CN107041285A - A kind of method of the live seedling of Dendrobidium huoshanness ball stem - Google Patents
A kind of method of the live seedling of Dendrobidium huoshanness ball stem Download PDFInfo
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- CN107041285A CN107041285A CN201710066494.7A CN201710066494A CN107041285A CN 107041285 A CN107041285 A CN 107041285A CN 201710066494 A CN201710066494 A CN 201710066494A CN 107041285 A CN107041285 A CN 107041285A
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- seedbed
- dendrobidium huoshanness
- ball stem
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
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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
- A01H4/001—Culture apparatus for tissue culture
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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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
- A01G24/22—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing plant material
- A01G24/23—Wood, e.g. wood chips or sawdust
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Abstract
The present invention relates to a kind of method of the live seedling of Dendrobidium huoshanness ball stem, comprise the following steps：Sow configuration, the preparation of nutrient solution, the selection of Dendrobidium huoshanness Fruit pod and the sterilization of culture medium, aseptic seeding, seed sprout culture, greenhouse construction, seedbed construction, matrix is prepared and sterilization, the preparation of ball stem suspension, broadcast sowing, the control of gentle light, control, the management of fertilizer and disease, the transplanting of moisture and humidity, the advantage of the invention is that：A kind of advantage of the method for live seedling of Dendrobidium huoshanness ball stem that the present invention is provided is：Reduce the input of tissue culture, eliminate hardening, the process of domestication in traditional handicraft, simplify the production process of nursery, reduce production cost, the survival rate of seedling cultivation is added, the survival rate of the kind transplantation of seedlings obtained by this method reduces cultivation risk up to 100%, the threshold of nursery is low, workable.
Description
Technical field
The present invention relates to domestication of plants technical field, more particularly to a kind of side of the live seedling of Dendrobidium huoshanness ball stem
Method.
Background technology
Dendrobidium huoshanness (scientific name:Dendrobium huoshanense C. Z. Tang et S. J. Cheng) it is commonly called as rice
Dry measure used in former times, is the herbaceous plant of orchid family Dendrobium.Because the compositions such as the alkaloid contained by it, Thick many candies, amino acid, trace element have by force
The effects such as improving eyesight of kidney voiceless sound, liver protection shield throat, nourishing Yin and clearing heat, nourishing the stomach to improve the production of body fluid, suppression growth of tumour cell, strengthen immunity, always
It is considered as first of nine big " celestial grass ".
Under wild state, Dendrobidium huoshanness growth is very slow, wherein extremely tiny, only one simple embryo and individual layer are thin
The aliform kind skin of born of the same parents is constituted, no endosperm, in its natural state, few Germination And Seedling, because excessive felling is utilized, growth is delayed
Slowly, the deterioration of ecological environment and the influence of the low reason of breeding potential, Dendrobidium huoshanness wild resource are endangered.
The method for obtaining Dendrobidium huoshanness regeneration plant by the method for Plant Tissue Breeding has two：（1）Use Dendrobidium huoshanness
Aseptic seeding is carried out after ripe seed disinfection, produced after bud induction, stem induction and root induction, last strong seedling culture and plants
Seedling；（2）Protocorm or Multiple Buds are intended by suitable explant induced synthesis, constantly rised in value（Produce certain amount）,
Then bud induction, stem induction and root induction are carried out, seedling is produced after finally carrying out strong seedling culture.
Tissue culture is related to culture matrix manufacturing, sterile access explant, sterile turn point of protocorm, differentiation during this
Sterile turn point of bud, sterile turn point of seedling, middle sterile turn point of seedling, sterile turn point of seedlings, culturing room management etc. step, be thus related to
The recruitment and management of seedling separation personnel, the construction of culture medium atelier, the construction in sterile working workshop, the construction of culturing room；And
The recruitment and management of related personnel, intelligent greenhouse and the construction for taming greenhouse are further related to during hardening, domestication.To sum up institute
State, require that enterprise there will be sizable input in human and material resources, financial resources in terms of prior art, thereby result in entreprise cost
Greatly, weight is born.
With continuing to develop for Dendrobidium huoshanness industrialized scale, the demand to Dendrobidium huoshanness seedling also increases constantly
Plus, the Dendrobidium huoshanness seedling of low-cost and high-quality is development trend.
The content of the invention
In view of this, it is necessary to which a kind of method of the live seedling of Dendrobidium huoshanness ball stem is provided.
The present invention is achieved in that a kind of method of the live seedling of Dendrobidium huoshanness ball stem, comprises the following steps：
Step one：Sow the configuration of culture medium.
Sowing culture medium includes formula as below：450 ~ 550mg/L of potassium nitrate, 200 ~ 300mg/L of ammonium sulfate, magnesium sulfate 200 ~
300mg/L, 200 ~ 300mg/L of potassium dihydrogen phosphate, 600 ~ 800mg/L of calcium nitrate, 0.81 ~ 0.85mg/L of KI, boric acid 6.0 ~
6.4mg/L, 22.1 ~ 22.5mg/L of manganese sulfate, 8.2 ~ 9.0mg/L of zinc sulfate, 0.2 ~ 0.3mg/L of sodium molybdate, copper sulphate 0.02 ~
0.03mg/L, 0.02 ~ 0.03mg/L of cobalt chloride, 80 ~ 120mg/L of inositol, 0.2 ~ 0.8mg/L of nicotinic acid, 1 ~ 3mg/L of glycine, salt
Allithiamine element 0.1mg/L, 0.2 ~ 0.8mg/L of puridoxine hydrochloride, 37.1 ~ 37.5mg/L of disodium ethylene diamine tetraacetate, ferrous sulfate
26.8 ~ 28.8mg/L, 2,4- dichlorphenoxyacetic acids, 0.1 ~ 0.3mg/L, 0.4 ~ 0.8mg/L of 6- benzyl purines, methyl α-naphthyl acetate 0.1mg/
L, gibberellin 0.2mg/L, sucrose 30g/L and agar 4g/L, by agar after high-temperature digestion, the medicine configured is fallen one by one
Enter, temperature maintains 55 ~ 65 DEG C and dispensed, then by 121 DEG C of moist heat sterilizations, a length of 20min during sterilizing.
Step 2：The preparation of nutrient solution.
Nutrient solution includes formula as below：450 ~ 550mg/L of potassium nitrate, 200 ~ 300mg/L of ammonium sulfate, magnesium sulfate 200 ~
300mg/L, 200 ~ 300mg/L of potassium dihydrogen phosphate, 600 ~ 800mg/L of calcium nitrate, 0.81 ~ 0.85mg/L of KI, boric acid 6.0 ~
6.4mg/L, 22.1 ~ 22.5mg/L of manganese sulfate, 8.2 ~ 9.0mg/L of zinc sulfate, 0.2 ~ 0.3mg/L of sodium molybdate, copper sulphate 0.02 ~
0.03mg/L, 0.02 ~ 0.03mg/L of cobalt chloride, 80 ~ 120mg/L of inositol, 0.2 ~ 0.8mg/L of nicotinic acid, 1 ~ 3mg/L of glycine, salt
Allithiamine element 0.1mg/L, 0.2 ~ 0.8mg/L of puridoxine hydrochloride, 37.1 ~ 37.5mg/L of disodium ethylene diamine tetraacetate, ferrous sulfate
26.8 ~ 28.8mg/L, 2,4- dichlorphenoxyacetic acids, 0.3 ~ 0.7mg/L, 6- benzyl purines 0.1mg/L, 0.6 ~ 1mg/L of methyl α-naphthyl acetate and
0.6 ~ 1mg/L of cycocel, is directly poured into medicine with purified water one by one, can be used after constant volume, sterilizing cooling, the reason for sterilizing
It is to try to be infected by miscellaneous bacteria when reduction ball stem is live.
Step 3：The selection and sterilization of Dendrobidium huoshanness Fruit pod.
Selection warming Dendrobidium huoshanness fruit ripe then, surface disease-free spot, pityriasis simplex enter the Dendrobidium huoshanness Fruit pod of harvesting
Row manual cleanup, removes the limb having Fruit pod surface and withered part more, and is rinsed repeatedly with clear water, is then placed in 75%
In alcohol untill flooding, 30min is soaked, is then placed in sterile inoculation disk, is dried on aseptic operating platform, it is standby.
Step 4：Aseptic seeding.
On sterile behaviour's platform, 1ml is extracted from seed suspension liquid with the liquid-transfering gun after sterilizing, is put into sowing culture medium,
Bottle cap is covered slowly to rock, until the uniform suspension in bottle be laid in media surface untill, be then placed in culture frame in.
Step 5：Seed sprouts culture.
After sowing terminates, seed-bearing culture medium will be put and be placed on progress under conditions of illumination 2000lux, 25 ~ 29 DEG C of temperature
Culture, incubation time be 2 months, you can see green expand the seed sprouted, the germination rate of seed more than 95%, than
General tissue culture formula improves 20% or so to the germination rate of seed.
Step 6：Greenhouse is built.
Using common steelframe booth, canopy high request is not less than 3 meters, above plastic covering film and sunshade rate reach 80 ﹪ with
On sunshade net, for rainwater-proof and stop sunray.
Step 7：Seedbed is built.
Due to being heavy seeding, seedbed need not be too many, therefore uses simple seedbed, but must make somebody a mere figurehead 45 ~ 55cm, and
With certain bearing capacity, beneficial to matrix storage and bottom ventilation, 1-1.5 meters of bedside, length is defined by canopy length, then by sponge
It is laid in that the seedbed made is long, the thickness of sponge is 8 ~ 12 centimetres, and beneficial to the humidity for maintaining seedbed, sponge will pass through permanganic acid
Potassium soaking disinfection.
Step 8：Matrix is prepared and sterilized.
By pine bark, rapeseed dregs by weight proportion 9：1 mixing, the rotten thoroughly rear screen cloth for crossing 3 mesh of fermentation, is soaked with potassium permanganate
24 hours, matrix is taken out, dry and be placed on after state of not dripping above the sponge seedbed made, thickness is no less than 5cm, so
After water, make matrix and seedbed completely drench.
Step 9：The preparation of ball stem suspension.
On desinfection chamber superclean bench face, in the culture medium bottle that sterile nutrient solution is poured into sprouting, then by ball
Stem and nutrient solution are poured out together, are placed on the malicious containers for future use that disappeared.
Step 10：Broadcast sowing.
Start to broadcast sowing after the Clear and Bright after outside air temperature turns to warm up, the Dendrobidium huoshanness ball stem after dilution is put into watering pot
In, rock uniform, be sprinkled upon on ready seedbed, note wanting Bian Sabian to rock.
Step 11：The control of gentle light.
After broadcasting sowing in 2 months, temperature of shed is controlled at 26 ~ 30 DEG C, most appropriate with 28 DEG C, and the first month broadcasted sowing hides
Light processing, can 20 centimeters of places are vacantly above the seedbed plus one layer of brushed fabric plays a part of insulation, controls wet and control light, hidden with double-deck
Screened postive controls light intensity, and illumination, illumination not more than 20000lux are gradually increased after budding.
Step 12：The control of moisture and humidity.
Latter every two days are broadcasted sowing with time nutrient solution of atomizers spray, needed for first makes ball stem have growth above the seedbed
Growth regulatory substance required for sufficient nutrition and differentiation, second makes seedbed holding surface moistening, prevents ball stem to be dehydrated, air
Humidity maintains 80 ﹪ or so.
Step 13：The management of fertilizer and disease.
Dendrobidium huoshanness ball stem sows later every other day sprinkling nutrient solution, and once, maintenance Dendrobidium huoshanness germinating growth must
The nutrition supply of palpus, until untill extracting leaf out.
After Dendrobidium huoshanness extracts two panels leaf out, the flower piece foliar fertilizer for spraying a 0.5-1 ‰ in every 15 days, daily disease
It is less, by spraying the thiophanate methyl of 750 times every month in growth period.
Periodically carried out disinfection week about to seedling raising greenhouse, using fumigant, such as protect mushroom king.
Step 14：Transplant.
The spring of Second Year, by possessed complete root, stem, leaf stem of noble dendrobium seedling pull up, by 4 ~ 6 plants one clump, 10 cm x
10 centimetres of ranks are away from being transplanted.
Further, in the step one, sowing culture medium includes formula as below：Potassium nitrate 500mg/L, ammonium sulfate
250mg/L, magnesium sulfate 250mg/L, potassium dihydrogen phosphate 250mg/L, calcium nitrate 700mg/L, KI 0.83mg/L, boric acid
6.2mg/L, manganese sulfate 22.3mg/L, zinc sulfate 8.6mg/L, sodium molybdate 0.25mg/L, copper sulphate 0.025mg/L, cobalt chloride
0.025mg/L, inositol 100mg/L, nicotinic acid 0.5mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride
0.5mg/L, disodium ethylene diamine tetraacetate 37.3mg/L, ferrous sulfate 27.8mg/L, 2,4- dichlorphenoxyacetic acids 0.2mg/L, 6-
Benzyl purine 0.6mg/L, methyl α-naphthyl acetate 0.1mg/L, gibberellin 0.2mg/L, sucrose 30g/L and agar 4g/L.
Further, in the step 2, nutrient solution includes formula as below：Potassium nitrate 500mg/L, ammonium sulfate 250mg/
L, magnesium sulfate 250mg/L, potassium dihydrogen phosphate 250mg/L, calcium nitrate 700mg/L, KI 0.83mg/L, boric acid 6.2mg/L, sulphur
Sour manganese 22.3mg/L, zinc sulfate 8.6mg/L, sodium molybdate 0.25mg/L, copper sulphate 0.025mg/L, cobalt chloride 0.025mg/L, flesh
Alcohol 100mg/L, nicotinic acid 0.5mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride 0.5mg/L, ethylenediamine
Tetraacethyl disodium 37.3mg/L, ferrous sulfate 27.8mg/L, 2,4- dichlorphenoxyacetic acid 0.5mg/L, 6- benzyl purines 0.1mg/
L, methyl α-naphthyl acetate 0.8mg/L and cycocel 0.8mg/L.
A kind of advantage of the method for live seedling of Dendrobidium huoshanness ball stem that the present invention is provided is：
1st, the input of tissue culture is reduced, hardening, the process of domestication in traditional handicraft is eliminated, simplifies the production process of nursery,
Reduce production cost.
2nd, the survival rate of seedling cultivation is added, the survival rate of the kind transplantation of seedlings obtained by this method subtracts up to 100%
Risk is cultivated less.
3rd, the threshold of nursery is low, workable.
Dendrobidium huoshanness high quality seed directly can be sprouted to form ball stem by this method by aseptic seeding, by using nutrition
Liquid is directly broadcast on the seedbed through handling well to the dilution of ball stem, by the training orientation of science, is moved again after direct seedling
Plant, entreprise cost can be effectively reduced, and seed planting percent is up to the survival rate 100% transplanted after more than 80%, seedling.
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
Embodiment：
A kind of method of live seedling of Dendrobidium huoshanness ball stem, comprises the following steps：
Step one：Sow the configuration of culture medium.
Sowing culture medium includes formula as below：Potassium nitrate 500mg/L, ammonium sulfate 250mg/L, magnesium sulfate 250mg/L, phosphoric acid
Potassium dihydrogen 250mg/L, calcium nitrate 700mg/L, KI 0.83mg/L, boric acid 6.2mg/L, manganese sulfate 22.3mg/L, zinc sulfate
8.6mg/L, sodium molybdate 0.25mg/L, copper sulphate 0.025mg/L, cobalt chloride 0.025mg/L, inositol 100mg/L, nicotinic acid 0.5mg/
L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride 0.5mg/L, disodium ethylene diamine tetraacetate 37.3mg/L, sulphur
Sour ferrous iron 27.8mg/L, 2,4- dichlorphenoxyacetic acid 0.2mg/L, 6- benzyl purines 0.6mg/L, methyl α-naphthyl acetate 0.1mg/L, gibberellin
0.2mg/L, sucrose 30g/L and agar 4g/L, by agar after high-temperature digestion, the medicine configured is poured into one by one, temperature dimension
Hold and dispensed at 55 ~ 65 DEG C, then by 121 DEG C of moist heat sterilizations, a length of 20min during sterilizing.
Step 2：The preparation of nutrient solution.
Nutrient solution includes formula as below：Potassium nitrate 500mg/L, ammonium sulfate 250mg/L, magnesium sulfate 250mg/L, biphosphate
Potassium 250mg/L, calcium nitrate 700mg/L, KI 0.83mg/L, boric acid 6.2mg/L, manganese sulfate 22.3mg/L, zinc sulfate
8.6mg/L, sodium molybdate 0.25mg/L, copper sulphate 0.025mg/L, cobalt chloride 0.025mg/L, inositol 100mg/L, nicotinic acid 0.5mg/
L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride 0.5mg/L, disodium ethylene diamine tetraacetate 37.3mg/L, sulphur
Sour ferrous iron 27.8mg/L, 2,4- dichlorphenoxyacetic acid 0.5mg/L, 6- benzyl purines 0.1mg/L, methyl α-naphthyl acetate 0.8mg/L and short strengthen
Plain 0.8mg/L, is directly poured into medicine with purified water one by one, be can be used after constant volume, sterilizing cooling, is to try to the reason for sterilizing
Reduction ball stem is infected when live by miscellaneous bacteria；
Step 3：The selection and sterilization of Dendrobidium huoshanness Fruit pod.
Selection warming Dendrobidium huoshanness fruit ripe then, surface disease-free spot, pityriasis simplex enter the Dendrobidium huoshanness Fruit pod of harvesting
Row manual cleanup, removes the limb having Fruit pod surface and withered part more, and is rinsed repeatedly with clear water, is then placed in 75%
In alcohol untill flooding, 30min is soaked, is then placed in sterile inoculation disk, is dried on aseptic operating platform, it is standby.
Step 4：Aseptic seeding.
On sterile behaviour's platform, 1ml is extracted from seed suspension liquid with the liquid-transfering gun after sterilizing, is put into sowing culture medium,
Bottle cap is covered slowly to rock, until the uniform suspension in bottle be laid in media surface untill, be then placed in culture frame in.
Step 5：Seed sprouts culture.
After sowing terminates, seed-bearing culture medium will be put and be placed on progress under conditions of illumination 2000lux, 25 ~ 29 DEG C of temperature
Culture, incubation time be 2 months, you can see green expand the seed sprouted, the germination rate of seed more than 95%, than
General tissue culture formula improves 20% or so to the germination rate of seed.
Step 6：Greenhouse is built.
Using common steelframe booth, canopy high request is not less than 3 meters, above plastic covering film and sunshade rate reach 80 ﹪ with
On sunshade net, for rainwater-proof and stop sunray.
Step 7：Seedbed is built.
Due to being heavy seeding, seedbed need not be too many, therefore uses simple seedbed, but must make somebody a mere figurehead 45 ~ 55cm, and
With certain bearing capacity, beneficial to matrix storage and bottom ventilation, 1-1.5 meters of bedside, length is defined by canopy length, then by sponge
It is laid in that the seedbed made is long, the thickness of sponge is 8 ~ 12 centimetres, and beneficial to the humidity for maintaining seedbed, sponge will pass through permanganic acid
Potassium soaking disinfection.
Step 8：Matrix is prepared and sterilized.
By pine bark, rapeseed dregs by weight proportion 9：1 mixing, the rotten thoroughly rear screen cloth for crossing 3 mesh of fermentation, is soaked with potassium permanganate
24 hours, matrix is taken out, dry and be placed on after state of not dripping above the sponge seedbed made, thickness is no less than 5cm, so
After water, make matrix and seedbed completely drench.
Step 9：The preparation of ball stem suspension.
On desinfection chamber superclean bench face, in the culture medium bottle that sterile nutrient solution is poured into sprouting, then by ball
Stem and nutrient solution are poured out together, are placed on the malicious containers for future use that disappeared.
Step 10：Broadcast sowing.
Start to broadcast sowing after the Clear and Bright after outside air temperature turns to warm up, the Dendrobidium huoshanness ball stem after dilution is put into watering pot
In, rock uniform, be sprinkled upon on ready seedbed, note wanting Bian Sabian to rock.
Step 11：The control of gentle light.
After broadcasting sowing in 2 months, temperature of shed is controlled at 26 ~ 30 DEG C, most appropriate with 28 DEG C, and the first month broadcasted sowing hides
Light processing, can 20 centimeters of places are vacantly above the seedbed plus one layer of brushed fabric plays a part of insulation, controls wet and control light, hidden with double-deck
Screened postive controls light intensity, and illumination, illumination not more than 20000lux are gradually increased after budding.
Step 12：The control of moisture and humidity.
Latter every two days are broadcasted sowing with time nutrient solution of atomizers spray, needed for first makes ball stem have growth above the seedbed
Growth regulatory substance required for sufficient nutrition and differentiation, second makes seedbed holding surface moistening, prevents ball stem to be dehydrated, air
Humidity maintains 80 ﹪ or so.
Step 13：The management of fertilizer and disease.
Dendrobidium huoshanness ball stem sows later every other day sprinkling nutrient solution, and once, maintenance Dendrobidium huoshanness germinating growth must
The nutrition supply of palpus, until untill extracting leaf out.
After Dendrobidium huoshanness extracts two panels leaf out, the flower piece foliar fertilizer for spraying a 0.5-1 ‰ in every 15 days, daily disease
It is less, by spraying the thiophanate methyl of 750 times every month in growth period.
Periodically carried out disinfection week about to seedling raising greenhouse, using fumigant, such as protect mushroom king.
Step 14：Transplant.
The spring of Second Year, by possessed complete root, stem, leaf stem of noble dendrobium seedling pull up, by 4 ~ 6 plants one clump, 10 cm x
10 centimetres of ranks are away from being transplanted.
A kind of advantage of the method for live seedling of Dendrobidium huoshanness ball stem that the present invention is provided is：
1st, the input of tissue culture is reduced, hardening, the process of domestication in traditional handicraft is eliminated, simplifies the production process of nursery,
Reduce production cost.
2nd, the survival rate of seedling cultivation is added, the survival rate of the kind transplantation of seedlings obtained by this method subtracts up to 100%
Risk is cultivated less.
3rd, the threshold of nursery is low, workable.
Dendrobidium huoshanness high quality seed directly can be sprouted to form ball stem by this method by aseptic seeding, by using nutrition
Liquid is directly broadcast on the seedbed through handling well to the dilution of ball stem, by the training orientation of science, is moved again after direct seedling
Plant, entreprise cost can be effectively reduced, and seed planting percent is up to the survival rate 100% transplanted after more than 80%, seedling.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
Any modifications, equivalent substitutions and improvements made within refreshing and principle etc., should be included in the scope of the protection.
Claims (3)
1. a kind of method of the live seedling of Dendrobidium huoshanness ball stem, it is characterised in that comprise the following steps：
Step one：Sow the configuration of culture medium；
Sowing culture medium includes formula as below：450 ~ 550mg/L of potassium nitrate, 200 ~ 300mg/L of ammonium sulfate, magnesium sulfate 200 ~
300mg/L, 200 ~ 300mg/L of potassium dihydrogen phosphate, 600 ~ 800mg/L of calcium nitrate, 0.81 ~ 0.85mg/L of KI, boric acid 6.0 ~
6.4mg/L, 22.1 ~ 22.5mg/L of manganese sulfate, 8.2 ~ 9.0mg/L of zinc sulfate, 0.2 ~ 0.3mg/L of sodium molybdate, copper sulphate 0.02 ~
0.03mg/L, 0.02 ~ 0.03mg/L of cobalt chloride, 80 ~ 120mg/L of inositol, 0.2 ~ 0.8mg/L of nicotinic acid, 1 ~ 3mg/L of glycine, salt
Allithiamine element 0.1mg/L, 0.2 ~ 0.8mg/L of puridoxine hydrochloride, 37.1 ~ 37.5mg/L of disodium ethylene diamine tetraacetate, ferrous sulfate
26.8 ~ 28.8mg/L, 2,4- dichlorphenoxyacetic acids, 0.1 ~ 0.3mg/L, 0.4 ~ 0.8mg/L of 6- benzyl purines, methyl α-naphthyl acetate 0.1mg/
L, gibberellin 0.2mg/L, sucrose 30g/L and agar 4g/L, by agar after high-temperature digestion, the medicine configured is fallen one by one
Enter, temperature maintains 55 ~ 65 DEG C and dispensed, then by 121 DEG C of moist heat sterilizations, a length of 20min during sterilizing；
Step 2：The preparation of nutrient solution；
Nutrient solution includes formula as below：450 ~ 550mg/L of potassium nitrate, 200 ~ 300mg/L of ammonium sulfate, 200 ~ 300mg/L of magnesium sulfate,
200 ~ 300mg/L of potassium dihydrogen phosphate, 600 ~ 800mg/L of calcium nitrate, 0.81 ~ 0.85mg/L of KI, 6.0 ~ 6.4mg/L of boric acid,
22.1 ~ 22.5mg/L of manganese sulfate, 8.2 ~ 9.0mg/L of zinc sulfate, 0.2 ~ 0.3mg/L of sodium molybdate, 0.02 ~ 0.03mg/L of copper sulphate,
0.02 ~ 0.03mg/L of cobalt chloride, 80 ~ 120mg/L of inositol, 0.2 ~ 0.8mg/L of nicotinic acid, 1 ~ 3mg/L of glycine, thiamine hydrochloride
0.1mg/L, 0.2 ~ 0.8mg/L of puridoxine hydrochloride, 37.1 ~ 37.5mg/L of disodium ethylene diamine tetraacetate, ferrous sulfate 26.8 ~
28.8mg/L, 2,4- dichlorphenoxyacetic acids, 0.3 ~ 0.7mg/L, 6- benzyl purines 0.1mg/L, 0.6 ~ 1mg/L of methyl α-naphthyl acetate and short strong
0.6 ~ 1mg/L of element, is directly poured into medicine with purified water one by one, be can be used after constant volume, sterilizing cooling, is to the greatest extent the reason for sterilizing
Amount reduction ball stem is infected when live by miscellaneous bacteria；
Step 3：The selection and sterilization of Dendrobidium huoshanness Fruit pod；
Selection warming Dendrobidium huoshanness fruit ripe then, the Dendrobidium huoshanness Fruit pod of harvesting is carried out hand by surface disease-free spot, pityriasis simplex
Work is cleared up, the limb having and withered part more on removal Fruit pod the surface, and is rinsed repeatedly with clear water, is then placed in 75% alcohol
In untill flooding, soak 30min, be then placed in sterile inoculation disk, dried on aseptic operating platform, it is standby；
Step 4：Aseptic seeding；
On sterile behaviour's platform, 1ml is extracted from seed suspension liquid with the liquid-transfering gun after sterilizing, is put into sowing culture medium, covers
Bottle cap is slowly rocked, until the uniform suspension in bottle be laid in media surface untill, be then placed in culture frame in；
Step 5：Seed sprouts culture；
After sowing terminates, it will put under conditions of seed-bearing culture medium is placed on illumination 2000lux, 25 ~ 29 DEG C of temperature and be cultivated,
Incubation time is 2 months, you can see that green expands the seed sprouted, the germination rate of seed is more than 95%, than general
Tissue culture is formulated improves 20% or so to the germination rate of seed；
Step 6：Greenhouse is built；
Using common steelframe booth, canopy high request is not less than 3 meters, above plastic covering film and sunshade rate reach 80 more than ﹪'s
Sunshade net, for rainwater-proof and stop sunray；
Step 7：Seedbed is built；
Due to being heavy seeding, seedbed need not be too many, therefore uses simple seedbed, but must make somebody a mere figurehead 45 ~ 55cm, and has
Certain bearing capacity, beneficial to matrix storage and bottom ventilation, 1-1.5 meters of bedside, length is defined by canopy length, and then sponge is tiled
Long in the seedbed made, the thickness of sponge is 8 ~ 12 centimetres, and beneficial to the humidity for maintaining seedbed, sponge will be by potassium permanganate leaching
Bubble sterilization；
Step 8：Matrix is prepared and sterilized；
By pine bark, rapeseed dregs by weight proportion 9：1 mixing, the rotten thoroughly rear screen cloth for crossing 3 mesh of fermentation is small with potassium permanganate immersion 24
When, matrix is taken out, dries and is placed on after state of not dripping above the sponge seedbed made, thickness is no less than 5cm, then pours
Water, makes matrix and seedbed drench completely；
Step 9：The preparation of ball stem suspension；
On desinfection chamber superclean bench face, in the culture medium bottle that sterile nutrient solution is poured into sprouting, then by ball stem and
Nutrient solution is poured out together, is placed on the malicious containers for future use that disappeared；
Step 10：Broadcast sowing；
Start to broadcast sowing after the Clear and Bright after outside air temperature turns to warm up, the Dendrobidium huoshanness ball stem after dilution be put into watering pot,
Rock uniform, be sprinkled upon on ready seedbed, note wanting Bian Sabian to rock；
Step 11：The control of gentle light；
After broadcasting sowing in 2 months, temperature of shed is controlled at 26 ~ 30 DEG C, and most appropriate with 28 DEG C, the first month broadcasted sowing is used at shading
Reason, can 20 centimeters of places are vacantly above the seedbed plus one layer of brushed fabric plays a part of insulation, controls wet and control light, use double-layered sunshade net
Light intensity is controlled, illumination, illumination not more than 20000lux are gradually increased after budding；
Step 12：The control of moisture and humidity；
Broadcast sowing latter every two days makes ball stem have the abundance needed for growing on seedbed with time nutrient solution of atomizers spray, first
Growth regulatory substance required for nutrition and differentiation, second makes seedbed holding surface moistening, prevents ball stem to be dehydrated, air humidity
Maintain 80 ﹪ or so；
Step 13：The management of fertilizer and disease；
Dendrobidium huoshanness ball stem is sowed every other day sprays nutrient solution once later, necessary to maintenance Dendrobidium huoshanness germinating growth
Nutrition supply, until untill extracting leaf out；
When Dendrobidium huoshanness extract out two panels leaf after, the flower piece foliar fertilizer for spraying a 0.5-1 ‰ in every 15 days, daily disease compared with
It is few, by spraying the thiophanate methyl of 750 times every month in growth period；
Periodically carried out disinfection week about to seedling raising greenhouse, using fumigant, such as protect mushroom king；
Step 14：Transplant；
The spring of Second Year, by possessed complete root, stem, leaf stem of noble dendrobium seedling pull up, by 4 ~ 6 plants one clump, 10 10 lis of cm x
The ranks of rice are away from being transplanted.
2. the method for a kind of live seedling of Dendrobidium huoshanness ball stem according to claim 1, it is characterised in that described
In step one, sowing culture medium includes formula as below：Potassium nitrate 500mg/L, ammonium sulfate 250mg/L, magnesium sulfate 250mg/L, phosphorus
Acid dihydride potassium 250mg/L, calcium nitrate 700mg/L, KI 0.83mg/L, boric acid 6.2mg/L, manganese sulfate 22.3mg/L, sulfuric acid
Zinc 8.6mg/L, sodium molybdate 0.25mg/L, copper sulphate 0.025mg/L, cobalt chloride 0.025mg/L, inositol 100mg/L, nicotinic acid
0.5mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride 0.5mg/L, disodium ethylene diamine tetraacetate
37.3mg/L, ferrous sulfate 27.8mg/L, 2,4- dichlorphenoxyacetic acid 0.2mg/L, 6- benzyl purines 0.6mg/L, methyl α-naphthyl acetate
0.1mg/L, gibberellin 0.2mg/L, sucrose 30g/L and agar 4g/L.
3. the method for a kind of live seedling of Dendrobidium huoshanness ball stem according to claim 1, it is characterised in that described
In step 2, nutrient solution includes formula as below：Potassium nitrate 500mg/L, ammonium sulfate 250mg/L, magnesium sulfate 250mg/L, di(2-ethylhexyl)phosphate
Hydrogen potassium 250mg/L, calcium nitrate 700mg/L, KI 0.83mg/L, boric acid 6.2mg/L, manganese sulfate 22.3mg/L, zinc sulfate
8.6mg/L, sodium molybdate 0.25mg/L, copper sulphate 0.025mg/L, cobalt chloride 0.025mg/L, inositol 100mg/L, nicotinic acid 0.5mg/
L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride 0.5mg/L, disodium ethylene diamine tetraacetate 37.3mg/L, sulphur
Sour ferrous iron 27.8mg/L, 2,4- dichlorphenoxyacetic acid 0.5mg/L, 6- benzyl purines 0.1mg/L, methyl α-naphthyl acetate 0.8mg/L and short strengthen
Plain 0.8mg/L.
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| CN108293863A (en) * | 2018-02-11 | 2018-07-20 | 绍兴儒林生物科技有限公司 | A kind of dendrobium candidum ecosystem is from the implantation methods that germinate |
| CN108575626A (en) * | 2018-03-30 | 2018-09-28 | 安徽省蓝渡农业科技有限公司 | The implantation methods of sealwort |
| CN111990254A (en) * | 2020-08-31 | 2020-11-27 | 南京农业大学 | Dendrobium nobile culture medium and application thereof |
| CN114793898A (en) * | 2022-04-22 | 2022-07-29 | 华南农业大学 | Seeding method for dendrobium officinale protocorm |
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| CN103004445A (en) * | 2013-01-06 | 2013-04-03 | 戴亚峰 | Method for directly seedling dendrobe seedlings by using plastic greenhouse |
| CN104542289A (en) * | 2014-12-31 | 2015-04-29 | 安徽康久生物科技有限公司 | Method for performing tissue culture of dendrobium candidum seedlings |
| CN105104209A (en) * | 2015-09-24 | 2015-12-02 | 戴亚峰 | Method for cultivating tissues to obtain seedlings of dendrobium huoshanense at one step and formula of nutrient liquid for method |
| CN105961199A (en) * | 2013-09-16 | 2016-09-28 | 周杰 | Tissue culture method of dendrobium officinale |
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| CN105961199A (en) * | 2013-09-16 | 2016-09-28 | 周杰 | Tissue culture method of dendrobium officinale |
| CN104542289A (en) * | 2014-12-31 | 2015-04-29 | 安徽康久生物科技有限公司 | Method for performing tissue culture of dendrobium candidum seedlings |
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| CN108293863A (en) * | 2018-02-11 | 2018-07-20 | 绍兴儒林生物科技有限公司 | A kind of dendrobium candidum ecosystem is from the implantation methods that germinate |
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| CN111990254A (en) * | 2020-08-31 | 2020-11-27 | 南京农业大学 | Dendrobium nobile culture medium and application thereof |
| CN114793898A (en) * | 2022-04-22 | 2022-07-29 | 华南农业大学 | Seeding method for dendrobium officinale protocorm |
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