CN104145794A - Temporary planting and seedling hardening method of dendrobium tissue culture seedlings - Google Patents
Temporary planting and seedling hardening method of dendrobium tissue culture seedlings Download PDFInfo
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Abstract
The invention discloses a temporary planting and seedling hardening method of dendrobium tissue culture seedlings, and relates to the technical field of dendrobium cultivation. The method includes the following steps that (1) by means of moderate microclimate conditions under protected cultivation frame overhead planting bed, ground under the bed serves as a dendrobium temporary planting seedbed; (2) culture medium main raw materials are fermented; (3) culture media are prepared and put into a seedling hole plate; (4) the dendrobium tissue culture seedlings of the right age are selected for indoor seedling hardening; (5) the tissue culture seedlings are placed on the temporary planting seedbed after temporary planting on the seedling hole plate; (6) management after temporary planting is carried out. After the method is adopted, the temporary planting survival rate of the dendrobium tissue culture seedlings is high, the cultured hole plate seedlings are strong and uniform, seedling survival is easy after planting, growth is quick, the method is suitable for seedlings for dendrobium protected cultivation and bionic cultivation large-scale production, and the utilization rate of equipment can be improved.
Description
Technical field
The invention belongs to dendrobe cultivation technical field, be specifically related to a kind of hardening off method of heeling in of dendrobe tissue culture seedling.
Background technology
The stem of noble dendrobium (Dendrobium) has another name called dendrobium, for the perennial herbaceous plant that grows nonparasitically upon another plant of the orchid family (Orchidaceace) Dendrobium (Dendrobium), it is important medicinal and ornamental plants, the species that belong to < < CITS > > appendix II, original producton location is mainly distributed in Tropical Asian and subtropics, Australia and Pacific Islands, approximately there is kind more than 1500 in the whole world, China approximately has 76 kinds, major part is distributed in southwest, south China, the ground such as Taiwan, nearly wherein there are 40 kinds to there is medical value.The kind of China's stem of noble dendrobium only accounts for global 5% left and right, but its medicinal development and utilization is taken its place in the front ranks of the world, the stem of noble dendrobium is China's tradition rare traditional Chinese medicine, applicating history is long, have nourishing Yin and clearing heat, beneficial stomach promotes the production of body fluid, moisten the lung and relieve the cough, anti-ageing, improve immunity, promoting blood circulation and removing blood stasis, improve cardio and vascular function, the effect of promoting longevity, more famous have a dendrobium candidum (D.officinale), Dendrobidium huoshanness (D.huosanness), HERBA DENDROBII (D.nobile), Dendrobium fimbriatum Hook (D.fimbiratum), Dendrobium Chrysotoxum Lindl (D.chrysotourn) etc., they are all used as medicine with fresh or dry stem.Some medicinal dendrobium kind exports for a long time, enjoys great prestige Southeast Asia and some American-European areas.The stem of noble dendrobium has the title of " soft gold ", " the celestial grass in the human world " in China, be called as in history first of China's " nine immortal grass ", pharyngolaryngitis, gastrointestinal disease, cataract, angiocardiopathy, diabetes and inhibition tumor growth are had to significant curative effect, particularly lung carcinoma cell is had to good inhibitory action, for eliminating the side effect after cancer radiation, chemotherapy and recovering physical efficiency, effect is fairly obvious.So people are growing to the demand of Dendrobium Plants, market demand constantly expands.
Wild Dendrodium is mainly grown in the torrid zone, subtropics virgin forest and similarly warm and moist environment, happiness is shady and cool moistening, foggy microclimate ventilates, avoid high temperature, dry, high light, grow in 900～1500 meters of height above sea level more, 12 ℃～18 ℃ of average temperatures of the whole year, relative moisture 60%～75%, light transmittance 60% left and right, 20 ℃～28 ℃ of season of growth temperature, 9 ℃～12 ℃ of winter temperatures, frostless foggy, in the evergreen broad-leaved forest that annual rainfall is 500～1000 millimeters, often grow nonparasitically upon another plant on half dark and damp thick forest trunk or rock, and normal and liver moss, the association of the unicellular lower eukaryote such as Shi Wei and lichens, in be mainly grown in that lofty mountains and steep hills overhanging cliff and rock crevice, growth conditions requires warm, moistening, ventilate, ventilative, leak.
Although the stem of noble dendrobium can result and fruit and seed amount large, 1 mellow fruit pod contains the seed of up to ten thousand, Natural seed setting rate is very low, for example the Natural seed setting rate of dendrobium candidum only 0.31%.The seed of the stem of noble dendrobium is very tiny, and embryonic development is incomplete, lacks endosperm tissue, needs could sprout with some fungi (blue bacterium) symbiosis under natural conditions, and germination rate is extremely low, for example the seed germination rate less than 5% of dendrobium candidum.Traditional artificial propagation mode is as plant division, cuttage etc., and reproduction speed is slow, and the rate of increase is very low, can not meet the requirement that commercialization is produced far away.
Due to the limitation of the stem of noble dendrobium to the specific (special) requirements of growing environment and distribution, under natural environmental condition, self-reproduction rate is very low, and the growing slowly of the stem of noble dendrobium, so wild Dendrodium quantity is rare, wild resource is very limited.China relies on for a long time excavates wild resource and meets the need of market, owing to excessively excavating and developing, cause Dendrobium Plants heavy damage, cause natural resources day by day rare, the famous and precious stem of noble dendrobium wild resource that the tradition such as dendrobium candidum, Dendrobidium huoshanness, HERBA DENDROBII are used as medicine has been on the verge of exhaustion in China, natural production can not be met the need of market far away, and this has not only seriously restricted the supply of a large amount of relevant medicines, and ecological and bio-safety have been caused to great harm.The < < Chinese Plants Red Data Book > > promulgating in August, 1998 lists 70 kinds of above Dendrobium Sws in protective plant, protected by the conservation of wild plant regulations > > of the < < People's Republic of China (PRC).The protection of reinforcement to Dendrobium Plants, the sustainable use that realizes Dendrobium Plants is extremely urgent, and the method for quickly breeding of the research stem of noble dendrobium and artificial cultivation technique are protection stem of noble dendrobium species and the important channel of meeting the need of market.
From 20 century 70s, the tissue rapid propagation technology of the stem of noble dendrobium and the research of artificial cultivation technique have just been started both at home and abroad.The stem of noble dendrobium is stricter to the requirement of environmental condition, require half warm, moistening, to have sufficient scattered light cloudy half light environment, its growth thermophilic is 15 ℃～30 ℃, 20 ℃ of left and right of optimum growth temperature, at 5 ℃, stop growing below, and seedling is easily endured cold below at 10 ℃.Relative air humidity requires maintenance more than 70%～80%, avoids dry and ponding, particularly when sprouting starts to sprout with new root formation, needs sufficient moisture.Illumination requires shading summer and autumn 70%, shading in winter 30%～50%.Therefore, adopting the facility cultivations such as booth is that stem of noble dendrobium large-scale production is preferably selected.Meanwhile, the stem of noble dendrobium is epiphyte, mainly grows under field conditions (factors) on some high megaphanerophytes dark and damp trunk or limestone, thereby also can simulating natural environment condition carry out Emulational culture under the condition of artificial propagation seedling.Emulational culture refers in the wild original producton location of the stem of noble dendrobium and selects under nearly natural forests, in simulating natural environment condition, the ecosystem environment of stem of noble dendrobium growth carries out tame method, attached master's difference during according to Emulational culture, can be divided into tree cultivation, stone cultivation and 3 kinds of forms of humus soil cultivation.
Along with development and the application of tissue culture technique, the tissue culture technology of the stem of noble dendrobium is mature on the whole at present.Utilize tissue culture technique to breed seedling and become the tame main source of seedling of the current stem of noble dendrobium, by tissue, cultivating the laggard pedestrian's work of amount reproduction group training seedling facility cultivation is the Main Means of meeting the need of market, tissue cultured test-tube seedling successfully carries out field-transplanting and growth, is to realize the key that stem of noble dendrobium industrialization is produced.But because the stem of noble dendrobium requires harsh to growing environment; the industrialization that has many factors still to restrict the stem of noble dendrobium is produced; wherein; owing to organizing culture environment and artificial cultivation environment to exist larger difference; group training seedling need to be adapted to extraneous natural environment comprehensively, by heterotrophism, is transitioned into autotrophy by gnotobasis, and a little less than the resistance because of group training seedling, survival ability is poor; group training transplantation of seedlings survival rate is low is a bottleneck of puzzlement stem of noble dendrobium large-scale production always, is restricting the artificial cultivation scale of dendrobe tissue culture seedling.When particularly the stem of noble dendrobium carries out Emulational culture, survival rate is lower; the survival rate of for example directly cultivating Emulational culture base after Dendrobidium huoshanness group training seedling bottle outlet is about 30% left and right; the transplanting survival rate of group training seedling be lowly always restriction Dendrobidium huoshanness large-scale development principal element (Xu Guangtao. improve the research of Dendrobidium huoshanness Emulational culture survival rate method. Anhui forestry science and technology; 2011,37 (6): 62-64.).Group training transplantation of seedlings survival rate is low not only has very large relation with cultivation matrix, and has very large relation with illumination, temperature, humidity in planting environment.For example, at artificially controlling temperature, control wet beam-connected greenhouse in officinal dendrobium stem plantation survival rate can reach more than 95%, if cultivated in comparatively modern booth, can be without centre heel in hardening link, directly field planting after group training seedling takes out from bottle, also can obtain higher survival rate and higher output.But in the wild during Emulational culture or while cultivating in simple plastic greenhouse, the direct transplanting survival rate of candidum tissue culturing seedling is very low, generally lower than 50%, and poor growth (Zheng Kuanyu, Deng Jun wave, Zhao Hui. the research of dendrobium officinale test-tube plantlet culture technique. Crop Diseases in Yunnan science and technology, 2010 (3): 21-22.; Song Shun, Xu Yi, Li Jingyang. the tissue of dendrobium candidum is cultivated and Fast-propagation progress. Chinese agronomy circular, 2013,29 (33): 286-290.).The research of money galaxy of literary talent etc. is thought, utilize the cultivation matrix that water retention property is good and gas permeability is strong can make the transplanting survival rate of Dendrobidium huoshanness group training seedling improve, and the planting effect of layering matrix is better, the suitable matrix of transplanting is layering matrix: 3/10 bark+top, bottom, 7/10 wood chip, higher (the money galaxy of literary talent of its transplanting survival rate and rudiment number, Zhang Jianxia, Wu Kunlin, Deng. Dendrobidium huoshanness sapling multiplication and cultivation research. tropical and subtropical plant journal, 2013, 21 (3): 240-246.), but this research is used the operation of layering substrate culture more complicated, and transplant the survival rate only 78% of latter 70 days statistics.So, from test-tube plantlet bottle outlet to field production, preferably there is a transition stage of heeling in hardening centre to the stem of noble dendrobium, make group training seedling after having adapted to natural environment again field planting just can obtain good cultivation effect, otherwise can have a strong impact on transplanting survival rate, growth rate and stem of noble dendrobium output.
In order to improve the dendrobe tissue culture seedling adaptability of environment to external world, the bottleneck problem of low this restriction industrialized development of solution group training transplantation of seedlings survival rate, Kong Dedong etc. filter out the matrix formulations of peat+perlite+vermiculite (volume ratio 3 ︰ 1 ︰ 1), application point disk seedling growing technology, make the survival rate of candidum tissue culturing seedling surpass 95%, and seedling growth is healthy and strong, there is promotional value (Kong Dedong, Huang Chongping, Zhou Jianhua, Deng. the optimization screening of dendrobium candidum plug seedling substrate. agricultural science and technology communication, 2010 (8): 70-72.).Xiao Ying etc. select tongue moss ︰ mud charcoal soil ︰ perlite 3 ︰ 1 ︰ 1 to heel in for matrix, make candidum tissue culturing seedling heel in survival rate and reach 95% (transplanting after 60 days), and the growth of stem is (Xiao Ying very significantly, Zhang Yaozhou. the impact that matrix hardening is heeled in dendrobium officinale test-tube plantlet season. Xinyang agricultural higher junior college journal, 2013,23 (2): 93-94.).The key technology that Xu Guangtao improves Dendrobidium huoshanness cultivation survival rate is before Emulational culture, to heel in the hardening of carrying out a period of time in hardening matrix after dendrobe tissue culture seedling bottle outlet.Hardening matrix is with the preparations by a certain percentage such as broken bark bits, meter Xin Shi and aobvious acid northeast peat soil in original producton location, the matrix of this material can not only meet the needs of hardening, but also be conducive to the stem of noble dendrobium seedling hardening phase, is transplanted on the stone matter base of Emulational culture after finishing.Through the hardening phase of 45～60 days, make the cultivation survival rate of Dendrobidium huoshanness from 30% original left and right, bring up to more than 90% (Xu Guangtao. improve the research of Dendrobidium huoshanness Emulational culture survival rate method. Anhui forestry science and technology, 2011,37 (6): 62-64.).The pine bark of 5～10 millimeters of pulverizing, 2～4 millimeters of perlites for Liu Shihui etc., the cultivation matrix mixing according to 4 ︰ 1 ratios, with 50 hole dishes, heel in hardening, candidum tissue culturing seedling stem stalk after domestication is sturdy, leaf look dark green, root system is slightly white, and adaptive capacity and resistance enhancing, transplanting survival rate can reach more than 97% (Liu Shihui, yellow lead maple, Yao Yongkang, Deng. the research of candidum tissue culturing seedling greenhouse acclimatization technology. Agriculture of Anhui science, 2013,41 (17): 7477-7478,7495.).The main contributions of these 4 researchs is, being generally used in the point disk seedling growing technology of grow seedlings of vegetable or cultivation that heeling in technology is applied to the stem of noble dendrobium, the transplanting survival rate of dendrobe tissue culture seedling is significantly improved.Its defect is, all using the primary raw material of peat as matrix for first 3, and the peat resource of China is fewer, and be non-renewable natural resources, although the research of Liu Shihui etc. does not use peat as the primary raw material of matrix, but because of the nutrition of this matrix seldom, heel in and rear fertilizing management is had relatively high expectations, require " thin fertile duty is executed ", not only need repeatedly to spray Liquid Fertilizer, but also to apply particle slow release fertilizer, could help dendrobium candidum to grow up strong and sturdy; Simultaneously, these 4 researchs all need to have special facility to heel in hardening, because the adaptability of group training seedling is poor, to having relatively high expectations of environmental condition, so also higher to the level of the requirement of facility and cultivation management, if heel in the simpler and cruder or extensive management of the facility of hardening, group training seedling to heel in survival rate still lower.Yet, the mode of the current stem of noble dendrobium facility cultivation of China is all generally to adopt built on stilts cultivation bed to cultivate, the flowers of many facility cultivations, particularly some high-grade cymbidium varieties are as Moth orchid etc., also be to adopt built on stilts cultivation bed to cultivate, ground under cultivation bed and Spatial General 6 R are all idle useless, and this is to one of cultivating facility waste greatly.During the substrate culture of stem of noble dendrobium facility, the stem of noble dendrobium seedling General Requirements intensity of illumination of producing cultivation is 8000～15000 luxs, and the stem of noble dendrobium seedling (group training seedling) of heeling in the hardening phase is more suitable when intensity of illumination is 3000～10000 lux, first 2 months intensities of illumination after particularly heeling in can not be too high, and suitable intensity of illumination is generally 3000～7000 luxs.We are 60～80 centimetres utilizing overhead height, width is 1.2～1.5 meters, between bed and bed, leave and while carrying out dendrobe cultivation in the facility in wide 0.5～1 meter of passageway, observe discovery, when meeting intensity of illumination being 8000～15000 lux on built on stilts cultivation bed, the intensity of illumination on the lower ground of bed of built on stilts cultivation bed is in the scope of 3000～10000 luxs, and under relative air humidity bed, space is general higher by 3%～5% than built on stilts cultivation bed superjacent air space, the lower space of the temperature changing amplitude of every day (maximum temperature and minimum temperature poor) bed than built on stilts cultivation bed superjacent air space general low 1 ℃～3 ℃, find, the flowers of many facility cultivations, particularly some high-grade cymbidium varieties are as Moth orchid etc. simultaneously, and the microclimate conditions that the lower microclimate conditions of built on stilts cultivation bed and the above-mentioned stem of noble dendrobium be built on stilts under cultivating bed is substantially the same.Because the stem of noble dendrobium seedling during carrying out hardening vacation requires stricter than seedlings to planting environment, require intensity of illumination lower, Du Genggao shades, relative air humidity is larger, the variation of temperature, humidity is relatively little, above-mentioned built on stilts cultivation bed space and ground below just in time can meet these conditions, is to carry out dendrobe tissue culture seedling to heel in the more satisfactory place of hardening.
Summary of the invention
The object of the present invention is to provide a kind of hardening off method of heeling in of dendrobe tissue culture seedling.The method is heeled in the hardening phase to the requirement of environmental condition and growth and development characteristics according to dendrobe tissue culture seedling, utilize the common built on stilts cultivation bed of stem of noble dendrobium facility cultivation, or be suitable for carrying out the microclimate conditions that the stem of noble dendrobium is heeled in hardening under the bed of the built on stilts cultivation bed of some facility flowers, using ground under bed as temporary seedbed, by preparing suitable cultivation matrix, strengthen heeling in the management of hardening phase, make group training seedling as early as possible by gnotobasis reform of nature environment, resistance and the survival ability of raising group training seedling, guarantee heeling in survival rate and improving transplanting survival rate of dendrobe tissue culture seedling, thereby the bottleneck of low this puzzlement stem of noble dendrobium large-scale production of solution group training transplantation of seedlings survival rate, realize the tame large-scale production of dendrobe tissue culture seedling, increase considerably the supply of the artificial cultivation stem of noble dendrobium, reach this precious species of the protection stem of noble dendrobium and the object that meets the growing market demand.
Dendrobe tissue culture seedling heel in a hardening off method, comprise the following steps:
(1) utilize the lower ground of built on stilts cultivation bed as stem of noble dendrobium temporary seedbed;
(2) fermentation process of cultivation matrix primary raw material:
By pine bark, maple wood sawdust, dry cow dung, be that fermentation process is carried out in 4～5 ︰ 4～5 ︰ 0.5～1 mixing by volume;
(3) preparation of cultivation matrix and seedling culture hole plate sabot;
First the cultivation matrix primary raw material after step (2) fermentation process is processed with carbendazim or thiophanate methyl medicine liquid spray, perlite drains open fire after soaking with clear water, again by cultivation matrix primary raw material and perlite by volume 3～4 ︰ 1 evenly mix, then pack in the hole of seedling culture hole plate;
(4) select of the right age dendrobe tissue culture seedling to carry out indoor hardening adaptation;
(5) after group training seedling is heeled in seedling culture hole plate, put on temporary seedbed;
(6) management after heeling in.
The described built on stilts cultivation bed of step (1) comprises: the built on stilts cultivation bed of stem of noble dendrobium facility cultivation or the built on stilts cultivation bed of flowers facility cultivation.
The described stem of noble dendrobium of step (1) comprises: dendrobium candidum (D.officinale), Dendrobidium huoshanness (D.huosanness), HERBA DENDROBII (D.nobile), Dendrobium fimbriatum Hook (D.fimbiratum) or Dendrobium Chrysotoxum Lindl (D.chrysotourn).
The particle size of cultivation matrix primary raw material pine bark is 0.5～1.2 centimetre.
The fermentation process of step (2) is specially: pine bark first soaks 1 day with clear water, then mix with maple wood sawdust and dry cow dung in proportion, regulate water content to 65%～70%, pile and in heapsly with plastic film, cover and to ferment, yeast phase 2～3 months, between yeast phase, need turning 5～6 times, every 10～15 days, open film turning 1 time, after having fermented, matrix is spread out dry standby.
Step (3) is that first with 800～1000 times of carbendazim or thiophanate methyl medicine liquid spray, to mix the cultivation matrix primary raw material after fermentation process thoroughly water content be 60%～65%, bank out with plastic film and cover 3～5 days, perlite drains open fire with clear water immersion after 1～2 day, again by cultivation matrix primary raw material and perlite by volume 3～4 ︰ 1 evenly mix, then pack in the hole of 50 holes or 32 hole seedling culture hole plates.
The standard of the of the right age dendrobe tissue culture seedling that step (4) is described is: every clump has 3～5 strains, and every strain has 3～5 of roots, long 2～3 centimetres of root, every strain have 4～6, leaf, stem thick 0.3 centimetre above, high 4～7 centimetres.
The described indoor hardening of step (4) adapts to: of the right age group of training seedling moved to from group training chamber and in booth or greenhouse, carry out the adaptation of normal temperature hardening, temperature is controlled at 15 ℃～28 ℃, relative moisture 50%～80%, shading 70% in 1 week, it is the sunshade net that ceiling all standing degree of shading is 70%, after 1 week, by reducing the size of ceiling covered with sunshade net, be adjusted to shading 50%～60% in canopy, keep ventilated in canopy, allow group training seedling from the environment of closed stablity to opening the environment transition changing, the hardening time is 2～4 weeks, with the leaf color jade green of dendrobe tissue culture seedling or dark green, when slightly hardening, the plump sturdy and stem of stem section is advisable, group training seedling bottle outlet opens wide bottleneck for first 2～3 days gradually, first only unscrews bottle cap, and unlimited degree strengthens gradually, bottle cap is all opened in last 1 day, during bottle outlet, group training seedling is taken out from bottle, with clear water, clean and stick to the medium on root, after draining the water, with 1000 times of carbendazim or thiophanate methyl, soak whole strain seedling 10 minutes, the root that then dries in the air at shady and cool ventilation place is waited to heel in half a day after root turns white.
The suitable time of heeling in described in step in said method (5) be mid-March～by the end of May or mid-September～mid-October.
Described in step (5), heel in is the group after indoor hardening adaptation, bottle outlet processing to be trained to seedling heel in ready seedling culture hole plate, take 3～5 individual plants as 1 clump, every cave is heeled in 1 clump, after heeling in, seedling culture hole plate is neatly put on temporary seedbed, with sprayer, on seedling culture hole plate, spray 3000 times of agricultural streptomycins immediately and 1000 times of carbendazim carry out disinfection, make seedling and matrix combine closely simultaneously.
Management after the heeling in of step (6) is: in 1 week after heeling in every day regularly spray clear water 1～2 time, make air humidity keep 80%～90%, do not water; After heeling in 1 week, reduce spraying number of times, make air humidity keep 70%～80%; If matrix is dry after heeling in 2 weeks, with watering can, waters and execute matrix to water content 60%～70%; Heel in after 20 days, within every 1～2 week, spray 1 time foliage fertilizer, the flower of the potassium dihydrogen phosphate with 0.2%～0.3% and 1500～2000 times of liquid the more alternately sprays for No. 1, only executes a kind of foliage fertilizer at every turn, and each amount of application is 30～45 kilograms of every mu of areas; After entering autumn, reduce gradually the number of times water and apply fertilizer, after the Beginning of Winter, stem of noble dendrobium growth stops substantially, and it requires lowly to liquid manure, need not apply fertilizer, and water content of substrate is not less than 30% and does not water, and generally heels in hardening and after 5～12 months, goes out garden field planting.
In said method, the root of dendrobe tissue culture seedling to be heeled in is soaked in the ABT3 root-inducing powder of 100 mg/litre to 15 minutes, and then heels in, to improve, heel in survival rate.
The inventive method need to be implemented in plastic tunnel or attached-greenhouse.The cultivation matrix that the present invention heels in for the stem of noble dendrobium is also the good matrix of dendrobe cultivation.
The present invention has following good effect: the adaptability of dendrobe tissue culture seedling is poor, requirement to planting environment condition is stricter, the requirement Du Genggao that shades, relative air humidity is larger, the variation of environmental condition is relatively little, according to dendrobe tissue culture seedling to the requirement of environmental condition and growth and development characteristics, select the common built on stilts cultivation bed of stem of noble dendrobium facility cultivation, or the lower ground of the bed of the built on stilts cultivation bed of some facility flowers is as temporary seedbed, do not need separately to build special cultivating facility, not only can improve the availability of facility, and can create relatively adapt circumstance condition for dendrobe tissue culture seedling growing after heeling in, thereby can raising group training seedling heel in survival rate.The gas permeability of dendrobe cultivation matrix and moisture capacity are the keys that obtains high transplanting survival rate, and the nutrition of matrix is the limiting factor that after transplant survival, impact is grown, too low or too high growing after being all unfavorable for surviving, the cultivation matrix of this method preparation is loose ventilative, drainage is good, contained fertilizer nutrient is appropriate, water conservation, fertilizer-preserving ability are strong, while heeling in dendrobe tissue culture seedling, survival rate is high, after surviving, grow very fast, and base starting material easily obtains, cheap, do not use the non-renewable natural resourcess such as peat.This method is easy and simple to handle, practical, can make dendrobe tissue culture seedling as early as possible by gnotobasis reform of nature environment, resistance and the survival ability of raising group training seedling, the survival rate of heeling in of group training seedling generally reaches more than 96%, the cave dish seedling robust growth of cultivating, neat and consistent, after field planting, slow seedling is fast, while adopting facility substrate culture, planting survival rates reaches more than 98% (than directly improving more than 15% by the contrast survival rate of group training seedling field planting), the planting survival rates of Emulational culture also can reach more than 92%, than after the Dendrobidium huoshanness group training seedling bottle outlet of report, directly carry out Emulational culture survival rate 30% left and right (Xu Guangtao. improve the research of Dendrobidium huoshanness Emulational culture survival rate method. Anhui forestry science and technology, 2011, 37 (6): 62-64.) raising is very obvious, be applicable to carrying out growing seedlings of stem of noble dendrobium facility cultivation and Emulational culture large-scale production, not only can solve the bottleneck of low this puzzlement stem of noble dendrobium large-scale production of dendrobe tissue culture transplantation of seedlings survival rate, and adopt and to heel in the cave direct field planting of comparable group of training seedling of the dish seedling field planting commodity stem of noble dendrobium of gathering for 1 year ahead of time, can realize the tame large-scale production of dendrobe tissue culture seedling.
Embodiment
There is following embodiment to be described in detail essence of the present invention, and can not limit the present invention.
Embodiment 1
(1) selection of temporary seedbed: the mode of the current stem of noble dendrobium facility cultivation of China is all generally to adopt built on stilts cultivation bed to cultivate, the overhead height of built on stilts cultivation bed is generally 60～80 centimetres, 1.2～1.5 meters of width, the matrix that can load 8～10 cm thicks in bed.The flowers of many facility cultivations, particularly some high-grade cymbidium varieties as Moth orchid etc., are also to adopt built on stilts cultivation bed to cultivate.Because the stem of noble dendrobium seedling during vacation hardening (group training seedling) requires intensity of illumination lower than seedlings, Du Genggao shades, relative air humidity is larger, the variation of temperature, humidity is relatively little, and above-mentioned built on stilts cultivation bed space and ground below just in time can meet these conditions, to carry out dendrobe tissue culture seedling to heel in the more satisfactory place of hardening, so select the lower ground of bed of the built on stilts cultivation bed of stem of noble dendrobium facility cultivation or the built on stilts cultivation bed of some facility flowers as temporary seedbed.
(2) cultivation matrix primary raw material and fermentation process thereof: the cultivation matrix of the stem of noble dendrobium should be take the good raw material of permeability as main, require loose ventilative, draining is good, fertilizer is appropriate, water conservation, fertilizer-preserving ability are strong, within whole vegetative period, be all the time the state that partly becomes thoroughly decomposed, could obtain best upgrowth situation like this.The gas permeability of dendrobe cultivation matrix and moisture capacity are to improve the key of transplanting survival rate, and the nutrition of matrix is the limiting factor of sprouting Germination and growth after transplant survival, the too low or too high Germination and growth that is all unfavorable for surviving rear sprouting.Existing result of study shows, it is primary raw material that more suitable dendrobe cultivation matrix generally be take peat and/or liver moss, and peat is non-renewable resources, and reserves are limited, the destruction that a large amount of exploitations can cause ecotope.Good and cheap in order to obtain, be applicable to local optimum dendrobe cultivation matrix formulations, select local pine bark cheap and easy to get, weed tree sawdust, maple wood sawdust, mushroom bacterium slag, flat mushroom bacterium slag and dry cow dung etc. are primary raw material, be equipped with the perlite being easy to get on market, 15 different matrix formulations have been prepared altogether, through updating of the formulation screening tests of 3 years and matrix formulations, summing up the relatively applicable applicable stem of noble dendrobium heels in the matrix formulations of hardening and is: (pine bark, maple wood sawdust, volume ratio=4～5 ︰ 4～5 ︰ 0.5～1 of dry cow dung), perlitic volume ratio=3～4 ︰ 1.Meanwhile, the spontaneous fermentation processing and the retort sterilization that have contrasted cultivation matrix primary raw material are processed 2 kinds of processing methods, and filtering out fermentation process is practical matrix treatments method.The concrete operations of cultivation matrix primary raw material and fermentation process thereof are: select particle size be pine bark, maple wood sawdust and the dry cow dung of 0.5～1.2 centimetre as the primary raw material of cultivation matrix, the constituent of cultivation matrix primary raw material is that the volume ratio of pine bark, maple wood sawdust, dry cow dung is 4～5 ︰ 4～5 ︰ 0.5～1.Pine bark first soaks 1 day with clear water, then mix with maple wood sawdust and dry cow dung in proportion, regulate water content to 65%～70%, pile heaps high 1.2～1.5 meters, wide 2.5～3.0 meters, with plastic film, cover and ferment, yeast phase is generally 2～3 months, needs turning 5～6 times between yeast phase, generally every 10～15 days, open film turning 1 time, during turning, as found that matrix becomes dry, should suitably add water.When summer high temperature ferments season, yeast phase is shorter, and yeast phase is answered proper extension when low temperature season ferments.After having fermented, matrix is spread out to dry rear pack standby.
(3) preparation of cultivation matrix: with the above-mentioned cultivation matrix primary raw material through fermentation process and perlite by volume 3～4 ︰ 1 be mixed with the cultivation matrix of heeling in hardening.First with 800～1000 times of carbendazim or thiophanate methyl medicine liquid spray, mixing the cultivation matrix primary raw material after fermentation process thoroughly water content is 60%～65%, bank out with plastic film and cover vexed 3～5 days, perlite drains open fire with clear water immersion after 1～2 day, again by cultivation matrix primary raw material and perlite by volume 3～4 ︰ 1 evenly mix, then just can sabot.
(4) medium in tray: seedling culture hole plate can be selected commercially available common plastics seedling culture hole plate, generally selecting specification is long 54 centimetres, wide 25 centimetres, 50 holes of high 5 centimetres or the seedling culture hole plate in 32 holes.During sabot, first the matrix preparing is evenly poured in seedling culture hole plate into shake seedling culture hole plate, allow matrix be filled in the hole of seedling culture hole plate, then with the little plank of 1 cm thick, 50 centimeter length, the matrix compacting on seedling culture hole plate surface is struck off, heel in first 1 day and water the water of revealing the exact details, dendrobe tissue culture seedling to be heeled in.
(5) select of the right age dendrobe tissue culture seedling to carry out indoor hardening adaptation: the stem of noble dendrobium that research institute uses comprises dendrobium candidum (D.officinale), Dendrobidium huoshanness (D.huosanness), HERBA DENDROBII (D.nobile), Dendrobium fimbriatum Hook (D.fimbiratum), Dendrobium Chrysotoxum Lindl (D.chrysotourn) etc.When every clump of dendrobe tissue culture seedling has 3～5 strains, every strain has 3～5 of roots, long 2～3 centimetres of root, 4～6, leaf, stem thick 0.3 centimetre above, high 4～7 centimetres time, by group training seedling from group training chamber move to the booth of planting environment term harmonization or greenhouse in carry out the adaptation of normal temperature hardening.Temperature is controlled at 15 ℃～28 ℃, relative moisture 50%～80%, intensity of illumination 3000～7000 luxs, it is shading 50%～70%, and increase gradually intensity of illumination, and keep ventilated in canopy, allow group training seedling from the environment of closed stablity to opening the environment transition changing, make its environment of reform of nature light scattering and alternating temperature gradually, the hardening time is 2～4 weeks.70%, 1 week backsight weather condition of shading shading 50%～60% in 1 week during concrete operations, is advisable while slightly hardening with the leaf color jade green of dendrobe tissue culture seedling or the plump sturdy and stem of dark green, stem section.Group training seedling bottle outlet opens wide bottleneck for front 2～3 days gradually, and bottle cap is only unscrewed by elder generation, and unlimited degree strengthens gradually, bottle cap is all opened in last 1 day, allows temperature, the damp condition of its reform of nature.During group training seedling bottle outlet, by hand side, clap bottle, make between medium and root loosening, then with finger or tweezers, from bottle limit by group training seedling, press from both sides out, with clear water, wash away gently the medium sticking on root, will wash agar off especially, in order to avoid agar is mouldy, cause rotten, then change clear water and clean 1 time.After draining the water, with 1000 times of carbendazim or thiophanate methyl, soak whole strain seedling 10 minutes, prevent the stem of noble dendrobium seedling butt rot after heeling in.Then the root that dries in the air at shady and cool ventilation place is waited to heel in half a day after root turns white.
(6) heeling in of group training seedling: when temperature is not less than 15 ℃, start to heel in, be generally the suitable period of heeling in mid-March～by the end of May or mid-September～mid-October.While heeling in, conventionally take 3～5 individual plants as 1 clump, every cave is heeled in 1 clump, after heeling in, seedling culture hole plate is neatly put on temporary seedbed, with sprayer, on seedling culture hole plate, sprayed 3000 times of agricultural streptomycins immediately and 1000 times of carbendazim carry out disinfection, make seedling and matrix combine closely simultaneously.In order improving, to heel in survival rate, the root of dendrobe tissue culture seedling to be heeled in can be soaked in the ABT3 root-inducing powder (production of Ai Bidi bio tech ltd, Beijing) of 100 mg/litre to 15 minutes with root induction, then to heel in.
(7) management after heeling in: dendrobe tissue culture seedling is heeled in the phase to the requirement of environmental condition generally: 20 ℃～30 ℃ of temperature daytimes, 10 ℃～20 ℃ of nights, intensity of illumination is 3000～7000 luxs, relative air humidity 60%～80%, usings the lower ground of bed of the built on stilts cultivation bed of stem of noble dendrobium facility cultivation or the built on stilts cultivation bed of some facility flowers and generally can meet these conditions during as temporary seedbed.Because the dendrobe tissue culture seedling of just having heeled in is very tender and lovely, the clear water 1～2 time of will regularly spraying initial several days every days, after heeling in, air humidity maintenance 80%～90% in 1 week, does not water.After heeling in 1 week, plant starts Fa Xingen, can reduce spraying number of times, and air humidity keeps 70%～80%.Matrix keeps alternation of wetting and drying to be conducive to root of hair sprouting, and the water management of matrix should be grasped and parch drenched principle, if the dry available watering can of matrix waters and executes matrix to drench (water content 65% left and right) after heeling in 2 weeks.Heel in after 20 days, within every 1～2 week, spray 1 time foliage fertilizer, the more No. 1 (a kind of plant from imported from America is general balanced fertile for the flower of the potassium dihydrogen phosphate with 0.2%～0.3% and 1500～2000 times of liquid, basic recipe: nitrogen, phosphorus, potassium content are 20-20-20, adds the M-77 trace element combination of world patent) alternately spray.After entering autumn and winter along with the reduction of temperature, stem of noble dendrobium growth of seedling slows down even and stops, the requirement of moisture is also reduced gradually, so should reduce gradually the number of times of water spray and fertilising, in the not obvious lack of water of matrix (water content of substrate lower than 30% time) in the situation that, do not water, generally select fine day 9:00～10:00 in the morning to water for every 15～20 days 1 time winter, after water spray, suitably ventilates.Generally heel in hardening and can go out garden field planting in 5～12 months.
Adopt the hardening off method of heeling in of above-mentioned dendrobe tissue culture seedling, not only can improve the availability of facility, and easy and simple to handle, practical.Group training seedling is heeled in the survival rate average out to 96.7% after 60 days, the cave dish seedling robust growth of cultivating, neat and consistent, after field planting, slow seedling is fast, heel in after the field planting of cave dish seedling the direct field planting of the comparable group of training seedling commodity stem of noble dendrobium of gathering for 1 year ahead of time, the survival rate average out to 98.6% (directly with the average survival of group training seedling field planting contrast be 83.4%) of field planting after 60 days while adopting facility substrate culture, the survival rate of Emulational culture field planting after 60 days on average also can reach 93.5% (in Table 1), than after the Dendrobidium huoshanness group training seedling bottle outlet of report, directly carry out Emulational culture survival rate 30% left and right (Xu Guangtao. improve the research of Dendrobidium huoshanness Emulational culture survival rate method. Anhui forestry science and technology, 2011, 37 (6): 62-64.) improve more than 60%, be applicable to carrying out growing seedlings of stem of noble dendrobium facility cultivation and Emulational culture large-scale production, can solve the bottleneck of low this puzzlement stem of noble dendrobium large-scale production of dendrobe tissue culture transplantation of seedlings survival rate, realize the tame large-scale production of dendrobe tissue culture seedling.
Table 1 the inventive method dendrobe tissue culture seedling heel in survival rate and the planting survival rates of heeling in dish seedling in cave after hardening
Claims (10)
- Dendrobe tissue culture seedling heel in a hardening off method, it is characterized in that, comprise the following steps:(1) utilize the lower ground of built on stilts cultivation bed as stem of noble dendrobium temporary seedbed;(2) fermentation process of cultivation matrix primary raw material:By pine bark, maple wood sawdust, dry cow dung, be that fermentation process is carried out in 4～5 ︰ 4～5 ︰ 0.5～1 mixing by volume;(3) preparation of cultivation matrix and seedling culture hole plate sabot;First the cultivation matrix primary raw material after step (2) fermentation process is processed with carbendazim or thiophanate methyl medicine liquid spray, perlite drains open fire after soaking with clear water, again by cultivation matrix primary raw material and perlite by volume 3～4 ︰ 1 evenly mix, then pack in the hole of seedling culture hole plate;(4) select of the right age dendrobe tissue culture seedling to carry out indoor hardening adaptation;(5) after group training seedling is heeled in seedling culture hole plate, put on temporary seedbed;(6) management after heeling in.
- A kind of dendrobe tissue culture seedling according to claim 1 heel in hardening off method, it is characterized in that, the described built on stilts cultivation bed of step (1) comprises: the built on stilts cultivation bed of stem of noble dendrobium facility cultivation or the built on stilts cultivation bed of flowers facility cultivation.
- A kind of dendrobe tissue culture seedling according to claim 1 heel in hardening off method, it is characterized in that, the particle size of pine bark is 0.5～1.2 centimetre.
- 4. according to the hardening off method of heeling in of a kind of dendrobe tissue culture seedling described in claim 1 or 3, it is characterized in that, the fermentation process of step (2) is specially: pine bark first soaks 1 day with clear water, then mix with maple wood sawdust and dry cow dung in proportion, regulate water content to 65%～70%, pile and in heapsly with plastic film, cover and to ferment, yeast phase 2～3 months, between yeast phase, need turning 5～6 times, every 10～15 days, open film turning 1 time, after having fermented, matrix is spread out dry standby.
- 5. a kind of dendrobe tissue culture seedling according to claim 1 heels in hardening off method, it is characterized in that, step (3) is that first with 800～1000 times of carbendazim or thiophanate methyl medicine liquid spray, to mix the cultivation matrix primary raw material after fermentation process thoroughly water content be 60%～65%, bank out with plastic film and cover 3～5 days, perlite drains open fire with clear water immersion after 1～2 day, again by cultivation matrix primary raw material and perlite by volume 3～4 ︰ 1 evenly mix, then pack in the hole of 50 holes or 32 hole seedling culture hole plates.
- 6. a kind of dendrobe tissue culture seedling according to claim 1 heels in hardening off method, it is characterized in that, the standard of the of the right age dendrobe tissue culture seedling that step (4) is described is: every clump has 3～5 strains, every strain has long 2～3 centimetres of 3～5 of roots, root, every strain have 4～6, leaf, stem thick 0.3 centimetre above, high 4～7 centimetres.
- 7. according to the hardening off method of heeling in of a kind of dendrobe tissue culture seedling described in claim 1 or 6, it is characterized in that, the described indoor hardening of step (4) adapts to: of the right age group of training seedling moved to from group training chamber and in booth or greenhouse, carry out the adaptation of normal temperature hardening, temperature is controlled at 15 ℃～28 ℃, relative moisture 50%～80%, shading 70% in 1 week, it is the sunshade net that ceiling all standing degree of shading is 70%, after 1 week, by reducing the size of ceiling covered with sunshade net, be adjusted to shading 50%～60% in canopy, keep ventilated in canopy, allow group training seedling from the environment of closed stablity to opening the environment transition changing, the hardening time is 2～4 weeks, with the leaf color jade green of dendrobe tissue culture seedling or dark green, when slightly hardening, the plump sturdy and stem of stem section is advisable, group training seedling bottle outlet opens wide bottleneck for first 2～3 days gradually, first only unscrews bottle cap, and unlimited degree strengthens gradually, bottle cap is all opened in last 1 day, during bottle outlet, group training seedling is taken out from bottle, with clear water, clean and stick to the medium on root, after draining the water, with 1000 times of carbendazim or thiophanate methyl, soak whole strain seedling 10 minutes, the root that then dries in the air at shady and cool ventilation place is waited to heel in half a day after root turns white.
- A kind of dendrobe tissue culture seedling according to claim 1 heel in hardening off method, it is characterized in that, the suitable time of heeling in described in step (5) be mid-March～by the end of May or mid-September～mid-October.
- A kind of dendrobe tissue culture seedling according to claim 1 heel in hardening off method, it is characterized in that, the management after the heeling in of step (6) is: after heeling in 1 week every day regularly spray clear water 1～2 time, make air humidity maintenance 80%～90%, do not water; After heeling in 1 week, reduce spraying number of times, make air humidity keep 70%～80%; If matrix is dry after heeling in 2 weeks, with watering can, waters and execute matrix to water content 60%～70%; Heel in after 20 days, within every 1～2 week, spray 1 time foliage fertilizer, the flower of the potassium dihydrogen phosphate with 0.2%～0.3% and 1500～2000 times of liquid the more alternately sprays for No. 1, only executes a kind of foliage fertilizer at every turn, and each amount of application is 30～45 kilograms of every mu of areas; After entering autumn, reduce gradually the number of times water and apply fertilizer, after the Beginning of Winter, stem of noble dendrobium growth stops substantially, and it requires lowly to liquid manure, need not apply fertilizer, and water content of substrate is not less than 30% and does not water, and generally heels in hardening and after 5～12 months, goes out garden field planting.
- 10. a kind of dendrobe tissue culture seedling according to claim 1 heels in hardening off method, it is characterized in that, the root of dendrobe tissue culture seedling to be heeled in is soaked in the ABT3 root-inducing powder of 100 mg/litre to 15 minutes, and then heels in, to improve, heel in survival rate.
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