CN106613940B - A kind of method for in-vitro rapid propagation of Billbergiapyramidalis - Google Patents
A kind of method for in-vitro rapid propagation of Billbergiapyramidalis Download PDFInfo
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- CN106613940B CN106613940B CN201610872185.4A CN201610872185A CN106613940B CN 106613940 B CN106613940 B CN 106613940B CN 201610872185 A CN201610872185 A CN 201610872185A CN 106613940 B CN106613940 B CN 106613940B
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- 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
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Abstract
A kind of method for in-vitro rapid propagation of Billbergiapyramidalis, include the following steps: to obtain explant and sterilize, acquisition adventitious bud will be cultivated in disinfection explant access initial culture base, initial culture base are as follows: on improvement White medium base, the concentration of ferric sulfate and sodium sulphate is first adjusted, then adds aspartic acid and heteroauxin；Adventitious bud is transferred to culture in subculture medium and obtains squamous subculture seedling, subculture medium are as follows: on improvement White medium base, adjusts the concentration of molybdenum oxide and boric acid, in addition adds vitamin K1, heteroauxin and methyl α-naphthyl acetate again；Squamous subculture seedling is transferred to culture in root media and obtains Billbergiapyramidalis seedling, root media are as follows: in B5On medium base, ammonium molybdate and gibberellin are in addition added again；Last hardening and transplanting.The present invention effectively improves the breeding coefficient of Billbergiapyramidalis, commercially produces for Billbergiapyramidalis and provides sufficient seedling source, and tissue culture method can effectively shorten growing-seedling period compared with seed propagation.
Description
Technical field
The present invention relates to a kind of tissue culture technique, the method for in-vitro rapid propagation of specifically a kind of Billbergiapyramidalis.
Background technique
Billbergiapyramidalis (Billbergia pyramidalis Lindl.) pineapple family perennial evergreen draft succulent, stem
It is very short.There is serration at the wealthy lanceolar of leaf, anxious point, edge, and hard leather matter, emerald green, there is thick cuticula on surface and absorbs scale.Spike
Inflorescence is upright, is higher by leafage, and bract is pink, corolla vermilion, rolls up outside petal, marginal belt purple.Blade revolves folded from rhizome
Shape is grown thickly, and base portion is in rosette-stape, and center is cylindrical in shape.It blooms more than winter-spring season.Blade leathery；Verdant and gloss, grows thickly into lotus throne
Shape is dignified beautiful；The mutual obvolvent of leaf base makes plant center at tubular, and interior to be filled with water without leaking, shape is like water tower, therefore " water of gaining the name
Tower flower ".Like warm, wet, half shade environment.It can not resist cold.It is slightly drought-enduring, originate in Brazil.Billbergiapyramidalis strain Cong Qingcui, pattern is gorgeous, is
Good pot flowers.Billbergiapyramidalis in full bloom is the good merchantable brand for interspersing balcony, room, plants foliage plant for good indoor room.It is former
American torrid zone is produced, is grown nonparasitically upon another plant on the tree of Tropical forests or in humus.There are cultivation, especially southern area in Chinese greenhouse more.So
And the traditional nursery propagation method yield of Billbergiapyramidalis reduces, virus accumulation is serious, variety deterioration, quality and yield significantly under
Drop, planting benefit decline.Group culturation rapid propagating technology on many plants as economic, efficient propagation method application, the period is short,
Breeding coefficient is high, at low cost, is not restricted by time and space.
However, the nursery propagation method yield reduction that short pearl is traditional, virus accumulation is serious, variety deterioration, quality and production
Measure sharp fall, planting benefit decline.Group culturation rapid propagating technology is answered as economic, efficient propagation method on many plants
With the period is short, breeding coefficient is high, at low cost, is not restricted by time and space.
Summary of the invention
The present invention in view of the deficiencies of the prior art, meets the needs of artificial fast breeding, large area high-yield cultivating, provides one
The method for in-vitro rapid propagation of kind Billbergiapyramidalis.
To achieve the goals above, the invention adopts the following technical scheme:
A kind of method for in-vitro rapid propagation of Billbergiapyramidalis, includes the following steps:
(1) it obtains explant and sterilizes
The young tender sprout of healthy and strong plant new life is chosen, originally water washing twig surface smut dust, first alcohol impregnates, then rises
Mercury disinfection obtains disinfection explant after last aseptic water washing is clean,
(2) Initial culture
Disinfection explant is divided into the stem with bud of 0.6~1.0cm, then accesses and cultivates 32~36 in initial culture base
It obtains adventitious bud,
Initial culture condition are as follows: temperature is 28 ± 2 DEG C, and humidity is 55~60%, and light application time is 8~10 hours/day, light
According to intensity be 1800~20001x, Initial culture the 1st~7 day, daily apply ultrasonic field 3~4 hours, ultrasonic field power be 10~
15W/cm2, ultrasonic field frequencies range is 25~28KHz, Initial culture the 8th~15 day, is applied ultrasonic field 4~6 hours daily, ultrasonic field
Power is 20~22W/cm2, ultrasonic field frequencies range is 30~35KHz；
Initial culture base are as follows: on improvement White medium base, the concentration of ferric sulfate is first adjusted to 3.0~
The concentration of sodium sulphate is adjusted to 160~180mg/L, then adds 6~8mg/L of aspartic acid by 3.2mg/L, adds heteroauxin 2
~3mg/L；
(3) squamous subculture
Adventitious bud is transferred in subculture medium and cultivates 40~45 days acquisition squamous subculture seedlings,
Squamous subculture condition are as follows: temperature is 30 ± 2 DEG C, and humidity is 65~70%, and light application time is 12~15 hours/day,
Intensity of illumination is 3000~35001x,
Subculture medium are as follows: on improvement White medium base, the concentration of boric acid is first adjusted to 2.0~2.2mg/
The concentration of molybdenum oxide is adjusted to 0.0003~0.0005mg/L by L, in addition adds 0.5~0.8mg/L of vitamin K1 again, addition
1~1.2mg/L of heteroauxin adds 1.2~1.5mg/L of methyl α-naphthyl acetate；
(4) culture of rootage
Squamous subculture seedling is transferred to cultivate 20~25 days in root media and obtains Billbergiapyramidalis seedling,
Culture of rootage condition are as follows: temperature is 30 ± 2 DEG C, and humidity is 40~50%, and light application time is 12~15 hours/day,
Intensity of illumination is 5000~60001x,
Root media are as follows: in B5On medium base, 0.01~0.02mg/L of ammonium molybdate is in addition added again, is added red mould
2~3mg/L of element, adjusting pH with limewash is 6.5；
(5) hardening and transplanting
After culture of rootage, culture bottle cap is opened, carries out opening experienced seedling 5~7 days under natural light, then by Billbergiapyramidalis
The seedling multiple wash and remove residual culture medium of clear water, with clear water is clear again after the carbendazim immersion root 30min of mass fraction 3%
It is moved into the small basin of plastics after washing, the small basin of plastics is placed in the greenhouse of shading rate 60%~70% 2~3 weeks, temperature 25 in greenhouse
~28 DEG C, it is 75~85% that spray sprinkler, which keeps humidity, and greenhouse hardening is transplanted after one month into field planting ground.
Compared with prior art, the present invention have the utility model has the advantages that
The breeding coefficient of Billbergiapyramidalis is effectively improved, is commercially produced for Billbergiapyramidalis and sufficient seedling source is provided.With sowing
Breeding is compared, and tissue culture method can effectively shorten growing-seedling period, largely can continuously be produced indoors, realizes batch production, specially
Industry and scale improve the culture survival rate of seedling, improve the yield and quality.
Specific embodiment
Technical solution of the present invention is further elaborated below by embodiment.
Embodiment 1
A kind of method for in-vitro rapid propagation of Billbergiapyramidalis, includes the following steps:
(1) it obtains explant and sterilizes
The young tender sprout of healthy and strong plant new life is chosen, originally water washing twig surface smut dust, first alcohol impregnates, then rises
Mercury disinfection obtains disinfection explant after last aseptic water washing is clean,
(2) Initial culture
Disinfection explant is divided into the stem with bud of 1.0cm, then accesses in initial culture base and cultivates acquisition in 36 days not
Normal bud,
Initial culture condition are as follows: temperature is 30 DEG C, and humidity 60%, light application time is 10 hours/day, and intensity of illumination is
20001x Initial culture the 1st~7 day, applies ultrasonic field 4 hours daily, and ultrasonic field power is 15W/cm2, ultrasonic field frequencies range is
28KHz Initial culture the 8th~15 day, applies ultrasonic field 6 hours daily, and ultrasonic field power is 22W/cm2, ultrasonic field frequencies range is
35KHz；
Initial culture base are as follows: on improvement White medium base, the concentration of ferric sulfate is first adjusted to 3.2mg/L, it will
The concentration of sodium sulphate is adjusted to 180mg/L, then adds aspartic acid 8mg/L, adds heteroauxin 3mg/L；
(3) squamous subculture
Adventitious bud is transferred in subculture medium and cultivates 45 days acquisition squamous subculture seedlings,
Squamous subculture condition are as follows: temperature is 32 DEG C, and humidity 70%, light application time is 15 hours/day, and intensity of illumination is
35001x,
Subculture medium are as follows: on improvement White medium base, the concentration of boric acid is first adjusted to 2.2mg/L, by oxygen
The concentration for changing molybdenum is adjusted to 0.0005mg/L, in addition adds vitamin K1 0.8mg/L again, adds heteroauxin 1.2mg/L, adds
Add methyl α-naphthyl acetate 1.5mg/L；
(4) culture of rootage
Squamous subculture seedling is transferred to cultivate 25 days in root media and obtains Billbergiapyramidalis seedling,
Culture of rootage condition are as follows: temperature is 32 DEG C, and humidity 50%, light application time is 15 hours/day, and intensity of illumination is
60001x,
Root media are as follows: in B5On medium base, ammonium molybdate 0.02mg/L is in addition added again, adds gibberellin 3mg/
L, adjusting pH with limewash is 6.5；
(5) hardening and transplanting
After culture of rootage, culture bottle cap is opened, carries out opening experienced seedling 5~7 days under natural light, then by Billbergiapyramidalis
The seedling multiple wash and remove residual culture medium of clear water, with clear water is clear again after the carbendazim immersion root 30min of mass fraction 3%
It is moved into the small basin of plastics after washing, the small basin of plastics is placed in the greenhouse of shading rate 70% 3 weeks, and 28 DEG C of temperature, spill by spraying in greenhouse
It is 85% that water, which keeps humidity, and greenhouse hardening is transplanted after one month into field planting ground.
Embodiment 2
A kind of method for in-vitro rapid propagation of Billbergiapyramidalis, includes the following steps:
(1) it obtains explant and sterilizes
The young tender sprout of healthy and strong plant new life is chosen, originally water washing twig surface smut dust, first alcohol impregnates, then rises
Mercury disinfection obtains disinfection explant after last aseptic water washing is clean,
(2) Initial culture
Disinfection explant is divided into the stem with bud of 0.6cm, then accesses in initial culture base and cultivates acquisition in 32 days not
Normal bud,
Initial culture condition are as follows: temperature is 26 DEG C, and humidity 55%, light application time is 8 hours/day, and intensity of illumination is
18001x Initial culture the 1st~7 day, applies ultrasonic field 3 hours daily, and ultrasonic field power is 10W/cm2, ultrasonic field frequencies range is
25KHz Initial culture the 8th~15 day, applies ultrasonic field 4 hours daily, and ultrasonic field power is 20W/cm2, ultrasonic field frequencies range is
30KHz；
Initial culture base are as follows: on improvement White medium base, the concentration of ferric sulfate is first adjusted to 3.0mg/L, it will
The concentration of sodium sulphate is adjusted to 160mg/L, then adds aspartic acid 6mg/L, adds heteroauxin 2mg/L；
(3) squamous subculture
Adventitious bud is transferred in subculture medium and cultivates 40 days acquisition squamous subculture seedlings,
Squamous subculture condition are as follows: temperature is 28 DEG C, and humidity 65%, light application time is 12 hours/day, and intensity of illumination is
30001x,
Subculture medium are as follows: on improvement White medium base, the concentration of boric acid is first adjusted to 2.0mg/L, by oxygen
The concentration for changing molybdenum is adjusted to 0.0003mg/L, in addition adds vitamin K1 0.5mg/L again, adds heteroauxin 1mg/L, addition
Methyl α-naphthyl acetate 1.2mg/L；
(4) culture of rootage
Squamous subculture seedling is transferred to cultivate 20~25 days in root media and obtains Billbergiapyramidalis seedling,
Culture of rootage condition are as follows: temperature is 28 DEG C, and humidity 40%, light application time is 12 hours/day, and intensity of illumination is
50001x,
Root media are as follows: in B5On medium base, ammonium molybdate 0.01mg/L is in addition added again, adds gibberellin 2mg/
L, adjusting pH with limewash is 6.5；
(5) hardening and transplanting
After culture of rootage, culture bottle cap is opened, carries out opening experienced seedling 5~7 days under natural light, then by Billbergiapyramidalis
The seedling multiple wash and remove residual culture medium of clear water, with clear water is clear again after the carbendazim immersion root 30min of mass fraction 3%
It is moved into the small basin of plastics after washing, the small basin of plastics is placed in the greenhouse of shading rate 60%~70% 2~3 weeks, temperature 25 in greenhouse
~28 DEG C, it is 75~85% that spray sprinkler, which keeps humidity, and greenhouse hardening is transplanted after one month into field planting ground.
Claims (1)
1. a kind of method for in-vitro rapid propagation of Billbergiapyramidalis, which comprises the steps of:
(1) it obtains explant and sterilizes
The young tender sprout of healthy and strong plant new life is chosen, originally water washing twig surface smut dust, first alcohol impregnates, then mercuric chloride disappears
Poison obtains disinfection explant after last aseptic water washing is clean,
(2) Initial culture
Disinfection explant is divided into the stem with bud of 0.6~1.0cm, then accesses in initial culture base to cultivate 32~36 days and obtain
Adventitious bud is obtained,
Initial culture condition are as follows: temperature is 28 ± 2 DEG C, and humidity is 55~60%, and light application time is 8~10 hours/day, and illumination is strong
Degree is 1800~20001x, Initial culture the 1st~7 day, is applied ultrasonic field 3~4 hours daily, and ultrasonic field power is 10~15W/
cm2, ultrasonic field frequencies range is 25~28KHz, Initial culture the 8th~15 day, is applied ultrasonic field 4~6 hours daily, ultrasonic field power
For 20~22W/cm2, ultrasonic field frequencies range is 30~35KHz；
Initial culture base are as follows: on improvement White medium base, the concentration of ferric sulfate is first adjusted to 3.0~3.2mg/L,
The concentration of sodium sulphate is adjusted to 160~180mg/L, then adds 6~8mg/L of aspartic acid, adds 2~3mg/L of heteroauxin；
(3) squamous subculture
Adventitious bud is transferred in subculture medium and cultivates 40~45 days acquisition squamous subculture seedlings,
Squamous subculture condition are as follows: temperature is 30 ± 2 DEG C, and humidity is 65~70%, and light application time is 12~15 hours/day, illumination
Intensity is 3000~35001x,
Subculture medium are as follows: on improvement White medium base, the concentration of boric acid is first adjusted to 2.0~2.2mg/L, it will
The concentration of molybdenum oxide is adjusted to 0.0003~0.0005mg/L, in addition adds 0.5~0.8mg/L of vitamin K1 again, adds indoles
1~1.2mg/L of acetic acid adds 1.2~1.5mg/L of methyl α-naphthyl acetate；
(4) culture of rootage
Squamous subculture seedling is transferred to cultivate 20~25 days in root media and obtains Billbergiapyramidalis seedling,
Culture of rootage condition are as follows: temperature is 30 ± 2 DEG C, and humidity is 40~50%, and light application time is 12~15 hours/day, illumination
Intensity is 5000~60001x,
Root media are as follows: in B5On medium base, 0.01~0.02mg/L of ammonium molybdate is in addition added again, addition gibberellin 2~
3mg/L, adjusting pH with limewash is 6.5；
(5) hardening and transplanting
After culture of rootage, culture bottle cap is opened, carries out opening experienced seedling 5~7 days under natural light, then by Billbergiapyramidalis seedling
With the multiple wash and remove residual culture medium of clear water, after clear water cleans again after impregnating root 30min with the carbendazim of mass fraction 3%
It moves into the small basin of plastics, the small basin of plastics is placed in the greenhouse of shading rate 60%~70% 2~3 weeks, temperature 25~28 in greenhouse
DEG C, it is 75~85% that spray sprinkler, which keeps humidity, and greenhouse hardening is transplanted after one month into field planting ground.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101385430A (en) * | 2008-09-25 | 2009-03-18 | 天津滨海大顺花卉科技发展股份有限公司 | Pineapple flower forcing and nourishing method |
| CN101857505A (en) * | 2009-04-09 | 2010-10-13 | 梁春 | Method for preparing nutrient fertilizer for billbergia pyramidalis |
| CN104542306A (en) * | 2015-02-02 | 2015-04-29 | 广西壮族自治区药用植物园 | Fast propagation method of longans |
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2016
- 2016-10-05 CN CN201610872185.4A patent/CN106613940B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101385430A (en) * | 2008-09-25 | 2009-03-18 | 天津滨海大顺花卉科技发展股份有限公司 | Pineapple flower forcing and nourishing method |
| CN101857505A (en) * | 2009-04-09 | 2010-10-13 | 梁春 | Method for preparing nutrient fertilizer for billbergia pyramidalis |
| CN104542306A (en) * | 2015-02-02 | 2015-04-29 | 广西壮族自治区药用植物园 | Fast propagation method of longans |
Non-Patent Citations (2)
| Title |
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| 斑马水塔花的组织培养和快速繁殖;梁国平等;《云南热作科技》;20021231;第25卷(第4期);第26-28页 |
| 红藻凤梨的离体快速繁殖;王春荣等;《植物生理学通讯》;20020228;第38卷(第1期);第46页，尤其是第1-4节 |
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