CN117256476A - Non-symbiotic germination and rapid breeding method for cymbidium yunnanensis seeds - Google Patents
Non-symbiotic germination and rapid breeding method for cymbidium yunnanensis seeds Download PDFInfo
- Publication number
- CN117256476A CN117256476A CN202311454865.0A CN202311454865A CN117256476A CN 117256476 A CN117256476 A CN 117256476A CN 202311454865 A CN202311454865 A CN 202311454865A CN 117256476 A CN117256476 A CN 117256476A
- Authority
- CN
- China
- Prior art keywords
- medium
- culture
- germination
- pods
- symbiotic germination
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 230000035784 germination Effects 0.000 title claims abstract description 49
- 241000732800 Cymbidium Species 0.000 title claims abstract description 18
- 238000009395 breeding Methods 0.000 title claims description 13
- 239000001963 growth medium Substances 0.000 claims abstract description 27
- 238000000034 method Methods 0.000 claims abstract description 16
- 235000013399 edible fruits Nutrition 0.000 claims abstract description 10
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims abstract description 8
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims abstract description 8
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims abstract description 4
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims abstract description 4
- 235000011130 ammonium sulphate Nutrition 0.000 claims abstract description 4
- YYRMJZQKEFZXMX-UHFFFAOYSA-L calcium bis(dihydrogenphosphate) Chemical compound [Ca+2].OP(O)([O-])=O.OP(O)([O-])=O YYRMJZQKEFZXMX-UHFFFAOYSA-L 0.000 claims abstract description 4
- 229910000389 calcium phosphate Inorganic materials 0.000 claims abstract description 4
- 238000003306 harvesting Methods 0.000 claims abstract description 4
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims abstract description 4
- 235000019341 magnesium sulphate Nutrition 0.000 claims abstract description 4
- 229940099596 manganese sulfate Drugs 0.000 claims abstract description 4
- 235000007079 manganese sulphate Nutrition 0.000 claims abstract description 4
- 239000011702 manganese sulphate Substances 0.000 claims abstract description 4
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims abstract description 4
- 235000019691 monocalcium phosphate Nutrition 0.000 claims abstract description 4
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 4
- 235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 4
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims abstract description 4
- 235000010333 potassium nitrate Nutrition 0.000 claims abstract description 4
- 239000004323 potassium nitrate Substances 0.000 claims abstract description 4
- 239000002609 medium Substances 0.000 claims description 24
- 238000005286 illumination Methods 0.000 claims description 23
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical class [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 20
- 238000004140 cleaning Methods 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 claims description 6
- 238000012136 culture method Methods 0.000 claims description 6
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 claims description 6
- 230000001954 sterilising effect Effects 0.000 claims description 6
- 239000012869 germination medium Substances 0.000 claims description 5
- 239000012879 subculture medium Substances 0.000 claims description 5
- 244000061456 Solanum tuberosum Species 0.000 claims description 4
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 4
- 229920001817 Agar Polymers 0.000 claims description 3
- 244000060011 Cocos nucifera Species 0.000 claims description 3
- 235000013162 Cocos nucifera Nutrition 0.000 claims description 3
- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 claims description 3
- 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
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
- 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 claims description 3
- 239000005708 Sodium hypochlorite Substances 0.000 claims description 3
- 229930006000 Sucrose Natural products 0.000 claims description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 3
- 229930003451 Vitamin B1 Natural products 0.000 claims description 3
- 239000008272 agar Substances 0.000 claims description 3
- 235000019301 disodium ethylene diamine tetraacetate Nutrition 0.000 claims description 3
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 3
- 239000011790 ferrous sulphate Substances 0.000 claims description 3
- 229960000367 inositol Drugs 0.000 claims description 3
- 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 claims description 3
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 3
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 3
- 229960003512 nicotinic acid Drugs 0.000 claims description 3
- 235000001968 nicotinic acid Nutrition 0.000 claims description 3
- 239000011664 nicotinic acid Substances 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- RADKZDMFGJYCBB-UHFFFAOYSA-N pyridoxal hydrochloride Natural products CC1=NC=C(CO)C(C=O)=C1O RADKZDMFGJYCBB-UHFFFAOYSA-N 0.000 claims description 3
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 claims description 3
- 238000002791 soaking Methods 0.000 claims description 3
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims description 3
- 239000005720 sucrose Substances 0.000 claims description 3
- 229960003495 thiamine Drugs 0.000 claims description 3
- 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 claims description 3
- 235000010374 vitamin B1 Nutrition 0.000 claims description 3
- 239000011691 vitamin B1 Substances 0.000 claims description 3
- 235000019158 vitamin B6 Nutrition 0.000 claims description 3
- 239000011726 vitamin B6 Substances 0.000 claims description 3
- 229940011671 vitamin b6 Drugs 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 2
- 239000002245 particle Substances 0.000 claims description 2
- 238000004659 sterilization and disinfection Methods 0.000 claims description 2
- 239000000758 substrate Substances 0.000 claims description 2
- 239000002352 surface water Substances 0.000 claims description 2
- 238000004383 yellowing Methods 0.000 claims description 2
- 241000196324 Embryophyta Species 0.000 abstract description 13
- 241000233855 Orchidaceae Species 0.000 description 14
- 238000009331 sowing Methods 0.000 description 9
- 235000004584 Yucca mohavensis Nutrition 0.000 description 8
- 244000110633 Yucca schidigera Species 0.000 description 8
- 235000006012 Yucca schidigera Nutrition 0.000 description 8
- 230000001488 breeding effect Effects 0.000 description 8
- 230000004083 survival effect Effects 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 238000010586 diagram Methods 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 230000010152 pollination Effects 0.000 description 4
- 230000007226 seed germination Effects 0.000 description 4
- 238000011031 large-scale manufacturing process Methods 0.000 description 3
- 235000011299 Brassica oleracea var botrytis Nutrition 0.000 description 2
- 235000017647 Brassica oleracea var italica Nutrition 0.000 description 2
- 240000003259 Brassica oleracea var. botrytis Species 0.000 description 2
- 241000259759 Cassida nobilis Species 0.000 description 2
- 244000247747 Coptis groenlandica Species 0.000 description 2
- 235000002991 Coptis groenlandica Nutrition 0.000 description 2
- 241000283070 Equus zebra Species 0.000 description 2
- 206010016807 Fluid retention Diseases 0.000 description 2
- 244000014908 Gaultheria leucocarpa var. yunnanensis Species 0.000 description 2
- 241000146448 Hedychium gracile Species 0.000 description 2
- 239000003205 fragrance Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- 241001083841 Aquatica Species 0.000 description 1
- 241000234305 Hedychium Species 0.000 description 1
- 241000947931 Hedyotis yunnanensis Species 0.000 description 1
- 241000906682 Hemsleya Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241001451963 Holcoglossum rupestre Species 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- 241000736285 Sphagnum Species 0.000 description 1
- 244000290333 Vanilla fragrans Species 0.000 description 1
- 235000009499 Vanilla fragrans Nutrition 0.000 description 1
- 235000012036 Vanilla tahitensis Nutrition 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000010154 cross-pollination Effects 0.000 description 1
- 230000000249 desinfective effect Effects 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 238000010413 gardening Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000000474 nursing effect Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000001699 photocatalysis Effects 0.000 description 1
- 238000007146 photocatalysis Methods 0.000 description 1
- 238000004382 potting Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000003716 rejuvenation Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
- A01C1/08—Immunising seed
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/002—Culture media for tissue culture
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Environmental Sciences (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Soil Sciences (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention belongs to the technical field of plant rapid propagation, and particularly discloses a non-symbiotic germination and rapid propagation method for a cymbidium yunnanensis seed, which comprises the following steps of: fruit clamp harvesting, pod pretreatment and non-symbiotic germination culture, wherein a culture medium used for the non-symbiotic germination culture is an improved VW culture medium, and the culture medium comprises the following components: 525mg/L potassium nitrate, 500mg/L ammonium sulfate, 250mg/L potassium dihydrogen phosphate, 250mg/L magnesium sulfate, 200mg/L calcium superphosphate, 7.5mg/L manganese sulfate and the like.
Description
Technical Field
The invention belongs to the technical field of plant rapid propagation, and particularly relates to a non-symbiotic germination and rapid propagation method for a cymbidium yunnanensis seed.
Background
The herba Hedyotidis Diffusae (Holcoglossum rupestre (hand-Mazz.) Garay) is a plant of the genus Hedychium (Holcoglossaum) of the family Orchidaceae, and is usually attached to the trunk of forest or on the stones below the forest. The special species of China are distributed in the northwest of Yunnan and are 6 months in the flowering period. The plant stems are very short, and the base parts grow a plurality of air roots; she Rouzhi is cylindrical, and the front end is tapered; the inflorescence is inclined, 6-10 flowers are provided, sepals and petals are white, the petals are nearly oval, the front ends are round and blunt, 3 veins are provided, the outside veins are branched, and the lips are red and 3 cracks. The flower shape is elegant, the leaf shape is delicate, the flower color is gorgeous, the flower has elegant fragrance, the flowering performance is good, the flower can bloom each year, and the flower has higher gardening value.
The plant of the genus zebra is not listed in the IUCN red list nor in the CITES appendix, but in practical investigation, it was found that the plant of the genus zebra belongs to an endangered group whose main causes of endangerment are loss of habitat, slow growth, uniaxial branching characteristics and artificial overdrawing. Because the number of living bodies in the field is less and the population is endangered to be extinct, all species of the genus Vanilla in 2023 are to be listed the Yunnan province focused on protecting wild plant catalogues. The number of surviving bodies in the field of the yunnan cymbidium is very small, the distribution is narrow, and the insects are attracted by the strong flower fragrance, so that pollinators can be reported back from secretion in the middle distance. The study shows that the tortoise beetle with legs is the only pollinator of the yunnan, but the tortoise beetle with legs only accounts for 2% of all visitors, and the yunnan has lower setting rate under natural condition, slower natural updating and aggravated endangered condition.
The study and report of the glossostrea aquatica at home and abroad are few, the basic study is weak, and the study on the aspect of pollination biology is mainly concentrated, and the nursing study work is blank. The flower market and the network find that a plurality of plants of the genus hynchophylla are sold, and all the plants come from field picking and digging. Artificial breeding, population regression rejuvenation and reconstruction are effective measures for the conservation of orchid plants, but supervision outside a protection area is difficult, and the reconstructed population can still be dug, so that the purpose of conservation cannot be achieved. In order to solve the problem of serious digging of wild yucca schidigera, the artificial rapid breeding technology is utilized to mould and breed a large amount of seedlings, the domesticated yucca schidigera with high cost performance can impact wild digging, consumers can buy plants of the yucca schidigera in the market and on the network at a lower price than the wild price, the consumers can naturally lose interest in the wild yucca schidigera which is difficult to keep alive, the wild yucca schidigera with long time loses market value, and the dug people can forgo searching and digging, so that the aim of utilizing and promoting protection is really achieved. The key point of the utilization of the yunnan is the artificial rapid propagation technology, so that the detoxified seedlings of the yunnan can be produced in a large scale in a short period and put on the market, the ornamental requirement of human beings is met, the artificial propagation is realized to replace natural digging, and the endangered condition of the yunnan can be effectively alleviated.
Disclosure of Invention
The invention mainly aims to provide an artificial rapid propagation technology of a cymbidium yunnanensis, and a rapid propagation method capable of realizing large-scale production is obtained through a seed non-symbiotic germination method and a culture medium formula so as to solve market demands.
In order to achieve the above object, the present invention provides the following technical solutions:
a non-symbiotic germination culture method of a cymbidium yunnanensis seed comprises the following steps:
(1) Harvesting fruit clips: picking pods with yellowing surfaces but not splitting;
(2) Pretreatment of fruit pods: cleaning and sterilizing;
(3) Non-symbiotic germination culture: the non-symbiotic medium used was modified VW medium: 525mg/L potassium nitrate, 500mg/L ammonium sulfate, 250mg/L potassium dihydrogen phosphate, 250mg/L magnesium sulfate, 200mg/L calcium superphosphate, 7.5mg/L manganese sulfate, 28mg/L ferrous sulfate, 10.0mg/L vitamin B1, 1.0mg/L vitamin B6, 1.0mg/L nicotinic acid, 1g/L inositol, 20g/L sucrose, 7g/L agar, 37.3mg/L disodium ethylenediamine tetraacetate and 10g/L coconut powder, and the balance being water.
Further, in the step (2), the cleaning and disinfecting method specifically comprises the following steps: cleaning with sterilized water in ultra-clean environment, wiping the surface of the pod with 75% absolute ethanol, soaking with 10% sodium hypochlorite solution for sterilization for 20 min, cleaning with sterilized water, and sucking water on the surface with sterile filter paper.
Further, the non-symbiotic germination culture conditions in step (3) are: culturing at 23+ -2deg.C under illumination with illumination intensity of 800-1000Lx and illumination time of 4h/d.
The invention also provides a quick breeding method of the cymbidium yunnanense seedlings, which comprises the following steps:
s1 germination culture: obtaining germinated seeds by adopting the non-symbiotic germination culture method;
s2, subculture: the subculture medium is modified VW medium, and 0.5 mg/L6-BA, 0.2mg/LNAA and 0.5g/L activated carbon powder are added;
s3, strong seedling cultivation: the used strong seedling culture medium is prepared by adding 0.2mg/L6-BA, 0.5mg/LNAA, 0.5g/L activated carbon powder and 100g/L potato homogenate on the basis of the improved VW culture medium;
s4, transplanting and planting seedlings.
Further, the pH value of the non-symbiotic germination medium, the secondary culture medium and the strong seedling medium is 5.4, and the culture medium is cooled for standby after being autoclaved at 121 ℃ for 20 minutes.
Further, the subculture conditions in step S2 are: the culture temperature is 23+/-2 ℃, the illumination intensity is 1000-1200Lx, and the illumination time is 8h/d.
Further, the strong seedlings in the step S3 are cultivated at the temperature of 23+/-2 ℃, the illumination intensity is 1200-1600Lx, and the illumination time is 16h/d.
Further, the seedling transplanting and planting in the step S4 is carried out, the used cultivation matrix is bark, and the average grain size of the bark is 3-12mm.
Preferably, the bark has an average particle size of 3-5mm.
Further, the seedling transplanting planting conditions are as follows: the temperature is 15-25 ℃, the relative humidity is 60-80%, and the illumination intensity is 1500-2000Lx.
The invention achieves the technical effects that:
according to the technical scheme provided by the invention, under the conditions of a culture medium formula, germination and breeding method, the yucca schidigera seeds can germinate, the average germination rate is as high as 96%, the protocorms are not brown, the period from sowing to bottle discharge only needs 6-7 months, the breeding period is greatly shortened, and the non-symbiotic germination and artificial rapid breeding technology can be directly used for large-scale production of yucca schidigera seedlings.
Drawings
FIG. 1 is a schematic diagram of the pod of the Sichuan cymbidium;
FIG. 2 is a schematic diagram of the pod treatment of the Sichuan cymbidium;
FIG. 3 schematically shows germination of a seed of Gaultheria Yunnanensis (a. VW is modified VW medium, b. VW+C is modified VW medium with C);
FIG. 4 shows a schematic representation of a subculture of Yunnan Sichuan orchid;
FIG. 5 shows a schematic diagram of the cultivation of strong seedlings of Tachyrhizus Yunnanensis;
FIG. 6 is a schematic diagram of the aseptic seedling emergence planting of the Tamarindi Indicae.
Detailed Description
The present invention is further described below in conjunction with embodiments, which are merely some, but not all embodiments of the present invention. All other embodiments, which can be made by those skilled in the art without making any inventive effort, are intended to be within the scope of the present invention.
Example 1
1. Artificial pollination of herba Hedyotidis Diffusae
Selecting a Hedychium gracile plant introduced from Yunnan Xinping to be stored in a orchid greenhouse in a orchid mass resource storage center of orchid, carrying out artificial cross pollination of different plants, and recording pollination time.
2. Harvesting of the pod of the Hedychium gracile
After 206 days of artificial pollination, the pods whose surfaces have turned yellow but have not been cracked are picked as sowing material for non-symbiotic germination of the seeds of the glossostrea yunnanensis.
3. Pretreatment of yunnan cymbidium pods
Cleaning picked fruit pods, cleaning the fruit pods with a large amount of sterilized water in an ultra-clean environment, wiping the surfaces of the fruit pods with 75% absolute ethyl alcohol, soaking and sterilizing the fruit pods for 20 minutes with 10% sodium hypochlorite solution, cleaning the fruit pods with a large amount of sterilized water, and then sucking surface water with sterile filter paper for later use.
4. Culture medium formula for artificial rapid propagation of herba Hedyotidis Diffusae
The non-symbiotic germination medium formula is as follows: modified VW (Vacin & Went Orchid Medium) medium: 525mg/L potassium nitrate, 500mg/L ammonium sulfate, 250mg/L potassium dihydrogen phosphate, 250mg/L magnesium sulfate, 200mg/L calcium superphosphate, 7.5mg/L manganese sulfate, 28mg/L ferrous sulfate, 10.0mg/L vitamin B1, 1.0mg/L vitamin B6, 1.0mg/L nicotinic acid, 1g/L inositol, 20g/L sucrose, 7g/L agar, 37.3mg/L disodium ethylenediamine tetraacetate, 10g/L coconut powder, and the balance being water.
Subculture medium: 0.5 mg/L6-BA, 0.2mg/LNAA and 0.5g/L activated carbon powder were added on the basis of the modified VW (Vacin & Went Orchid Medium) medium.
Seedling strengthening culture medium: 0.2mg/L6-BA, 0.5mg/LNAA, 0.5g/L activated carbon powder and 100g/L potato homogenate were added on the basis of modified VW (Vacin & Went Orchid Medium) medium.
The pH value of the non-symbiotic germination medium, the secondary culture medium and the strong seedling medium is 5.4, and the culture medium is cooled for standby after being autoclaved at 121 ℃ for 20 minutes.
5. Non-symbiotic germination culture of yunnan cymbidium seeds (figures 1 and 2)
The pretreated pod of the Hemsleya yunnanensis is transversely cut by a sterile knife to expose the seeds, the seeds are uniformly sown on the surface of a modified VW (Vacin & Went Orchid Medium) culture medium, about 100 seeds are sown in each bottle, and 5 bottles are sown in each pod. After sowing, placing the culture flask on a culture rack at 23+/-2 ℃ for illumination culture, wherein the illumination intensity is 800-1000Lx, and the illumination time is 4h/d. And (5) observing the germination condition of the seeds every 5 days from 15 days after sowing, recording, and counting the germination time and the germination rate of the seeds.
6. Seed germination time and germination rate of the herba Hedyotidis Diffusae (figure 3)
The seed of the herba hynchi Oleracei begins to expand and break through the seed coat 20 days after sowing, the white protocorm turns green 30 days later, and the protocorm is differentiated to leave 40 days later, so that the seedling begins to be formed. The average germination rate of the yunnan cymbidium seeds is 96% by taking 30 days after sowing as the time for uniformly counting the germination rate. The improved VW (Vacin & Went Orchid Medium) culture medium is very suitable for germination of the Hedyotis yunnanensis seeds, and the mature and full seeds also provide necessary nutrition for embryo germination.
7. Effect of light culture and dark culture on seed germination of Gaultheria Yunnanensis
The seed of different genus has great difference in the demand of light in the germination process, some seeds can accelerate the germination of the seed and improve the germination rate under the induction of light, and some seeds can inhibit the germination of the seed under the light, and the seeds can only germinate under the full dark culture. The seed of the yunnan cymbidium can shorten the time required by germination under proper photocatalysis, can lead the protocorm to differentiate leaves earlier, and greatly shortens the time required by breeding. Meanwhile, the germination rate counted after 30 days of sowing in the culture under light was also higher than that in the dark culture (Table 1). In the subsequent large-scale production, the culture method adopted for the non-symbiotic germination of the Yunnan bracket seeds is light culture.
TABLE 1 Effect of light dark culture on non-symbiotic germination of Philippine Coulosa seeds
| Average germination time | Average germination rate/% | |
| Light culture | 20 | 96 |
| Dark culture | 28 | 82 |
8. Influence of the addition of activated carbon on the germination of the seed of the Philippine Coptis
In the non-symbiotic germination process of the yunnan broccoli seeds, the addition of activated carbon was found to have an effect on the time and germination rate of the seed germination, and as can be seen from table 2, the addition of activated carbon in the modified VW (Vacin & Went Orchid Medium) medium inhibited the germination of the seed, and in order to obtain more seedlings and shorten the breeding time, no activated carbon was added to the medium during the non-symbiotic germination stage of the seed.
TABLE 2 Effect of addition of activated carbon on non-symbiotic germination of Philippine Console seeds
| Average germination time | Average germination rate/% | |
| No active carbon is added | 20 | 96 |
| Adding 0.2g/L activated carbon powder | 25 | 88 |
9. Substruction of Yunnan Ganlian (fig. 4)
The original bulbs of the herba Hedyotidis Diffusae seeds begin to differentiate leaves 40 days after sowing, and the leaves are transferred to a secondary culture medium for continuous light culture, wherein the secondary culture medium is added with 0.5 mg/L6-BA, 0.2mg/L NAA and 0.5g/L activated carbon powder on the basis of a modified VW (Vacin & Went Orchid Medium) culture medium. The culture temperature is 23+/-2 ℃, the illumination intensity is 1000-1200Lx, and the illumination time is 8h/d. The protocorms/seedlings of the broccoli are rapidly differentiated and grown in a subculture medium, and are subjected to subculture once every 30 days for 3 times in total, so that the growth of the seedlings and roots can be accelerated, and the phenomenon of browning and withering does not occur.
10. Zhuanxiyan strong seedling culture (figure 5)
After the yunnan cymbidium is subjected to subculture in the subculture medium for three times, 3-4 leaves are grown, 2-3 leaves are grown at about 2cm, and at the moment, the young seedlings are transferred into the strong seedling culture medium for strong seedling culture. The culture temperature is 23+/-2 ℃, the illumination intensity is 1200-1600Lx, and the illumination time is 16h/d. The strong seedling culture medium is prepared by adding 0.2mg/L6-BA, 0.5mg/LNAA, 0.5g/L activated carbon powder and 100g/L potato homogenate based on the modified VW (Vacin & Went Orchid Medium) culture medium. The seedlings of the herba Hedyotidis Diffusae grow continuously in the strong seedling culture medium, after two months, the leaves are 5-6, the length of the leaves is 8-10cm, the length of the leaves is 3-4, and the length of the roots is 4-5cm, and the seedlings can be planted in the bottles. Under the culture medium and the culture method, the seed germination rate of the cymbidium yunnanensis is high, the time from sowing to bottle-out planting is about 6-7 months, the breeding period is greatly shortened, and the aim of rapid breeding can be achieved.
11. Seedling transplanting planting of the Yunnan Ganlian (fig. 6)
The seedling of the herba hynchi Oleracei which can be planted out of the bottle is firstly subjected to seedling hardening, the cleaning culture medium can be taken out after the seedling hardening is carried out for 10 days and the seedling is adapted to the greenhouse environment, and the seedling is planted into a culture medium which is prepared in advance after the moisture is dried. Three groups of control tests are respectively arranged for screening matrixes suitable for planting the herba Hedyotidis Diffusae, wherein the cultivation matrixes are fine bark (diameter: 3-5 mm), sphagnum moss and bark (diameter: 9-12 mm), and the survival rate of seedlings of the herba Hedyotidis Diffusae is counted after 60 days of planting. Greenhouse planting conditions: the temperature is 15-25 ℃, the relative humidity is 60-80%, the illumination intensity is 1500-2000Lx, the result is shown in Table 3, the survival rate difference is relatively large after 60 days of planting, the survival rate of the yucca schidigera planted by the bark (3-5 mm) is up to 98.33%, the survival rate in the bark (9-12 mm) is 70.67%, and the survival rate in the sphagna is 52.67% at the lowest. Thicker bark has poor water retention, plants are easy to dry and die, and frequent watering is needed. The water retention of the sphagna is too good, the water can wet for a long time to cause root rot after once watering, the thin bark (3-5 mm) is adopted for potting planting in the subsequent large-scale planting of the sphagna, and the plants grow up and are transferred into the bark of 9-12mm for planting after half a year of planting. 3-5g of slow release fertilizer is applied to each pot, and the bark is thoroughly dried and whitened before watering.
TABLE 3 influence of the cultivation substrate on the seedling survival of the Philippine Coptis (60D)
The foregoing disclosure is merely illustrative of the preferred embodiments of the present invention and is not intended to limit the scope of the claims herein, as equivalent changes may be made in the claims herein without departing from the scope of the invention.
Claims (10)
1. The non-symbiotic germination culture method for the cymbidium yunnanensis seeds is characterized by comprising the following steps of:
(1) Harvesting fruit clips: picking pods with yellowing surfaces but not splitting;
(2) Pretreatment of fruit pods: cleaning and sterilizing;
(3) Non-symbiotic germination culture: the non-symbiotic germination medium used was modified VW medium: 525mg/L potassium nitrate, 500mg/L ammonium sulfate, 250mg/L potassium dihydrogen phosphate, 250mg/L magnesium sulfate, 200mg/L calcium superphosphate, 7.5mg/L manganese sulfate, 28mg/L ferrous sulfate, 10.0mg/L vitamin B1, 1.0mg/L vitamin B6, 1.0mg/L nicotinic acid, 1g/L inositol, 20g/L sucrose, 7g/L agar, 37.3mg/L disodium ethylenediamine tetraacetate and 10g/L coconut powder, and the balance being water.
2. The method according to claim 1, wherein in the step (2), the cleaning and sterilizing method comprises the steps of cleaning the surfaces of the pods with sterilized water in an ultra-clean environment, wiping the surfaces of the pods with 75% absolute ethanol, soaking the pods in 10% sodium hypochlorite solution for 20 minutes for sterilization, cleaning the pods with sterilized water, and sucking the surface water with sterile filter paper for later use.
3. The method of claim 1, wherein the non-symbiotic germination culture conditions in step (3) are: culturing at 23+ -2deg.C under illumination with illumination intensity of 800-1000Lx and illumination time of 4h/d.
4. The quick breeding method of the cymbidium yunnanense seedlings is characterized by comprising the following steps of:
s1 germination culture: employing the non-symbiotic germination culture method of any one of claims 1-3;
s2, subculture: the subculture medium is modified VW medium, and 0.5 mg/L6-BA, 0.2mg/LNAA and 0.5g/L activated carbon powder are added;
s3, strong seedling cultivation: the used strong seedling culture medium is prepared by adding 0.2mg/L6-BA, 0.5mg/LNAA, 0.5g/L activated carbon powder and 100g/L potato homogenate on the basis of the improved VW culture medium;
s4, transplanting and planting seedlings.
5. The method of claim 1 or 4, wherein the non-symbiotic germination medium, the secondary medium and the strong medium have a pH of 5.4 and are autoclaved at 121 ℃ for 20 minutes and cooled for later use.
6. The method according to claim 4, wherein the subculture conditions in step S2 are: the culture temperature is 23+/-2 ℃, the illumination intensity is 1000-1200Lx, and the illumination time is 8h/d.
7. The method according to claim 4, wherein the strong seedlings are cultivated in the step S3 at a cultivation temperature of 23+/-2 ℃ and an illumination intensity of 1200-1600Lx for 16h/d.
8. The method according to claim 4, wherein the seedling transplanting in step S4 is performed by using bark as a cultivation substrate, and the average grain size of the bark is 3-12mm.
9. The method according to claim 8, wherein the bark has an average particle size of 3-5mm.
10. The method according to claim 8 or 9, wherein the seedling transplanting planting conditions are: the temperature is 15-25 ℃, the relative humidity is 60-80%, and the illumination intensity is 1500-2000Lx.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311454865.0A CN117256476A (en) | 2023-11-03 | 2023-11-03 | Non-symbiotic germination and rapid breeding method for cymbidium yunnanensis seeds |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311454865.0A CN117256476A (en) | 2023-11-03 | 2023-11-03 | Non-symbiotic germination and rapid breeding method for cymbidium yunnanensis seeds |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117256476A true CN117256476A (en) | 2023-12-22 |
Family
ID=89216164
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311454865.0A Pending CN117256476A (en) | 2023-11-03 | 2023-11-03 | Non-symbiotic germination and rapid breeding method for cymbidium yunnanensis seeds |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117256476A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN118104567A (en) * | 2024-04-29 | 2024-05-31 | 云南省林业和草原科学院 | Long-term-preservation short-distance living body regeneration and efficient propagation method for glossostrea planata |
-
2023
- 2023-11-03 CN CN202311454865.0A patent/CN117256476A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN118104567A (en) * | 2024-04-29 | 2024-05-31 | 云南省林业和草原科学院 | Long-term-preservation short-distance living body regeneration and efficient propagation method for glossostrea planata |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105191727A (en) | Dragon fruit planting method | |
| CN115104536B (en) | Nuphar apapratus seed germination, breeding and cultivation method | |
| CN107926715A (en) | A kind of eggplant or/and the engrafting and cultivating method of capsicum or/and tomato | |
| CN105103714A (en) | Manual germination accelerating method and seedling raising method of Bletilla sfriata (Thunb.)Reiehb.f. | |
| Abbaszadeh et al. | An effective nutrient media for asymbiotic seed germination and in vitro seedling development of phalaenopsis ‘Bahia Blanca’ | |
| CN109287487B (en) | Seed germination rate improving method and cultivation method for paphiopedilum makino | |
| CN112154916B (en) | Culture medium for culture and breeding method of lilac daphne flower bud explants and culture and breeding method of lilac daphne flower bud explants | |
| CN111134019B (en) | Method for directly generating somatic embryos and regenerating plants of ilex denticulata | |
| CN101564010B (en) | Method for rapidly propagating tupelos | |
| CN115589948B (en) | Rhizoma bletillae non-symbiotic germination medium and propagation method | |
| CN109247235B (en) | Rapid breeding and seedling raising method for cymbidium faberi Rolfe | |
| CN114532225B (en) | Tissue culture rapid propagation and cultivation method for paphiopedilum delbrueckii | |
| CN106922397B (en) | Method for cultivating easily-branched poinsettia | |
| CN110999914B (en) | Chinese medicinal material seed pretreatment agent and application thereof | |
| CN111202002B (en) | Tissue culture and rapid propagation method of clerodendrum japonicum | |
| CN117256476A (en) | Non-symbiotic germination and rapid breeding method for cymbidium yunnanensis seeds | |
| CN101720671B (en) | Method for tissue culture of witch hazel | |
| CN111727802A (en) | Novel cultivation method of Chinese toon sprouts | |
| CN112088776A (en) | Tissue culture rapid propagation method for high-value tree species albizia julibrissin | |
| Della Rahayu et al. | Asymbiotic seed germination and plantlet development of Dendrobium spectabile (Blume) Miq. | |
| KR102597761B1 (en) | Method of mass propagation of Maesa japonica | |
| CN113796315B (en) | Quick propagation method of endangered wild plant cymbidium floribundum | |
| NL2029503B1 (en) | Hybrid seed production method based on subgynoecious cucumber as female parent | |
| CN112913692B (en) | Method for efficient breeding by taking senna chinensis seeds as explants | |
| KR100505930B1 (en) | In vitro masspropagation medium of Sarracenia species and method for mass production thereof using the same |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |