off culture base
Culture medium holder is combined as, i.e., aseptic filter paper bridge and solid carbon dioxide tongue are pressed 1：4 volume ratio, which is soaked in deionized water, makes it
Swells and be made.
The 4th, the above-mentioned disinfectant being prepared, culture medium and culture medium holder are used in the tissue-culturing rapid propagation side of flower plants and nursery stock
In method, following steps are specifically included：
(1) make explant from the disease-free stem apex sprouted then of robust growth, after scrubbing clean, use detergent solution
Explant surface is scrubbed with banister brush after immersion 5min, the dust on its surface and part thalline are removed, then rushed with running water
Volumetric concentration is placed in after wash clean to soak 5s in 70% alcohol, with being placed into after aseptic water washing 2 times equipped with above-mentioned disinfectant
Container in immersion 50min, immersion terminate after with aseptic water washing 2 times, the globule on surface is finally dried with aseptic filter paper, it is standby
With；
(2) first 1/5 holder obtained above for accounting for blake bottle volume is added in blake bottle, adds above-mentioned system
The bud inducement cultivation base or proliferated culture medium or Rooting and hardening-off culture base obtained, first the explant after sterilization is successively inoculated into successively
Cultivated in bud inducement cultivation base and proliferated culture medium, each nurturing period provides illumination using cold-cathode fluorescence lamp
1500LUX, temperature are 23 DEG C, and relative humidity is 65%；Inoculate and carried out in Rooting and hardening-off culture base obtained above
Cultivate, and be filled with carbon dioxide, provide illumination 3000LUX using cold-cathode fluorescence lamp, temperature is 23 DEG C, and relative humidity is
65%, the nursery stock that takes root is finally obtained, it is standby；
(3) big Tanaka is transplanted to after selecting the nursery stock progress hardening of taking root of well-grown and stalwartness.
Embodiment 2
1st, the preparation of disinfectant
Disinfectant is mainly made up of following raw material：25 parts of garlic, 30 parts of tealeaves, 17 parts of madder, 25 parts of folium artemisiae argyi, reed
18.5 parts of luxuriant growth, 18.5 parts of Sophora alopecuroide, 16 parts of cogongrass cliff berry, 16 parts of garden burnet, windproof 18.5 parts, 6 parts of gelatinized corn starch, gummy 4.5 parts, castor-oil plant
4 parts of 2.5 parts of oil, 2.5 parts of Chinese juniper oil, 4 parts of spaonin powder and tea seed cake powder.
The expectation of said ratio is prepared into disinfectant using following methods, specifically includes following steps：
(a) by fresh garlic, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry, garden burnet and it is windproof removal of impurities and it is clear
After washing, it is put into physiological saline and soaks 9min, then with rinsed with sterile water, it is standby；
(b) by the garlic handled well, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry, garden burnet and windproof chopping, then
According to 1:35 ratio mixes with sterilized water decocts 35min, is removed the gred with filtered through gauze, obtains filtered fluid, standby；
(c) gelatinized corn starch, natural gum, spaonin powder and tea seed cake powder are added in warm water dissolve it is well mixed after, pour into step (2)
Filtered fluid in, add castor oil and Chinese juniper oil, be uniformly mixed, obtain the disinfectant.
2nd, prepared by culture medium
Bud inducement cultivation base or proliferated culture medium or Rooting and hardening-off culture base are to the addition of albumen in MS minimal mediums
Peptone, caseinhydrolysate, glucose, bananas juice, potato juice, natrium nitrosum, azanol, dihydro zeatin, isopentenyl gland purine,
Benzac, urea, BNOA, sodium phenate and activated carbon, after being uniformly mixed, pH is adjusted to 5.8, then in height
Temperature is lower to sterilize, and cools down and produces.
3rd, prepared by culture medium holder
Aseptic filter paper bridge and solid carbon dioxide tongue phase are employed in bud inducement cultivation base, proliferated culture medium and Rooting and hardening-off culture base
Culture medium holder is combined as, i.e., aseptic filter paper bridge and solid carbon dioxide tongue are pressed 1：4.5 volume ratio, which is soaked in deionized water, to be made
Its swells and be made.
The 4th, the above-mentioned disinfectant being prepared, culture medium and culture medium holder are used in the tissue-culturing rapid propagation side of flower plants and nursery stock
In method, following steps are specifically included：
(1) make explant from the disease-free stem apex sprouted then of robust growth, after scrubbing clean, use detergent solution
Explant surface is scrubbed with banister brush after immersion 6min, the dust on its surface and part thalline are removed, then rushed with running water
Volumetric concentration is placed in after wash clean to soak 8s in 73% alcohol, with being placed into after aseptic water washing 3 times equipped with above-mentioned disinfectant
Container in immersion 55min, immersion terminate after with aseptic water washing 3 times, the globule on surface is finally dried with aseptic filter paper, it is standby
With；
(2) first 2/5 holder obtained above for accounting for blake bottle volume is added in blake bottle, adds above-mentioned system
The bud inducement cultivation base or proliferated culture medium or Rooting and hardening-off culture base obtained, first the explant after sterilization is successively inoculated into successively
Cultivated in bud inducement cultivation base and proliferated culture medium, each nurturing period provides illumination using cold-cathode fluorescence lamp
1700LUX, temperature are 25 DEG C, and relative humidity is 70%；Inoculate and carried out in Rooting and hardening-off culture base obtained above
Cultivate, and be filled with carbon dioxide, provide illumination 3300LUX using cold-cathode fluorescence lamp, temperature is 25 DEG C, and relative humidity is
70%, the nursery stock that takes root is finally obtained, it is standby；
(3) big Tanaka is transplanted to after selecting the nursery stock progress hardening of taking root of well-grown and stalwartness.
Embodiment 3
1st, the preparation of disinfectant
Disinfectant is mainly made up of following raw material：30 parts of garlic, 35 parts of tealeaves, 20 parts of madder, 30 parts of folium artemisiae argyi, reed
20 parts of luxuriant growth, 20 parts of Sophora alopecuroide, 17 parts of cogongrass cliff berry, 17 parts of garden burnet, windproof 20 parts, 7 parts of gelatinized corn starch, 6 parts of natural gum, 3 parts of castor oil, Chinese juniper
5 parts of 3 parts of pitch, 5 parts of spaonin powder and tea seed cake powder.
The expectation of said ratio is prepared into disinfectant using following methods, specifically includes following steps：
(a) by fresh garlic, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry, garden burnet and it is windproof removal of impurities and it is clear
After washing, it is put into physiological saline and soaks 10min, then with rinsed with sterile water, it is standby；
(b) by the garlic handled well, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry, garden burnet and windproof chopping, then
According to 1:40 ratio mixes with sterilized water decocts 40min, is removed the gred with filtered through gauze, obtains filtered fluid, standby；
(c) gelatinized corn starch, natural gum, spaonin powder and tea seed cake powder are added in warm water dissolve it is well mixed after, pour into step (2)
Filtered fluid in, add castor oil and Chinese juniper oil, be uniformly mixed, obtain the disinfectant.
2nd, prepared by the culture medium of each cultivation stage
Bud inducement cultivation base or proliferated culture medium or Rooting and hardening-off culture base are to the addition of albumen in MS minimal mediums
Peptone, caseinhydrolysate, glucose, bananas juice, potato juice, natrium nitrosum, azanol, dihydro zeatin, isopentenyl gland purine,
Benzac, urea, BNOA, sodium phenate and activated carbon, after being uniformly mixed, pH is adjusted to 6.5, then in height
Temperature is lower to sterilize, and cools down and produces.
3rd, prepared by culture medium holder
Aseptic filter paper bridge and solid carbon dioxide tongue phase are employed in bud inducement cultivation base, proliferated culture medium and Rooting and hardening-off culture base
Culture medium holder is combined as, i.e., aseptic filter paper bridge and solid carbon dioxide tongue are pressed 1：5 volume ratio, which is soaked in deionized water, makes it
Swells and be made.
4th, the above-mentioned disinfectant being prepared, the culture medium of each cultivation stage and culture medium holder are used in flower plants and nursery stock
Tissue culture and rapid propagation method on, specifically include following steps：
(1) make explant from the disease-free stem apex sprouted then of robust growth, after scrubbing clean, use detergent solution
Explant surface is scrubbed with banister brush after immersion 7min, the dust on its surface and part thalline are removed, then rushed with running water
Volumetric concentration is placed in after wash clean to soak 10s in 75% alcohol, with being placed into after aseptic water washing 3 times equipped with above-mentioned disinfectant
Container in immersion 60min, immersion terminate after with aseptic water washing 3 times, the globule on surface is finally dried with aseptic filter paper, it is standby
With；
(2) first 1/5 holder obtained above for accounting for blake bottle volume is added in blake bottle, adds above-mentioned system
The bud inducement cultivation base or proliferated culture medium or Rooting and hardening-off culture base obtained, first the explant after sterilization is successively inoculated into successively
Cultivated in bud inducement cultivation base and proliferated culture medium, each nurturing period provides illumination using cold-cathode fluorescence lamp
2000LUX, temperature are 27 DEG C, and relative humidity is 75%；Inoculate and carried out in Rooting and hardening-off culture base obtained above
Cultivate, and be filled with carbon dioxide, provide illumination 3500LUX using cold-cathode fluorescence lamp, temperature is 27 DEG C, and relative humidity is
75%, the nursery stock that takes root is finally obtained, it is standby；
(3) big Tanaka is transplanted to after selecting the nursery stock progress hardening of taking root of well-grown and stalwartness.
The applicant has done lot of experiments during experiment, now arranges part test as follows：
1st group：Breeding for flower plants and nursery stock is carried out according to the method for plant tissue culture of routine.
2nd group：The raw material of disinfectant is garlic, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry, garden burnet and prevented
Wind, other processing modes are identical with the embodiment of the present invention 1.
3rd group：The raw material of disinfectant is garlic, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry, garden burnet and prevented
Wind, gelatinized corn starch and natural gum, other processing modes are identical with the embodiment of the present invention 1.
4th group：The raw material of disinfectant is garlic, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry, garden burnet and prevented
Wind, castor oil and Chinese juniper oil, other processing modes are identical with the embodiment of the present invention 1.
5th group：The raw material of disinfectant is garlic, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry, garden burnet and prevented
Wind, spaonin powder and tea seed cake powder, other processing modes are identical with the embodiment of the present invention 1.
The growing state of observation flower plants and nursery stock daily, and record, statistical result is as shown in table 1.
Shown from table 1,2-3 groups use thimerosal of the plant extracts as explant, reduce the wound to explant
Evil, rootage duration, bud induction rate, rooting rate and the survival rate of flower plants and nursery stock are better than the 1st group, but the 2nd group of contamination rate
Flower plants and nursery stock high than the 1st group, that embodiment 1-3 is cultivated, contamination rate, rootage duration, bud induction rate, rooting rate and is survived
Rate is optimal；In summary, the tissue culture and rapid propagation method of flower plants and nursery stock of the invention is the most scientific and effective, antibacterial efficiency high, to explant
Body tissue fanout free region, it is pollution-free, the bud induction rate, rooting rate and survival rate of flower plants and nursery stock are effectively improved, when reduction is taken root
Between, promote flower plants and nursery stock breeds production.
The growing state of the flower plants and nursery stock of table 1
| Group | Contamination rate % | Rootage duration/day | Bud induction rate % | Rooting rate % | Survival rate % |
| 1st group | 10 | 35 | 65 | 67 | 71 |
| 2nd group | 10.3 | 33 | 70 | 72 | 73 |
| 3rd group | 9.3 | 27 | 76 | 78 | 76 |
| 4th group | 9.6 | 30 | 74 | 75 | 75 |
| 5th group | 8.9 | 25 | 83 | 82 | 83 |
| Embodiment 1 | 5.2 | 20 | 89 | 91 | 92 |
| Embodiment 2 | 4.7 | 18 | 93 | 94 | 95 |
| Embodiment 3 | 4.1 | 16 | 95 | 96 | 97 |
Described above is the detailed description for the present invention preferably possible embodiments, but embodiment is not limited to this hair
Bright patent claim, the equal change completed or modification change under the technical spirit suggested by all present invention, all should belong to
Cover the scope of the claims in the present invention.
Claims (8)
1. a kind of tissue culture and rapid propagation method of flower plants and nursery stock, it is characterised in that comprise the following steps：
(1) make explant from the disease-free stem apex sprouted then of robust growth, after scrubbing clean, soaked with detergent solution
Explant surface is scrubbed with banister brush after 5-7min, the dust on its surface and part thalline are removed, then rinsed with running water
Volumetric concentration is placed in after clean to soak 5-10s in 70-75% alcohol, with being placed into after aseptic water washing 2-3 time equipped with sterilizing
Immersion 50-60min in the container of agent, immersion use aseptic water washing 2-3 times after terminating, the water on surface are finally dried with aseptic filter paper
Pearl, it is standby；
The disinfectant is mainly made up of following raw material：Garlic, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry, garden burnet,
Windproof, gelatinized corn starch, natural gum, castor oil, Chinese juniper oil, spaonin powder and tea seed cake powder；
(2) first holder is added in blake bottle, the bud inducement cultivation base that adds after sterilizing or proliferated culture medium or taken root
Strong seedling culture base, then the explant after sterilization is inoculated into culture medium, successively carries out bud inducement cultivation, Multiplying culture successively
And Rooting and hardening-off culture, the nursery stock that takes root is finally obtained, it is standby；
(3) big Tanaka is transplanted to after selecting the nursery stock progress hardening of taking root of well-grown and stalwartness.
2. the tissue culture and rapid propagation method of flower plants and nursery stock according to claim 1, it is characterised in that in step (1), the sterilization
Agent is mainly made up of following raw material：Garlic 20-30 parts, tealeaves 25-35 parts, madder 15-20 parts, folium artemisiae argyi 20-30 parts, reed
Luxuriant growth 17-20 parts, Sophora alopecuroide 17-20 parts, cogongrass cliff berry 15-17 parts, garden burnet 15-17 parts, windproof 17-20 parts, gelatinized corn starch 5-7 parts, tree
Glue 4-6 parts, castor oil 2-3 parts, Chinese juniper oil 2-3 parts, spaonin powder 3-5 parts and tea seed cake powder 3-5 parts.
3. the tissue culture and rapid propagation method of flower plants and nursery stock according to claim 2, it is characterised in that the disinfectant is according to above-mentioned
Prepared by proportioning, specifically include following steps：
(a) after fresh garlic, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry, garden burnet and windproof removal of impurities and cleaning,
It is put into physiological saline and soaks 8-10min, then with rinsed with sterile water, it is standby；
(b) by the garlic handled well, tealeaves, madder, folium artemisiae argyi, aloe, Sophora alopecuroide, cogongrass cliff berry, garden burnet and windproof chopping, according still further to
1:30-40 ratio mixes with sterilized water decocts 30-40min, is removed the gred with filtered through gauze, obtains filtered fluid, standby；
(c) gelatinized corn starch, natural gum, spaonin powder and tea seed cake powder are added in warm water dissolve it is well mixed after, pour into the mistakes of step (2)
In filtrate, castor oil and Chinese juniper oil are added, is uniformly mixed, obtain the disinfectant.
4. the tissue culture and rapid propagation method of flower plants and nursery stock according to claim 1, it is characterised in that：Step is supported described in (2)
Thing employs aseptic filter paper bridge and solid carbon dioxide tongue is combined, i.e., aseptic filter paper bridge and solid carbon dioxide tongue are soaked in deionized water makes its swollen
It is swollen to be made.
5. the tissue culture and rapid propagation method of flower plants and nursery stock according to claim 4, it is characterised in that：Step is supported described in (2)
The addition of thing is the 1/5-2/5 of blake bottle volume, and wherein the volume ratio of aseptic filter paper bridge and solid carbon dioxide tongue is 1：4-5.
6. the tissue culture and rapid propagation method of flower plants and nursery stock according to claim 1, it is characterised in that：Bud lures described in step (2)
It is that peptone, water are with the addition of in MS minimal mediums to lead culture medium or the proliferated culture medium or the Rooting and hardening-off culture base
Solve casein, glucose, bananas juice, potato juice, natrium nitrosum, azanol, dihydro zeatin, isopentenyl gland purine, more chlorine
Benzoic acid, urea, BNOA, sodium phenate and activated carbon.
7. the tissue culture and rapid propagation method of flower plants and nursery stock according to claim 1, it is characterised in that：Step is taken root described in (2)
Strong seedling culture base is also filled with carbon dioxide.
8. the tissue culture and rapid propagation method of flower plants and nursery stock according to claim 1, it is characterised in that：Rank is respectively cultivated in step (2)
Duan Jun provides illumination using cold-cathode fluorescence lamp, and temperature is 23-27 DEG C, and relative humidity is 65-75%, and pH is 5.0-
6.5, wherein intensity of illumination is 1500-2000LUX in the incubation of the bud inducement cultivation base or the proliferated culture medium,
Intensity of illumination is 3000-3500LUX in the incubation of the Rooting and hardening-off culture base.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710519637.5A CN107372105A (en) | 2017-06-30 | 2017-06-30 | A kind of tissue culture and rapid propagation method of flower plants and nursery stock |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710519637.5A CN107372105A (en) | 2017-06-30 | 2017-06-30 | A kind of tissue culture and rapid propagation method of flower plants and nursery stock |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107372105A true CN107372105A (en) | 2017-11-24 |
Family
ID=60334644
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710519637.5A Pending CN107372105A (en) | 2017-06-30 | 2017-06-30 | A kind of tissue culture and rapid propagation method of flower plants and nursery stock |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107372105A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110250225A (en) * | 2019-06-28 | 2019-09-20 | 大连大学 | A kind of explant sterilizing reagent and preparation method thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105418234A (en) * | 2015-12-07 | 2016-03-23 | 广德县先雨农业有限公司 | Nutrition rooting powder for cuttage of illicium anisatum |
| CN105557516A (en) * | 2015-12-14 | 2016-05-11 | 广西壮族自治区药用植物园 | Method for eliminating bacterial pollution of tissue culture seedlings |
| CN105961205A (en) * | 2016-07-28 | 2016-09-28 | 广西陆川县乌坭坡珍珠番石榴专业合作社 | Tissue culture method for increasing survival rate of psidium guajava L. |
| CN105993961A (en) * | 2016-07-28 | 2016-10-12 | 大连大学 | Plant tissue culture support |
| CN106305419A (en) * | 2016-08-19 | 2017-01-11 | 广西壮族自治区农业科学院甘蔗研究所 | Open cultivation method of sugarcane tissue culture seedling |
-
2017
- 2017-06-30 CN CN201710519637.5A patent/CN107372105A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105418234A (en) * | 2015-12-07 | 2016-03-23 | 广德县先雨农业有限公司 | Nutrition rooting powder for cuttage of illicium anisatum |
| CN105557516A (en) * | 2015-12-14 | 2016-05-11 | 广西壮族自治区药用植物园 | Method for eliminating bacterial pollution of tissue culture seedlings |
| CN105961205A (en) * | 2016-07-28 | 2016-09-28 | 广西陆川县乌坭坡珍珠番石榴专业合作社 | Tissue culture method for increasing survival rate of psidium guajava L. |
| CN105993961A (en) * | 2016-07-28 | 2016-10-12 | 大连大学 | Plant tissue culture support |
| CN106305419A (en) * | 2016-08-19 | 2017-01-11 | 广西壮族自治区农业科学院甘蔗研究所 | Open cultivation method of sugarcane tissue culture seedling |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110250225A (en) * | 2019-06-28 | 2019-09-20 | 大连大学 | A kind of explant sterilizing reagent and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101116424B (en) | Highly effective lily bulblet inducement culture method | |
| CN103875515B (en) | A kind of blueberry tissue culture outside sprout-cultivating-bottle radication method | |
| CN107455261B (en) | A kind of method of Acer saccharum tissue culture quick breeding | |
| CN109258460A (en) | Micro-stem tip culture combines the breeding method of heat treatment acquisition Zengcheng honey chrysanthemum detoxic seedling | |
| CN101595824B (en) | Rapid in-vitro seedling raising method by utilizing sandalwood seed embryo | |
| CN107439315A (en) | A kind of cuttage breeding method of podocarpus | |
| CN108293878A (en) | A kind of tissue culture method of snakegourd tender leaf | |
| CN105557486A (en) | Preparation method of light media for dendrobium officinale | |
| CN106359087A (en) | Tissue culture quick-breeding seedling raising method for radix asparagi | |
| CN107996242A (en) | A kind of Moringa container seedling culture method | |
| CN103070014B (en) | Method for breeding summer truffle root seedling through inoculation of suspension liquid | |
| CN106941904A (en) | A kind of efficiently quick Cut Flower Chrysanthemum Morifolium straight cutting cultural method | |
| CN106922536A (en) | A kind of method that fast energy-saving cultivates bletilla seedling | |
| CN109042330A (en) | A kind of method for tissue culture of spindle tree | |
| CN105917913A (en) | Method for breeding mycorrhizal seedlings by truffles and quercus acutissima | |
| CN108782247A (en) | A kind of method for tissue culture of late cherry " Yu Yihuang " kind of Japan | |
| CN110604060B (en) | Method for obtaining regeneration plant by in vitro culture of Zhejiang red camellia anther | |
| CN102301958B (en) | Method for culturing in vitro tissues of Semiliquidambar cathayensis | |
| CN107188700A (en) | A kind of cuttage and seedling culture method of osmanthus fragrans | |
| CN106489737A (en) | A kind of culture medium of Hybrid Tea tissue cultures and method | |
| CN107372105A (en) | A kind of tissue culture and rapid propagation method of flower plants and nursery stock | |
| CN106069780B (en) | A technique for tissue culture breeding is carried out using bletilla stem tuber | |
| CN110122335A (en) | The abductive approach of Xu Xiang Kiwifruit Tissue Culture seedling salt-tolerant mutant | |
| CN105660709B (en) | A kind of method for the accelerating agent of Chinese yew root growth and its for crossbreeding | |
| CN107372120A (en) | The cultural method that dove tree branch is quickly bred |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20171124 |