CN103477977A - High-quality test-tube plantlet root restriction cultivation method and container of dendrobium officinale - Google Patents
High-quality test-tube plantlet root restriction cultivation method and container of dendrobium officinale Download PDFInfo
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Abstract
The invention discloses a final singling and root restriction cultivation method and container which are suitable for preventing root systems from being difficultly output from a flask due to winding during the tissue culture based rooting and strengthening cultivation of orchid, represented by dendrobium officinale. The method is characterized in that a conventional culture flask is internally separated into equal areas, i.e., a culture medium is separated into equal parts; corresponding plantlets are planted into the separated areas during final singling of rapid propagation, and thus, the root system developed during plantlet strengthening and rooting just can grow in corresponding areas, and the plantlets just like grow in a seedling raising dish; the root winding only appears in the separated areas sometimes, mutual root winding of all plantlets in the whole flask is avoided, and thus stronger test-tube plantlets can be cultivated; the grown-up plantlets can be output from the flask conveniently, therefore, the amount of labor for outputting from the flask can be greatly saved, simultaneously the root system and stem leaves cannot be damaged, and as a result, the output efficiency as well as survival rate of transplanting can be greatly improved.
Description
Technical field
the present invention relates to a kind of orchid (particularly Dendrobium, Paphiopedilum, Cymbidium etc.) tissue cultivating and seedling method, the final singling cultivation, the strengthening seedling and rooting stage that are specially in Plant Tissue Breeding are limit root to cultivate, be a kind of high-tech seedling growing process novel method, belong to the technical field of bioengineering field of plant tissue culture technique.
Background technology
the orchid family is the first section in monocotyledon, and wherein numerous species has important ornamental value, medical value and edibility.The asexual tillering propagation reproduction coefficient of orchid is low, be unsuitable for human wants, only for one deck kind suitcase, wrap up in cells,primordial group during its seed maturity, without complete embryonic knob structure, under natural conditions, the sexual propagation coefficient is also very low, for the demand that meets human lives and production, only have the method for cultivating by tissue to carry out vegetative propagation and (get stem, bud, root, flower waits the organ tissue culturing breeding) or the seed bottle broadcast and breed seedling, all tissue culture processes all will experience one protocorm or seedling are inoculated on the strengthening seedling and rooting medium and carry out making strong sprout its formation to have the healthy and strong test-tube plantlet of healthy and strong root system and certain altitude plant, at this moment in order to guarantee the test-tube plantlet quality, in culture vessel, educable seedling number is relatively fixing, this process is called final singling and cultivates or cry strong seedling culture.
the current domestic orchid with the stem of noble dendrobium class headed by dendrobium candidum, pocket orchid and Cymbidium that has that carries out factorial seedling growth by tissue culture technique, tradition final singling cultural method is that seedling is inoculated in a blake bottle entirely, along with seedling is grown up gradually, the root system of test-tube plantlet is wrapped in the blake bottle bottom, deep-rooted root is intricate to combine together, traditional orchid special culture bottle, the large bottle of 630ml, the bottom external diameter is 9.6cm, and top seedling outlet external diameter is 4.9cm; Adopt the 370ml tissue culture bottle of straight barrel type, seedling outlet is slightly larger, but strong seedling culture after a period of time, and root system is formed one in bottom plate, very difficult during bottle outlet, can cause very large damage to root, stem, the leaf of test-tube plantlet, has reduced greatly transplanting survival rate; Simultaneously in order to reduce bottle outlet as far as possible, the damage operating personnel of test-tube plantlet are obtained to careful bottle outlet; The work of cleaning in addition medium after bottle outlet also the careful careful root system that do not damage, the time of expending like this increase, reduced the operating efficiency of bottle outlet.
adopt limit root cultural method can overcome that test-tube plantlet grows vigorously and bottle outlet between contradiction, test-tube seedling transplanting is simple and convenient, test-tube plantlet sustains damage little, after transplanting, transplanting seedling time is few, transplanting survival rate improves greatly.
up to the present nobody proposes limit root cultural method and design specialized container in tissue is cultivated.
Summary of the invention
about the objective of the invention is in order to solve in the dendrobium candidum factorial seedling growth, after final singling culture of rootage a period of time, seedling is grown up gradually, the root system of test-tube plantlet is wrapped in the blake bottle bottom, deep-rooted root is intricate to combine together, and hinders root during bottle outlet, hinders leaf, the time-consuming bad phenomenon of requiring great effort.The present invention has also designed the special culture bottle that orchid limit root is cultivated.
purpose of the present invention can be achieved through the following technical solutions.
dendrobium candidum high-quality test-tube plantlet limit root cultural method, major technique of the present invention is proposing " limit root cultivation " these new ideas, and has designed special-purpose culture vessel, and concrete steps are as follows:
1, sowing: Seeds of Dendrobium Candidum is seeded on germination medium, induces to sprout to obtain protocorm.Seed germination and differential medium MS+NAA (0.6-0.4) mg/L+6-BA (0.15-0.05) mg/L+ sucrose 30g/L+agar powder 7.2g/L+ coconut milk 100 ml/L.
2, seedling is cultivated.Protocorm is proceeded on differential medium, cultivate 3-4 week, form and have the long root of 2-3 bar 0.5 cm, stem differentiates 2-3 joint, has 3-4 sheet leaflet, the seedling of the high 0.6-0.8cm of plant.
3, the limit root is cultivated.Protocorm that will be larger or differentiation seedling are colonizated on the strengthening seedling and rooting medium be contained in limit root culture vessel is limit root to cultivate, the strengthening seedling and rooting medium is MS+NAA (1.0-0.4) mg/L+6-BA (0.1-0.05) mg/L+ sucrose 30g/L+agar powder 7.2 g/L+ banana puree 60 g/L, cultivating seedling after 8 to 12 weeks grows up, root system is healthy and strong but mutually be not wound around, during bottle outlet, easily operate, very convenient.
4, test-tube seedling transplanting.The high-quality seedling that limit root strong seedling culture is obtained proceeds in the seeding room of natural lighting and temperature or booth and takes exercise 7 days, then open bottle cap 3-5 days, take out test-tube plantlet and clean the root medium, make surface water second to half-dried rear cultivation, take the China fir fragment as cultivation matrix, keep suitable ventilation and air humidity, when temperature is suitable, the seedling seedling-slowing stage is two weeks, recover growth after the 3rd week, transplanting survival rate is greater than 95%.
container for above-mentioned cultivation: the bottle of 650ml is divided into 25 zones, and the jar of 370ml evenly is divided into 16 zones.Region height is 2.5-3 cm.According to circumstances, for interregional material exchange, on interlayer, can design the circular aperture that 4-9 diameter is 0.1-0.15 cm.
culture vessel, its profile is conventional various blake bottles, while being mainly making, in its bottom, uses the material identical with bottle to make corresponding tool aperture and separates, and has not only limited the growth scope of root but also do not hinder the favourable material exchange of growth between each seedling root.
the present invention is not only applicable to iron sheet stone solution, also is applicable to the various stems of noble dendrobium that in group training factorial seedling growth, root system easily is wound around, pocket is blue, state is blue, hybrid orchid.
the technology of the present invention has the following advantages:
1, in blake bottle, the seedling of different subregions can be cultivated the long period and look more strong, take subregion during bottle outlet as flat clip goes out, and not there will be the situation that root system is pulled apart, stem fractures, leaf is injured.
2, improve the efficiency of bottle outlet, facilitated the cleaning of root system, reduced the amount of labour used.
3, the long 2.5-3cm of vertical that is of root system conveniently cultivates.
4, the seedling damage is little, and transplanting seedling time reduces, and recovers growth soon, and survival rate is improving greatly.
Claims (7)
1. dendrobium candidum high-quality test-tube plantlet is limit the root cultivation, the pollination capsule of latter 5 months is taken off, sowing is on germination medium, after 3 weeks, visible seed germination produces protocorm, protocorm that will be closeer in 7 to 10 weeks uses the oese subculture in germination medium, in 14 to 19 weeks, will after protocorm or differentiation, with the seedling of 1-3 sheet leaflet, be inoculated in special-purpose bottle and be limit root to cultivate.
2., when the limit root is cultivated, in each zone, field planting seedling number is stem of noble dendrobium class 1-3 seedling, due in its incubation easily the differentiation lateral bud become seedling, so finally can be formed with the test-tube plantlet of growing thickly of 4-6 seedling, this with cultivation in the quantity of every Cong Yaoqiu just in time suitable; Blue each the zonule field planting seedling number of class of pocket is the 1-2 seedling, because the blue class test-tube seedling transplanting of pocket adopts single seedling cultivation more.
3. because seed in a dendrobium candidum fruit reaches nearly 100,000, after planting germination rate is more than 95%, after seed germination protocorm seedling, the seedling quality is higher than artificial fecundation protocorm seedling quality, so as far as possible only with the protocorm or the seedling final singling that after planting form first, if induce Protocorm Multiplication, Subculture times is no more than three times.
4. according to the described culture vessel of right 2, its profile is conventional various blake bottles, while being mainly making, in its bottom, use the material identical with bottle to make corresponding tool aperture and separate, not only limited the growth scope of root but also do not hinder the favourable material exchange of growth between each seedling root.
5. culture vessel is separated: the bottle of 650ml is divided into 25 zones, and the jar of 370ml evenly is divided into 16 zones.
6. separated region interlayer height used is 2.5-3cm.
7. according to circumstances, can design 4-9 diameter on interlayer is 0.1-0.15 cm circular aperture, and the effect of aperture has 2 points, and first facilitates interregional material exchange, second utilizes the difference of aperture level height, can be according to the height of medium in the different adjustment interlayer of cultivating plant.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105145298A (en) * | 2015-07-30 | 2015-12-16 | 黄旭胜 | Dendrobium officinale pot culturing method |
| CN106171956A (en) * | 2016-07-15 | 2016-12-07 | 萍乡市祥国农业科技发展有限公司 | Red bar Herba Dendrobii artificial hybridization purification and rejuvenation propagation method |
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| US3962822A (en) * | 1974-04-08 | 1976-06-15 | Canadian Patents And Development Limited | Planting containers |
| US20100077662A9 (en) * | 2005-06-29 | 2010-04-01 | Ifa Nurseries, Inc. | Device for protecting container-grown seedling root systems and method for its manufacture |
| CN101933456B (en) * | 2009-06-29 | 2012-06-27 | 西双版纳增靓生物科技有限公司 | Method for quickly breeding seedlings of dendrobium officinale capsule |
| CN202285679U (en) * | 2011-10-26 | 2012-07-04 | 郑炜 | Growing zone dividing board |
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2013
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| US3962822A (en) * | 1974-04-08 | 1976-06-15 | Canadian Patents And Development Limited | Planting containers |
| US20100077662A9 (en) * | 2005-06-29 | 2010-04-01 | Ifa Nurseries, Inc. | Device for protecting container-grown seedling root systems and method for its manufacture |
| CN101933456B (en) * | 2009-06-29 | 2012-06-27 | 西双版纳增靓生物科技有限公司 | Method for quickly breeding seedlings of dendrobium officinale capsule |
| CN202285679U (en) * | 2011-10-26 | 2012-07-04 | 郑炜 | Growing zone dividing board |
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| 周丽等: "铁皮石斛胚培养与快速繁殖", 《黔西南民族师范高等专科学校学报》, no. 3, 30 September 2006 (2006-09-30), pages 88 - 90 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105145298A (en) * | 2015-07-30 | 2015-12-16 | 黄旭胜 | Dendrobium officinale pot culturing method |
| CN106171956A (en) * | 2016-07-15 | 2016-12-07 | 萍乡市祥国农业科技发展有限公司 | Red bar Herba Dendrobii artificial hybridization purification and rejuvenation propagation method |
| CN106171956B (en) * | 2016-07-15 | 2018-03-16 | 萍乡市祥国农业科技发展有限公司 | Red bar dendrobium candidum artificial hybridization purification and rejuvenation propagation method |
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