CN107182788A - The propagation method of bletilla seedling - Google Patents

The propagation method of bletilla seedling Download PDF

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Publication number
CN107182788A
CN107182788A CN201710559079.5A CN201710559079A CN107182788A CN 107182788 A CN107182788 A CN 107182788A CN 201710559079 A CN201710559079 A CN 201710559079A CN 107182788 A CN107182788 A CN 107182788A
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Prior art keywords
seedling
bottle
bletilla
culture
hardening
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梁琼
何艳
陈燚格
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Hubei Jingfeng Agriculture Co ltd
Wuhan Botanical Garden of CAS
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Hubei Jingfeng Agriculture Co ltd
Wuhan Botanical Garden of CAS
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Priority to CN201710559079.5A priority Critical patent/CN107182788A/en
Publication of CN107182788A publication Critical patent/CN107182788A/en
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/008—Methods for regeneration to complete plants
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/001—Culture apparatus for tissue culture

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  • Life Sciences & Earth Sciences (AREA)
  • Developmental Biology & Embryology (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Botany (AREA)
  • Environmental Sciences (AREA)
  • Cultivation Of Plants (AREA)

Abstract

The invention discloses a kind of propagation method of bletilla seedling, belong to Chinese medicine planting technology field.This method is：The breeding of seedling uses the type of rearing of tissue culture propagation；1. the collection of seed；2. culture medium is selected；3. nursery：The tissue cultures program of induction → induced bud differentiation → root induction → greenhouse hardening is sprouted using seed；4. bottle seedling is emerged；5. bottle seedling takes off bottle and processing；6. seedling cultivation step is tamed；7. domestication seedling is emerged.Compared with the prior art, the invention provides a set of bletilla sapling multiplication method, it is to avoid the big shortcoming of stem tuber breeding consumption kind amount, improves the survival rate of bletilla tissue-cultured seedling；Seedling cultivation step is tamed there is provided a set of bletilla, the survival rate of domestication seedling is improved；Specification bletilla culture medium prescription ratio, bottle seedling and domestication seedling are emerged standard, efficiently solve the problem of technology and standard lack in large-scale production.

Description

The propagation method of bletilla seedling
Technical field
The invention belongs to Chinese medicine planting technology field, more particularly to a kind of propagation method of bletilla seedling.
Background technology
Bletilla [Bletilla striata(Thunb. ex A. Murray) Rchb. f.] belong to for many years for orchid family bletilla Raw herbaceous plant, the high 18-60cm of plant；Pseudobulb oblate spheroid, annulus as upper mask water chestnut, rich viscosity；Stem is sturdy, rigidness； 4-6 pieces of leaf, long and narrow circular or lanceolar, long 8-29cm, wide 1.5-4cm, tip is tapering, and base portion, which is received, narrow into sheath and embraces stem；Inflorescence Have 3-10 flower, often not branch or extremely rare branch；Rachis is more or less tortuous in " it " shape；Petal length of a film round shape drapes over one's shoulders pin Shape, long 2-2.5cm is bloomed and withered and fallen often；Hua great, aubergine or pink；Sepal and petal are closely isometric, long and narrow circle, long 25- 30mm, wide 6-8mm, the anxious point of tip；Petal is slightly wider compared with sepal；Lip is slightly shorter compared with sepal and petal, obovate elliptic, long 23- 28mm, white band aubergine, has purple arteries and veins；5 typhloscle pieces of mask on lip disk, extend the nearly top of middle sliver, only in from base portion It is wavy above sliver；Stamen 18-20mm of column length, column has narrow wing, slightly bow；The month at florescence 4-5.
Bletilla Chang Yesheng covers thick grass or sylvan life wetland in hills, low mountain stream side and the moon, is distributed on the south China Qinling Mountains The ground such as provinces and regions and Japan；Be used as medicine with stem tuber, with tonifying lung hemostasis, detumescence and promoting granulation the effects such as, while be also widely used in medicine, The industries such as beauty, coating and printing and dyeing；In recent years, because demand is big, artificial wild bletilla is by serious private digging coyoting, resource Amount is drastically reduced, and has been included in world's endangered plant protection species, and by《Chinese Plants Red Data Book-rare extinction plants》 1st is included, while being also written into《Endangered species of wild fauna and flora international trade pact》（CITES）Protected species.
In actual production, artificial bletilla cultivation is chronically at the original of " harvest-reserve seed for planting-cultivated again on the spot on the spot on the spot " Recurrent state, bletilla quality is degenerated year by year.In recent years, due to bletilla industry development demand, bletilla seedling tissue culture technology starts emerging Rise and progressively turn into the important sources that bletilla seedling is supplied, but the bletilla tissue-cultured seedling death rate is higher, the presence of invalid seedling phenomenon, with And production cycle longer grade generates certain restriction to alleviating the extensive industrialization plantation of bletilla using tissue-cultured seedling, optimizes bletilla group Training technology, promotes high quality seedling supply, particularly significant by what is developed in a healthy way to bletilla industry.
CN201410666249.6 discloses " a kind of scale high yield cultivating method of bletilla striata ", main numerous including seedling Grow, hardening, field-transplanting, growing season management, the management in growth period next year and harvesting.Meanwhile, by microbial technique and eco-species Planting technology is combined, and can be secreted using effective microorganisms(EM) and be synthesized substantial amounts of organic acid, amino acid, enzyme, active hormones, antioxidative enzyme Alto, quickly promotes the growth of plant, has decomposed the residual of medicine, accelerate photosynthesis and the metabolism of plant, shorten The cycle of plantation and the yield for improving the bletilla striata.
CN20151067431.7 discloses " a kind of high yield cultivating method of the nuisanceless bletilla striata ", main to utilize orchid family root nodule The beneficial symbiosis bacterial nutrient solution of bacterium or Orchid Mycorrhizae bacterium after artificial culture carries out that battalion is made after breeding fermentation with fertile soil Soil is supported, after Nutrition Soil is mixed with the soil in kind planting ditch, the field for carrying out tissue-cultured seedling is transplanted, and then it is new to be effectively promoted the bletilla striata The generation of root and the hypertrophy of stem tuber, while completely avoid the generation of pest and disease damage using EM insect prevention liquid.
CN201410364717.4 discloses a kind of " bletilla striata tissue culture method ", mainly accurate by environment disinfected, explant Standby, culture medium preparation, sowing, culturing room's culture, greenhouse hardening, sylvan life hardening, the nine step compositions in transplanting crop field and harvest.Its It is mainly characterized by compost and is divided into bottom compost and upper strata compost, bottom plants producing for material：Take the humic in pine forests Soil, addition farm manure and phosphoric acid man and phosphate fertilizer so that farm manure and potassium phosphate and phosphate fertilizer mixing retting form fermentation fertilizer for one month As bottom compost, culture plate installs after 3 centimeters of bottom composts coconut shell flour being layered on above bottom compost thick 3 centimeters i.e. For compost.
CN201510733897.3 discloses " a kind of method of bletilla striata tissue culture ", the main preparation including explant, Culture and transplant agar 5g/L, white sugar 205g/L, 6- benzyl amino in three steps, its culture medium that explant is put into culture medium Purine is 1.5mg/L, and methyl α-naphthyl acetate is 0.1mg/L；Condition of culture is intensity of illumination 1600-2000lux, and light application time is 15h/d, Cultivation temperature is 24-28 DEG C.
CN201610436028.9 discloses " a kind of bletilla striata tissue culture and rapid propagation method and utilization ", mainly by the sterilizing of explant Method, bletilla seed asepsis sprouting, embryo form and embryo be prominent to break in the seed coat and process observation, the tissue-culturing rapid propagation of the bletilla striata and uses TTC methods Determine seed vitality.Activated carbon 0.2g/L, agar 7.0g/L, Ph5.8, carbon source addition are added in using MS as minimal medium For（30 g/L、60g/L）, it is high with the method fast, proliferation rate that sprouts, while improving transplanting survival rate.
CN201611244428.6 discloses " a kind of bletilla tissue culture seedlings transplanting acclimatization technology ", mainly by the way that sandy soil are used Cauldron is made popular to more than 80 DEG C to kill parasitic ovum, and the sandy soil after cooling are sprinkled into grow to the high bletilla striata groups of 5cm or so Train in seedling bottle, then culture 15-30d in tissue culture room is to complete domestication again, this mode can protect children without cleaning tissue-cultured seedling Seedling root, is also saved artificial.
The content of the invention
The purpose of the present invention, which is that, provides a kind of propagation method of bletilla high quality seedling, effectively improves bletilla seedling survival Rate；This method has simple and practical, cost low and manages convenient advantage, it is adaptable to which large-scale promotion, is a kind of production cycle The bletilla sapling multiplication method that short, breeding potential is high and survival rate is high.
To achieve these goals, the present invention is adopted the following technical scheme that, specifically includes following steps.
The propagation method of bletilla seedling uses the type of rearing of tissue culture propagation；
1. the collection of seed
A, provenance selection, B, the selection of Fruit pod, C, harvesting, D, preservation；
2. culture medium is selected
A, seed sprout inducing culture, B, bletilla differential medium and protocorm induction medium, C, Rooting and hardening-off culture base；
3. nursery
The tissue cultures program of induction → induced bud differentiation → root induction → greenhouse hardening is sprouted using seed；
A, seed sprout induction, B, differentiation and proliferation culture, C, Rooting and hardening-off culture, D, greenhouse hardening；
4. bottle seedling is emerged
5. bottle seedling takes off bottle and processing
A, hardening, B, seedling taking, wash seedling and seedling separation at C, D, sterilization, and E, dry in the air seedling；
6. seedling cultivation step is tamed
A, the warm canopy construction of domestication, B, seedbed are arranged, C, soil configuration, D, lid shading screen, E, cultivation seedling, F, moisture control, G, insect pest Preventing and treating, H, weeding；
7. domestication seedling is emerged
Tame the standard of emerging of seedling.
Compared with prior art, the present invention has the advantages that：
1. a set of bletilla sapling multiplication method is provided（Tissue culture）, it is to avoid the big shortcoming of stem tuber breeding consumption kind amount, simultaneously Bletilla seed is sprouted into induction, the condition of culture of differentiation and proliferation and strong plantlets and rootage has carried out perfect respectively, improves bletilla group Train the survival rate of seedling；
2. provide a set of bletilla and tame seedling cultivation step, and propose the domestication canopy construction of domestication seedling, soil and configure, plant at seedling A whole set of perfect implantation methods such as reason, moisture control and pest and disease damage processing, improve the survival rate of domestication seedling；
3. specification bletilla culture medium prescription ratio, bottle seedling and domestication seedling are emerged standard, efficiently solve skill in large-scale production The problem of art and standard lack.
Brief description of the drawings
Fig. 1 .1 are that bottle seedling is emerged（In bottle）,
Fig. 1 .2 are that bottle seedling is emerged（Bottle is outer）；
Fig. 2 .1 are that the warm canopy of domestication builds one of picture,
Fig. 2 .2 are that the warm canopy of domestication builds the two of picture,
Fig. 2 .3 are that the warm canopy of domestication builds the three of picture；
Fig. 3 is that domestication Wen Peng seedbeds are arranged；
Fig. 4 .1 are the warm canopy soil configurations of domestication（In bag）,
Fig. 4 .2 are the warm canopy soil configurations of domestication（Bag is outer）；
Fig. 5 .1：Domestication seedling is emerged（In basin）,
Fig. 5 .2：Domestication seedling is emerged（Outside basin）.
Embodiment
Describe in detail with reference to the accompanying drawings and examples：
The breeding of seedling uses the type of rearing of tissue culture propagation；
1. the collection of seed
A, provenance selection：Selection 3 years or the plot of more than 3 years planting seasons, florescence growth selection key is strong, tuber growth compared with Fast plant, rejects plant flower that is sick and weak and opening variegated flower, retains seed collecting plant flower；
B, Fruit pod selection：The Fruit pod for selecting completely filled fruit, maturity high, maturity is advisable with the micro- yellowish-brown of surface color, shallow Not enough, dark brown then post-mature, the Fruit pod split can not be used for green then maturity；
C, harvesting：Fruit pod is gently cut with scissors, is eluted with water, natural airing；
D, preservation：By the Fruit pod vial or plastic bottle packing after airing, it is to avoid extruding rupture, -20 DEG C of Cord bloods；
2. culture medium is selected
A, seed sprout inducing culture
Unit：mg/L；
A great number of elements：NH4NO3(1650), KNO3(1900), CaCl2•2H2O(440）, MgSO4•7H2O (370), KH2PO4 (170)；
Trace element：KI (0.83), H3BO3(6.2), MnSO4•4H2O (22.3), ZnSO4•7H2O (8.6), Na2MoO4•2H2O (0.25), CuSO4•5H2O (0.025), CoCl2•6H2O (0.025)；
Molysite：FeSO4•7H2O (27.8), Na2-EDTA•2H2O(37.3)；
Organic matter：Inositol (100), nicotinic acid (0.5), puridoxine hydrochloride (0.5), thiamine hydrochloride (0.1), glycine (2.0)；
Additive：Sucrose (5%), pH value (5.5-5.8), agar powder（0.4%）, peptone（0.1% peptone）；
B, differential medium and protocorm induction medium
Unit：mg/L；
A great number of elements：NH4NO3(1650), KNO3(1900), CaCl2•2H2O(440）, MgSO4•7H2O (370), KH2PO4 (170)；
Trace element：KI (0.83), H3BO3(6.2), MnSO4•4H2O (22.3), ZnSO4•7H2O (8.6), Na2MoO4•2H2O (0.25), CuSO4•5H2O (0.025), CoCl2•6H2O(0.025)；
Molysite：FeSO4•7H2O (27.8), Na2-EDTA•2H2O(37.3)；
Organic matter：Inositol (100), nicotinic acid (0.5), puridoxine hydrochloride（Vitamin B6）(0.5), thiamine hydrochloride（Vitamin B1） (0.1), glycine (2.0)；
Hormone：6-BA (0.5), NAA (0.5)；
Additive：Sucrose (5%), pH value (5.5-5.8), carbon dust (0.05%), agar powder（0.6%）；
C：Rooting and hardening-off culture base
Unit：mg/L；
A great number of elements：H4NO3(1650), KNO3(1900), CaCl2•2H2O (440), MgSO4•7H2O (370), KH2PO4(170)；
Trace element：KI (0.83), H3BO3(6.2), MnSO4•4H2O (22.3), ZnSO4•7H2O (8.6), Na2MoO4•2H2O (0.25), CuSO4•5H2O (0.025), CoCl2•6H2O(0.025)；
Molysite： FeSO4•7H2O (27.8), Na2-EDTA•2H2O(37.3)；
Organic matter：Inositol (100), nicotinic acid (0.5), puridoxine hydrochloride（Vitamin B6）(0.5), thiamine hydrochloride (0.1), sweet Propylhomoserin (2.0)；
Additive：Sucrose (3%), pH value (5.5-5.8), carbon dust (0.05%), agar powder（0.6%）,（30-40g/L murphy juices）；
3. nursery
The tissue cultures program of induction → induced bud differentiation → root induction → greenhouse hardening is sprouted using seed；
A, seed sprout induction
The ripe bletilla capsule collected is scrubbed in bromogeramine solution and removes dirt, puts and 0.1% mercuric chloride is used on superclean bench 10min is sterilized, then with aseptic water washing 3-5 time, by it with after the filter paper suck dry moisture sterilized, fruit top is cut with scalpel Portion, then longitudinally splitted from middle part of fruit, by seed uniform broadcasting to the culture medium prepared, note light seeding, it is too close to be unfavorable for Later stage cultivates；It is subsequently placed under conditions of 28 DEG C of room temperature, intensity of illumination 1500-2000lux, daily light application time 12h and is trained Support；
B, differentiation and proliferation culture
Fiber differentiation 40d bletilla seedling is transferred on differentiation strong seedling culture base in time, in 30 DEG C of room temperature, intensity of illumination Under the conditions of 1500-2000lux, daily light application time 12h, 60d is cultivated, the material of culture of rootage is used as；
C, Rooting and hardening-off culture
Differentiation seedling is transferred on Rooting and hardening-off culture base, in 30-35 DEG C of room temperature, intensity of illumination 1500-2000lux, daily Continue to cultivate under the conditions of light application time 15h, after a week, seedling is mushroomed out, and pseudobulb also increases therewith, false squama after 60d Stem is ripe, height of seedling 3-5cm, blade 3-4 pieces；
D, greenhouse hardening
The good bletilla seedling band bottle of culture of rootage is placed in greenhouse, 15-20d, the shading of plastic house 75% are adapted at room temperature The shading screen of rate, then opens bottle cap and continues hardening 3-5d；
4. bottle seedling is emerged（See Fig. 1 .1-1.2）
The bottle outlet standard of bottle seedling：Old leaf and yellow leaf ratio 5%-20%, plant height 5-10cm, plant fresh weight are not less than 0.2g, the number of blade >=3, leaf long 2cm-5cm, leaf width 0.5cm, the cm of pseudobulb diameter >=0.3；Radical >=2.
5. bottle seedling takes off bottle and processing
A, hardening：Bottle seedling reaches and greenhouse is first put into after cultivation base, and even bottle is placed directly in native table, the screening that one layer 75% of plastic house Screened negative, first opens bottle cap hardening 3d before transplanting, contribute to bottle seedling to adapt to external environment；
B, seedling taking：It is careful during seedling taking from bottle, it can gently be taken out with the small hook of tweezers or iron wire, rhizome seedling should not be hurt （Note：If easy sticker glass dregs in small, broken bits are difficult to clean on broken bottle seedling taking, seedling, easily hand is scratched when planting seedling）；Seedling is washed after taking out with clear water The culture medium of net root residual, notices that seedling must not exceed 20min in clear water, when washing seedling based on rinsing, avoids hindering small as far as possible Seedling；
C, wash seedling and seedling separation：Seedling is washed while seedling separation, during seedling separation it is noted that：Due in tissue culture bottle the culture environment of high-moisture and Limited growing space causes tissue-cultured seedling root tender and crisp easily broken and is intertwined, to avoid injuring seedling, more difficult separated during seedling separation Should not separate by force, several plants with planting, after domestication 2-3 months, seedling can nature separate, play after seedling, domestication seedling root pine It is soft, it is easily separated；
D, sterilization：Sterilized in the Bravo or carbendazim decoction of 800 times of the immersion at once of washed seedling, note only needing root immersion to disappear In venom, and it is immersed in the time in thimerosal and must not exceed 15 seconds.
E, dry in the air seedling：Seedling pendulum after sterilization is placed on shady place on hardboard or newspaper and put, and puts the summary of 30min or so moisture It is dry to transplant；
6. seedling cultivation step is tamed
A, the warm canopy construction of domestication：The warm canopy of domestication selects GP-C825 type assembled galvanized steel pipe greenhouses, a width of 8m of greenhouse, length 40m, can suitably be adjusted according to specific landform；Greenhouse is built using " vertical four card " structure, i.e., one longitudinal tie adds four cards Groove；The integral layout of greenhouse considers south-north direction first, considers further that and is laid out along contour；It is specifically shown in Fig. 2 .1- Fig. 2 .3；
B, seedbed are arranged：5 width are arranged for 1.2m in the warm canopy of domestication, thick 10cm furrow, 6 depth are 20cm, and width is 30cm trench；It is specifically shown in Fig. 3；
C, soil configuration：Soil property is sandy loam, by soil deep ploughing several times, smashes the bulky grain in ground, bulk diameter is no more than 3cm；50% fine earth, 40% sawdust, 10% wood shavings are uniformly mixed, thick 10cm or so seedbed is ultimately formed；It is specifically shown in figure 4.1- Fig. 4 .2；
D, lid shading screen：Bletilla bottle seedling or domestication seedling, which were just transplanted in the 3-5 behind crop field months, is unable to strong illumination, therefore bottle seedling Shade net is got out before transplanting；One layer 75% of shade net must be covered by transplanting in first 3 months, can optionally be gone after 3 months Fall shade net；
E, cultivation seedling：Plant before seedling and spread phoxim by the amount of every mu of 7 bags, every bag 500 grams of granules on moisture in the soil face, bottle seedling dense planting is tamed, Seeding row spacing is 8-10cm, and 0.07 hectare about plants ten thousand plants of 16-18；Bottle seedling field planting is appropriate to the occasion shallow unsuitable deep, root is buried, pseudobulb is deep About 0.5-1cm in soil, gently with fortifield village, unsuitable finger pressure；The deep ditches of -5cm can be drawn on seedbed, seedling is drained into ditch Interior, property blinding, had both improved plantation efficiency again, was not easy to hurt root；
F, moisture control：No more than 1h it is to pour field planting water after transplanting, field planting water should not be irrigated once, can be divided 2-3 times and be irrigated；Then Covered, in order to avoid solar glare hinders seedling, optionally rewatered every 2d or so immediately with one layer 75% of shade net；Next note seeing Examine table soil and keep moistening, now seedling surrounding soil environment will be based on moistening；20d or so rewaters once after transplanting, At this moment it is just basicly stable into the seedling survived, water is suitably controlled after 20d, to promote to give birth to new root, sprouting is sent out, and appropriate arid is favourable Go deep into soil layer in new root and draw nutrition；Without watering after 30d, upper soll layer moistening is kept；Such as rainy season set, it should be noted that see Blade and the disease incidence of root are examined, such as note abnormalities to be handled in time；
G, pest control：After bottle seedling is transplanted, due to environment moistening, snail, slug easily occur, and " snail is net " or " spiral shell are spread in advance Good fortune "（Effective ingredient is the insecticide of the methaldehyde）；The more difficult preventing and treating of insect pest is cutworm and mole cricket, every mu of use 90% Trichlorphon 200g stirs the rice bran or wheat bran 10kg of perfume, is sprinkling upon field trapping；
H, weeding：Bletilla bottle seedling must note crop smothering during taming, because bletilla plant is also small, cannot keep under control weeds, therefore rainy season comes Face it is noted that uprooting weed, it is to avoid farmland running to weeds；
Tame successfully within 3-5 months after transplantation of seedlings, you can transplant planting；
Note：Bottle seedling is transplanted can not apply fertilizer in latter month, because seedling is also weak, easily produce salting-out phenomenon（It is commonly called as burn seedlings）, two Foliar fertilizer, or Water soluble fertilizer can be suitably sprayed after individual month, preferably the Water soluble fertilizer containing trace element；First tested again during fertilising big Area is applied, in order to avoid burn seedlings.
7. domestication seedling is emerged（It is specifically shown in Fig. 5 .1-5.2）
Tame seedling bottle outlet standard：Plant height >=18cm, plant fresh weight is not less than 2.0g, the number of blade >=3, leaf length >=14cm, and leaf width >= 1.5cm, pseudobulb diameter >=0.5cm；Radical >=7.

Claims (8)

1. a kind of propagation method of bletilla seedling, it is characterised in that：
The breeding of seedling uses the type of rearing of tissue culture propagation；
1. the collection of seed
A, provenance selection, B, the selection of Fruit pod, C, harvesting, D, preservation；
2. culture medium is selected
A, seed sprout inducing culture, B, bletilla differential medium and protocorm induction medium, C, Rooting and hardening-off culture base；
3. nursery
The tissue cultures program of induction → induced bud differentiation → root induction → greenhouse hardening is sprouted using seed；
A, seed sprout induction, B, differentiation and proliferation culture, C, Rooting and hardening-off culture, D, greenhouse hardening；
4. bottle seedling is emerged
5. bottle seedling takes off bottle and processing
A, hardening, B, seedling taking, wash seedling and seedling separation at C, D, sterilization, and E, dry in the air seedling；
6. seedling cultivation step is tamed
A, the warm canopy construction of domestication, B, seedbed are arranged, C, soil configuration, D, lid shading screen, E, cultivation seedling, F, moisture control, G, insect pest Preventing and treating, H, weeding；
7. domestication seedling is emerged
Tame the standard of emerging of seedling.
2. a kind of propagation method of bletilla seedling as described in claim 1, it is characterised in that：
The collection of described step 1. seed：
A, provenance selection：Selection 3 years or the plot of more than 3 years planting seasons, florescence growth selection key is strong, tuber growth compared with Fast plant, rejects plant flower that is sick and weak and opening variegated flower, retains seed collecting plant flower；
B, Fruit pod selection：The Fruit pod for selecting completely filled fruit, maturity high, maturity is advisable with the micro- yellowish-brown of surface color, shallow Not enough, dark brown then post-mature, the Fruit pod split can not be used for green then maturity；
C, harvesting：Fruit pod is gently cut with scissors, is eluted with water, natural airing；
D, preservation：By the Fruit pod vial or plastic bottle packing after airing, it is to avoid extruding rupture, -20 DEG C of Cord bloods.
3. a kind of propagation method of bletilla seedling as described in claim 1, it is characterised in that：
Described step 2. select by culture medium：
A, seed sprout inducing culture
Unit：mg/L；
A great number of elements：NH4NO3(1650), KNO3(1900), CaCl2•2H2O(440）, MgSO4•7H2O (370), KH2PO4 (170)；
Trace element：KI (0.83), H3BO3(6.2), MnSO4•4H2O (22.3), ZnSO4•7H2O (8.6), Na2MoO4•2H2O (0.25), CuSO4•5H2O (0.025), CoCl2•6H2O (0.025)；
Molysite：FeSO4•7H2O (27.8), Na2-EDTA•2H2O(37.3)；
Organic matter：Inositol (100), nicotinic acid (0.5), puridoxine hydrochloride (0.5), thiamine hydrochloride (0.1), glycine (2.0)；
Additive：Sucrose (5%), pH value (5.5-5.8), agar powder（0.4%）, peptone（0.1% peptone）；
B, differential medium and protocorm induction medium
Unit：mg/L；
A great number of elements：NH4NO3(1650), KNO3(1900), CaCl2•2H2O(440）, MgSO4•7H2O (370), KH2PO4 (170)；
Trace element：KI (0.83), H3BO3(6.2), MnSO4•4H2O (22.3), ZnSO4•7H2O (8.6), Na2MoO4•2H2O (0.25), CuSO4•5H2O (0.025), CoCl2•6H2O(0.025)；
Molysite：FeSO4•7H2O (27.8), Na2-EDTA•2H2O(37.3)；
Organic matter：Inositol (100), nicotinic acid (0.5), puridoxine hydrochloride（Vitamin B6）(0.5), thiamine hydrochloride（Vitamin B1） (0.1), glycine (2.0)；
Hormone：6-BA (0.5), NAA (0.5)；
Additive：Sucrose (5%), pH value (5.5-5.8), carbon dust (0.05%), agar powder（0.6%）；
C：Rooting and hardening-off culture base
Unit：mg/L；
A great number of elements：H4NO3(1650), KNO3(1900), CaCl2•2H2O (440), MgSO4•7H2O (370), KH2PO4(170)；
Trace element：KI (0.83), H3BO3(6.2), MnSO4•4H2O (22.3), ZnSO4•7H2O (8.6), Na2MoO4•2H2O (0.25), CuSO4•5H2O (0.025), CoCl2•6H2O(0.025)；
Molysite： FeSO4•7H2O (27.8), Na2-EDTA•2H2O(37.3)；
Organic matter：Inositol (100), nicotinic acid (0.5), puridoxine hydrochloride（Vitamin B6）(0.5), thiamine hydrochloride (0.1), sweet Propylhomoserin (2.0)；
Additive：Sucrose (3%), pH value (5.5-5.8), carbon dust (0.05%), agar powder（0.6%）,（30-40g/L murphy juices）.
4. a kind of propagation method of bletilla seedling as described in claim 1, it is characterised in that：
The 3. nursery of described step：
The tissue cultures program of induction → induced bud differentiation → root induction → greenhouse hardening is sprouted using seed；
A, seed sprout induction
The ripe bletilla capsule collected is scrubbed in bromogeramine solution and removes dirt, puts and 0.1% mercuric chloride is used on superclean bench 10min is sterilized, then with aseptic water washing 3-5 time, by it with after the filter paper suck dry moisture sterilized, fruit top is cut with scalpel Portion, then longitudinally splitted from middle part of fruit, by seed uniform broadcasting to the culture medium prepared, note light seeding, it is too close to be unfavorable for Later stage cultivates；It is subsequently placed under conditions of 28 DEG C of room temperature, intensity of illumination 1500-2000lux, daily light application time 12h and is trained Support；
B, differentiation and proliferation culture
Fiber differentiation 40d bletilla seedling is transferred on differentiation strong seedling culture base in time, in 30 DEG C of room temperature, intensity of illumination Under the conditions of 1500-2000lux, daily light application time 12h, 60d is cultivated, the material of culture of rootage is used as；
C, Rooting and hardening-off culture
Differentiation seedling is transferred on Rooting and hardening-off culture base, in 30-35 DEG C of room temperature, intensity of illumination 1500-2000lux, daily Continue to cultivate under the conditions of light application time 15h, after a week, seedling is mushroomed out, and pseudobulb also increases therewith, false squama after 60d Stem is ripe, height of seedling 3-5cm, blade 3-4 pieces；
D, greenhouse hardening
The good bletilla seedling band bottle of culture of rootage is placed in greenhouse, 15-20d, the shading of plastic house 75% are adapted at room temperature The shading screen of rate, then opens bottle cap and continues hardening 3-5d.
5. a kind of propagation method of bletilla seedling as described in claim 1, it is characterised in that：
Described step 4. emerge by bottle seedling：
The bottle outlet standard of bottle seedling：Old leaf and yellow leaf ratio 5%-20%, plant height 5-10cm, plant fresh weight are not less than 0.2g, the number of blade >=3, leaf long 2cm-5cm, leaf width 0.5cm, the cm of pseudobulb diameter >=0.3；Radical >=2.
6. a kind of propagation method of bletilla seedling as described in claim 1, it is characterised in that：
5. bottle seedling takes off bottle and processing to described step：
A, hardening：Bottle seedling reaches and greenhouse is first put into after cultivation base, and even bottle is placed directly in native table, the screening that one layer 75% of plastic house Screened negative, first opens bottle cap hardening 3d before transplanting, contribute to bottle seedling to adapt to external environment；
B, seedling taking：It is careful during seedling taking from bottle, it can gently be taken out with the small hook of tweezers or iron wire, rhizome seedling should not be hurt （Note：If easy sticker glass dregs in small, broken bits are difficult to clean on broken bottle seedling taking, seedling, easily hand is scratched when planting seedling）；Seedling is washed after taking out with clear water The culture medium of net root residual, notices that seedling must not exceed 20min in clear water, when washing seedling based on rinsing, avoids hindering small as far as possible Seedling；
C, wash seedling and seedling separation：Seedling is washed while seedling separation, during seedling separation it is noted that：Due in tissue culture bottle the culture environment of high-moisture and Limited growing space causes tissue-cultured seedling root tender and crisp easily broken and is intertwined, to avoid injuring seedling, more difficult separated during seedling separation Should not separate by force, several plants with planting, after domestication 2-3 months, seedling can nature separate, play after seedling, domestication seedling root pine It is soft, it is easily separated；
D, sterilization：Sterilized in the Bravo or carbendazim decoction of 800 times of the immersion at once of washed seedling, note only needing root immersion to disappear In venom, and it is immersed in the time in thimerosal and must not exceed 15 seconds；
E, dry in the air seedling：Seedling pendulum after sterilization is placed on shady place on hardboard or newspaper and put, put 30min or so moisture slightly it is dry i.e. It can transplant.
7. a kind of propagation method of bletilla seedling as described in claim 1, it is characterised in that：
6. described step tames seedling cultivation step：
A, the warm canopy construction of domestication：The warm canopy of domestication selects GP-C825 type assembled galvanized steel pipe greenhouses, a width of 8m of greenhouse, length 40m, can suitably be adjusted according to specific landform；Greenhouse is built using " vertical four card " structure, i.e., one longitudinal tie adds four cards Groove；The integral layout of greenhouse considers south-north direction first, considers further that and is laid out along contour；
B, seedbed are arranged：5 width are arranged for 1.2m in the warm canopy of domestication, thick 10cm furrow, 6 depth are 20cm, and width is 30cm trench；It is specifically shown in Fig. 3；
C, soil configuration：Soil property is sandy loam, by soil deep ploughing several times, smashes the bulky grain in ground, bulk diameter is no more than 3cm；50% fine earth, 40% sawdust, 10% wood shavings are uniformly mixed, thick 10cm or so seedbed is ultimately formed；It is specifically shown in figure 4.1- Fig. 4 .2；
D, lid shading screen：Bletilla bottle seedling or domestication seedling, which were just transplanted in the 3-5 behind crop field months, is unable to strong illumination, therefore bottle seedling Shade net is got out before transplanting；One layer 75% of shade net must be covered by transplanting in first 3 months, can optionally be gone after 3 months Fall shade net；
E, cultivation seedling：Plant before seedling and spread phoxim by the amount of every mu of 7 bags, every bag 500 grams of granules on moisture in the soil face, bottle seedling dense planting is tamed, Seeding row spacing is 8-10cm, and 0.07 hectare about plants ten thousand plants of 16-18；Bottle seedling field planting is appropriate to the occasion shallow unsuitable deep, root is buried, pseudobulb is deep About 0.5-1cm in soil, gently with fortifield village, unsuitable finger pressure；The deep ditches of -5cm can be drawn on seedbed, seedling is drained into ditch Interior, property blinding, had both improved plantation efficiency again, was not easy to hurt root；
F, moisture control：No more than 1h it is to pour field planting water after transplanting, field planting water should not be irrigated once, can be divided 2-3 times and be irrigated；Then Covered, in order to avoid solar glare hinders seedling, optionally rewatered every 2d or so immediately with one layer 75% of shade net；Next note seeing Examine table soil and keep moistening, now seedling surrounding soil environment will be based on moistening；20d or so rewaters once after transplanting, At this moment it is just basicly stable into the seedling survived, water is suitably controlled after 20d, to promote to give birth to new root, sprouting is sent out, and appropriate arid is favourable Go deep into soil layer in new root and draw nutrition；Without watering after 30d, upper soll layer moistening is kept；Such as rainy season set, it should be noted that see Blade and the disease incidence of root are examined, such as note abnormalities to be handled in time；
G, pest control：After bottle seedling is transplanted, due to environment moistening, snail, slug easily occur, and " snail is net " or " spiral shell are spread in advance Good fortune "；The more difficult preventing and treating of insect pest is cutworm and mole cricket, and every mu stirs fragrant rice bran or wheat bran with 90% trichlorphon 200g 10kg, is sprinkling upon field trapping；
H, weeding：Bletilla bottle seedling must note crop smothering during taming, because bletilla plant is also small, cannot keep under control weeds, therefore rainy season comes Face it is noted that uprooting weed, it is to avoid farmland running to weeds；
Tame successfully within 3-5 months after transplantation of seedlings, you can transplant planting；
Note：Bottle seedling is transplanted can not apply fertilizer in latter month, because seedling is also weak, easily produce salting-out phenomenon, after two months can be with Foliar fertilizer, or Water soluble fertilizer suitably are sprayed, preferably the Water soluble fertilizer containing trace element；Large area is first tested again during fertilising to apply, with Unburned seedling.
8. a kind of propagation method of bletilla seedling as described in claim 1, it is characterised in that：
7. described step is tamed seedling and is emerged：
Tame seedling bottle outlet standard：Plant height >=18cm, plant fresh weight is not less than 2.0g, the number of blade >=3, leaf length >=14cm, and leaf width >= 1.5cm, pseudobulb diameter >=0.5cm；Radical >=7.
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CN108094209A (en) * 2017-12-28 2018-06-01 临沧市云瑞堂生物科技有限公司 A kind of bletilla striata quickly breeds the tissue culture method of seedling
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CN110771443B (en) * 2019-11-22 2021-12-21 阳山县三连阳生态农林开发有限公司 Yangshan bletilla striata single culture medium formula sterile seeding seedling production process
CN113207354A (en) * 2021-06-08 2021-08-06 北京市园林科学研究院 Method for treating seeds of beautiful evening primrose
CN114667857A (en) * 2022-02-08 2022-06-28 贵州省农作物品种资源研究所 Bletilla striata seedling domestication optimization method and domesticated bletilla striata seedlings

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Application publication date: 20170922