CN102657097A - In-vitro culture method for tender stem segments of dendrobium nobile - Google Patents
In-vitro culture method for tender stem segments of dendrobium nobile Download PDFInfo
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- CN102657097A CN102657097A CN2012101799012A CN201210179901A CN102657097A CN 102657097 A CN102657097 A CN 102657097A CN 2012101799012 A CN2012101799012 A CN 2012101799012A CN 201210179901 A CN201210179901 A CN 201210179901A CN 102657097 A CN102657097 A CN 102657097A
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Abstract
The invention provides an in-vitro culture method for tender stem segments of dendrobium nobile. The in-vitro culture method is characterized by comprising the following specific steps of: step 1, selecting newly-sprouted tender branches, peeling off leaves and sheaths, cleaning, disinfecting, washing by sterile water, and absorbing water by sterile filter paper; step 2, cutting the tender branches obtained in the step 1 into stem segments, and inducing axillary buds to germinate and grow; step 3, cutting off the buds obtained in the step 2, and inducing clustered buds to grow; step 4, cutting the clustered buds into single buds, inoculating the single buds on a third culture medium, and then performing rooting culture to obtain rooted test-tube seedlings; and step 5, transplanting the rooted test-tube seedlings in a moss matrix, placing in a plastic plug and then culturing into 'moss plug seedlings', watering enough after transplanting, starting to spraying clean water on leaf surfaces the next day, dressing nutrient solution after half month, transplanting in a flowerpot after more than three months of culture, and then culturing as potted flowers or plastic shed big seedlings. The seedlings of dendrobium nobile obtained by the culture method provided by the invention are fast in propagation speed and high in propagation rate.
Description
Technical field
The present invention relates to a kind of dendrobium in-vitro rapid culture method for tender stem segments, utilize shoot induced bundle behind cleaning and sterilizing of taking out about living length 10cm to sprout, obtain complete test-tube plantlet through cultivating the back again, belong to dendrobium cultural method technical field.
Background technology
Dendrobium is that the wet high-grade famous and precious torrid zone of a kind of happiness temperature happiness is blue, and spring dendrobium wherein is one of main kind, and adaptability is stronger; In a great variety gorgeous, the florescence reaches the several months, and ornamental value is high; Its seedling leans on cottage propagation, and the low and easy poison in spite of illness of reproduction rate can't satisfy the need of producing with seedling.
Summary of the invention
The purpose of this invention is to provide the spring dendrobium seedling cultural method that a kind of reproduction speed is fast, reproduction rate is high.
For achieving the above object, the invention provides a kind of in-vitro rapid culture method for tender stem segments of spring dendrobium, it is characterized in that concrete steps are:
The first step: choosing and newly taking out living length is 5～15 centimetres spray, peels off blade and leaf sheath, cleans with liquid detergent; Elder generation is with alcohol solution dipping sterilization 0.5～1.5min of 75vol% on superclean bench; Again with clean your quick solution soaking sterilization 15min, clean you quick and volume ratio water are 1: 50 in described clean your the quick solution, use the mercuric chloride solution soaking disinfection 15min of 0.1vol% at last; Use aseptic water washing, use the aseptic filter paper wipe dry;
Second step: the spray that the first step is obtained is cut into the long stem section of 0.5～1.5cm, and each stem section is with 1 stipes, is inoculated in and induces the axillary bud sprouting growth in first medium;
The 3rd step: downcut the sprouting that second step obtained, be inoculated in second medium induced bundle growth of sprouting;
The 4th step: after the bud of will growing thickly is cut into simple bud, is inoculated into and carries out culture of rootage, the test-tube plantlet that obtains taking root on the 3rd medium;
The 5th step: the test-tube plantlet that will take root moves in the liver moss matrix, breeds " liver moss cave dish seedling " in the plastics cave dish of packing into, waters sufficient water after the transplanting; Second day beginning foliage spray clear water; Every day 2 times, impose nutrient solution behind the first quarter moon, semimonthly; After cultivating more than three months, move into and do the potted flower cultivation in the flowerpot or do the cultivation of plastic tunnel seedlings.
Preferably; First medium in described second step obtains by in the MS medium, adding the basic element of cell division, gibberellin, methyl and banana puree; The concentration of the basic element of cell division is 1mg/L in first medium; The concentration of gibberellin is 0.2mg/L, and the concentration of methyl is 0.1mg/L, and the concentration of banana puree is 50g/L.
Preferably; Second medium in said the 3rd step obtains by in the MS medium, adding the basic element of cell division, gibberellin, methyl and banana puree; The concentration of the basic element of cell division is 1-2mg/L in second medium; The concentration of gibberellin is 0.2mg/L, and the concentration of methyl is 0.1-0.2mg/L, and the concentration of banana puree is 50g/L.
Preferably, the 3rd medium in said the 4th step is obtained by interpolation indolebutyric acid and methyl in the 1/2MS medium, and the concentration of indolebutyric acid is 0.5mg/L in the 3rd medium, and methyl concentration is 0.1mg/L.
Preferably, the composition of the nutrient solution in said the 5th step is: the aqueous solution that contains 0.1vol% potassium dihydrogen phosphate and 0.1vol% ammonium nitrate.
Compared with prior art, advantage of the present invention is: resulting spring dendrobium sapling multiplication speed is fast, reproduction rate is high.
Embodiment
Specify the present invention below in conjunction with embodiment.
Embodiment
A kind of in-vitro rapid culture method for tender stem segments of spring dendrobium, concrete steps are:
The first step: choosing and newly taking out living length is 10 centimetres spray, peels off blade and leaf sheath, cleans two times with liquid detergent; Elder generation is with the alcohol solution dipping sterilization 1min of 75vol% on superclean bench; Again with clean your quick solution soaking sterilization 15min, clean you quick and volume ratio water are 1: 50 in described clean your the quick solution, use the mercuric chloride solution soaking disinfection 15min of 0.1vol% at last; With aseptic water washing three times, use the aseptic filter paper wipe dry;
Second step: the spray that the first step is obtained is cut into the long stem section of 1cm, and each stem section is with 1 stipes, is inoculated in and induces the axillary bud sprouting growth in first medium; First medium obtains by in the MS medium, adding the basic element of cell division, gibberellin, methyl and banana puree; The concentration of the basic element of cell division is 1mg/L in first medium; The concentration of gibberellin is 0.2mg/L, and the concentration of methyl is 0.1mg/L, and the concentration of banana puree is 50g/L;
The 3rd step: downcut the sprouting that second step obtained, be inoculated in second medium induced bundle growth of sprouting; Second medium obtains by in the MS medium, adding the basic element of cell division, gibberellin, methyl and banana puree; The concentration of the basic element of cell division is 1.5mg/L in second medium; The concentration of gibberellin is 0.2mg/L, and the concentration of methyl is 0.15mg/L, and the concentration of banana puree is 50g/L;
The 4th step: after the bud of will growing thickly is cut into simple bud, is inoculated into and carries out culture of rootage, the test-tube plantlet that obtains taking root on the 3rd medium; The 3rd medium is obtained by interpolation indolebutyric acid and methyl in the 1/2MS medium, and the concentration of indolebutyric acid is 0.5mg/L in the 3rd medium, and methyl concentration is 0.1mg/L; 23 ℃ of culturing room's temperature, illumination 12 hours every days, intensity of illumination 3000LX.
The 5th step: the test-tube plantlet that will take root moves in the liver moss matrix, and the plastics cave dish of packing into is bred " liver moss cave dish seedling " in (72 hole), water sufficient water after the transplanting; Second day beginning foliage spray clear water; Every day 2 times, impose nutrient solution behind the first quarter moon, semimonthly; After cultivating more than three months, move into and do the potted flower cultivation in the flowerpot.The composition of nutrient solution is: contain the aqueous solution of 0.1vol% potassium dihydrogen phosphate and 0.1vol% ammonium nitrate, applied amount is each plastics cave dish (72 hole) 1000ml.
Wherein, The blastogenesis of growing thickly of the 3rd one-step inducing is long, every 35-45 days propagation once, each growth coefficient 1: 3-5; With 10 calculating of year propagation; 1 bud of growing thickly can obtain 50000-60000 simple bud (deduction microbial contamination rate about 5%) in 1 year, promptly changed for the 4th step over to can obtain 55000 strain test-tube plantlets (rooting rate 96%) during culture of rootage, proved absolutely superiority of the present invention.
Claims (5)
1. the in-vitro rapid culture method for tender stem segments of a spring dendrobium is characterized in that, concrete steps are:
The first step: choosing and newly taking out living length is 5～15 centimetres spray, peels off blade and leaf sheath, cleans with liquid detergent; Elder generation is with alcohol solution dipping sterilization 0.5～1.5min of 75vol% on superclean bench; Again with clean your quick solution soaking sterilization 15min, clean you quick and volume ratio water are 1: 50 in described clean your the quick solution, use the mercuric chloride solution soaking disinfection 15min of 0.1vol% at last; Use aseptic water washing, use the aseptic filter paper wipe dry;
Second step: the spray that the first step is obtained is cut into the long stem section of 0.5～1.5cm, and each stem section is with 1 stipes, is inoculated in and induces the axillary bud sprouting growth in first medium;
The 3rd step: downcut the sprouting that second step obtained, be inoculated in second medium induced bundle growth of sprouting;
The 4th step: after the bud of will growing thickly is cut into simple bud, is inoculated into and carries out culture of rootage, the test-tube plantlet that obtains taking root on the 3rd medium;
The 5th step: the test-tube plantlet that will take root moves in the liver moss matrix, breeds " liver moss cave dish seedling " in the plastics cave dish of packing into, waters sufficient water after the transplanting; Second day beginning foliage spray clear water; Every day 2 times, impose nutrient solution behind the first quarter moon, semimonthly; After cultivating more than three months, move into and do the potted flower cultivation in the flowerpot or do the cultivation of plastic tunnel seedlings.
2. the in-vitro rapid culture method for tender stem segments of spring dendrobium as claimed in claim 1; It is characterized in that; First medium in described second step obtains by in the MS medium, adding the basic element of cell division, gibberellin, methyl and banana puree, and the concentration of the basic element of cell division is 1mg/L in first medium, and the concentration of gibberellin is 0.2mg/L; The concentration of methyl is 0.1mg/L, and the concentration of banana puree is 50g/L.
3. the in-vitro rapid culture method for tender stem segments of spring dendrobium as claimed in claim 1; It is characterized in that; Second medium in said the 3rd step obtains by in the MS medium, adding the basic element of cell division, gibberellin, methyl and banana puree, and the concentration of the basic element of cell division is 1-2mg/L in second medium, and the concentration of gibberellin is 0.2mg/L; The concentration of methyl is 0.1-0.2mg/L, and the concentration of banana puree is 50g/L.
4. the in-vitro rapid culture method for tender stem segments of spring dendrobium as claimed in claim 1; It is characterized in that; The 3rd medium in said the 4th step is obtained by interpolation indolebutyric acid and methyl in the 1/2MS medium; The concentration of indolebutyric acid is 0.5mg/L in the 3rd medium, and methyl concentration is 0.1mg/L.
5. the in-vitro rapid culture method for tender stem segments of spring dendrobium as claimed in claim 1 is characterized in that, the composition of the nutrient solution in said the 5th step is: the aqueous solution that contains 0.1vol% potassium dihydrogen phosphate and 0.1vol% ammonium nitrate.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103314852A (en) * | 2013-06-28 | 2013-09-25 | 上海市农业科学院 | Method for efficiently propagating dendrobium by using roots and culture medium of dendrobium |
| CN103371100A (en) * | 2012-04-17 | 2013-10-30 | 上海市农业科学院 | Tissue culture and rapid propagation method of nobile-type dendrobium seedlings |
| CN103416303A (en) * | 2013-07-10 | 2013-12-04 | 浙江建设职业技术学院 | Inducing method for spring dendrobium protoemm-like body |
| CN103548681A (en) * | 2013-10-31 | 2014-02-05 | 浙江农林大学天目学院 | Culture medium for rapidly inducing adventitious buds in tissue culture of dendrobium nobile |
| CN103583358A (en) * | 2013-10-24 | 2014-02-19 | 湖南农业大学 | Method for in vitro culturing of regenerated plant of dendrobium officinale |
| CN106069751A (en) * | 2016-06-14 | 2016-11-09 | 中国林业科学研究院林业研究所 | A kind of method of spring Herba Dendrobii kind ' firebird ' tissue-culturing rapid propagation |
| CN108739321A (en) * | 2018-05-28 | 2018-11-06 | 四川省植物工程研究院 | A kind of wild Dendrodium propagation method |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103371100A (en) * | 2012-04-17 | 2013-10-30 | 上海市农业科学院 | Tissue culture and rapid propagation method of nobile-type dendrobium seedlings |
| CN103371100B (en) * | 2012-04-17 | 2014-09-17 | 上海市农业科学院 | Tissue culture and rapid propagation method of nobile-type dendrobium seedlings |
| CN103314852A (en) * | 2013-06-28 | 2013-09-25 | 上海市农业科学院 | Method for efficiently propagating dendrobium by using roots and culture medium of dendrobium |
| CN103416303A (en) * | 2013-07-10 | 2013-12-04 | 浙江建设职业技术学院 | Inducing method for spring dendrobium protoemm-like body |
| CN103583358A (en) * | 2013-10-24 | 2014-02-19 | 湖南农业大学 | Method for in vitro culturing of regenerated plant of dendrobium officinale |
| CN103583358B (en) * | 2013-10-24 | 2016-03-30 | 湖南农业大学 | A kind of method of dendrobium cultured in vitro regeneration plant |
| CN103548681A (en) * | 2013-10-31 | 2014-02-05 | 浙江农林大学天目学院 | Culture medium for rapidly inducing adventitious buds in tissue culture of dendrobium nobile |
| CN103548681B (en) * | 2013-10-31 | 2016-02-24 | 浙江农林大学天目学院 | The substratum of rapid evoking adventive bud in a kind of spring stem of noble dendrobium tissue culture |
| CN106069751A (en) * | 2016-06-14 | 2016-11-09 | 中国林业科学研究院林业研究所 | A kind of method of spring Herba Dendrobii kind ' firebird ' tissue-culturing rapid propagation |
| CN108739321A (en) * | 2018-05-28 | 2018-11-06 | 四川省植物工程研究院 | A kind of wild Dendrodium propagation method |
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Application publication date: 20120912 |