CN1258970C - Method for tissue culture quick breeding of rattlesnake plantain - Google Patents
Method for tissue culture quick breeding of rattlesnake plantain Download PDFInfo
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- CN1258970C CN1258970C CN 200410061424 CN200410061424A CN1258970C CN 1258970 C CN1258970 C CN 1258970C CN 200410061424 CN200410061424 CN 200410061424 CN 200410061424 A CN200410061424 A CN 200410061424A CN 1258970 C CN1258970 C CN 1258970C
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Abstract
The present invention discloses a method of tissue culture and rapid propagation of rattlesnake plantains. Plant stem segments are used as explants, budlets of rattlesnake plantains are inducted by using different culture medium prescriptions, the budlets are propagated, strengthened and rooted in a caespitose bud mode, and the purpose of rapid propagation is achieved. The method of tissue culture and rapid propagation of rattlesnake plantains has the advantages of simplicity, feasibility, convenient operation and rapid propagation speed.
Description
Technical field
The invention belongs to field of plant tissue culture technique, more specifically relate to a kind of method of the variegated leaf orchid being carried out tissue-culturing rapid propagation by minimal medium and additional different components.
Background technology
Variegated leaf orchid (Goodyera schlechtendaliana) has another name called leaflet green grass or young crops, Herba goodyearae repentis, belongs to the orchid family variegated leaf Cymbidium herbaceos perennial, and is high about 20 centimetres, the hillside or the sylvan life of cheuch evergreen broad-leaved of being born in height above sea level 500-2800 rice, and happiness covers moist environment.The variegated leaf orchid has higher medical value, and is cold in nature with all herbal medicine, lightly seasoned; Removing heat from the lung to relieve cough, removing toxicity for detumescence, promoting blood circulation and stopping pain, softening and resolving hard mass; In proved recipes such as the cough of clinical treatment pulmonary tuberculosis, bronchitis, venomous snake bite, carbuncle furuncle sore, nose furuncle, all be used.In addition, the blue blade of variegated leaf is very beautiful, very exquisite, and the title of Saphire Brown is arranged, and can little potted plant planting be put on the case, or does interspersing of artificial hillock, gardens and serve as a contrast or foil plant.Because this plant species is of many uses, drug effect is remarkable, the medicinal herb grower does not add the protection digging without restraint, makes it to be in condition in imminent danger.Offshoot is generally adopted in the breeding of variegated leaf orchid, because under field conditions (factors) reproduction coefficient is extremely low, general year reproduction coefficient is about 2, and it is required to satisfy market far away.
Summary of the invention
The object of the present invention is to provide the method for the blue tissue-culturing rapid propagation of a kind of variegated leaf, method is simple, easy to operate; adopt this kind method can breed the blue seedling of a large amount of variegated leafs fast; with effective protection wild resource, solve the deficient problem in traditional Chinese medicine source, satisfy people's medication and view and admire demand.
In order to achieve the above object, the present invention adopts following technical measures: with the blue plants stems section of variegated leaf as explant, at first plant was washed 1 hour with flowing water, clean, be cut into the long stem section of 1～2cm, with 75% alcohol-pickled 10～30 seconds, aseptic water washing 3 times, again with 0.1% mercuric chloride sterilization 10～15 minutes, with aseptic water washing 4～5 times, the stem section that disinfects is inserted in the MS medium for preparing, this medium comprises the material of MS minimal medium and additional different components, additional different material is: 6-benzyl aminopurine (6-Benzyl aminopurine is called for short BA) 1.0～4.0mg/L, methyl (I-NaphthyIacetic acid is called for short NAA) 0.1～0.5mg/L, agar 5000～6000mg/L, sugar 20000～30000mg/L, coconut milk 5%～10%; Illumination cultivation (indoor), illumination every day 12～14 hours, light intensity 2000～3000Lx, temperature is 20～25 ℃, cultivates 30～60 days, eustipes part begins to sprout young shoot; Then the young shoot that grows is changed in the proliferated culture medium, this medium comprises the material of MS minimal medium and additional different components, additional different material is: 6-benzyl aminopurine (6-Benzylaminopurine is called for short BA) 2.0～6.0mg/L, methyl (I-NaphthyIacetic acid is called for short NAA) 0.5～1.0mg/L, agar 5000～6000mg/L, sugar 20000～30000mg/L, coconut milk 5%～10%; Illumination cultivation, light intensity 2000～3000Lx, illumination every day 12～14 hours, temperature is 20～25 ℃, cultivates 30～60 days, young shoot can grow the bud of growing thickly, and expands once numerous in later per 30 days; The 3rd is strong seedling culture, when breeding to requirement, thereby making bud grow up, the strong seedling culture base that utilization prepares reaches the purpose in strong sprout, this medium comprises the material of 1/2MS minimal medium and additional different components, additional different material is: 6-benzyl aminopurine (6-Benzyl aminopurine is called for short BA) 0.5～2.0mg/L, methyl (I-NaphthyIacetic acid is called for short NAA) 0.1～0.5mg/L, agar 5000～6000mg/L, sugar 20000～30000mg/L, bananas juice 5%～10%; Illumination cultivation, light intensity 2000～3000Lx, illumination every day 12～14 hours, temperature is 20～25 ℃, cultivates 30～60 days, treats that seedling grows to 3～4cm and can carry out culture of rootage when high; The 4th is culture of rootage, the seedling that 3～4cm is high inserts in the root media for preparing, this root media comprises the material of 1/2MS minimal medium and additional different components, additional different material is: methyl (I-NaphthyIacetic acid is called for short NAA) 0.2～0.8mg/L, agar 5000～6000mg/L, sugar 20000～30000mg/L, active carbon 0.3～0.5%; Illumination cultivation, light intensity 2000～3000Lx, illumination every day 12～14 hours, temperature is 20～25 ℃, cultivates 20～30 days, can grow 2～3 roots, and root is sturdy, and about 2cm is long.
The present invention compared with prior art, have the following advantages and effect: method is simple, and is easy to operate, reproduction speed is fast, 30 days propagation multiple reaches 3～4 times, bud can be bred 1,000,000 strain seedlings in 1 year.
Embodiment
The concrete steps of the blue breeding of variegated leaf are as follows:
(1) chooses the blue plant of the variegated leaf that newly grows as explant,, clean with flowing water flushing 1 hour, be cut into the long stem section of 1～2cm, in 75% alcohol, soak 10 or 20 or 30 seconds, aseptic water washing 3 times, with 0.1% mercuric chloride sterilization 10 or 12 or 15 minutes, use aseptic water washing 4～5 times again; Next is that the stem section that will disinfect inserts in the prepared culture medium, this medium comprises the material of MS minimal medium and additional different components, wherein additional composition is 6-benzyl aminopurine 2.0mg/L, methyl 0.2mg/L, agar powder 5500mg/L, sugared 30000mg/L, and coconut milk 10% is mixed; The 3rd is illumination cultivation, light intensity 2000Lx, and illumination every day 12 hours, temperature is 25 ℃, cultivates 40 days, eustipes part begins to sprout young shoot.
The composition of MS minimal medium (unit: mg/L):
Nitric acid ammonia (NH
4NO
3) 1650
Potassium nitrate (KNO
3) 1900
Calcium chloride dihydrate (CaCl
22H
2O) 440
Epsom salt (MgSO
47H
2O) 370
Potassium dihydrogen phosphate (KH
2PO
4) 170
Potassium iodide (KI) 0.83
Boric acid (H
3BO
3) 6.2
Four water manganese sulphate (MnSO
44H
2O) 22.3
White vitriol (ZnSO
47H
2O) 8.6
Sodium Molybdate Dihydrate (Na
2MoO
42H
2O) 0.25
Cupric sulfate pentahydrate (CuSO
45H
2O) 0.025
CoCL2 (CoCl
26H
2O) 0.025
Ferrous sulfate heptahydrate (FeSO
47H
2O) 27.8
Disodium ethylene diamine tetraacetate (Na
2EDTA2H
2O) 37.3
Inositol 100
Nicotinic acid 0.5
Puridoxine hydrochloride 0.5
Thiamine hydrochloride 0.1
Glycine 2
(2) enrichment culture: the young shoot that grows is changed in the proliferated culture medium, this medium comprises the material of MS minimal medium and additional different components, MS minimal medium composition is the same, additional composition is BA5.0mg/L, NAA1.0mg/L, agar 5500mg/L, sugared 30000mg/L, coconut milk 10%; Illumination cultivation (indoor), light intensity 2000Lx, illumination every day 12 hours, temperature is 25 ℃, cultivates 30 days, young shoot may have grown into the bud of growing thickly, and expands once numerous in later per 30 days;
(3) strong seedling culture: when breeding to requirement, thereby making bud grow up, the strong seedling culture base that utilization prepares reaches the purpose in strong sprout, this medium comprises the material of 1/2MS minimal medium and additional different components, the 1/2MS minimal medium is that MS minimal medium composition consumption is reduced by half, MS minimal medium composition is the same, added substance is: BA 1.0mg/L, NAA0.2mg/L, agar 5500mg/L, sugar 30000mg/L, bananas juice 10% illumination cultivation, light intensity 2000Lx, illumination every day 12 hours, temperature is 25 ℃, cultivates 30 days, treats that seedling grows to 3-4cm and can carry out culture of rootage when high;
(4) culture of rootage: the seedling that 3～4cm is high inserts in the root media for preparing, this root media comprises the material of 1/2MS minimal medium and additional different components, the 1/2MS minimal medium is that MS minimal medium composition consumption is reduced by half, MS minimal medium composition is the same, added substance is: NAA 0.6mg/L, agar 5500mg/L, sugared 20000mg/L, active carbon 0.5%; Illumination cultivation, light intensity 2000Lx, illumination every day 12 hours, temperature is 25 ℃, cultivates 25 days, can grow 2～3 roots, and root is sturdy, and about 2cm is long.
Claims (1)
1, the method for the blue tissue-culturing rapid propagation of a kind of variegated leaf comprises the following steps:
A, with the blue plants stems section of variegated leaf as explant, plant is cleaned, be cut into the long stem section of 1-2cm, with 75% alcohol-pickled 10～30 seconds, with 0.1% mercuric chloride sterilization 10～15 minutes, with aseptic water washing 4～5 times, the stem section that disinfects is inserted in the MS medium for preparing, medium is the material of MS minimal medium and additional different components, additional different material is: 6-benzyl aminopurine 1.0～4.0mg/L, methyl 0.1～0.5mg/L, agar 5000～6000mg/L, sugar 20000～30000mg/L, coconut milk 5%～10%; Illumination cultivation, illumination every day 12～14 hours, light intensity 2000～3000Lx, temperature is 20～25 ℃;
B, enrichment culture, the young shoot that grows is changed in the proliferated culture medium, this medium is the material of MS minimal medium and additional different components, and additional different material is: 6-benzyl aminopurine 2.0～6.0mg/L, methyl 0.5～1.0mg/L, agar, sugar, Coconut Juice content and illumination condition are with step A;
C, strong seedling culture, the strong seedling culture base is the material of 1/2MS minimal medium and additional different components, additional different material is: 6-benzyl aminopurine 0.5～2.0mg/L, methyl 0.1～0.5mg/L, bananas juice 5%～10%, and agar, sugared content and illumination condition are with step A;
D, culture of rootage, the seedling that 3～4cm is high inserts in the root media for preparing, medium is the material of 1/2MS minimal medium and additional different components, additional different material is: methyl 0.2～0.8mg/L, active carbon 0.3～0.5%, agar, sugared content and illumination condition are with step A.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410061424 CN1258970C (en) | 2004-12-24 | 2004-12-24 | Method for tissue culture quick breeding of rattlesnake plantain |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410061424 CN1258970C (en) | 2004-12-24 | 2004-12-24 | Method for tissue culture quick breeding of rattlesnake plantain |
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|---|---|
| CN1647616A CN1647616A (en) | 2005-08-03 |
| CN1258970C true CN1258970C (en) | 2006-06-14 |
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| CN 200410061424 Expired - Fee Related CN1258970C (en) | 2004-12-24 | 2004-12-24 | Method for tissue culture quick breeding of rattlesnake plantain |
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Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1319437C (en) * | 2004-12-31 | 2007-06-06 | 浙江省农业科学院 | Group culture medium of Spotleaf-orchis and rapid breeding method thereof |
| CN102524062B (en) * | 2011-12-05 | 2013-06-05 | 天津市华泰兰园种养殖专业合作社 | Culture medium for tissue culture of orchid |
| CN102577973B (en) * | 2012-03-12 | 2013-07-31 | 云南山里红生物科技有限公司 | Tissue culture method for Cymbidium floribundum |
| CN103755461B (en) * | 2013-12-23 | 2015-09-16 | 佛山市顺德区今日景艺生物科技有限公司 | The medium that Orchid Tissue is cultivated and cultural method thereof |
| CN104770303B (en) * | 2015-04-23 | 2016-11-30 | 福建农林大学 | A kind of organic additive induces leafy Herba goodyearae repentis plant regeneration and efficient propagation method |
| CN105165612B (en) * | 2015-09-25 | 2017-08-25 | 临沂大学 | A kind of scolopendrifolious cleisostoma herb tissue culture and rapid propagation method |
| CN106376460B (en) * | 2016-08-25 | 2018-06-15 | 中国科学院昆明植物研究所 | The in-vitro conservation method of stigma aspidistra |
| CN106386513A (en) * | 2016-12-23 | 2017-02-15 | 信阳师范学院 | Tissue-culture and rapid-propagation method for Goodyera biflora |
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