CN101926259B - Orchid germchit propagating method - Google Patents
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- 241000233855 Orchidaceae Species 0.000 title claims abstract description 41
- 230000001902 propagating Effects 0.000 title claims abstract description 12
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- 230000007226 seed germination Effects 0.000 claims abstract description 13
- 239000002609 media Substances 0.000 claims description 25
- OKTJSMMVPCPJKN-UHFFFAOYSA-N carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 18
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 229920001817 Agar Polymers 0.000 claims description 9
- CZMRCDWAGMRECN-UGDNZRGBSA-N D-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 9
- CZMRCDWAGMRECN-GDQSFJPYSA-N Sucrose Natural products O([C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O1)[C@@]1(CO)[C@H](O)[C@@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-GDQSFJPYSA-N 0.000 claims description 9
- 239000008272 agar Substances 0.000 claims description 9
- 239000002775 capsule Substances 0.000 claims description 9
- 239000005720 sucrose Substances 0.000 claims description 9
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- 239000001963 growth media Substances 0.000 claims description 5
- 241000894006 Bacteria Species 0.000 claims description 4
- 238000004140 cleaning Methods 0.000 claims description 4
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- FBPFZTCFMRRESA-KAZBKCHUSA-N D-Mannitol Natural products OC[C@@H](O)[C@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KAZBKCHUSA-N 0.000 claims description 3
- FBPFZTCFMRRESA-KVTDHHQDSA-N Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims description 3
- 241001090911 Rhizoctonia sp. CLB111 Species 0.000 claims description 3
- 238000004500 asepsis Methods 0.000 claims description 3
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- 102100010936 AIF1L Human genes 0.000 claims description 2
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- 230000003203 everyday Effects 0.000 description 1
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- 239000005556 hormone Substances 0.000 description 1
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- 229960002523 mercuric chloride Drugs 0.000 description 1
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- 230000002062 proliferating Effects 0.000 description 1
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Abstract
The invention relates to an orchid germchit propagating method, belonging to an orchid seedling propagating technology, in particular to a propagating technology for seedlings cultured based on tissue culture and mycorrhiza fungi symbiosis. In the orchid germchit propagating method, the germchit can be obtained in the steps of seed germination, successive multiplication of protocorm and rooting culture. The method is characterized in that the rooted seedling can be further acclimatized and the mycorrhiza fungi are mycorrhized after the germchit is obtained by using the propagating method. By using the mycorrhized culture technology based on sterile germination and mycorrhiza fungi symbiosis of the orchid seed, fine varieties can be propagated, fine orchid plants can be selected from the seedlings of seeds, and the propagating speed of the orchid can be quickened, thereby overcoming the defects of slow growth, long cycle and even nonflowering of the pure tissue culture seedling, and having important utility values in the large-area industrialized propagation of the orchid.
Description
Technical field
The invention belongs to a kind of orchid seedling propagating technology, the propagation technique that especially a kind of tissue culture and mycorrhizal fungi symbiosis culture are grown seedlings.
Background technology
Orchid is meant the general name of whole orchid (Orchidaceae).In China, traditional " orchid " notion only refers to originate in the orchid family, Cymbidium (Cymbidium) plant of China, claims the state orchid, and give birth on ground, so it is blue to claim ground to give birth to again.Orchid is a class ornamental value and the high rare species of economic worth.Wherein, the Yunnan orchid just has 100 to belong to, and kind more than 530 is the hat in the whole nation.
But up to the present, China orchid is develop and useedd and still is in the junior stage, and fails to solve prolific problem, so imbalance between supply and demand is outstanding.Be because the driving of economic interests on the one hand, the peasant goes up a hill and excavates wild orchid in a large number, the wild orchid resource is subjected to heavy damage, and especially famousr and precious Chinese cymbidium (Cymbidium sinense), Chunlan (Cymbidiumgoeringii), cold blue (Cymbidium kanran), spring sword (Cymbidium longibracteatun) and company lobe orchid (Cymbidiumlianpan) are destroyed the most serious.Next is that area of woods reduces significantly, and the change of orchid living environment not only forms threat to the continuity of wild orchid population, also makes the numerous species orchid disappear in some areas.In recent years; China has strengthened the protection work of wild plant resource; the administrative regulation " People's Republic of China's conservation of wild plant regulations " of first special protection wild plant of China was implemented from January 1st, 1997; regulation according to these regulations; draft and through having approved " national key protected wild plants register ", all kinds of the orchid family all are listed in the register.But it still is far from being enough only adopting legal form protection orchid resource, when strengthening management, also should pay attention to developing the propagation technique of orchid, could solve the purpose of protection orchid species at all.
The propagation technique of orchid mainly contains two kinds at present, plant division and tissue culture.Traditional division propagation speed is slow, and reproduction rate is low, and the cycle is long, and easily hinders maternal plant, and seed germination rate under field conditions (factors) is extremely low, is difficult to satisfy needs of scale production.And tissue culture, transplanting aseptic seedling survival rate is low, grows slowly, blooms late, does not even bloom, and reason mainly is the mycorrhizal fungi that lacks symbiosis with it.Fungi provides growth and breeding necessary nutriment for orchid.It is natural a kind of universal phenomenon that orchid and mycorrhizal fungi form mycorhiza; so mycorrhizal fungi is the key that solves the artificial cultivation orchid for orchid provides nutrition to grow, also be China's orchid resource and the subject matter of utilizing protection to be faced.
Summary of the invention
To be solved by this invention is exactly to only depend on plant division and tissue culture propagating Chunlan can not satisfy the problem of China's orchid plant protection of resources.The present invention separates endogenetic fungus from wild Chunlan root, make itself and Chunlan tissue cultivating seedling symbiosis culture, filter out and with the effective symbiosis of Chunlan tissue cultivating seedling and can obviously promote effective mycorrhizal fungi that it grows, the Chunlan tissue cultivating seedling is carried out Mycorrhizal cultivate, fundamentally overcome the test-tube plantlet breeding, lack mycorrhizal fungi, it is low to transplant the aseptic seedling survival rate, poor growth, it is slow to bloom, and spends little shortcoming of even not blooming.
A kind of orchid germchit propagating method of the present invention, its seedling obtains mainly to comprise seed germination, protocorm shoot proliferation and process of rooting culture, it is characterized in that this propagation method after seedling obtains, also the take root hardening and the nursery stock Mycorrhizal process of seedling, be specially:
Hardening: the seedling of will taking root takes out according to uncork lid transition 1～1.5 week back under the condition at indoor natural light, clean attach at the medium of seedling root standby;
Nursery stock Mycorrhizal: the seedling that takes out is planted in the dish of cave with the matrix parcel that contains mycorrhizal fungi, under mild temperature, place the cultivation of shady and cool ventilation place, during this time to leaf portion spray water, promote new root growth, 3 week backs move into carries out normal water, fertilizer, pencil reason in the greenhouses, get final product after 9.5 months the Mycorrhizal seedling; Described matrix be by weight respectively with add in 1/4 part of peat, 1/4 part of perlite, 1/4 part of dried detritus soil and the 1/4 part of laterite be mixed with bacterium liquid (Rhizoctonia sp.CLB111) robur leaf three-class strain a little and get.
Above-mentioned seed germination process mainly may further comprise the steps:
A) selection of explant: the capsule of selecting Chunlan not split is an explant;
B) seed treatment: handle with taking out powder shape seed asepsis after the Chunlan capsule aseptic process;
C) seed germination: with the even sowing of treated Chunlan seed incubation growth on the seed germination medium is protocorm; Employed culture medium prescription is: 1/2MS+NAA0.5～1.0mg/L+IBA2～3mg/L+100ml/L coconut palm breast+mannitol 1g/L+ activated carbon 0.5g/L+ sucrose 25g/L+ agar 3g/L, the medium pH value is 5.0～6.0.
Above-mentioned protocorm shoot proliferation process be with the protocorm of initial culture and bud respectively on medium shoot proliferation cultivated 50～60 days, grow up to sprouting or protocorm, continuous cutting protocorm subculture gets final product geometric growth later on; Employed culture medium prescription is: 1/2MS+NAA0.1～0.5mg/L+6-BA0.5～0.8mg/L+ activated carbon 0.5g/L+ sucrose 25g/L+ agar 3g/L, the medium pH value is 5.0～6.0.
Above-mentioned process of rooting culture is the underdeveloped seedling of bigger root system to be changed on the root media cultivate, and rooting rate is reached more than 95%, and plant strain growth is vigorous, the seedling of 7 weeks back formation, 5～7cm; Employed culture medium prescription is: 1/2MS+NAA0.5mg/L+ activated carbon 0.5g/L+ sucrose 25g/L+ agar 3g/L, the medium pH value is 5.0～6.0.
The present invention can effectively utilize the symbiosis of orchid mycorhiza to promote the characteristics of growth by adjusting the symbiosis culture of hormone concentration, proportioning and root fungus fungi, and the breeding cycle of shortening orchid tissue cultivating seedling, survival rate has reached more than 95%.The present invention adopts the Mycorrhizal cultivation technology of Chunlan seed asepsis sprouting and mycorrhizal fungi symbiosis, both can breed improved seeds, from seedling from seed, select good blue strain, can improve Chunlan fast again and breed speed, overcome pure tissue cultivating seedling poor growth, cycle is long, and the shortcoming of even not blooming expands numerous important value that has to the large tracts of land industrialization of Chunlan.
Embodiment
Embodiment 1: select for use on the fine individual plant in the Baoshan, Yunnan Province, adopted in 2003 the Chunlan seed, carry out hardening in June, 2004 in the green house at Xi'nan College of Forestry, this propagation method orchid germchit survival rate has reached more than 95%, and concrete sapling multiplication process is as follows:
(1) seed germination: open-air ripe 180 days capsule is adopted back, refrigerated for 2 weeks, wash with flowing water that to get the capsule that Chunlan do not split in 20 minutes be explant in 4 degree refrigerator kinds.Surface sterilization is 30～50 seconds in 70～75% alcohol, and rinsed with sterile water was once sterilized 20 minutes with 2% clorox again, used aseptic water washing 4～5 times, sterilized 10～15 minutes with 0.1% mercuric chloride solution again, used aseptic water washing at last 5 times.The ripe capsule of cleaning is placed suck dry moisture on the sterilization filter paper, the capsule of sterilization is placed sterilized culture dish, vertically cut the Chunlan capsule with the aseptic operation cutter; Take out Chunlan powder shape seed with tweezers again and put into preprepared aseptic little filter paper bag, handle taking-up in 8～10 minutes in the NaOH solution of immersion 0.15～0.2mol/L, in sterile water, wash 4～5 times, change the preceding dried filter paper extruding filter paper bag suck dry moisture of water at every turn with the bacterium of going out; Then treated Chunlan seed is sowed equably on the seed germination medium, changed over to illumination every day 12～14 hours, intensity of illumination 1000～1500lux, 25 ℃ of temperature after secretly cultivating for 2 weeks.Cultivate and to be grown to protocorm in 160～180 days.The prescription of its medium is 1/2MS+NAA0.5～1.0mg/L+IBA2-3mg/L+100ml/L coconut palm breast+mannitol 1g/L+ activated carbon 0.5g/L+ sucrose 25g/L+ agar 3g/L, and the medium pH value is 5.0～6.0.
(2) protocorm shoot proliferation: with the protocorm and the bud shoot proliferation cultivation on medium respectively of initial culture, the protocorm growth rate is fast, and cultivation effect is good, grows up to sprouting or protocorm in 50～60 days, constantly cut the protocorm subculture later on, get final product geometric growth.The prescription of its medium is 1/2MS+NAA0.1～0.5mg/L+6-BA0.5～0.8mg/L+ activated carbon 0.5g/L+ sucrose 25g/L+ agar 3g/L, and the medium pH value is 5.0～6.0.
(3) culture of rootage: the underdeveloped seedling of root system that will be bigger changes on the root media to be cultivated, and plant strain growth is vigorous, the seedling of 7 weeks back formation, 5～7cm, well developed root system.The prescription of its medium is 1/2MS+NAA0.5mg/L+ activated carbon 0.5g/L+ sucrose 25g/L+ agar 3g/L, and the medium pH value is 5.0～6.0.
(4) hardening: with 8～10 centimetres high take root seedling after indoor natural light is according to uncork lid 1～1.5 week of transition under the condition, take out clean attach at the medium of seedling root standby.
(5) nursery stock Mycorrhizal: the seedling that takes out is planted in the dish of cave with the matrix parcel that contains mycorrhizal fungi, keep mild temperature in managing and protecting, place the cultivation of shady and cool ventilation place, only need leaf portion spray water during this time, help new root growth, move in the greenhouse after 3 weeks, carry out normal water, fertile management, survival rate can reach more than 95%, get final product after 9.5 months the Mycorrhizal seedling.Its matrix be by weight respectively with add in 1/4 part of peat, 1/4 part of perlite, 1/4 part of dried detritus soil and the 1/4 part of laterite be mixed with bacterium liquid (Rhizoctonia sp.CLB111) robur leaf three-class strain a little and get.
Claims (1)
1. orchid germchit propagating method, its seedling obtains mainly to comprise seed germination, protocorm shoot proliferation and process of rooting culture, it is characterized in that this propagation method after seedling obtains, also take root seedling hardening and with the Mycorrhizal process of mycorrhizal fungi, be specially:
(1) the seed germination process mainly may further comprise the steps:
A) selection of explant: the capsule of selecting Chunlan not split is an explant;
B) seed treatment: handle with taking out Powdered seed asepsis after the Chunlan capsule aseptic process;
C) seed germination: with the even sowing of treated Chunlan seed incubation growth on the seed germination medium is protocorm;
The employed culture medium prescription of seed germination is: 1/2MS+NAA0.5～1.0mg/L+IBA2～3mg/L+100ml/L coconut palm breast+mannitol 1g/L+ activated carbon 0.5g/L+ sucrose 25g/L+ agar 3g/L, and the medium pH value is 5.0～6.0;
(2) protocorm shoot proliferation process be with the protocorm of initial culture and bud respectively on medium shoot proliferation cultivated 50～60 days, grow up to sprouting or protocorm, constantly cut the protocorm subculture later on, get final product geometric growth, employed culture medium prescription is: 1/2MS+NAA0.1～0.5mg/L+6-BA0.5～0.8mg/L+ activated carbon 0.5g/L+ sucrose 25g/L+ agar 3g/L, and the medium pH value is 5.0～6.0;
(3) process of rooting culture is the underdeveloped seedling of bigger root system to be changed on the root media cultivate, rooting rate is reached more than 95%, plant strain growth is vigorous, the seedling of 7 weeks back formation, 5～7cm, the prescription of employed medium is 1/2MS+NAA0.5mg/L+ activated carbon 0.5g/L+ sucrose 25g/L+ agar 3g/L, and the medium pH value is 5.0～6.0;
(4) the hardening process is that the seedling of taking root is taken out according to uncork lid transition 1～1.5 week back under the condition at indoor natural light, clean attach at the medium of seedling root standby;
(5) nursery stock Mycorrhizal process is that the seedling that will take out is planted in the dish of cave with the matrix parcel that contains mycorrhizal fungi, under mild temperature, place the cultivation of shady and cool ventilation place, during this time to leaf portion spray water, promote new root growth, 3 week backs move into carries out normal water, fertile management in the greenhouse, get final product after 9.5 months the Mycorrhizal seedling; Described matrix be by weight respectively with add in 1/4 part of peat, 1/4 part of perlite, 1/4 part of dried detritus soil and the 1/4 part of laterite be mixed with Rhizoctonia sp.CLB111 bacterium liquid robur leaf three-class strain a little and get.
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| CN2010101277884A CN101926259B (en) | 2010-03-19 | 2010-03-19 | Orchid germchit propagating method |
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| CN2010101277884A CN101926259B (en) | 2010-03-19 | 2010-03-19 | Orchid germchit propagating method |
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| CN101926259A CN101926259A (en) | 2010-12-29 |
| CN101926259B true CN101926259B (en) | 2011-12-14 |
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Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102726288A (en) * | 2012-07-22 | 2012-10-17 | 云南农业大学 | Method for culturing and rapidly propagating large-flowered cymbidiums |
| CN103314862B (en) * | 2013-07-16 | 2015-12-02 | 深圳市夺天工园林建设有限公司 | A kind of method of efficient acquisition Chunlan detoxification seedling |
| ES2551658B2 (en) | 2015-10-22 | 2016-03-30 | Agrotecnologias Naturales, S.L. | Adhesive composition for stably bonding fungal spores to the surface of vegetable seeds and method for quantifying adhesion capacity. |
| CN106069778A (en) * | 2016-07-31 | 2016-11-09 | 张玉薇 | The method for tissue culture of Herba Cymbidii Goeringii |
| CN106234218A (en) * | 2016-07-31 | 2016-12-21 | 张玉薇 | Herba Cymbidii Goeringii tissue culture culture medium |
| CN107006375A (en) * | 2017-06-02 | 2017-08-04 | 合肥华盖生物科技有限公司 | A kind of orchid fast propagating culture medium |
| CN107439259A (en) * | 2017-09-26 | 2017-12-08 | 上海应用技术大学 | A kind of preparation method of potted orchids special active cultivation matrix |
| CN108668893B (en) * | 2018-04-23 | 2021-08-20 | 福建省农业科学院农业生物资源研究所 | Rapid seedling raising method for spiranthes sinensis seeds |
| CN109845609B (en) * | 2019-04-12 | 2021-06-18 | 江苏里下河地区农业科学研究所 | Household balcony cymbidium culture method |
| CN113079999B (en) * | 2021-03-16 | 2022-03-15 | 南京鑫宇农业发展有限公司 | Cultivation method of multi-arrow butterfly orchid |
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| CN1169426C (en) * | 2002-11-07 | 2004-10-06 | 中国科学院武汉植物研究所 | Cymbidium quick-acting breeding method |
| CN100342778C (en) * | 2005-12-16 | 2007-10-17 | 江苏阳光生态农林开发股份有限公司 | Method for cultivating cymbidium goeringii seed by tissue culture |
| CN101622922B (en) * | 2009-08-07 | 2011-08-17 | 昆明市林业科学研究所 | Symbiotic germination method of Chinese cymbidium hybrid seeds and bacteria |
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