CN103828716A - Tissue culture method of dianthus deltoids - Google Patents
Tissue culture method of dianthus deltoids Download PDFInfo
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Abstract
The invention relates to a tissue culture method of dianthus deltoids. The tissue culture method comprises the steps of taking stem apex of dianthus deltoids twigs as an explants; subjecting the stem apex to sterilization; and culturing by inoculating the stem apex to a primary induction medium composed of MS, 0.01 mg/L 6-BA and 0.05 mg/L NAA, a subculture medium composed of MS, 0.01 mg/L 6-BA and 0.05 mg/L NAA and a rooting medium composed of 1/2 MS and 0.05 mg/L IBA successively under culture conditions that a culture temperature is 25+/-1 DEG C; illumination intensity is 2,500 Lx and light is supplemented for 4 hours/day at night except natural light in the daytime. The culture method can obtain large-scale indoor bred dianthus deltoids seedlings in a short time, has high production rate and has potential ecologic benefits and social benefits.
Description
Technical field
The invention belongs to plant biotechnology field, particularly, relate to the method for tissue culture of a kind of maiden China pink, this cultural method is specially adapted to maiden China pink, be much better than the method for tissue culture of this medium for other China pinks, comprise the selection of China pink explant, stem-tip tissue cultivate in subculture medium and cultivate the selection of algebraically, the content such as the selection of condition of culture in tissue culture procedures.
Background technology
China pink is the showy flowers of herbaceous plants of perennial work one, biennial cultivation, and abundant because of its pattern, the florescence is long, and cultivation management is extensive, is important ornamental flower.The whole world approximately has more than 600 to plant, and is widely distributed in north temperate zone, is mainly distributed in Eurasia, especially Mediterranean Region, and minority is produced North America and north African.Record according to " Chinese Plants will ", China Carnation has 17 kinds, 1 subspecies, 9 mutation, is distributed in Northern grassland and hill country more, is mostly born in dry area without shade, and some kind is born in border or sylvan life, desert and half-desert.Inner Mongol water resource famine, the few water of multiplex sunlight is wanted in landscape planting, should popularize drought resisting green plants in an all-round way, and be grown in the China pink of Inner Mongol arid, semiarid zone, there is indomitable vitality, can survive the winter smoothly in the wild, few at rainwater, without pouring in the situation that also can normal growth, its root system is huge, can prevent erosion and energy anti-harmful gas and damage by disease and insect, there is important hygienic environment-protecting function, can be used as the important component part in gardens, also can be used for afforesting and beautifying of the multiple places such as the roadbed such as highway, sky way, Lu Po.But due to the threat from mankind's activity or natural transition two aspects, its population quantity is existed in retrogression of nature problem and tissue culture expanding propagation kind and occur that sparkling phenomenon causes factorial seedling growth limited amount.Solution is exactly the existing method for tissue culture to China pink of improvement, and this is Fast-propagation, detoxification rejuvenation and the effective ways that reduce sparkling.Under laboratory condition, reducing its sparkling and be one needs practice go the new problem of groping and inquiring into, and data that can reference in the time formulating technical scheme is few, all must settle one by one by test, and key will solve explant and draw materials position, period; The selection of each cultivation period medium; Fast breeding technique etc. after condition of culture and seedling.The present invention, to develop, reduces costs as target, forming batch production, industrialization is produced as object, formulates corresponding work plan and implementing method.
Tissue is cultivated under isolated culture condition, and the histocyte of different plants and kindred plant different parts, to nutritional requirement difference, only has and met their specific (special) requirements separately, could grow better.Preparation and the screening technique of grasping medium are to obtain one of successful key link of group training.
Plant Tissue Breeding is very important animal nutrition.The cell of plant, stem section, blade etc. all can be cultivated.Get cell, the Organ and tissue of plant, insert in the container of appropriate culture medium, under certain condition of culture, can be differentiated to form whole plant.Cultivate as the tissue of biotechnology powerful measure, day by day come into one's own, bring out in detoxifying fast breeding, the sudden change of farm-forestry crop, the aspect such as cell engineering and gene engineering all plays an important role.
In the tissue of pinkwort is cultivated, often select stem section, blade, petal and stem apex etc. to obtain regeneration plant as explant.Stem section is also one of common used material in pinkwort Fast-propagation, and the advantage such as draw materials conveniently because it has, quantity is many is widely adopted.Watad etc. have studied with stem section and have made explant, adopt that gel entrapment culture base is cultivated, liquid shake is cultivated and liquid nutrient medium on floating interfacial film is auxiliary the impact of 3 kinds of different training methods on carnation regeneration frequency such as cultivates, result shows, the regeneration frequency of the third training method can improve two to three times than other two kinds of modes, and this can be greatly cost-saving in plant fast propagation process.Godo etc. cultivate and are studied LychnisFlos-Cuculi stem section, and the regeneration frequency of explant on 1/2MS+0.2mg/L BA and 1/2MS medium is respectively 30% and 70%.On MS+10mg/L BA medium, each stem section node on average can generate 8 and can utilize bud, and the genetic stability quite stable of bud.
The blade of pinkwort is cultivated and can be obtained regeneration plant by direct or indirect Regeneration Ways.Jethwani etc. add growth hormone PAA to break up research to the callus of Chinese phyllostachys nuda leaf sheet in medium again.Found that, the suitableeest Calli Differentiation medium is MS+BA (2.0,5.0mg/L)+PAA (0.5,1.0mg/L).No matter be used alone or in combination BA, NAA, 2,4-D, callus all can not break up generation bud again.In addition, they also compare the callus induction situation of the 1st of Chinese China pink the leaf (from top) and the 2nd leaf, research shows that the 1st leaf and the 2nd callus that leaf base portion induces have regeneration capacity, and the callus of the 2nd leaf remainder induction does not have regeneration capacity.The research reports such as Kantia: Chinese China pink leaf explant is substantially cultivated on tomb at the MS of 3mg/L BA+0.5mg/L NAA and 3mg/L BA+1mg/L NAA and cultivated, and all can directly produce indefinite bud; And at 0.5mg/L BA+1mg/L2, though also can produce indefinite bud on the MS medium of 4-D, being attended by the generation of callus, the indefinite bud of generation will could be grown on the medium containing BA and NAA.Paresk etc. are take the blade of carnation, Chinese China pink and sweetwilliam as explant, adopt MS+1mg/L2,4-D liquid nutrient medium has induced somatic embryo, somatic embryo develops into seedling on MS+1mg/L GA3 solid culture medium, has set up first the direct somatic embryo inducement regenerating system of pinkwort.
Nakano etc. are take Carnation Petal, blade and stem section as explant induction indefinite bud.Result only has petal to have higher regeneration frequency, and suitable medium is MS+1.1-2.2mg/L BA and MS+0.9mg/L NAA.Fzsiter etc. have studied the impact of two kinds of training methods on China pink shoot regeneration frequency take Carnation Petal as explant.Result shows, China pink petal liquid medium within is than the regeneration rate on solid culture medium high (the former can reach 100%, and the latter only has 50%); Each position of petal all can form regeneration bud, and each explant can produce at most 7 indefinite buds.Casanova etc. also induce regeneration bud by Carnation Petal, and inductivity reaches 84.4%, and the regeneration bud inductivity of rolC Transgenic carnation petal is up to 90.4%.
Stem apex is the conventional position of drawing materials during pinkwort tissue is cultivated, except for Fast-propagation, also for detoxification cultivation, germ plasm resource preservation etc.Since early 1990s, report the achievement in research of this respect both at home and abroad.Carnation is because the cottage propagation adopting in producing causes quality degradation, and therefore, its Shoot Tip Culture has important production practices meaning.Can etc. have studied different growth hormone, the basic element of cell division and gibberellin combination to the fast numerous impact of fringed pink stem apex, and result shows: shoot apical meristem in MS+5.0mg/L BA+1.0mg/L IAA medium than to contain the Multiple Buds producing in the medium of other several combination of regulators many.Onamu etc. reported the dosage of TDZ and the processing time very large on the impact of carnation stem apex in-vitro propagate efficiency.Their research shows, the concentration of TDZ is 1-5mg/L, and the processing time is 3-10d, and when carnation stem apex is carried out to in-vitro propagate, efficiency can reach 100%.
Up to now, both at home and abroad to cultivate successful Caryophyllaceae kind a lot of for tissue, but what be more common in report is production and the cultivation for carnation, less to maiden China pink Study on tissue culture.Sum up the data in literature over nearly 20 years, it is that stem section, blade are more successful that maiden's Caryophyllaceae kind tissue is cultivated majority, about caning be counted on one's fingers especially of maiden China pink Shoot Tip Culture.The artificial propagation of maiden China pink still mainly adopts seeding method and cuttage, breeds limited amount, time length and very easily causes indefinite bud sparkling, disease to infect and quality deterioration.Can be used at present separating the tissue culture method breeding of shortening seedling raise period, batch production productions, seedling Vitrification Occurred, rejuvenating and study lessly, therefore carrying out maiden China pink Study on tissue culture in a deep going way has become one of work that researcher urgently carries out.Carry out kind innovation with the wild resource of China's abundant, apply diversified breeding technique, paying attention on the basis of conventional cross-breeding, carry out breeding of new variety in conjunction with modern biotechnology means, carry out the batch production exploitation of kind of ball using detoxication and tissue culture seedling as yielding ability original seed, be the developing direction that maiden China pink tissue is cultivated simultaneously.Tissue culture method breeding is drawn materials conveniently; be not subject to the restriction of natural conditions; can constantly carry out amount reproduction; maiden China pink Fast-propagation, the most effectual way that reduces vitrifying, detoxification rejuvenation and rearing new variety; can obtain in a short time a large amount of seedlings; solved the vitrifying of maiden China pink, shortened seedling raise period, the less problem of provenance in cultivation, for the research from now on of maiden China pink and scale, industrialization development utilization provide theoretical foundation.
The complete procedure that stem-tip tissue is cultivated is generally divided into several sport technique segments such as rooting culture of formulating culture scheme, explant selection and processing, inoculation, first culture, shoot proliferation and expanding numerous cultivation, strong sprout and culture of rootage, test-tube plantlet, and front to have mentioned preparation and the screening technique of grasping medium be to obtain to organize to train successful key link.The present invention filters out the optimal medium of cultivating different times for maiden China pink tissue, adjust the proportioning to hormone in different times medium, medium according to different formulations preparation has met the nutritional need in each period of maiden China pink and has grown, and has solved the long problem of a large amount of vitrification phenomenons of maiden China pink seedling and growing-seedling period simultaneously.
Before mentioned domestic and international many scholars the tissue of China pink cultivated and is also studied, be applied at present in the production of carnation China pink, but what be more common in report is that cultivar produces for fresh cut-flowers, medium and method are not also suitable for maiden China pink, and the present invention has systematically studied selection, the optimal culture condition of the explant of maiden China pink, optimal medium and the propagation of different phase is cultivated algebraically.
The present invention is easy and simple to handle, with low cost, has a good application prospect.The present invention has systematically studied the optimal medium of different phase in the group training of maiden China pink, the suitableeest condition of culture and training method, solved maiden China pink vitrifying, shortened seedling raise period, the difficult problem of the less problem of provenance in cultivation, increase kind and there is stronger practicality for realizing drought resisting landscape planting new concept, for reliable basis has been established in the batch production production that realizes China pink.
Summary of the invention
In embodiment of the present invention, the method for tissue culture that comprises a kind of maiden China pink, is characterized in that it is made up of following steps:
Step 1: explant is taken from the stem apex of maiden China pink spray;
Step 2: explant is inoculated in just for inducing culture MS+6-BA0.01mg/L+NAA0.05mg/L above, and induction differentiates Multiple Buds;
Step 3: the Multiple Buds of first generation induction was cultivated for 3 generations on subculture medium MS+6-BA0.01mg/L+NAA0.05mg/L, then its Multiple Buds is cultivated to 2 generations on the MS medium of improvement, finally above-mentioned Multiple Buds was cultivated after 1 generation on subculture medium, transferred to the upper cultivation of root media 1/2MS+IBA0.05mg/L;
Step 4: the seedling of taking root of the maiden China pink after above-mentioned root induction is transplanted.
The condition of culture of each step is: 25 ± 1 ℃ of cultivation temperature, intensity of illumination 2500Lx, except natural lighting on daytime, night light filling 4 hours/day.
Wherein, the MS medium of improvement is the ionic equilibrium solution that changes part mineral salt and ion concentration in MS medium, the minimal medium of cultivating for maiden China pink shoot proliferation, and it specifically consists of:
Macroelement: KNO
31900mg/L; NH
4nO
3413mg/L; CaCl
22H
2o440mg/L; MgSO
47H
2o370mg/L; KH
2pO
4310mg/L
Trace element: KI0.83mg/L; H
3bO
36.2mg/L; MnSO
44H
2o22.3mg/L; ZnSO
47H
2o8.6mg/L; Na
2moO
42H
2o0.25mg/L; CuSO
45H
2o0.25mg/L; CoCl
26H
2o0.025mg/L
Molysite: FeSO
47H
2o27.8mg/L; Na
2-EDTA2H2O37.3mg/L
Organic substance: inositol 100mg/L; Nicotinic acid 0.5mg/L; Puridoxine hydrochloride (vitamin B6) 0.5mg/L; Thiamine hydrochloride (vitamin B1) 0.1mg/L; Glycine 2.0mg/L
6-BA1.0mg/L；NAA0.5mg/L。
In embodiment of the present invention, the sterilization treatment process of organizing maiden China pink explant in incubation step 1 of maiden China pink is: on super-clean bench, put into sterile chamber, in every liter drips 75% the alcohol that 2-3 drips " Tween ", soak 10 seconds, process 5 minutes with 0.15% mercuric chloride solution again, aseptic washing 2 times, suck dry moisture on last aseptic filter paper.
In embodiment of the present invention, the organizing in incubation step 2 seeded process of maiden China pink, maiden China pink spray stem apex leaf primordium otch inserts medium.
The present invention also passes through comparative test, filters out each stage optimal medium of maiden China pink group training:
A. be just the first for inducing culture of maiden China pink the best for inducing culture: MS+6-BA0.01mg/L+NAA0.05mg/L, not only inductivity is high, and the newly-increased bud number of each explant is many, and differentiation rate is the highest, and its differentiation rate can reach 200%.
B. shoot proliferation medium: MS+6-BA0.01mg/L+NAA0.05mg/L, the 6-BA of high concentration has inhibitory action to maiden China pink propagation, excessive concentration, can suppress the growth of bud, along with increasing of 6-BA concentration, in the time that 6-BA concentration exceedes 0.2mg/L, grow thickly and downgrade obviously, easily produce vitrifying seedling.
C. root media: 1/2MS+IBA0.05mg/L, low salt concn is favourable to taking root of maiden China pink.
Maiden China pink, no matter at first culture or in subculture is cultivated, in MS+6-BA0.01mg/L+NAA0.05mg/L cultivates, has facilitation to the growth of Multiple Buds, compare other combinations even more ideal, Multiple Buds quantity is many, and seedling is larger, transplants emergence rate best results.
In specific embodiments of the present invention, also comprise the acclimatization and transplants stage after maiden China pink takes root, result shows, vermiculite+sheep excrement+diammonium phosphate, as cultivation matrix best results, is applicable to the growth of seedling root system very much.
Embodiment
Choosing and the impact of method of operating on adventitious bud inducing of embodiment 1. maiden China pink explants
The maiden China pink of experiment use is adopted in cover careless drought resisting from He Xin garden, the Inner Mongol and afforests the test nursery of drought-resistant plant research institute of limited company Experimental Base, samples and samples and supplement at any time according to test progress, different times.Adopt the stem apex that is about 0.5-1.0cm on maiden China pink children shoot, after flowing water rinses, with 70% alcohol-pickled 10sec, then with 0.1% mercuric chloride sterilization 5min, aseptic water washing 2 times, each 5min, the first culture base (MS) that is inoculated in improvement is upper, sets up test-tube plantlet clone.The Multiple Buds of choosing subculture 10d left and right, Multiple Buds cuts off base portion; The stem apex of getting Multiple Buds, is about 0.5-1.0cm, as the explant of regeneration.
Wherein, the first of improvement is the ionic equilibrium solution that changes part mineral salt and ion concentration in MS medium for MS medium, and for the just minimal medium of culture of maiden China pink, it specifically consists of:
Macroelement: KNO
3950mg/L; NH
4nO
3825mg/L; CaCl
22H
2o220mg/L; MgSO
47H
2o185mg/L; KH
2pO
485mg/L
Trace element: KI0.83mg/L; H
3bO
36.2mg/L; MnSO
44H
2o22.3mg/L; ZnSO
47H
2o8.6mg/L; Na
2moO
42H
2o0.25mg/L; CuSO
45H
2o0.25mg/L; CoCl
26H
2o0.025mg/L
Molysite: FeSO
47H
2o27.8mg/L; Na
2-EDTA2H2O37.3mg/L
Organic substance: inositol 100mg/L; Nicotinic acid 0.5mg/L; Puridoxine hydrochloride (vitamin B6) 0.5mg/L; Thiamine hydrochloride (vitamin B1) 0.1mg/L; Glycine 2.0mg/L
6-BA2.5mg/L; NAA1.0mg/L; Active carbon 1g/L; Hydrolysis network albumen 0.5g/L; Antibiotic 50mg/L.
The screening that embodiment 2. maiden China pink tissues are cultivated each stage optimal medium
1, the statistical computation formula of screening effect
Survival rate=(the explant sum of viable explant number/inoculation) × 100%
Just for bud induction rate=(the explant sum of the explant number/inoculation of sprouting) × 100%
Shoot proliferation multiple=(sum of the Multiple Buds number/access individual plant bud inducing) × 100%
Rooting rate=(the explant sum of the explant number/inoculation of differentiation root) × 100%
2, China pink tissue is cultivated the screening of each stage optimal medium
Take MS as minimal medium, make curing agent with the carragheen of 3.5g/L, 3% edible sugar replaces sucrose as carbon source, replaces distilled water with running water, and the basic element of cell division and the growth hormone that add respectively different proportionings carry out the screening of medium.
2.1, the selection of inductive differentiation medium
The concentration of 6-BA is established to 0.01mg/L, 0.05mg/L, 0.1mg/L, 0.2mg/L, five concentration gradients of 0.5mg/L, the concentration of NAA is established 0.01mg/L, 0.05mg/L, 0.1mg/L, tetra-concentration gradients of 0.2mg/L, carry out different assembly, every bottle graft 2-3 maiden China pink stem apex (stem apex is about 0.5-1cm) when inoculation, observe stem apex on different culture media indefinite bud a situation arises.
As seen from Table 1,7 kinds of hormone combinations all can be induced maiden China pink differentiation Multiple Buds, but the differentiation rate difference of each combination, No. 1 MS+6-BA0.01mg/L+NAA0.05mg/L is that maiden China pink Shoot Tip Culture base is best inducing culture, not only inductivity is high, and the newly-increased bud number of each explant is many, differentiation rate is the highest, and its differentiation rate can reach 200%.
Table 1 hormone concentration and the impact of combination on maiden China pink stem apex inducing clumping bud thereof
2.2, the selection of shoot proliferation medium
The Multiple Buds that is about 0.5-1.0cm differentiating is taken out, be inoculated on the shoot proliferation medium of MS+6-BA0.01mg/L+NAA0.05mg/L, additional edible white granulated sugar 40g/L, carragheen 3.5g/L, subculture cultivated for 3 generations, bred and maiden China pink seedling; Then the just above-mentioned Multiple Buds that is about 0.5-1.0cm differentiating takes out, and is inoculated in shoot proliferation on the MS medium of improvement and cultivates, additional edible white granulated sugar 40g/L, carragheen 3.5g/L, subculture cultivated for 2 generations, bred and maiden China pink seedling, can reduce the sparkling phenomenon of maiden China pink; Finally, the above-mentioned Multiple Buds that is about 0.5-1.0cm differentiating is taken out, be inoculated on the shoot proliferation medium of MS+6-BA0.01mg/L+NAA0.05mg/L, additional edible white granulated sugar 40g/L, carragheen 3.5g/L, subculture cultivated for 1 generation, bred and maiden China pink seedling.
Wherein, the MS medium of improvement is the ionic equilibrium solution that changes part mineral salt and ion concentration in MS medium, the minimal medium of cultivating for maiden China pink shoot proliferation, and it specifically consists of:
Macroelement: KNO
31900mg/L; NH
4nO
3413mg/L; CaCl
22H
2o440mg/L; MgSO
47H
2o370mg/L; KH
2pO
4310mg/L
Trace element: KI0.83mg/L; H
3bO
36.2mg/L; MnSO
44H
2o22.3mg/L; ZnSO
47H
2o8.6mg/L; Na
2moO
42H
2o0.25mg/L; CuSO
45H
2o0.25mg/L; CoCl
26H
2o0.025mg/L
Molysite: FeSO
47H
2o27.8mg/L; Na
2-EDTA2H2O37.3mg/L
Organic substance: inositol 100mg/L; Nicotinic acid 0.5mg/L; Puridoxine hydrochloride (vitamin B6) 0.5mg/L; Thiamine hydrochloride (vitamin B1) 0.1mg/L; Glycine 2.0mg/L
6-BA1.0mg／L；NAA0.5mg/L。
The indefinite bud that above-mentioned all inductions differentiate is inoculated on proliferated culture medium, 6-BA establishes 0.01mg/L, 0.05mg/L, 0.1mg/L, tetra-concentration gradients of 0.2mg/L, NAA establishes 0.01mg/L, 0.05mg/L, 0.1mg/L, tetra-concentration gradients of 0.2mg/L totally eight kinds of medium, 4 individual plants of every bottle graft, the differential growth situation of observed and recorded regenerated adventitious bud, adds up the rate of increase of bud under different hormone combinations condition for 15 days afterwards.
After induction is cultivated, the 0.5-1.0cm budlet producing in inducing culture is moved into respectively to (in table 2) in proliferated culture medium, the bastem portion of 2-3 days rear section materials starts observed and recorded, statistical results after 15 days while there is bud clump.Observation shows, the 6-BA of high concentration has inhibitory action to maiden China pink propagation, and excessive concentration, can suppress the growth of bud, along with increasing of 6-BA concentration, in the time that 6-BA concentration exceedes 0.2mg/L, grows thickly and downgrades obviously, easily produces vitrifying seedling.The optimum multiplication medium formula filtering out is MS+6-BA0.01mg/L+NAA0.05mg/L.
Table 2 hormone concentration and the impact of combination on maiden China pink adventitious bud proliferation thereof
Test shows, maiden China pink is no matter at first culture or in subculture is cultivated, in MS+6-BA0.01mg/L+NAA0.05mg/L cultivates, growth to Multiple Buds has facilitation, compares other combinations even more ideal, and Multiple Buds quantity is many, seedling is larger, transplants emergence rate best results.
2.3, the selection of root media
By good growing way in subculture medium seedling, be divided into individual plant and proceed in root media, every bottle graft enters 4 strains, and on observed and recorded different culture media, the situation of taking root of seedling, added up rooting rate after 10 days.
From the culture of rootage result of the test of 10 days, two medium all have inducing action to the formation of root, and wherein No. 2 root media rooting efficiencies are better, and rooting rate reaches 88.89%.The base portion of taking root can increase the Multiple Buds with root newly, on average can reach 3-4.Visible low salt concn is favourable to the growth of maiden China pink root, and suitable medium of taking root is 1/2MS+IBA0.05mg/L (table 3).
The impact that table 3 mineral salt are taken root on seedling
Above embodiment object is for illustrating content of the present invention, but should not be construed as limitation of the present invention.Without departing from the spirit and substance of the case in the present invention, the modification that the inventive method, step or condition are done or replacement, all belong to scope of the present invention.
Claims (5)
1. a method for tissue culture for maiden China pink, is characterized in that it is made up of following steps:
Step 1: adopt the stem apex that is about 0.5-1.0cm on maiden China pink children shoot, be inoculated in the MS medium of improvement after sterilization treatment, choose the Multiple Buds of subculture 10d left and right, Multiple Buds cuts off base portion; The stem apex of getting Multiple Buds, is about 0.5-1.0cm, as the explant of regeneration;
Step 2: explant is inoculated in just for inducing culture MS+6-BA0.01mg/L+NAA0.05mg/L above, and induction differentiates Multiple Buds;
Step 3: the Multiple Buds of first generation induction was cultivated for 3 generations on subculture medium MS+6-BA0.01mg/L+NAA0.05mg/L, then its Multiple Buds is cultivated to 2 generations on the MS medium of improvement, finally above-mentioned Multiple Buds is cultivated after a generation on subculture medium, transferred to the upper cultivation of root media 1/2MS+IBA0.05mg/L;
Step 4: the seedling of taking root of the maiden China pink after above-mentioned root induction is transplanted.
2. cultural method according to claim 1, is characterized in that, the condition of culture of each step is: 25 ± 1 ℃ of cultivation temperature, intensity of illumination 2500Lx, except natural lighting on daytime, night light filling 4 hours/day.
3. cultural method according to claim 1, it is characterized in that, the sterilization treatment process of the explant of step 1 is: on super-clean bench, put into sterile chamber, in every liter drips 75% the alcohol that 2-3 drips " Tween ", soak 10 seconds, process 5 minutes with 0.15% mercuric chloride solution again, aseptic washing 2 times, suck dry moisture on last aseptic filter paper.
4. cultural method according to claim 1, is characterized in that, in step 1 seeded process, maiden China pink spray stem apex leaf primordium otch inserts medium.
5. cultural method according to claim 1, is characterized in that, the matrix that step 3 is cultivated after transplanting is made up of vermiculite, sheep excrement, diammonium phosphate.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105325289A (en) * | 2014-08-11 | 2016-02-17 | 中国农业科学院蔬菜花卉研究所 | Method for overcoming vitrification in shoot tip culture of carnations by using high-molecular water-absorbent resin |
| CN107155604A (en) * | 2017-06-16 | 2017-09-15 | 内蒙古蒙草生态环境（集团）股份有限公司 | The cultivation management method of " girl in red carnation " |
| CN110810245A (en) * | 2019-11-29 | 2020-02-21 | 内蒙古蒙草生态环境（集团）股份有限公司 | Tissue culture method of floral rod |
| CN112400696A (en) * | 2020-12-28 | 2021-02-26 | 内蒙古蒙草生态环境（集团）股份有限公司 | Tissue culture method of evergreen common selfheal fruit-spike bamboo |
| CN112400695A (en) * | 2020-12-28 | 2021-02-26 | 内蒙古蒙草生态环境（集团）股份有限公司 | Culture medium for culturing evergreen common summer pink |
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| CN101836588A (en) * | 2010-03-29 | 2010-09-22 | 云南省农业科学院花卉研究所 | Method for cryopreservation and plant regeneration of Dianthus caryophyllus shoot tip |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105325289A (en) * | 2014-08-11 | 2016-02-17 | 中国农业科学院蔬菜花卉研究所 | Method for overcoming vitrification in shoot tip culture of carnations by using high-molecular water-absorbent resin |
| CN105325289B (en) * | 2014-08-11 | 2018-01-16 | 中国农业科学院蔬菜花卉研究所 | Overcome the vitrified method of carnation Shoot Tip Culture using hydroscopic high-molecular resin |
| CN107155604A (en) * | 2017-06-16 | 2017-09-15 | 内蒙古蒙草生态环境（集团）股份有限公司 | The cultivation management method of " girl in red carnation " |
| CN110810245A (en) * | 2019-11-29 | 2020-02-21 | 内蒙古蒙草生态环境（集团）股份有限公司 | Tissue culture method of floral rod |
| CN110810245B (en) * | 2019-11-29 | 2022-08-30 | 蒙草生态环境(集团)股份有限公司 | Tissue culture method of floral rod |
| CN112400696A (en) * | 2020-12-28 | 2021-02-26 | 内蒙古蒙草生态环境（集团）股份有限公司 | Tissue culture method of evergreen common selfheal fruit-spike bamboo |
| CN112400695A (en) * | 2020-12-28 | 2021-02-26 | 内蒙古蒙草生态环境（集团）股份有限公司 | Culture medium for culturing evergreen common summer pink |
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|---|---|
| CN103828716B (en) | 2015-11-25 |
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