CN102792890A - Rapid propagation method of tissue culture for zelkova schneideriana hand.-Mazz - Google Patents
Rapid propagation method of tissue culture for zelkova schneideriana hand.-Mazz Download PDFInfo
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- CN102792890A CN102792890A CN201210273106XA CN201210273106A CN102792890A CN 102792890 A CN102792890 A CN 102792890A CN 201210273106X A CN201210273106X A CN 201210273106XA CN 201210273106 A CN201210273106 A CN 201210273106A CN 102792890 A CN102792890 A CN 102792890A
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Abstract
The invention discloses a rapid propagation method of tissue culture for zelkova schneideriana hand.-Mazz. According to the method, selecting stem segments with buds as explants, and appropriate culture mediums for each stage for the tissue culture, a large number of nursery stocks, which maintain the qualified characteristics of a mother stock, can be obtained; and the genotype and all the qualified characteristics of an original plant can be 100% maintained, thereby greatly improving the propagation coefficient.
Description
Technical field
The invention belongs to the plant propagation technical field, be specifically related to the tissue culture and rapid propagation method of a kind of beech tree.
Background technology
The beech tree
Zelkova schneiderianaHand.-Mazz. be the Ulmaceae Zeldova.Be precious hard broad-leaved commerical tree species, the wood quality of beech tree is hard flexible, and beautiful texture is glossy, and does not have special odor, has become many families and has carried out interior decoration and the preferred material of making top-grade furniture, liked by consumers in general.Simultaneously, it is tree-like attractive in appearance that beech are set, and leaf can have very high landscape architecture and ornamental value with presenting various colors season.And beech tree damage by disease and insect is few, has very strong resistance against diseases; Tree crown and root system are all very huge, and the fallen leaves amount is many, have the good water and soil conservation and the effect of improving the soil, and play crucial effect in protection with in improving the ecological environment.But because too much felling and shortage protection, the wild resource of beech tree is quite deficient, and the beech tree is classified as Chinese Second Class Key Protected Plant.
Along with expanding economy and the people new pursuit to living standard, present quarter in all parts of the country, park, roadside greening are in the ascendant, and China market is to greening seedling, and particularly the demand of rare tree species is increasing, and imbalance between supply and demand is outstanding.Add that China's forest plantation seeds unification and " needleization " phenomenon are very serious, caused a series of problems such as the ecosystem is unbalance and low in economic efficiency, develop good hard broad-leaved forest plantation and caused the great attention of China forest department.In forest plantation seeds of new generation structural adjustment, select material good, fast natural disposition can and the stronger rare tree species of insect resistance capacity such as the beech tree afforestation common recognition that become brainstrust, the demand of high-quality speed being given birth to beech sapling wood constantly increases.But, to the research of beech trees, relatively lagging behind like the research of aspects such as introduction and acclimatization, breeding of new variety, breeding and popularization, the very different phenomenon of seed seedling-raising is very serious, and the science and technology support of production application and market supply can not be provided.Therefore, particularly important to the research of the quick propagating technology of beech trees.
Present research report about beech tree tissue culture mainly contains the tissue culture of terminal bud and passes through the tissue culture of blade evoked callus.The bud nursery stock uniformity of growing thickly that the tissue culture of terminal bud obtains is poor, reproduction coefficient low (27 ~ 125); Regeneration plant that the tissue culture of blade evoked callus produces and the uniformity of maternal plant are poor.
Summary of the invention
The present invention is intended to overcome the deficiency of prior art, and a kind of method of beech tree tissue-culturing quick-propagation is provided.
In order to achieve the above object, technical scheme provided by the invention is:
Beech tree quick breeding method for tissue culture comprises the steps:
(1) beech is set young shoot with alcohol disinfecting 25 ~ 35s of 75%, use aseptic water washing,, use aseptic water washing again with 0.1% mercuric chloride sterilization, 8 ~ 12min;
(2) will be cut into the stem section that long 0.8 ~ 1.2cm contains an axillalry bud at least through the branch after step (1) is handled, and implant in the inducing clumping bud medium and cultivated 28 ~ 32 days, obtain to grow thickly bud; Wherein, said inducing clumping bud medium is: WPM+1.0 ~ 3.0 mg.L
-1BA+0 ~ 1.0 mg.L
-1NAA, agar 6.5 g.L
-1, sucrose 20 g.L
-1, the pH value is 5.7;
The bud of growing thickly that (3) will behind step (2) inducing culture, grow changes over to cultivates 60 ~ 65 days in the subculture medium, carry out the propagation of bud; Wherein, said subculture medium is: WPM+2.0 ~ 3.0 mg.L
-1BA+1.0 ~ 2.0 mg.L
-1NAA, agar 6.5 g.L
-1, sucrose 20 g.L
-1, the pH value is 5.7;
(4) cutting the height that behind step (3) successive transfer culture, grows is the beech tree healthy seedling more than 2.0 cm, implants in the root media and cultivates 45 ~ 50 days, obtains regeneration plant; Wherein, said root media is: root media is: WPM+0.5 ~ 1.0 mg.L
-1IBA+2.0 g.L
-1AC, agar 7.0 mg.L
-1, sucrose 20 g.L
-1, the pH value is 5.7;
(5) will clean through the regeneration plant that step (4) obtains, and be transplanted to and refine seedling cultivation 7 ~ 10 days in the mixed-matrix of being made up of peat soil and perlite, outdoor again cultivation 90 ~ 100 days promptly can be moved to field production, and the week of shading during field production gets final product.
Preferably, to set young shoot be to give birth to then to be with bud children shoot spring to the said beech of step (1).
Preferably, it is that healthy and strong branch with the beech trees fallen leaves in winter that contain axillalry bud places the beaker that distilled water is housed that the said beech of step (1) are set young shoot, carries out water planting after 10 ~ 15 days under 25 ℃, the young shoot that is formed by axillary bud sprouting.
Preferably, peat soil and perlitic volume ratio are 1:1 in the described mixed-matrix of step (5).
Preferably, the condition of culture in the step (2) to (4) is: 24 ± 2 ℃ of room temperatures, intensity of illumination PPFD=65.5 μ .moL.m
-2.s
-1, 16 hours every days of light application time.
Wherein, grow thickly in the step of the present invention (2) inductivity of bud is 80 ~ 90%; The bud growth coefficient of bud enrichment culture is 7 ~ 10 in the step (3); The rooting rate of culture of rootage is 86 ~ 95% in the step (4); Transplant in the step (5) to the survival rate of field production be 95 ~ 100%.
Compared with prior art, beneficial effect of the present invention is:
1, the inventive method select tape leaf stem section is an explant, and selects each suitable stage medium, carries out tissue culture, and a large amount of nursery stocks of the maternal plant merit that can be maintained can the 100% former plant genotype of maintenance and all good characteristics thereof.
2, the present invention can realize factorial seedling growth, and produces the beech sapling wood of neat and consistent in enormous quantities, increases substantially reproduction coefficient (343 ~ 1000).The calculating of reproduction coefficient is: breeding a time is 4 months, can breed 3 times in 1 year 12 months, and growth coefficient once is 7 ~ 10, and the reproduction coefficient in 1 year is 7 * 7 * 7 ~ 10 * 10 * 10 (343 ~ 1000) so.
Embodiment
Below in conjunction with embodiment the present invention is done further detailed description.
Embodiment 1
(1) with the beech chosen tree give birth to then spring with bud children shoot with alcohol disinfecting 30 ~ 35s of 75%, with aseptic water washing 3 times,, use aseptic water washing again 3 times with the 0.1% mercuric chloride 8min that sterilizes;
(2) will be cut into the stem section that long 1cm contains an axillalry bud at least through the branch after step (1) is handled, and implant in the bud inducing culture and cultivated 28 ~ 30 days, obtain to grow thickly bud; Wherein, said bud inducing culture is: WPM+3.0 mg.L
-1BA, agar 6.5 g.L
-1, sucrose 20 g.L
-1, the pH value is 5.7, medium autoclaving 20min; The inductivity of bud of growing thickly is 84 ~ 90%;
The bud of growing thickly that (3) will behind step (2) inducing culture, grow changes in the subculture medium and cultivated 60 days, carries out the propagation of bud, and the bud growth coefficient is 8 ~ 10; Wherein, said subculture medium is: WPM+3.0 mg.L
-1BA+2.0 mg.L
-1NAA, agar 6.5 g.L
-1, sucrose 20 g.L
-1, the pH value is 5.7, medium autoclaving 20min;
(4) cutting the height that behind step (3) successive transfer culture, grows is the beech tree healthy seedling more than the 2.0cm, implants in the root media and cultivates 45 days, obtains regeneration plant, and rooting rate is 89 ~ 95%; Wherein, said root media is: root media is: WPM+0.5 mg.L
-1IBA+2.0 g.L
-1AC, agar 7.0 mg L
-1, sucrose 20 g.L
-1, the pH value is 5.7, medium autoclaving 20min;
(5) will clean through the regeneration plant that step (4) obtains; Be transplanted to by refining seedling in peat soil and the perlite mixed-matrix that 1:1 forms by volume and cultivated 7 days, outdoor again cultivation 90 days promptly can be moved to field production; Shading during field production gets final product in a week, and transplanting survival rate is 100%.
Condition of culture in the above-mentioned steps (2) to (4) is: 24 ± 2 ℃ of room temperatures, intensity of illumination PPFD=65.5 μ .moL.m
-2.s
-1, 16 hours every days of light application time.
Result of the test shows, a large amount of nursery stocks of the maternal plant merit that can be maintained can 100% keep former plant genotype and all good characteristics thereof, have higher reproduction coefficient (512 ~ 1000).
Embodiment 2
The healthy and strong branch that will contain the beech trees fallen leaves in winter of axillalry bud places the beaker that distilled water is housed, and carries out water planting after 15 ~ 30 days under 25 ℃, obtains the young shoot that axillary bud sprouting forms, and carries out following steps then;
(1) beech is set young shoot with alcohol disinfecting 25 ~ 30s of 75%,,, use aseptic water washing again 3 times with 0.1% mercuric chloride sterilization 12min with aseptic water washing 3 times;
(2) will be cut into the stem section that long 1.2cm contains an axillalry bud at least through the branch after step (1) is handled, and implant in the inducing clumping bud medium and cultivated 30 ~ 32 days, obtain to grow thickly bud; Wherein, said inducing clumping bud medium is: WPM+1.0 mg.L
-1BA+1.0 mg.L
-1NAA, agar 6.5 g.L
-1, sucrose 20 g.L
-1, the pH value is 5.7, medium autoclaving 20min;
The bud of growing thickly that (3) will behind step (2) inducing culture, grow changes over to cultivates the enrichment culture that carried out bud in 65 days in the subculture medium, the bud growth coefficient is 7 ~ 9; Wherein, said subculture medium is: WPM+2.0 mg.L
-1BA+1.0 mg.L
-1NAA, agar 6.5 g.L
-1, sucrose 20 g.L
-1, the pH value is 5.7, medium autoclaving 20min;
(4) cutting the height that behind step (3) successive transfer culture, grows is the beech tree healthy seedling more than 2.0 cm, implants in the root media and cultivates 50 days, obtains regeneration plant, and rooting rate is 86%; Wherein, said root media is: root media is: WPM+1.0 mg.L
-1IBA+2.0 g.L
-1AC, agar 7.0 mg L
-1, sucrose 20 g.L
-1, the pH value is 5.7, medium autoclaving 20min;
(5) will clean through the regeneration plant that step (4) obtains; Be transplanted to by refining seedling in peat soil and the perlite mixed-matrix that 1:1 forms by volume and cultivated 10 days, outdoor again cultivation 100 days promptly can be moved to field production; Shading during field production gets final product in a week, and transplanting survival rate is 95%.
Condition of culture in the above-mentioned steps (2) to (4) is: 24 ± 2 ℃ of room temperatures, intensity of illumination PPFD=65.5 μ .moL.m
-2.s
-1, 16 hours every days of light application time.
Result of the test shows, a large amount of nursery stocks of the maternal plant merit that can be maintained can 100% keep former plant genotype and all good characteristics thereof, have higher reproduction coefficient (343 ~ 729).
Claims (5)
1. a beech tree quick breeding method for tissue culture comprises the steps:
(1) beech is set young shoot with alcohol disinfecting 25 ~ 35s of 75%, use aseptic water washing,, use aseptic water washing again with 0.1% mercuric chloride sterilization, 8 ~ 12min;
(2) will be cut into the stem section that long 0.8 ~ 1.2cm contains an axillalry bud at least through the branch after step (1) is handled, and implant in the inducing clumping bud medium and cultivated 28 ~ 32 days, obtain to grow thickly bud; Wherein, said inducing clumping bud medium is: WPM+1.0 ~ 3.0 mg.L
-1BA+0 ~ 1.0 mg.L
-1NAA, agar 6.5 g.L
-1, sucrose 20 g.L
-1, the pH value is 5.7;
The bud of growing thickly that (3) will behind step (2) inducing culture, grow changes over to cultivates 60 ~ 65 days in the subculture medium, carry out the propagation of bud; Wherein, said subculture medium is: WPM+2.0 ~ 3.0 mg.L
-1BA+1.0 ~ 2.0 mg.L
-1NAA, agar 6.5 g.L
-1, sucrose 20 g.L
-1, the pH value is 5.7;
(4) cutting the height that behind step (3) successive transfer culture, grows is the beech tree healthy seedling more than the 2.0cm, implants in the root media and cultivates 45 ~ 50 days, obtains regeneration plant; Wherein, said root media is: WPM+0.5 ~ 1.0 mg.L
-1IBA+2.0 g.L
-1AC, agar 7.0 mg L
-1, sucrose 20 g.L
-1, the pH value is 5.7;
(5) will clean through the regeneration plant that step (4) obtains, and be transplanted to and refine seedling cultivation 7 ~ 10 days in the mixed-matrix of being made up of peat soil and perlite, outdoor again cultivation 90 ~ 100 days promptly can be moved to field production.
2. beech are set quick breeding method for tissue culture according to claim 1, it is characterized in that, it is to give birth to then to be with bud children shoot spring that the said beech of step (1) are set young shoot.
3. beech are set quick breeding method for tissue culture according to claim 1; It is characterized in that; It is the beech that contain axillalry bud to be set the healthy and strong branch of falling leaves winter place the beaker that distilled water is housed that the said beech of step (1) are set young shoot; Carry out water planting under 25 ℃ after 15 ~ 30 days, the young shoot that forms by axillary bud sprouting.
4. beech are set quick breeding method for tissue culture according to claim 1, it is characterized in that peat soil and perlitic volume ratio are 1:1 in the described mixed-matrix of step (5).
5. beech are set quick breeding method for tissue culture according to claim 1, it is characterized in that the condition of culture in the step (2) to (4) is: 24 ± 2 ℃ of room temperatures, intensity of illumination PPFD=65.5 μ .moL.m
-2.s
-1, 16 hours every days of light application time.
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103004591A (en) * | 2012-11-29 | 2013-04-03 | 绵阳师范学院 | Fagus pashanica tissue culture rapid propagation method |
CN103125385A (en) * | 2013-01-28 | 2013-06-05 | 浙江红叶园艺有限公司 | Cutting propagation method of strengthening antipollution factor zelkova schneideriana |
-
2012
- 2012-08-03 CN CN201210273106.XA patent/CN102792890B/en not_active Expired - Fee Related
Non-Patent Citations (3)
Title |
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金晓玲: "榉树的生物学特性和微繁技术研究", 《中国博士学位论文全文数据库基础科学辑》 * |
金晓玲等: "榉树茎尖的培养", 《中南林学院学报》 * |
钟飞霞等: "榉树无性繁殖技术研究进展", 《现代农业科技》 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103004591A (en) * | 2012-11-29 | 2013-04-03 | 绵阳师范学院 | Fagus pashanica tissue culture rapid propagation method |
CN103125385A (en) * | 2013-01-28 | 2013-06-05 | 浙江红叶园艺有限公司 | Cutting propagation method of strengthening antipollution factor zelkova schneideriana |
CN103125385B (en) * | 2013-01-28 | 2015-07-08 | 浙江红叶园艺有限公司 | Cutting propagation method of strengthening antipollution factor zelkova schneideriana |
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