CN107616094A - A kind of pocket orchid culture medium and its application method - Google Patents
A kind of pocket orchid culture medium and its application method Download PDFInfo
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- CN107616094A CN107616094A CN201710986165.4A CN201710986165A CN107616094A CN 107616094 A CN107616094 A CN 107616094A CN 201710986165 A CN201710986165 A CN 201710986165A CN 107616094 A CN107616094 A CN 107616094A
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- 239000001963 growth medium Substances 0.000 title claims abstract description 42
- 241000233855 Orchidaceae Species 0.000 title claims abstract description 39
- 238000000034 method Methods 0.000 title claims abstract description 20
- 239000000463 material Substances 0.000 claims abstract description 33
- 229920001213 Polysorbate 20 Polymers 0.000 claims abstract description 15
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims abstract description 15
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims abstract description 15
- 229930192334 Auxin Natural products 0.000 claims abstract description 14
- 241000196324 Embryophyta Species 0.000 claims abstract description 14
- 239000002363 auxin Substances 0.000 claims abstract description 14
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 claims abstract description 14
- 239000000758 substrate Substances 0.000 claims abstract description 14
- 238000004904 shortening Methods 0.000 claims abstract description 8
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 claims abstract description 7
- 239000000126 substance Substances 0.000 claims abstract description 7
- 239000011573 trace mineral Substances 0.000 claims abstract description 7
- 235000013619 trace mineral Nutrition 0.000 claims abstract description 7
- 238000002203 pretreatment Methods 0.000 claims abstract description 4
- 230000001954 sterilising effect Effects 0.000 claims description 9
- 238000012545 processing Methods 0.000 claims description 4
- 238000004659 sterilization and disinfection Methods 0.000 claims description 4
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 claims description 3
- 230000000844 anti-bacterial effect Effects 0.000 claims description 3
- 239000003899 bactericide agent Substances 0.000 claims description 3
- 230000015572 biosynthetic process Effects 0.000 claims description 3
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 claims description 3
- 229910052603 melanterite Inorganic materials 0.000 claims description 3
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 238000009395 breeding Methods 0.000 description 4
- 230000001488 breeding effect Effects 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241001516485 Cypripedium Species 0.000 description 2
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 229960000367 inositol Drugs 0.000 description 2
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 2
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 description 2
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 2
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 description 1
- 241000732800 Cymbidium Species 0.000 description 1
- 241000045439 Cypripedioideae Species 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 241000519406 Paphiopedilum Species 0.000 description 1
- 241000921313 Phyllopodium Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 229930003451 Vitamin B1 Natural products 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000035613 defoliation Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 239000007943 implant Substances 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000002073 mitogenetic effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 230000001863 plant nutrition Effects 0.000 description 1
- 235000010333 potassium nitrate Nutrition 0.000 description 1
- 239000004323 potassium nitrate Substances 0.000 description 1
- 230000001902 propagating effect Effects 0.000 description 1
- RADKZDMFGJYCBB-UHFFFAOYSA-N pyridoxal hydrochloride Natural products CC1=NC=C(CO)C(C=O)=C1O RADKZDMFGJYCBB-UHFFFAOYSA-N 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- 235000019190 thiamine hydrochloride Nutrition 0.000 description 1
- 239000011747 thiamine hydrochloride Substances 0.000 description 1
- 229960000344 thiamine hydrochloride Drugs 0.000 description 1
- 235000010374 vitamin B1 Nutrition 0.000 description 1
- 239000011691 vitamin B1 Substances 0.000 description 1
- 235000019158 vitamin B6 Nutrition 0.000 description 1
- 239000011726 vitamin B6 Substances 0.000 description 1
- 229940011671 vitamin b6 Drugs 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention discloses a kind of pocket orchid culture medium, it is characterised in that the culture medium includes culture medium substrate and auxin, and the culture medium substrate includes a great number of elements, trace element, molysite, organic substance；The auxin includes BA, Tween20 and TDZ；The dosage of the BA is 5ppm, and the concentration of the Tween20 is 100ppm；The concentration of the TDZ is 8mg/L.Cultural method includes pre-treatment, Initial culture, squamous subculture.The present invention had opened the plant shortening stem of flower to cultivate body material using pocket orchid, solved the blue cultivation body pollution of pocket in tissue cultures, browning, slow-growing, and bred the problems such as multiplying power is low, and provided a kind of pocket orchid rapid propagation method.
Description
Technical field
The present invention relates to plant and its culture field, more particularly to a kind of pocket orchid culture medium and its application method.
Background technology
Pocket is blue also known as celestial shoe blue (Lady ' sslipper) is orchid family (Orchidaceae), lady's slipper subfamily
(Cypripedioideae) general name of implants.Pocket orchid Cymbidium (Paphiopedilum) interior orchid is most to be passed through in lady's slipper subfamily
The crop for value of helping.After pocket orchid was listed in the species in Washington pact (CITES) annex one in 1989, inlet and outlet transaction is tight
International business's behavior is carried out by government control, complete prohibition Wild plant lattice.
Currently commercially propagation method is based on aseptic seeding, and Lan Miao is to most fast person of blooming after bottle outlet
Paph.Maudiae, need the time of 1 to 2 year 1 year；But some spend more strain, about more than 4-6 can just bloom.After bottle outlet to
Blooming about needs more than 3-4.Because pocket orchid seedling is irregular, and the technology of clone breeding seedling strain remains in plant division side
Method, it can not largely produce the plant with excellent character of blooming.Therefore, the mitogenetic propagating system for establishing pocket orchid is extremely urgent class
Topic.
At present for the research of pocket orchid tissue cultures, mostly using bottle seedling be material (woods, 1998；Flood, 2004；It is old,
2000；Kawase, 1995a；YasugiandYagi, 1995；Tanakaetal., 1995,1996), minority is using ripe strain
Shortening stem and bennet bud do breed material (Huang, 1988；Kawase, 1990,1994,1997；StewartandButton,
1975) the problem of bottle seedling speed of growth is slow, is so all faced.But it can face sterilizing in the blue tissue culture procedures of pocket be not easy, be brown
Change seriously with breeding the problem of multiplying power is slow.
The defects of in order to overcome prior art, is, it is necessary to design a kind of pocket orchid culture medium and its application method.
The content of the invention
The defects of in order to overcome in the prior art, there is provided a kind of pocket orchid culture medium and its application method.
The present invention is realized by following proposal：
A kind of pocket orchid culture medium, the culture medium include culture medium substrate and auxin, and the culture medium substrate includes a large amount of
Element, trace element, molysite, organic substance；
The auxin includes BA, Tween20 and TDZ；
The dosage of the BA is 5ppm, and the concentration of the Tween20 is 100ppm；The concentration of the TDZ is 8mg/L.
Calculated by mass concentration unit of mg/L, a great number of elements includes following component：
Calculated by mass concentration unit of mg/L, the trace element includes following component：
Calculated by mass concentration unit of mg/L, the molysite includes following component：
FeSO4·7H2O 27.8
Na2-EDTA·2H2O 37.3。
Calculated by mass concentration unit of mg/L, the organic substance includes following component：
A kind of application method of pocket orchid culture medium, the application method comprise the following steps：
The first step, pre-treatment
Before carrying out tissue cultures, body material is cultivated in selection, and first cultivation body material is cut off the water supply and handled 2 weeks, then removes root,
Remove and proceed by multiple sterilization treatment within second day after root；The cultivation body material of sterilizing is put into after shortening stem removes stem top,
Make in the NaClO that the dipped mass concentration in stem top is 0.005%, then move into pocket orchid culture medium and cultivated, and in the blue culture of pocket
Bactericide is added in base；
Second step, Initial culture
Auxin is added in culture medium substrate：BA and Tween20, the BA dosage are 5ppm, and the Tween20's is dense
Spend for 100ppm；After the cultivation body material darkroom that first step processing is completed is cultivated 3 weeks, and cultivating body material has sprout to be formed
When, remove darkroom irradiation culture 1 month, have jade-green expansion leaf into；
3rd step, squamous subculture
Body material is cultivated after Initial culture base transplanting comes, auxin is added in culture medium substrate：TDZ, the TDZ's
Concentration is 8mg/L；Cultivation body material after the completion of second step is handled, which removes, cultivates 8 weeks, plant height 2.85cm after blade, afterwards instead
Multiple operation, can reach required increment multiple.
In the first step, the plant of 7 leaves is selected as cultivation body material.
Beneficial effects of the present invention are：
The plant shortening stem that the present invention had opened flower using pocket orchid solves the blue cultivation body of pocket in tissue cultures as body material is cultivated
It is pollution, browning, slow-growing, and the problems such as breeding multiplying power is low, and provide a kind of pocket orchid rapid propagation method.
Embodiment
With reference to specific embodiment, the present invention is further described：
A kind of pocket orchid culture medium, the culture medium include culture medium substrate and auxin, and the culture medium substrate includes a large amount of
Element, trace element, molysite, organic substance；
The auxin includes BA, Tween20 and TDZ；
The dosage of the BA is 5ppm, and the concentration of the Tween20 is 100ppm；The concentration of the TDZ is 8mg/L.
Calculated by mass concentration unit of mg/L, a great number of elements includes following component：
Calculated by mass concentration unit of mg/L, the trace element includes following component：
Calculated by mass concentration unit of mg/L, the molysite includes following component：
FeSO4·7H2O 27.8
Na2-EDTA·2H2O 37.3。
Calculated by mass concentration unit of mg/L, the organic substance includes following component：
The alias of puridoxine hydrochloride in the present invention is vitamin B6, and the alias of thiamine hydrochloride is vitamin B1.In reality
In the application of border, inositol is usually to separate to be made into concentrate alone in addition, because it compares indissoluble, is typically made into 100 times, and
It is organic after removing inositol, it still can be made into 1000 times.
A kind of application method of pocket orchid culture medium, the application method comprise the following steps：
The first step, pre-treatment
Before carrying out tissue cultures, body material is cultivated in selection, and first cultivation body material is cut off the water supply and handled 2 weeks, then removes root,
Remove and proceed by multiple sterilization treatment within second day after root；The cultivation body material of sterilizing is put into after shortening stem removes stem top,
Make in the NaClO that the dipped mass concentration in stem top is 0.005%, then move into pocket orchid culture medium and cultivated, and in the blue culture of pocket
Bactericide is added in base；In the present invention, the plant of 7 leaves is selected as cultivation body material.
Second step, Initial culture
Auxin is added in culture medium substrate：BA and Tween20, the BA dosage are 5ppm, and the Tween20's is dense
Spend for 100ppm；After the cultivation body material darkroom that first step processing is completed is cultivated 3 weeks, and cultivating body material has sprout to be formed
When, remove darkroom irradiation culture 1 month, have jade-green expansion leaf into；The sprout of pocket orchid plant is present on the inside of leaf base,
And one phyllopodium position there was only an axillary bud, stem block is after culture, most axillary bud only long sprouting, and only a small number of axillary buds have point
Change the situation for forming polygerm to occur.
3rd step, squamous subculture
Body material is cultivated after Initial culture base transplanting comes, auxin is added in culture medium substrate：TDZ, the TDZ's
Concentration is 8mg/L；Cultivation body material after the completion of second step is handled, which removes, cultivates 8 weeks, plant height 2.85cm after blade, afterwards instead
Multiple operation, can reach required increment multiple.
The present invention divided the seedling strain of plant nutrition system successfully to be trained as cultivation body material using what the blue cenospecies of pocket had opened flower
Support the shortening stem of defoliation plant, and from axillary bud development seedling.By culture materials, first pretreatment dries 2 before sterilization in the present invention
In week, success rate can be improved.And sterilizing 9 minutes is shaken with the Tween20 containing 100ppm and 1% liquor natrii hypochloritis so that this hair
Bright cultivation body material survival rate is higher.Culture materials are higher with 7 persons of blade, survival rate to select.
The CuSO in pocket orchid culture medium4·5H2Concentration is improved to 0.75mg/l, can grow 1 bud.In liquid medium within
The newly-increased sprout number of culture is more compared with solid medium, and can reduce the formation of root.The H in pocket orchid culture medium3BO3Concentration drops
To original 2/3, sprout can be promoted to be formed ahead of time.When pocket orchid culture medium subtracts ammonium nitrate with potassium nitrate, addition yeast extraction
Thing substitution nitrogen source can increase newly.Although pocket orchid sprout hyperplasia can be promoted in the TDZ low concentrations person of high concentration (8 or 16mg/l),
It is still lower than adding BA50mg/l effect.In addition, disleaf processing can increase the formation of sprout and suppress to cultivate the long root of body.
The plant shortening stem that the present invention had opened flower using pocket orchid solves the blue cultivation body of pocket in tissue cultures as body material is cultivated
It is pollution, browning, slow-growing, and the problems such as breeding multiplying power is low, and provide a kind of pocket orchid rapid propagation method.
Although more detailed elaboration is done to technical scheme and has been enumerated, it will be appreciated that for ability
For field technique personnel, modification is made to above-described embodiment or uses equivalent alternative solution, this is to those skilled in the art
It is it is clear that these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to the present invention for member
Claimed scope.
Claims (7)
1. a kind of pocket orchid culture medium, it is characterised in that the culture medium includes culture medium substrate and auxin, the culture medium substrate
Including a great number of elements, trace element, molysite, organic substance；
The auxin includes BA, Tween20 and TDZ；
The dosage of the BA is 5ppm, and the concentration of the Tween20 is 100ppm；The concentration of the TDZ is 8mg/L.
A kind of 2. pocket orchid culture medium according to claim 1, it is characterised in that：Calculated by mass concentration unit of mg/L,
The a great number of elements includes following component：
A kind of 3. pocket orchid culture medium according to claim 1, it is characterised in that：Calculated by mass concentration unit of mg/L,
The trace element includes following component：
A kind of 4. pocket orchid culture medium according to claim 1, it is characterised in that：Calculated by mass concentration unit of mg/L,
The molysite includes following component：
FeSO4·7H2O 27.8
Na2-EDTA·2H2O 37.3。
A kind of 5. pocket orchid culture medium according to claim 1, it is characterised in that：Calculated by mass concentration unit of mg/L,
The organic substance includes following component：
6. a kind of application method of pocket orchid culture medium as claimed in claim 1, it is characterised in that the application method includes following
Step：
The first step, pre-treatment
Before carrying out tissue cultures, body material is cultivated in selection, and first cultivation body material is cut off the water supply and handled 2 weeks, is then removed root, is removed
Proceed by multiple sterilization treatment within second day after root；The cultivation body material of sterilizing is put into after shortening stem removes stem top, makes stem
Push up in the NaClO that dipped mass concentration is 0.005%, then move into pocket orchid culture medium and cultivated, and in pocket orchid culture medium
Add bactericide；
Second step, Initial culture
Auxin is added in culture medium substrate：BA and Tween20, the BA dosage are 5ppm, and the concentration of the Tween20 is
100ppm；After the cultivation body material darkroom that first step processing is completed is cultivated 3 weeks, and when cultivating body material and having sprout formation,
Remove darkroom irradiation culture 1 month, have jade-green expansion leaf into；
3rd step, squamous subculture
Body material is cultivated after Initial culture base transplanting comes, auxin is added in culture medium substrate：TDZ, the concentration of the TDZ
For 8mg/L；Cultivation body material after the completion of second step is handled is cultivated 8 weeks after removing blade, plant height 2.85cm, is grasped repeatedly afterwards
Make, can reach required increment multiple.
A kind of 7. application method of pocket orchid culture medium according to claim 6, it is characterised in that：In the first step, 7 are selected
The plant of piece leaf is as cultivation body material.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710986165.4A CN107616094B (en) | 2017-10-20 | 2017-10-20 | Paphiopedilum culture medium and using method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710986165.4A CN107616094B (en) | 2017-10-20 | 2017-10-20 | Paphiopedilum culture medium and using method thereof |
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| Publication Number | Publication Date |
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| CN107616094A true CN107616094A (en) | 2018-01-23 |
| CN107616094B CN107616094B (en) | 2020-02-21 |
Family
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109258463A (en) * | 2018-09-18 | 2019-01-25 | 广西壮族自治区林业科学研究院 | A kind of asexual reproduction method of paphiopedilum armeniacum |
| CN116671438A (en) * | 2023-05-29 | 2023-09-01 | 广西壮族自治区中国科学院广西植物研究所 | Method for prolonging quick seedling raising of stems of paphiopedilum |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109258463A (en) * | 2018-09-18 | 2019-01-25 | 广西壮族自治区林业科学研究院 | A kind of asexual reproduction method of paphiopedilum armeniacum |
| CN109258463B (en) * | 2018-09-18 | 2022-08-02 | 广西壮族自治区林业科学研究院 | Vegetative propagation method of paphiopedilum armeniacum |
| CN116671438A (en) * | 2023-05-29 | 2023-09-01 | 广西壮族自治区中国科学院广西植物研究所 | Method for prolonging quick seedling raising of stems of paphiopedilum |
| CN116671438B (en) * | 2023-05-29 | 2024-05-21 | 广西壮族自治区中国科学院广西植物研究所 | Method for prolonging quick seedling raising of stems of paphiopedilum |
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