CN104521752B - A kind of Lay bamboo grass tissue-culturing rapid propagation and the method for industrial seedling rearing - Google Patents
A kind of Lay bamboo grass tissue-culturing rapid propagation and the method for industrial seedling rearing Download PDFInfo
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- CN104521752B CN104521752B CN201410750097.8A CN201410750097A CN104521752B CN 104521752 B CN104521752 B CN 104521752B CN 201410750097 A CN201410750097 A CN 201410750097A CN 104521752 B CN104521752 B CN 104521752B
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Abstract
The invention belongs to agricultural biological technical field, be specifically related to a kind of Lay bamboo grass tissue-culturing rapid propagation and the method for industrial seedling rearing.The method comprises the following steps: the pretreatment of Lay bamboo grass, outside shade select and process, callus induction, proliferation and differentiation, root culture, transplanting seedling exercising, industrial seedling rearing.Selected by Lay bamboo grass pretreatment, outside shade and process, filter out callus, proliferation and differentiation, the culture medium prescription such as take root, set up regenerating system, and take suitable seedling exercising measure and industrial seedling rearing condition to control, it is achieved and the tissue-culturing rapid propagation of Lay bamboo grass and industrial seedling rearing, callus induction rate is 92 98%, growth coefficient 5.6, rooting rate 98%, transplants seedling exercising survival rate 99%, solves the difficult problem that Lai Zhucao can not pass through seminal propagation, cottage propagation power is low.
Description
Technical field
The invention belongs to agricultural biological technical field, be specifically related to a kind of Lay bamboo grass tissue-culturing rapid propagation and the method for industrial seedling rearing.
Background technology
Lay bamboo is introduced from Lesotho as a kind of bacterium glass, is under the jurisdiction of Arundo donax and belongs to.Lesotho is positioned at the southeast, Africa, and height above sea level 1400-3482m, is the country that mean sea level is the highest in the world, belongs to continental subtropical climate, average annual precipitation about 780mm, and long-term minimum temperature-10 DEG C, extreme minimum temperature is up to-18 DEG C.Lay bamboo all has distribution in Lesotho whole area, can normally survive the winter, drought-enduring, waterlogging, barren-resistant.Lay bamboo has whiplike, well developed root system, plant height 350-620cm, stem hollow, degree of lignification is high, because it resists cold, relatively Salt And Alkali Tolerance, and well developed root system, be suitable as in erosion zone, desert, desert, the ecologically fragile areas such as salt-soda soil carry out ecological management and a kind of bacterium glass of development bacterium glass industry.But because it does not has seed, extremely low by stipes cottage propagation power, therefore need to cultivate, by tissue-culturing rapid propagation mode, the careless Seedling that growth is consistent, quality is high with industrial breeding technique.Outside shade selects and pretreatment, optimal medium recipe determination, and high-efficiency regeneration system foundation and the control of industrial seedling rearing growth conditions are the keys of Lay bamboo tissue culture rapid breeding and industrial seedling rearing.
Summary of the invention
Present invention aims to the difficult problem that Lai Zhucao can not pass through seminal propagation, cottage propagation power is low, it is provided that a kind of Lay bamboo grass tissue-culturing rapid propagation and the method for industrial seedling rearing.The method makes the nursery of Lay bamboo grass not limited by regional, weather, can provide the high quality seedling that a large amount of specification is consistent in a short time, for erosion zone, desert, desert, the ecological management of the ecologically fragile areas such as salt-soda soil and the development of bacterium glass industry.
For achieving the above object, the present invention adopts the following technical scheme that
A kind of Lay bamboo grass tissue-culturing rapid propagation and the method for industrial seedling rearing, comprise the following steps: the pretreatment of Lay bamboo grass, outside shade select and process, callus induction, proliferation and differentiation, root culture, transplanting seedling exercising, industrial seedling rearing.
Described Lay bamboo grass pretreatment is: plant is used 0.2wt% urea liquid and 0.2wt% potassium dihydrogen phosphate, executed once every 1 day, execute 3 times altogether.
Described outside shade selects and is processed as: selects pretreated Lay bamboo grass blade joint and stem section, cleans, with 75% alcohol wipe surface, sterilize 45 seconds；Place into sterilization 10min in 0.12% mercuric chloride solution；Finally using aseptic water washing 5 times, excise the two ends of Lay bamboo grass blade joint, only retain lateral bud, lateral bud, stem section are cut into the segment of 2mm again.
Described callus induction is: half-light is cultivated, cultivation temperature 22 ~ 26 DEG C, incubation time 45 days；The formula of culture medium is: 1/2MS+2mg L-1BA+0.1mg·L-12,4-D, additional 3% sucrose, 0.6% agar, pH=6；The induction of the lateral bud rate 98%, stem section inductivity 92%.
Described proliferation and differentiation method particularly includes: illumination 16 hour/day, light intensity 2500 Lx, cultivation temperature 23 ~ 27 DEG C, incubation time 35 days；Culture medium prescription is: 1/2MS+2mg L-1BA+0.2mg·L-1NAA；Growth coefficient 5.6.
Described root culture actual conditions is: illumination 10 hour/day, light intensity 1500 Lx, cultivation temperature 21 ~ 25 DEG C, incubation time 15 days；Culture medium prescription 1/2MS+2mg L-1BA+0.1mg·L-1IAA；Rooting rate 98%.
Described transplanting seedling exercising method particularly includes: rinsed well by the root system of the regrowth that height of seedling is 4-6cm whole plant, moves to batch production canopy seedling exercising；Substrate used is the waste mushroom after bacterium glass cultivation Pleurotus ostreatus, adds sandy loam and the Calx of 3% of 30%, mixes rear fermentation reactor system 3-6 month thoroughly；Shoot survival percent 99%.
Industrial seedling rearing condition: transplant in latter 7 days, fine day 11:00-15:00 sunshade, temperature 24 ~ 28 DEG C, at 10:00 and 16:00 spraying humidification, relative air humidity controls at 80-85%；After transplanting 8-20 days, intensity of illumination 8000 ~ 10000Lx, temperature 25 ~ 29 DEG C, 9:00-18:00 was every 3 hours spraying humidifications, and relative air humidity controls at 85-90%；After transplanting 21-40 days, intensity of illumination 12000-18000 Lx, temperature 26 ~ 30 DEG C, 9:00-18:00 was every 2 hours spraying humidifications, relative air humidity controls at 85-95%, within the 25th day and the 35th day, sprays 0.2% carbamide, the foliage fertilizer of 0.2% potassium dihydrogen phosphate after transplanting.
The beneficial effects of the present invention is:
1) present invention is processed by Lay bamboo grass pretreatment, outside shade, filter out callus, proliferation and differentiation, the culture medium prescription such as take root, set up regenerating system, and take suitable seedling exercising measure and industrial seedling rearing condition to control, achieving tissue-culturing rapid propagation and the industrial seedling rearing of Lay bamboo grass, callus induction rate is 92-98%, growth coefficient 5.6, rooting rate 98%, transplants seedling exercising survival rate 99%；
2) present invention solves the difficult problem that Lai Zhucao can not pass through seminal propagation, cottage propagation power is low, the nursery making Lay bamboo grass is not limited by area, weather, be provided that the high quality seedling that a large amount of specification is consistent in a short time, for erosion zone, desert, desert, the ecological management of the ecologically fragile areas such as salt-soda soil and the development of bacterium glass industry.
Detailed description of the invention
Present invention the following example further illustrates the present invention, but protection scope of the present invention is not limited to the following example.
Embodiment
1
1) plant selects: growth selection phase 1-2, the Lay bamboo grass plant of robust growth；
2) pretreatment: the plant being intended to carry out group training being carried out strong strain accelerating germination 6 days, uses the carbamide of 0.2% and the potassium dihydrogen phosphate of 0.2%, executed once every 1 day, each amount of application is 150ml/ strain, reaches strong strain accelerating germination purpose；
3) outside shade selects: select to carry out the Lay bamboo grass blade joint of pretreatment and stem section；
4) outside shade processes: cleans Lay bamboo grass blade joint and stem section, with the alcohol wipe surface of 75%, sterilizes 45 seconds；Placing in the mercuric chloride solution of 0.12% and sterilize 10 minutes, finally use aseptic water washing 5 times, excision Lay bamboo grass blade saves two ends, only retains lateral bud, and lateral bud, stem section are cut into the segment of 2mm；
5) callus induction: culture medium prescription 1/2MS+2mg L-1BA+0.1mg·L-12,4-D, additional 3% sucrose, 0.6% agar, pH=6, half-light cultivation, cultivation temperature 25 DEG C, incubation time 45 days, the induction of the lateral bud rate 98%, stem section inductivity 92%；
6) proliferation and differentiation: culture medium prescription 1/2MS+2mg L-1BA+0.2mg·L-1NAA, illumination 16 hour/day, light intensity 2500 Lx, cultivation temperature 25 DEG C, incubation time 35 days, growth coefficient 5.6；
7) root culture: culture medium prescription 1/2MS+2mg L-1BA+0.1mg·L-1IAA, illumination 10 hour/day, light intensity 1500 Lx, cultivation temperature 23 DEG C, incubation time 15 days, rooting rate 98%；
8) seedling exercising is transplanted: the root system of the regrowth that height of seedling 4-6cm is grown up to whole plant is rinsed well, move to batch production canopy seedling exercising, substrate is the waste mushroom (seeing Publication No. CN 101121613A) after bacterium glass cultivation Pleurotus ostreatus, add sandy loam and the Calx of 3% of 30%, mix rear fermentation reactor system thoroughly 5 months, shoot survival percent 99%；
9) industrial seedling rearing condition: transplant in latter 7 days, fine day 11:00-15:00 sunshade, temperature 26 DEG C, at 10:00 and 16:00 spraying humidification, relative air humidity 80-85%；After transplanting 8-20 days, intensity of illumination 9000Lx, temperature 27 DEG C, 9:00-18:00 was every 3 hours spraying humidifications, relative air humidity 85-90%；After transplanting 21-40 days, intensity of illumination 15000Lx, temperature 28 DEG C, 9:00-18:00, every 2 hours spraying humidifications, relative air humidity 85-95%, executed 0.2% carbamide, the foliage fertilizer of 0.2% potassium dihydrogen phosphate in the 25th day and the 35th day after transplanting.
The foregoing is only presently preferred embodiments of the present invention, all impartial changes done according to scope of the present invention patent and modification, all should belong to the covering scope of the present invention.
Claims (3)
1. a Lay bamboo grass tissue-culturing rapid propagation and the method for industrial seedling rearing, it is characterised in that: comprise the following steps: the pretreatment of Lay bamboo grass, outer implant select and process, callus induction, proliferation and differentiation, root culture, transplanting seedling exercising, industrial seedling rearing；Industrial seedling rearing condition: transplant in latter 7 days, fine day 11:00-15:00 sunshade, temperature 24 ~ 28 DEG C, at 10:00 and 16:00 spraying humidification, relative air humidity controls at 80-85%；After transplanting 8-20 days, intensity of illumination 8000 ~ 10000Lx, temperature 25 ~ 29 DEG C, 9:00-18:00 was every 3 hours spraying humidifications, and relative air humidity controls at 85-90%；After transplanting 21-40 days, intensity of illumination 12000-18000 Lx, temperature 26 ~ 30 DEG C, 9:00-18:00 was every 2 hours spraying humidifications, relative air humidity controls at 85-95%, within the 25th day and the 35th day, sprays 0.2% carbamide, the foliage fertilizer of 0.2% potassium dihydrogen phosphate after transplanting；
Described Lay bamboo grass pretreatment is: plant being used 0.2wt% urea liquid and 0.2wt% potassium dihydrogen phosphate, executed once every 1 day, each amount of application is 150ml/ strain, executes 3 times altogether；
Described callus induction is: the formula of culture medium is: 1/2MS+2mg L-1BA+0.1mg·L-12,4-D, additional 3% sucrose, 0.6% agar, pH=6；Half-light is cultivated, cultivation temperature 22 ~ 26 DEG C, incubation time 45 days；
Described proliferation and differentiation method particularly includes: culture medium prescription is: 1/2MS+2mg L-1BA+0.2mg·L-1NAA；Illumination 16 hour/day, light intensity 2500 Lx, cultivation temperature 23 ~ 27 DEG C, incubation time 35 days；
Described root culture actual conditions is: culture medium prescription 1/2MS+2mgL-1BA+0.1mgL-1IAA；Illumination 10 hour/day, light intensity 1500 Lx, cultivation temperature 21 ~ 25 DEG C, incubation time 15 days.
Lay bamboo grass tissue-culturing rapid propagation the most according to claim 1 and the method for industrial seedling rearing, it is characterised in that: described outer implant selects and is processed as: selects pretreated Lay bamboo grass blade joint and stem section, cleans, with 75% alcohol wipe surface, sterilize 45 seconds；Place into sterilization 10min in 0.12% mercuric chloride solution；Finally using aseptic water washing 5 times, excise the two ends of Lay bamboo grass blade joint, only retain lateral bud, lateral bud, stem section are cut into the segment of 2mm again.
Lay bamboo grass tissue-culturing rapid propagation the most according to claim 1 and the method for industrial seedling rearing, it is characterised in that: described transplanting seedling exercising method particularly includes: the root system of the regrowth that height of seedling is 4-6cm whole plant is rinsed well, move to batch production canopy seedling exercising；Substrate used is the waste mushroom after bacterium glass cultivation Pleurotus ostreatus, adds sandy loam and the Calx of 3% of 30%, mixes rear fermentation reactor system 3-6 month thoroughly.
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| CN105248280B (en) * | 2015-10-30 | 2018-01-12 | 运城市绿碧源农林开发有限公司 | Oasis No.1 once-seedling forming tissue culture and rapid propagation method |
| CN110402857B (en) * | 2019-08-30 | 2021-12-03 | 三亚福联水产发展有限公司 | Rapid propagation method for grouper |
| CN113692935B (en) * | 2021-09-26 | 2022-07-26 | 开远云之澳花卉有限公司 | Method for improving variety of cut flower of Australian wintersweet into variety of potted flower |
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| US6389746B1 (en) * | 1999-10-12 | 2002-05-21 | David I. Bransby | Method of propagating fibercane (Arundo) |
| WO2002063023A2 (en) * | 2001-02-05 | 2002-08-15 | The University Of South Carolina Research Foundation | Sustained totipotent regenerable tissue culture of arundo donax (giant reed) and totipotent tissue and plants produced therefrom |
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