CN105165609A - Method for efficiently acquiring gerbera jamesonii detoxification seedling - Google Patents
Method for efficiently acquiring gerbera jamesonii detoxification seedling Download PDFInfo
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- CN105165609A CN105165609A CN201510569632.4A CN201510569632A CN105165609A CN 105165609 A CN105165609 A CN 105165609A CN 201510569632 A CN201510569632 A CN 201510569632A CN 105165609 A CN105165609 A CN 105165609A
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Abstract
The invention discloses a method for efficiently acquiring a gerbera jamesonii detoxification seedling. The method is characterized by comprising the following steps: (1) collecting gerbera jamesonii tender leaves, and de-differentiating the gerbera jamesonii to generate callus; (2) thermally treating the callus, continuously inducing the callus to form adventitious bud, and rooting the adventitious bud to form a strong tissue culture seedling; (3) further stripping a tissue culture seedling stem tip, and de-differentiating to generate callus; (4) thermally treating the callus, and inducing to form a seedling; (5) randomly extracting the tissue culture seedling in the step (4), carrying out the virus detection, if the seedling is qualified in detection, the seedling is the gerbera jamesonmii detoxification seedling, if the seedling is not qualified in the detection, repeating the steps (1) to (4) until the qualified gerbera jamesonii detoxification seedling is acquired. The callus is generated by virtue of the twice de-differentiation, and then the callus is thermally treated and de-differentiated to finally obtain the gerbera jamesonii detoxification seedling, so that the difficulty of the existing detoxification technology such as non-thoroughness in detoxification, complexity in operation and can meet the factory-like detoxification production requirement.
Description
Technical field
The present invention relates to a kind of method of efficient acquisition flameray gerbera detoxification seedling, be specifically related to a kind of produce callus by twice dedifferentiation and will break up again after callus thermal treatment, the final efficient method obtaining flameray gerbera detoxification seedling.
Background technology
Flameray gerbera (
gerberajamesoniibolus) calling African daisy, heronsbill, is composite family herbaceos perennial, and originate in South Africa De Wa Silan area, rear introducing Britain, extensively cultivates now all over the world.Flameray gerbera happiness is warm for winter, the weather that summer is nice and cool, if large, delicate and pretty tall and straight, the free-and-easy pretty ease of flower, pattern gorgeous and not bewitching, beautiful rosy clouds, coquettish elegance, the refreshing heart are pleasing, gives graceful soft, indifferent to fame or gain quiet sense.Flameray gerbera is commonly used to cut-flower plug bottle, if be equipped with the green ornamental foliage plants such as kidney Tibetan, asparagus fern, more fire one's imagination enchanted or fascinated, flameray gerbera is also very suitable is used as potted plant viewing and admiring, and is the good merchantable brand interspersing desk, show window, decoration parlor, ornamental value is higher, the four seasons can supply flower under facility condition, now having ranked among the row of world market situation of selling well cut-flower, is fresh flower kind famous and precious both at home and abroad, one of world-renowned five large cut-flowers, with the fashionable whole world of the glamour of its uniqueness.
Flameray gerbera is cross-pollinated plant, helerothallism, and its seed progeny will inevitably morph, and variation type common manifestation is that bennet attenuates, deliquescing, and flower deformation is little, and Cut-flower evaluation reduces greatly.The gerbera seeds life-span is extremely short, and germination rate is low, its germination rate barely 30%, and adopt seminal propagation, vegetative growth phase is long, blooms slow.Flameray gerbera commonly uses division propagation, but every strain is often only and can be separated 5-6 new strain, cannot meet the requirement that industrialization is produced, and use division propagation always, easily make virus by generation accumulation, thus cause flameray gerbera deterioration of variety serious.
Tissue culture technique is the main modes of reproduction of flameray gerbera merchandized handling, both can produce good seed in a large number in a short time, also can be used for producing detoxic seedling, prevents kind of matter from degenerating.After 20 century 70s, in succession expand the research to flameray gerbera tissue cultures and Fast-propagation both at home and abroad, flameray gerbera adopts in-vitro culture method to carry out numerous soon, just can obtain seedling neat in a large number in a short time, set up high-efficiency regeneration system, the preservation and the improved seeds that are conducive to seed resource are promoted; Plantlet in vitro also has the features such as growth potential is strong, high-quality, high yield, shortening breeding cycle, raising reproduction coefficient, convenient transport, and thus excised African chrysanthemum is numerous soon has wide DEVELOPMENT PROSPECT.
The rapid propagation in vitro technology of flameray gerbera is increasingly mature, but the detoxification technology of flameray gerbera need progressive, detoxification efficiency is low, complex operation has seriously hindered the raising of flameray gerbera quality and the demand of production, therefore study and set up and a kind ofly can keep kind good characteristic and higher reproduction coefficient, the effective ways of the virus causing disease in culture materials can be removed again, for the further diffusion preventing virus, the risk reducing grower has important practical significance.Current flameray gerbera is subject to causing harm of a lot of virus, and the existence of these viruses has a strong impact on production and the export trade of plantlet in vitro, and the complete detoxification problem therefore solving flameray gerbera plantlet in vitro is very important.At present effective way be there is no to the control of viral diseases of plants, and prevent vegetational zone virus from may be one of the most effective way from source, therefore carry out detoxification by stem-tip tissue cultivation in conjunction with heat treatment technics and be widely used in control virus disease, but detoxification efficiency need further raising.
Summary of the invention
The object of the invention is the deficiency existed for existing flameray gerbera detoxification technology; a kind of sustainability, easy to operate and can the method for efficient acquisition flameray gerbera detoxification seedling of large-scale production is provided; the detoxification this method solving existing detoxification technology not thoroughly, the difficult problem of complex operation, the needs that batch production detoxification is produced can be met.
The object of the invention is to solve by the following technical programs:
A method for efficient acquisition flameray gerbera detoxification seedling, is characterized in that: the method step is as follows:
1) gather flameray gerbera tender leaf, dedifferentiation produces callus;
2) callus is heat-treated, then callus is continued to induce indefinite bud, adventitious bud rooting is trained healthy and strong plantlet in vitro;
3) peel off plantlet in vitro stem apex further, dedifferentiation produces callus;
4) again callus is heat-treated rear induction seedling;
5) randomly draw the plantlet in vitro in step 4), carry out Viral diagnosis, if detect qualified i.e. flameray gerbera virus-elimination seedlings, need 1 be repeated if defective) to 4) step, until obtain qualified flameray gerbera virus-elimination seedlings.
The detailed step of the method is as follows:
1) gather flameray gerbera tender leaf, dedifferentiation produces callus:
Gather the tender leaf growing to the anosis flameray gerbera of the health in squaring period, running water is cleaned, running water 30min, with 70% alcohol-pickled 15s, in 0.1% mercuric chloride, addend drips Tween-20 solution and soaks 15min, and stir with glass bar, with aseptic water washing 4-5 time, filter paper blots, and punches with the card punch of bore dia 4mm, by the blade inoculation of laying to light culture under DEG C condition of 26-28 on calli induction media, within 15-20 days, produce the callus of light green grain of rice shape afterwards;
2) callus is heat-treated, then callus is continued to induce indefinite bud, adventitious bud rooting is trained healthy and strong plantlet in vitro:
To large grain of rice size be grown to, transfer to thermal treatment in incubator with the callus of a small amount of explant, after excising brownization block section, callus is transferred to adventitious bud induction culture base glazing to cultivate, after 20-30 days, Calli Differentiation goes out indefinite bud, and under the bud of more than height 1cm born being again forwarded to root media 26-28 DEG C condition, light is trained healthy and strong plantlet in vitro;
3) peel off plantlet in vitro stem apex further, dedifferentiation produces callus:
Treat that plantlet in vitro grows to 6cm-10cm, the shoot tip meristem stripping 0.4-0.7cm size is inoculated on Stem tip induction callus medium, and under 26-28 DEG C of condition, light culture 30-45 days, induces callus;
4) again callus is heat-treated rear induction seedling:
Again stem apex callus is heat-treated, callus is transferred on adventitious bud induction culture base after excising brownization block section, under 26-28 DEG C of condition, light is cultivated after 20-30 days and is induced indefinite bud, then transfers into root media, can obtain flameray gerbera seedling after 40-60 days;
5) randomly draw the plantlet in vitro in step 4), carry out Viral diagnosis, if detect qualified i.e. flameray gerbera virus-elimination seedlings, need 1 be repeated if defective) to 4) step, until obtain qualified flameray gerbera virus-elimination seedlings:
Randomly draw 8-10 only complete step 4) flameray gerbera blade, carry out the detection of RT-PCR retroviruse, if detect qualified i.e. flameray gerbera virus-elimination seedlings.
Flameray gerbera kind elects sunlight seashore or exquisite as.
Calli induction media for inducing flameray gerbera blade to produce callus in step 1) is: MS+2.0mg/L6-BA+0.5mg/LNAA+0.3g/LCH+3% sucrose+6.0g/L agar, pH is 5.8.
Step 2) in adventitious bud induction culture base that just callus continues to induce indefinite bud be: 1/2MS+5mg/L6-BA+0.1NAA+3% sucrose+5.0g/L plant gel; Described by the illumination condition of the medium of callus induction seedling is: intensity of illumination is 1500-2000Lux, and the photoperiod is that 14h light/10h is dark.
Step 2) and step 4) in heat treatment method be: to hocket 2-3 circulation at 38 DEG C of-42 DEG C of high temperature treatment 6h-8h, then 26 DEG C of-28 DEG C of low temperature treatment 10h-12h.
Culture medium prescription for inducing stem apex to produce callus in step 3) is: MS+2.0mg/L6-BA+1mg/LNAA+0.5mg/LKT+3% sucrose, pH is 5.8.
The adventitious bud induction culture based formulas in step 4), callus induction being gone out indefinite bud is: MS+0.5mg/L6-BA+0.5mg/LKT+2% sucrose, and pH is 5.8; Prescription of rooting medium is: 1/2MS+0.1mg/LIBA+1.5% sucrose, and pH is 5.8.
Be: intensity of illumination is 1500-2000Lux that the photoperiod is that 16h light/8h is dark by the illumination condition of stem apex callus induction indefinite bud in step 4).
The viral species of step 5) Viral diagnosis is tomato black ring virus, tobacco mosaic virus, cucumber mosaic virus and Tobacco rattle virus.
The method of a kind of efficient acquisition flameray gerbera detoxification seedling of the present invention, be the preferred result that inventor obtains through carefully studying, after lot of experiment validation, compared to existing technology, innovative point of the present invention is:
The present invention is by adopting comprehensive detoxification technology, the i.e. method that combines of twice callus induction, callus thermal treatment detoxification, stem apex detoxify, effectively overcome the difficult problems such as detoxification material is more, detoxification operating process is loaded down with trivial details, detoxification inefficiency, efficiently can obtain flameray gerbera detoxification seedling.
The invention have employed the heat treated poison-removing method of callus, after adopting callus thermal treatment of the present invention, cut away brownization part, the unconspicuous callus part of brownization degree after cultivation thermal treatment, effectively can get rid of the virus that callus carries, substantially increase detoxification efficiency, and avoid sterile working, simple to operate.
The method of efficient acquisition flameray gerbera detoxification seedling of the present invention adopts Stem tip induction to become callus; improve the kind of flameray gerbera, be stripped of flameray gerbera virus further again, the method flow process simply, easily operates; therefore be easy to the virus-elimination seedlings forming large-scale production, be suitable for promoting the use of.
Accompanying drawing explanation
Fig. 1 is the result figure that the dedifferentiation of sunlight seashore produces callus;
Fig. 2 is the result figure that sunlight seashore callus differentiates indefinite bud again;
Fig. 3 is the schematic diagram peeling off stem apex operation under aseptic condition;
Fig. 4 is the sunlight seashore culture of rootage result figure of 60 days;
Fig. 5 is the result figure that under this technology path, rapid propagation in vitro obtains a large amount of sunlight seashore detoxic seedling.
Embodiment
Below in conjunction with specific embodiment, the present invention is further illustrated.
A kind of method of efficient acquisition flameray gerbera detoxification seedling, the detailed step of the method is as follows: 1) gather flameray gerbera kind sunlight seashore or exquisite tender leaf, dedifferentiation produces callus: gather tender that grows to the anosis flameray gerbera of the health in squaring period, running water is cleaned, running water 30min, with 70% alcohol-pickled 15s, in 0.1% mercuric chloride, addend drips Tween-20 solution and soaks 15min, and stir with glass bar, with aseptic water washing 4-5 time, filter paper blots, punch with the card punch of bore dia 4mm, by the blade inoculation of laying, to calli induction media, (calli induction media formula is: MS+2.0mg/L6-BA+0.5mg/LNAA+0.3g/LCH+3% sucrose+6.0g/L agar, pH is 5.8) light culture under upper 26-28 DEG C of condition, within 15-20 days, produce the callus of light green grain of rice shape afterwards, 2) callus is heat-treated, then callus is continued to induce indefinite bud, adventitious bud rooting is trained healthy and strong plantlet in vitro: will large grain of rice size be grown to, transfer in incubator with the callus of a small amount of explant, 38 DEG C of-42 DEG C of heat treatment process 6h-8h, then 26 DEG C of-28 DEG C of low temperature treatment 10h-12h, the 2-3 that hockets circulate.Then (adventitious bud induction culture based formulas is: 1/2MS+5mg/L6-BA+0.1NAA+3% sucrose+5.0g/L plant gel after excising brownization block section, callus to be transferred to adventitious bud induction culture base, pH is 5.8) light is cultivated under upper 26-28 DEG C of condition, the intensity of illumination that light is cultivated controls as 1500-2000Lux, photoperiod control as 14h light/10h is dark, after 20-30 days, Calli Differentiation goes out indefinite bud, the bud of more than height 1cm born is again forwarded to root media and is trained healthy and strong plantlet in vitro, 3) plantlet in vitro stem apex is peeled off further, dedifferentiation produces callus: treat that plantlet in vitro grows to 6cm-10cm, the shoot tip meristem stripping 0.4-0.7cm size is inoculated on Stem tip induction callus medium that (Stem tip induction callus culture medium prescription is: MS+2.0mg/L6-BA+1mg/LNAA+0.5mg/LKT+3% sucrose, pH is 5.8), under 26-28 DEG C of condition, light culture 30-45 days, induces callus, 4) again callus is heat-treated rear induction seedling: again heat-treated by stem apex callus, heat treatment method is with step 2), (adventitious bud induction culture based formulas is: MS+0.5mg/L6-BA+0.5mg/LKT+2% sucrose after excising brownization block section, callus to be transferred to adventitious bud induction culture base, pH is 5.8) on, intensity of illumination is 1500-2000Lux, photoperiod is the illumination condition that 16h light/8h is dark, indefinite bud is induced after cultivating 20-30 days under the temperature condition of 26-28 DEG C, then transferring into root media, (prescription of rooting medium is: 1/2MS+0.1mg/LIBA+1.5% sucrose, pH is 5.8), flameray gerbera seedling can be obtained after 40-60 days, 5) plantlet in vitro in step 4) is randomly drawed, carry out Viral diagnosis, if detect qualified i.e. flameray gerbera virus-elimination seedlings, need 1 be repeated if defective) to 4) step, until obtain qualified flameray gerbera virus-elimination seedlings: randomly draw 8-10 only complete step 4) flameray gerbera blade, carry out the detection of RT-PCR retroviruse, the viral species of detection is tomato black ring virus, tobacco mosaic virus, cucumber mosaic virus and Tobacco rattle virus, if detect qualified i.e. flameray gerbera virus-elimination seedlings.
Embodiment one
A method for efficient acquisition flameray gerbera detoxification seedling, the detailed step of the method is as follows:
1) tender leaf of flameray gerbera kind sunlight seashore is gathered, dedifferentiation produces callus: gather the tender leaf growing to the anosis flameray gerbera of the health in squaring period, running water is cleaned, running water 30min, with 70% alcohol-pickled 15s, in 0.1% mercuric chloride, addend drips Tween-20 solution and soaks 15min, and stir with glass bar, with aseptic water washing 5 times, filter paper blots, punch with the card punch of bore dia 4mm, by the blade inoculation of laying, to calli induction media, (calli induction media formula is: MS+2.0mg/L6-BA+0.5mg/LNAA+0.3g/LCH+3% sucrose+6.0g/L agar, pH is 5.8) light culture at upper 27 DEG C, after 18 days, we observe the callus creating light green grain of rice shape, as shown in Figure 1,
2) callus is heat-treated, then callus is continued to induce indefinite bud, adventitious bud rooting is trained healthy and strong plantlet in vitro: will grow to large grain of rice size, callus with a small amount of explant is transferred in constant incubator, 40 DEG C of heat treatment process 7h, then 27 DEG C of low temperature treatment 11h, hocket and circulate for 3 times, then (adventitious bud induction culture based formulas is: 1/2MS+5mg/L6-BA+0.1NAA+3% sucrose+5.0g/L plant gel after excising brownization block section, callus to be transferred to adventitious bud induction culture base, pH is 5.8) light is cultivated at upper 27 DEG C, the intensity of illumination that light is cultivated controls as 1800Lux, photoperiod controls as 14h light/10h is dark, Calli Differentiation can be observed after 23 days and go out indefinite bud, as shown in Figure 2, the bud of more than height 1cm born is again forwarded to root media and is trained healthy and strong plantlet in vitro,
3) plantlet in vitro stem apex is peeled off further, dedifferentiation produces callus: treat that plantlet in vitro grows to 6cm-10cm, the shoot tip meristem stripping 0.4-0.7cm size is inoculated on Stem tip induction callus medium that (Stem tip induction callus culture medium prescription is: MS+2.0mg/L6-BA+1mg/LNAA+0.5mg/LKT+3% sucrose, pH is 5.8), stem apex strip operation as shown in Figure 3, then by stem apex light culture 30-45 days under condition at 27 DEG C, callus is induced;
4) again callus is heat-treated rear induction seedling: again heat-treated by stem apex callus, heat treatment method is with step 2), (adventitious bud induction culture based formulas is: MS+0.5mg/L6-BA+0.5mg/LKT+2% sucrose after excising brownization block section, callus to be transferred to adventitious bud induction culture base, pH is 5.8) on, intensity of illumination is 1800Lux, photoperiod is the illumination condition that 16h light/8h is dark, cultivate under 27 DEG C of temperature condition, adventitious bud inducing is had to occur after 22 days, then transferring into root media, (prescription of rooting medium is: 1/2MS+0.1mg/LIBA+1.5% sucrose, pH is 5.8), within about 40 days, induce early stage flameray gerbera seedling, after 60 days, seedling is induced together substantially, as Fig. 4,
5) plantlet in vitro in step 4) is randomly drawed, carry out Viral diagnosis: randomly draw 10 complete step 4) flameray gerbera blade, carry out the detection of RT-PCR retroviruse, the viral species detected is tomato black ring virus, tobacco mosaic virus, cucumber mosaic virus and Tobacco rattle virus, do not find that virus is residual, show the thoroughly detoxification of flameray gerbera seedling, this seedling is expanded numerous in vitro, obtain a large amount of detoxification seedling, as Fig. 5.
The present invention is by adopting comprehensive detoxification technology, the i.e. method that combines of twice callus induction, callus thermal treatment detoxification, stem apex detoxify, effectively overcome the difficult problems such as detoxification material is more, detoxification operating process is loaded down with trivial details, detoxification inefficiency, efficiently can obtain flameray gerbera detoxification seedling.The invention have employed the heat treated poison-removing method of callus, after adopting callus thermal treatment of the present invention, cut away brownization part, the unconspicuous callus part of brownization degree after cultivation thermal treatment, effectively can remove the virus that callus carries, substantially increase detoxification efficiency, and decrease sterile working, simple to operate.The method of efficient acquisition flameray gerbera detoxification seedling of the present invention adopts Stem tip induction to become callus; improve the kind of flameray gerbera, be stripped of flameray gerbera virus further again, the method flow process simply, easily operates; therefore be easy to the virus-elimination seedlings forming large-scale production, be suitable for promoting the use of.
Above embodiment is only and technological thought of the present invention is described, can not limit protection scope of the present invention with this, every technological thought proposed according to the present invention, and any change that technical scheme basis is done, all falls within scope; The technology that the present invention does not relate to all is realized by prior art.
Claims (10)
1. the efficient method obtaining flameray gerbera detoxification seedling, is characterized in that: the method step is as follows:
1) gather flameray gerbera tender leaf, dedifferentiation produces callus;
2) callus is heat-treated, then callus is continued to induce indefinite bud, adventitious bud rooting is trained healthy and strong plantlet in vitro;
3) peel off plantlet in vitro stem apex further, dedifferentiation produces callus;
4) again callus is heat-treated rear induction seedling;
5) randomly draw the plantlet in vitro in step 4), carry out Viral diagnosis, if detect qualified i.e. flameray gerbera virus-elimination seedlings, need 1 be repeated if defective) to 4) step, until obtain qualified flameray gerbera virus-elimination seedlings.
2. the method for efficient acquisition flameray gerbera detoxification seedling according to claim 1, is characterized in that: the detailed step of the method is as follows:
1) gather flameray gerbera tender leaf, dedifferentiation produces callus:
Gather the tender leaf growing to the anosis flameray gerbera of the health in squaring period, running water is cleaned, running water 30min, with 70% alcohol-pickled 15s, in 0.1% mercuric chloride, addend drips Tween-20 solution and soaks 15min, and stir with glass bar, with aseptic water washing 4-5 time, filter paper blots, and punches with the card punch of bore dia 4mm, by the blade inoculation of laying to light culture under DEG C condition of 26-28 on calli induction media, within 15-20 days, produce the callus of light green grain of rice shape afterwards;
2) callus is heat-treated, then callus is continued to induce indefinite bud, adventitious bud rooting is trained healthy and strong plantlet in vitro:
To large grain of rice size be grown to, transfer to thermal treatment in incubator with the callus of a small amount of explant, after excising brownization block section, callus is transferred to adventitious bud induction culture base glazing to cultivate, after 20-30 days, Calli Differentiation goes out indefinite bud, and under the bud of more than height 1cm born being again forwarded to root media 26-28 DEG C condition, light is trained healthy and strong plantlet in vitro;
3) peel off plantlet in vitro stem apex further, dedifferentiation produces callus:
Treat that plantlet in vitro grows to 6cm-10cm, the shoot tip meristem stripping 0.4-0.7cm size is inoculated on Stem tip induction callus medium, and under 26-28 DEG C of condition, light culture 30-45 days, induces callus;
4) again callus is heat-treated rear induction seedling:
Again stem apex callus is heat-treated, callus is transferred on adventitious bud induction culture base after excising brownization block section, under 26-28 DEG C of condition, light is cultivated after 20-30 days and is induced indefinite bud, then transfers into root media, can obtain flameray gerbera seedling after 40-60 days;
5) randomly draw the plantlet in vitro in step 4), carry out Viral diagnosis, if detect qualified i.e. flameray gerbera virus-elimination seedlings, need 1 be repeated if defective) to 4) step, until obtain qualified flameray gerbera virus-elimination seedlings:
Randomly draw 8-10 only complete step 4) flameray gerbera blade, carry out the detection of RT-PCR retroviruse, if detect qualified i.e. flameray gerbera virus-elimination seedlings.
3. the method for efficient acquisition flameray gerbera detoxification seedling according to claim 1 and 2, is characterized in that: flameray gerbera kind elects sunlight seashore or exquisite as.
4. the method for efficient acquisition flameray gerbera detoxification seedling according to claim 1 and 2, it is characterized in that: the calli induction media for inducing flameray gerbera blade to produce callus in step 1) is: MS+2.0mg/L6-BA+0.5mg/LNAA+0.3g/LCH+3% sucrose+6.0g/L agar, pH is 5.8.
5. the method for efficient acquisition flameray gerbera detoxification seedling according to claim 1 and 2, is characterized in that: step 2) in adventitious bud induction culture base that just callus continues to induce indefinite bud be: 1/2MS+5mg/L6-BA+0.1NAA+3% sucrose+5.0g/L plant gel; Described by the illumination condition of the medium of callus induction seedling is: intensity of illumination is 1500-2000Lux, and the photoperiod is that 14h light/10h is dark.
6. the method for efficient acquisition flameray gerbera detoxification seedling according to claim 1 and 2, is characterized in that: step 2) and step 4) in heat treatment method be: to hocket 2-3 circulation at 38 DEG C of-42 DEG C of high temperature treatment 6h-8h, then 26 DEG C of-28 DEG C of low temperature treatment 10h-12h.
7. the method for efficient acquisition flameray gerbera detoxification seedling according to claim 1 and 2, it is characterized in that: the culture medium prescription for inducing stem apex to produce callus in step 3) is: MS+2.0mg/L6-BA+1mg/LNAA+0.5mg/LKT+3% sucrose, pH is 5.8.
8. the method for efficient acquisition flameray gerbera detoxification seedling according to claim 1 and 2, it is characterized in that: the adventitious bud induction culture based formulas in step 4), callus induction being gone out indefinite bud is: MS+0.5mg/L6-BA+0.5mg/LKT+2% sucrose, pH is 5.8; Prescription of rooting medium is: 1/2MS+0.1mg/LIBA+1.5% sucrose, and pH is 5.8.
9. the method for efficient acquisition flameray gerbera detoxification seedling according to claim 1 and 2, it is characterized in that: in step 4) by the illumination condition of stem apex callus induction indefinite bud be: intensity of illumination is 1500-2000Lux, the photoperiod is that 16h light/8h is dark.
10. the method for efficient acquisition flameray gerbera detoxification seedling according to claim 1 and 2, is characterized in that: the viral species of step 5) Viral diagnosis is tomato black ring virus, tobacco mosaic virus, cucumber mosaic virus and Tobacco rattle virus.
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| CN111528092A (en) * | 2020-05-26 | 2020-08-14 | 浙江海丰花卉有限公司 | Culture method of chrysanthemum virus-free seedlings |
| CN114698550A (en) * | 2022-04-29 | 2022-07-05 | 安徽科技学院 | African daisy callus and cluster bud growth promoting culture medium, use method and application |
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