CN109122302A - A kind of method of sword-leaved cymbidium and Herba Renantherae coccineae generic cross cultivation orchid new varieties - Google Patents
A kind of method of sword-leaved cymbidium and Herba Renantherae coccineae generic cross cultivation orchid new varieties Download PDFInfo
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- CN109122302A CN109122302A CN201811220534.XA CN201811220534A CN109122302A CN 109122302 A CN109122302 A CN 109122302A CN 201811220534 A CN201811220534 A CN 201811220534A CN 109122302 A CN109122302 A CN 109122302A
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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
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
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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
Abstract
The invention discloses a kind of methods that sword-leaved cymbidium and Herba Renantherae coccineae generic cross rapidly and efficiently cultivate orchid new varieties.This method includes that pollen collecting and preservation, artificial pollination and hybridization fruit are cultivated, Hybrids embryo culture development is rhizomes, rhizomes Multiplying culture, rhizomes differentiation culture, adventitious bud rooting seedling culture and at transplantation of seedlings and cultivation.The present invention is using cryo-conservation and the sterile great-hearted pollen of tool solves the bottleneck problem of orchid generic cross flowering asynchronism, the hypogenetic critical issue of inter-genera distant hybridization hybrid embryo is solved using embryo rescue culture technique, it is developed by single hybrid embryo sterile culture to carry out corresponding rhizomes clonal expansion culture after rhizomes, the shortcomings that avoiding conventional hybridization breeding year limit for length, can quickly breeding go out orchid new varieties.
Description
Technical field
The present invention relates to technical field of plant propagation, and in particular to a kind of sword-leaved cymbidium is rapidly and efficiently trained with Herba Renantherae coccineae generic cross
Educate the method for orchid new varieties.
Background technique
Orchid is the rare flower in the fashionable world, is to cultivate most one of wide, the maximum flowers type of sales volume in the world, closely
Year over develop especially rapidly, increased every year with 10% or more speed, at present whole world orchid year the amount of consumption more than 4,000,000,000
Dollar.Holland, Thailand, Singapore, the U.S., South Korea, Japan etc. are the more developed countries of orchid industry, and Thailand is that cattleya is maximum
Exported country.The orchid produced at present is mostly the cattleyas such as iris, dendrobium and Bowring cattleya, and the state orchid with Chinese tradition characteristic is then
It is small.
Sword-leaved cymbidium is that state is blue a kind of, and state orchid is the tradition inscription flower in China, and ornamental values and the economic values are high, are often referred to orchid family
Partially non-hibernating eggs in epidendrum generally comprises sword-leaved cymbidium (Cymbidium ensifolium), Chunlan
(C.goeringii), orchid (C.faberi), Chinese cymbidium (C.sinense), cold orchid (C.kanran), Cymbidium lianpan (C.tortis modeling
Expect alum), 7 kinds of spring sword.The blue simple and elegant delicate fragrance of state, is known as " spending middle gentleman ", appreciates by the refined visitor of scholar, has two in China
More than thousand years cultivation histories, and unique blue culture is formed, state orchid is also all the fashion in country in Southeast Asia.Sword-leaved cymbidium is with a long history,
It is wide in variety, have good ornamental value and economic and cultural value.
Sword-leaved cymbidium (Cymbidium emiflohtm) is ground plant, and pseudobulb ovoid contains within phyllopodium.Leaf 2 to
6 pieces, band shape is glossy, 30~60 centimetres long, 1~2.5 centimetre wide.Hua Ting is issued from pseudobulb base portion, uprightly, generally shorter than
Leaf；Raceme has 3~9 flowers；Spend often it is aromatic, color changes greatly, usually chartreuse and have purple plague purpura；Bending is closely narrow
Oblong or narrow ellipse；The narrow ellipse of petal or narrow ovate-elliptic, it is 1.4~2.5 centimetres long, it is 5~8 centimetres wide, it is close open and flat；
Lip is closely oval, 1.5~2.3 centimetres long, and slightly 3 split.The narrow ellipse of calabash fruit, it is 5~6 centimetres long, it is about 2 centimetres wide.Florescence is usually 6
~October.
Flame Cymbidium (Reanathera) the plant whole world, to grow nonparasitically upon another plant or semiepiphyte, is distributed in the southeast there are about 21 kinds
Asia to Himalaya Region, including South China, China originate in 3 kinds.Wherein a kind of 3 kinds of Herba Renantherae coccineaes of China's production
Ren.Coccinea produce Guangdong, Hainan Region, plant it is tall and big growth it is healthy and vigorous, 2 kinds (Ren.imschootiana and
Ren.sinica Yunnan) is produced.Herba Renantherae coccineae growth is healthy and vigorous, and pattern is gorgeous, spend it is several numerous, it is remote see it is gorgeous such as flame, with very high
Horticultural applications value and Breeding value, be International Flower top grade flower all the fashion in the market, have in the world a large amount of
Fan.
Crossbreeding is to cultivate the effective means of orchid new varieties, and sword-leaved cymbidium is hybridized with Herba Renantherae coccineae, is expected to cultivate
There are the orchid new varieties of more color changes again with fragrance, but the two is not belong to, and the sword-leaved cymbidium flowering naturally phase is 6~10
Month, and the flowering naturally phase of Herba Renantherae coccineae is 3~May, hybridizes and successfully needs to solve flowering asynchronism and distant hybridization compatibility not
Genetic stability, the critical issue of consistency and specificity of high bottleneck problem and new varieties.Currently, domestic existing orchid
The paper report of flower crossbreeding, pollen preservation, embryo rescue etc., but have no that sword-leaved cymbidium and Herba Renantherae coccineae generic cross cultivate new varieties
Method report.
Summary of the invention
It is an object of the invention to develop one kind orchid new varieties that rapidly and efficiently cultivation sword-leaved cymbidium hybridizes with Herba Renantherae coccineae
Method solves the problems, such as that distant hybridization success rate is low in orchid crossbreeding, breeding cycle is long.
The present invention selects to have the high sword-leaved cymbidium of obvious characteristic ornamental value as hybridization female parent, 2~4 after the expansion of maternal flower
Artificial pollination is carried out in it, male parent is then the high fire of the obvious characteristic ornamental value of tool of the tool viability saved using sterile cryogenic
The pollen of flame orchid.The fruit successfully obtained of pollinating is developed into development to aseptic seeding, promotion hybrid embryo is carried out after a certain period of time
For single rhizomes, then its group expanded by the clonal expansion technology of single rhizomes, is transplanted and planted after seedling differentiation
It trains to the plant that blooms, while establishing the vegetative propagation technique system of new lines, obtain new varieties finally by the authorization of new varieties
Certificate, to realize the purpose of the present invention.
The method of the present invention carries out pollen to solve the skills such as cold and rewetting processing before saving orchid pollen, pollination using sterile cryogenic
Art is solved the problems, such as the bottleneck problem of the mildew and orchid generic cross flowering asynchronism when pollen saves, is saved and cultivated using embryo
Technology solves the hypogenetic critical issue of inter-genera distant hybridization hybrid embryo, is root shape by the development of single hybrid embryo sterile culture
The shortcomings that carrying out corresponding rhizomes clonal expansion culture after stem, avoiding conventional hybridization breeding year limit for length, can rapidly and efficiently train
Bring out orchid new varieties.
The present invention is achieved by the following technical programs:
A kind of method that sword-leaved cymbidium rapidly and efficiently cultivates orchid new varieties with Herba Renantherae coccineae generic cross, comprising the following steps:
(1) it pollen collecting and preservation: when male parent Herba Renantherae coccineae single flower petal is just unfolded, is picked out using the picking tool of sterilizing
The anther cap of orchid, then orchid massula is chosen into the container of cleaning sterile with the picking tool of sterilizing and is sealed, it is placed in 4~6
It DEG C is kept in dark place；
(2) artificial pollination and hybridization fruit are cultivated: when maternal sword-leaved cymbidium blooms, the massula that step (1) is saved is in room
Thaw 20~30min under the conditions of temperature, then adds sterile water and carries out rewetting 20~30min of processing, obtains having great-hearted flower
Powder；To have great-hearted pollen pollination in the cultivation on the flower column cap of maternal sword-leaved cymbidium, carrying out hybridization capsule；
(3) Hybrids embryo culture development is rhizomes: when hybridization capsule is developed to 180~200 days, picking hybridizes fruit, warp
Hybridization fruit is cut after disinfection, and powdered hybrid embryo is seeded in embryo rescue culture medium and is carried out sterile culture 80~100 days, is obtained
To rhizomes, the embryo saves every liter of culture medium and contains 160~200mg of ammonium nitrate, 80~100mg of potassium dihydrogen phosphate, potassium chloride
160~200mg, 80~100mg of magnesium sulfate, 80~100mg of calcium nitrate, 1~3mg of zinc sulfate, 1~3mg of boric acid, manganese sulfate 0.1
~0.5mg, 0.03~0.05mg of copper sulphate, 0.03~0.05mg of nickel chloride, 0.01~0.05mg of potassium iodide, ferrous sulfate 16~
20mg, 30~50mg of vitamin C, 8~10mg of niacin, 3~5mg of thiamine hydrochloride, 0.3~0.5 mg of puridoxine hydrochloride, paddy ammonia
0.3~0.5mg of acid, 0.5~1mg of cobalamin, 0.05~0.1mg of 6- benzyl purine, 0.2~0.5mg of methyl α-naphthyl acetate, coconut milk 50~
100mL, 50~80g of banana puree, 15~30g of sucrose, 6~7g of agar, surplus are water, pH 5.4~5.6；
(4) rhizomes is proliferated: the single rhizomes that step (3) obtains being transferred in proliferated culture medium and carries out clonal expansion
Culture 50~60 days, obtains the consistent rhizomes group of inheritance stability, described every liter of proliferated culture medium containing ammonium nitrate 160~
200 mg, 80~100mg of potassium dihydrogen phosphate, 160~200mg of potassium chloride, 80~100mg of magnesium sulfate, 80~100mg of calcium nitrate,
1~3mg of zinc sulfate, 1~3mg of boric acid, 0.1~0.5mg of manganese sulfate, 0.03~0.05mg of copper sulphate, nickel chloride 0.03~0.05
Mg, 0.01~0.05mg of potassium iodide, 16~20mg of ferrous sulfate, 30~50mg of vitamin C, 8~10mg of niacin, thiamine hydrochloride
3~5mg of element, 0.3~0.5mg of puridoxine hydrochloride, 0.3~0.5mg of glutamic acid, 0.5~1mg of cobalamin, 80~120 mg of inositol,
0.2~2.0mg of methyl α-naphthyl acetate, 0.5~3.0mg of benzyl aminoadenine, 1~2.0g of active carbon, 50~150g of banana, sucrose 10~
20g, 5~6g of agar, surplus are water, pH 5.5~5.7:
(5) rhizomes differentiation culture: the rhizomes group that step (4) obtains is transferred in differential medium and is broken up
Culture, until protocorms are differentiated to form the adventitious bud of high 3~5cm, described every liter of differential medium containing spend No. 1 1~2g of treasured,
Spend No. 2 1~2g of treasured, 20~40mg of ferrous sulfate, 20~40mg of disodium ethylene diamine tetraacetate, 0.5~2g of peptone, banana puree 50
~80g, 50~100mg of activated carbon, 80~120mg of inositol, 1.5~2.5mg of glycine, 0.05~0.2 mg of thiamine hydrochloride,
0.4~0.8mg of puridoxine hydrochloride, 0.4~0.8mg of niacin, 3.0~5.0mg of 6- benzyl purine, 0.4~0.5mg of methyl α-naphthyl acetate, sugarcane
20~30g of sugar, 6~7g of agar, surplus are water, pH 5.4~5.6；
(6) adventitious bud rooting seedling culture: the adventitious bud that step (5) obtains is transferred in Rooting and hardening-off culture base and is carried out
Seedling culture, described every liter of Rooting and hardening-off culture base containing spending No. 1 1~2g of treasured, spend treasured No. 2 1~2g, ferrous sulfate 20~40
Mg, 20~40mg of disodium ethylene diamine tetraacetate, 0.5~2g of peptone, 50~60g of mashed potatoes, 500~1000mg of activated carbon, flesh
80~120mg of alcohol, 1.5~2.5mg of glycine, 5~10mg of thiamine hydrochloride, 0.5~1.0mg of puridoxine hydrochloride, niacin 5~
10mg, 0.5~1.0mg of methyl α-naphthyl acetate, 15~30g of sucrose, 6~7g of agar, surplus are water, pH 5.4~5.6；
(7) at transplantation of seedlings and cultivation: when the seedling of the long supreme 5~8cm of adventitious bud, be transferred to natural light lower refining seedling 10~
It 20 days, then moves into Lan Shi: sawdust: being cultivated in the mixed-matrix that bark volume ratio is 1:1:1, obtain sword-leaved cymbidium and flame Cymbidium
Intermolecular hybrid kind.
The step (3) carries out sterile culture, condition of culture in embryo rescue culture medium are as follows: cultivation temperature 24~28
DEG C, 1500~2000lx of illuminance, illumination 12h/d.
The step (4) carries out Multiplying culture, condition of culture in proliferated culture medium are as follows: cultivation temperature 24~28
DEG C, 1500~2000lx of illuminance, illumination 12h/d.
The step (5) carries out differentiation culture, condition of culture in differential medium are as follows: cultivation temperature is 25~29
DEG C, 2000~3000lx of illuminance, illumination 12h/d.
The step (6) carries out seedling culture, condition of culture in Rooting and hardening-off culture base are as follows: cultivation temperature is
25~29 DEG C, 2000~3000lx of illuminance, illumination 12h/d.
The present invention also provides the asexual reproduction methods of a kind of sword-leaved cymbidium and Herba Renantherae coccineae intergeneric conjugal transfer, specifically, include with
Lower step:
(1) material is made with above-mentioned seedling, cuts the tender shoots of seedling strain, carries out disinfection, the tender shoots after disinfection is placed in
In rhizomes induced medium GB1,7~10d of dark culturing at 26 ± 2 DEG C, then in 1000~2000lx of intensity of illumination, light
According to time 10h/d, 26 ± 2 DEG C of cultivation temperature, cultivates 90~100 days, induce rhizomes, the rhizomes induced medium
Every liter of GB1 containing spend No. 1 1~2g of treasured, spend No. 2 1~2g of treasured, 20~40mg of ferrous sulfate, 20~40mg of disodium ethylene diamine tetraacetate,
0.5~2g of peptone, 50~100mL of coconut milk, 80~120mg of inositol, 1.5~2.5mg of glycine, thiamine hydrochloride 0.05~
0.2mg, 0.4~0.8mg of puridoxine hydrochloride, 0.4~0.8mg of niacin, 0.5~1.0 mg of 6- benzyl purine, methyl α-naphthyl acetate 1.0~
2.0mg, 0.5~1.0g of active carbon, 15~30g of sucrose, 6~7g of agar, surplus are water, pH 5.4~5.6；
(2) it after obtaining rhizomes, repeats above-mentioned sword-leaved cymbidium and Herba Renantherae coccineae generic cross rapidly and efficiently cultivates orchid new varieties
Rhizomes proliferation, rhizomes differentiation, adventitious bud rooting seedling in method, at the process of transplantation of seedlings and cultivation, complete new lines
Vegetative propagation.
Compared with prior art, novelty of the invention and beneficial effect are:
(1) present invention carries out pollen to solve the skills such as cold and rewetting processing before saving orchid pollen, pollination using sterile cryogenic
Art solves the problems, such as the bottleneck problem of the mildew and orchid generic cross flowering asynchronism when pollen saves.
(2) nutritional ingredient abundant and exogenous hormone that the present invention is saved in culture technique culture medium using embryo are able to satisfy miscellaneous
The needs of embryo development growth solve the depauperation of inter-genera distant hybridization hybrid embryo to (Hybrids embryo culture development is rhizomes)
Critical issue, promotion hybrid embryonic development be rhizomes.
(3) present invention carries out corresponding rhizomes clonal expansion after rhizomes by the development of single hybrid embryo sterile culture
Culture, the consistent quasi aromatic hybrid rice of a large amount of inheritance stability can be bred by the differentiation of rhizomes, seedling culture, is hybridized to
Cultivating orchid new varieties only needs 4~6 years, and traditional orchid hybridization is cultivated new varieties and generally required 8~10 years, avoids tradition miscellaneous
The shortcomings that handing over breeding to carry out the complicated program of backcrossing purifying, breeding year limit for length, can quickly breeding go out orchid new varieties.
(4) each hybrid embryo of the present invention can cultivate into a new varieties, and breeding efficiency is high.
Specific embodiment
The following examples are further illustrations of the invention, rather than limiting the invention.Used in embodiment
Spending precious No. 1 (HYPONeX 1), spending precious No. 2 (HYPONeX 2) is that TaiWan, China platform produced in USA and gardening enterprise stock are limited
Limited liability company dispenses product.
Embodiment 1
(1) Herba Renantherae coccineae pollen collecting and preservation:
A, the preparation of picking tool and storing utensil: the tooth for picking massula is put into respectively with the vial of 300mL
The 1mL plastic tube of label and storage massula, closes the lid, spare after sterilizing being carried out under the conditions of 121 DEG C 30 minutes.
B, Herba Renantherae coccineae (male parent) is bloomed at the beginning of 3 months, when its single flower petal just unfolds, is applied and has been sterilized when the morning 8~9
Toothpick gently pick out colored anther cap, then massula is sticked in toothpick and is put on toothpick point and rapidly by massula to clean nothing
In the 1mL plastic tube of bacterium, every pipe is put into 1 colored massula, and the plastic tube that placed massula is used a sterile glass again
Glass bottle concentration is loaded and is sealed, and is placed in 4 DEG C and is kept in dark place.
(2) artificial pollination and hybridization fruit are cultivated: when sword-leaved cymbidium (female parent) just blooms in August, by the fire of step (1) preservation
Flame orchid powder carries out solving the processing of cold and rewetting, to restore and improve the viability of pollen.First by the Herba Renantherae coccineae pollen of cryo-conservation
Thaw 20min at room temperature, then adds a few drop sterile waters to carry out rewetting processing in the plastic tube for placing massula again
20min finally uses triphenyltetrazolium chloride Determination Staining pollen viability, and rate of dyeing determines that pollen is vibrant up to 83%,
It is pollinated on the column cap that maternal sword-leaved cymbidium has opened 2~3 days flowers.Bagging after pollination removes bag after about 1 week, to hang up
Label has the information such as parent name, hybridization date and pollination people on label.Pay attention to observing its developmental state, statistics pollination
Success rate, setting percentage is up to 85% or more.After success carries out hybridization pollination, reinforces the fertilizer and water management of maternal sword-leaved cymbidium plant, guarantee
Hybridize the normal development of capsule.The developmental state for paying attention to observation hybridization capsule waits fruit developments to can acquire progress after 180 days
Aseptic seeding.
(3) Hybrids embryo culture development is rhizomes: after hybridization capsule is developed to 180 days, picking hybridization fruit is sterilized
(orchid capsule is impregnated into 15~30s in 70% ethanol solution of volume fraction, then is sterilized with 0.1% mercuric chloride solution of mass fraction
Aseptic water washing 4~5 times, the moisture on capsule surface is blotted with aseptic filter paper by 15~20min) hybridization fruit is cut afterwards, by powder
Shape hybrid embryo be seeded to embryo rescue culture medium in, under conditions of 24 DEG C of cultivation temperature, illuminance 1500lx, illumination 12h/d into
Visible hybrid embryo, which expands, after row sterile culture 60 days turns green, and single hybrid embryo can be sprouted to form single rhizomes when cultivating 80 days,
The embryo saves every liter of 160mg containing ammonium nitrate of culture medium, potassium dihydrogen phosphate 80mg, potassium chloride 160mg, magnesium sulfate 80mg, nitre
Sour calcium 80mg, zinc sulfate 1mg, boric acid 1mg, manganese sulfate 0.1mg, copper sulphate 0.03mg, nickel chloride 0.03mg, potassium iodide
0.01mg, ferrous sulfate (FeSO4.7H2O) 16mg, vitamin C 30mg, niacin 8mg, thiamine hydrochloride (VB1) 3mg, hydrochloric acid
Pyridoxol (VB6) 0.3mg, glutamic acid 0.3mg, cobalamin 0.5mg, 6- benzyl purine 0.05mg, methyl α-naphthyl acetate (NAA) 0.2mg,
Coconut milk 50mL, banana puree 50g, sucrose 15g, agar 6g, surplus are water, adjust pH 5.4, preparation method is by all the components
It is uniformly mixed by its content, adjusts pH value, sterilization.
(4) rhizomes be proliferated: by the single rhizomes that step (3) obtains be transferred in single rhizomes proliferated culture medium into
Row clonal expansion culture, subculture cycle are 50 days, and the separately culture of one, each rhizomes number carries out rhizomes proliferation, by 4
The rhizomes of~8 shoot proliferation cultures, each number can be proliferated out consistent shape of 1000~400000 inheritance stabilities
Stem group, described every liter of 160mg containing ammonium nitrate of rhizomes proliferated culture medium, potassium dihydrogen phosphate 80mg, 160 mg of potassium chloride, sulphur
Sour magnesium 80mg, calcium nitrate 80mg, zinc sulfate 1mg, boric acid 1mg, manganese sulfate 0.1mg, 0.03 mg of copper sulphate, nickel chloride 0.03mg,
Potassium iodide 0.01mg, ferrous sulfate 16mg, vitamin C 30mg, niacin 8mg, thiamine hydrochloride 3mg, puridoxine hydrochloride 0.3mg,
Glutamic acid 0.3mg, cobalamin 0.5mg, inositol 80mg, methyl α-naphthyl acetate 0.2mg, benzyl aminoadenine 0.5mg, active carbon 1g, banana
50g, sucrose 10g, agar 5g, surplus are water, and pH 5.5, condition of culture is 28 DEG C of cultivation temperature, illuminance 2000lx, illumination
12h/d。
(5) rhizomes differentiation culture: the rhizomes group that step (4) obtains is transferred in differential medium, is being cultivated
Temperature carries out differentiation culture 45 days under conditions of being 25 DEG C, illuminance 2000lx, illumination 12h/d after, rhizomes group is being cultivated
The adventitious bud of high 3~5cm is differentiated to form on base, this adventitious bud material number corresponds to its rhizomes number.The differentiation culture
Every liter of base containing spending No. 1 1g of treasured, spend treasured No. 2 1g, ferrous sulfate (FeSO4.7H2O) 20mg, disodium ethylene diamine tetraacetate (EDTA-
2Na) 20mg, peptone 0.5g, banana puree 50g, activated carbon 50mg, inositol 80mg, glycine 1.5mg, thiamine hydrochloride (VB1)
0.05mg, puridoxine hydrochloride (VB6) 0.4mg, niacin 0.4mg, 6- benzyl purine 3.0mg, methyl α-naphthyl acetate (NAA) 0.4mg, sucrose
20g, agar 6g, surplus are water, adjust pH 5.4, and preparation method is to be uniformly mixed all the components by its content, adjust pH value, go out
Bacterium disinfection.
(6) adventitious bud for high 3~5cm that step (5) obtains adventitious bud rooting seedling culture: is transferred to strong plantlets and rootage training
It supports in base, is carried out seedling culture 60 days under conditions of being 25 DEG C, illuminance 2000lx, illumination 12h/d in cultivation temperature, height of seedling can
Reach 5~8cm, radical 3~5, rooting rate is 100% or more.Seedling numbers corresponding its adventitious bud number.Described taking root is strong
Every liter of seedling culture medium containing spending No. 1 1g of treasured, spend treasured No. 2 1g, ferrous sulfate (FeSO4.7H2O) 20mg, disodium ethylene diamine tetraacetate
(EDTA-2Na) 20mg, peptone 0.5g, mashed potatoes 50g, activated carbon 500mg, inositol 80mg, glycine 1.5mg, hydrochloric acid sulphur
Amine element (VB1) 5mg, puridoxine hydrochloride (VB6) 0.5mg, niacin 5mg, methyl α-naphthyl acetate (NAA) 0.5mg, sucrose 15g, agar 6g, surplus
For water, pH 5.4 is adjusted, preparation method is to be uniformly mixed all the components by its content, adjusts pH value, sterilization.
(7) at transplantation of seedlings and cultivation: when the seedling of adventitious bud length to 5~8cm of height of seedling, culture bottle being transferred to natural light
Lower refining seedling 10 days, then take out it from vial, clean the culture medium of root, move into blue stone: sawdust: bark volume ratio is
In the mixed-matrix of 1:1:1, appropriate ventilation and enough humidity are kept, the survival rate of transplanting is built up to 90% or more
Blue and Herba Renantherae coccineae generic cross kind.Plant after 30d or so is survived can generate new root system, may move into greenhouse at this time and carry out just
Ordinary water, fertilizer, pencil reason.
Embodiment 2
(1) Herba Renantherae coccineae pollen collecting and preservation:
A, the preparation of picking tool and storing utensil: the tooth for picking massula is put into respectively with the vial of 300mL
The 1mL plastic tube of label and storage massula, closes the lid, spare after sterilizing being carried out under the conditions of 121 DEG C 30 minutes.
B, Herba Renantherae coccineae (male parent) is bloomed at the beginning of 3 months, when its single flower petal just unfolds, is applied and has been sterilized when the morning 8~9
Toothpick gently pick out colored anther cap, then massula is sticked in toothpick and is put on toothpick point and rapidly by massula to clean nothing
In the 1mL plastic tube of bacterium, every pipe is put into 1 colored massula, and the plastic tube that placed massula is used a sterile glass again
Glass bottle concentration is loaded and is sealed, and is placed in 6 DEG C and is kept in dark place.
(2) artificial pollination and hybridization fruit are cultivated: when sword-leaved cymbidium (female parent) just blooms in August, by the fire of step (1) preservation
Flame orchid powder carries out solving the processing of cold and rewetting, to restore and improve the viability of pollen.First by the Herba Renantherae coccineae pollen of cryo-conservation
Thaw 30min at room temperature, then adds a few drop sterile waters to carry out rewetting processing in the plastic tube for placing massula again
30min finally uses triphenyltetrazolium chloride Determination Staining pollen viability, and rate of dyeing determines that pollen is vibrant up to 83%,
It is pollinated on the column cap that maternal sword-leaved cymbidium has opened 2~3 days flowers.Bagging after pollination removes bag after about 1 week, to hang up
Label has the information such as parent name, hybridization date and pollination people on label.Pay attention to observing its developmental state, statistics pollination
Success rate, setting percentage is up to 85% or more.After success carries out hybridization pollination, reinforces the fertilizer and water management of maternal sword-leaved cymbidium plant, guarantee
Hybridize the normal development of capsule.The developmental state for paying attention to observation hybridization capsule waits fruit developments to can acquire progress after 200 days
Aseptic seeding.
(3) Hybrids embryo culture development is rhizomes: after hybridization capsule is developed to 200 days, picking hybridization fruit is sterilized
(orchid capsule is impregnated into 15~30s in 70% ethanol solution of volume fraction, then is sterilized with 0.1% mercuric chloride solution of mass fraction
Aseptic water washing 4~5 times, the moisture on capsule surface is blotted with aseptic filter paper by 15~20min) hybridization fruit is cut afterwards, by powder
Shape hybrid embryo be seeded to embryo rescue culture medium in, under conditions of 28 DEG C of cultivation temperature, illuminance 2000lx, illumination 12h/d into
Visible hybrid embryo, which expands, after row sterile culture 60 days turns green, and single hybrid embryo can be sprouted to form single rhizomes when cultivating 100 days,
Described embryo rescue every liter of 200mg containing ammonium nitrate of culture medium, potassium dihydrogen phosphate 100mg, potassium chloride 200mg, magnesium sulfate 100mg,
Calcium nitrate 100mg, zinc sulfate 3mg, boric acid 3mg, manganese sulfate 0.5mg, copper sulphate 0.05mg, nickel chloride 0.05mg, potassium iodide
0.05mg, ferrous sulfate (FeSO4.7H2O) 20mg, vitamin C 50mg, niacin 10mg, thiamine hydrochloride (VB1) 5mg, hydrochloric acid
Pyridoxol (VB6) 0.5mg, glutamic acid 0.5mg, cobalamin 1mg, 6- benzyl purine 0.1mg, methyl α-naphthyl acetate (NAA) 0.5mg, coconut
Juice 100mL, banana puree 80g, sucrose 30g, agar 7g, surplus are water, adjust pH 5.6, preparation method is by all the components by it
Content is uniformly mixed, and adjusts pH value, sterilization.
(4) rhizomes be proliferated: by the single rhizomes that step (3) obtains be transferred in single rhizomes proliferated culture medium into
Row clonal expansion culture, subculture cycle are 60 days, and the separately culture of one, each rhizomes number carries out rhizomes proliferation, by 4
The rhizomes of~8 shoot proliferation cultures, each number can be proliferated out consistent shape of 1000~400000 inheritance stabilities
Stem group, described every liter of 200mg containing ammonium nitrate of rhizomes proliferated culture medium, potassium dihydrogen phosphate 100mg, 200 mg of potassium chloride,
Magnesium sulfate 100mg, calcium nitrate 100mg, zinc sulfate 3mg, boric acid 3mg, manganese sulfate 0.5mg, 0.05 mg of copper sulphate, nickel chloride
0.05mg, potassium iodide 0.05mg, ferrous sulfate 20mg, vitamin C 50mg, niacin 10mg, thiamine hydrochloride 5mg, hydrochloric acid pyrrole are trembled
Alcohol 0.5mg, glutamic acid 0.5mg, cobalamin 1mg, inositol 120mg, methyl α-naphthyl acetate 2.0mg, benzyl aminoadenine 3.0mg, active carbon
2.0g, banana 150g, sucrose 20g, agar 6g, surplus are water, and pH 5.7, condition of culture is 24 DEG C of cultivation temperature, illuminance
1500lx, illumination 12h/d.
(5) rhizomes differentiation culture: the rhizomes group that step (4) obtains is transferred in differential medium, is being cultivated
Temperature carries out differentiation culture 45 days under conditions of being 29 DEG C, illuminance 3000lx, illumination 12h/d after, rhizomes group is being cultivated
The adventitious bud of high 3~5cm is differentiated to form on base, this adventitious bud material number corresponds to its rhizomes number.The differentiation culture
Every liter of base containing spending No. 1 2g of treasured, spend treasured No. 2 2g, ferrous sulfate (FeSO4.7H2O) 40mg, disodium ethylene diamine tetraacetate 40mg, albumen
Peptone 2g, banana puree 80g, activated carbon 100mg, inositol 120mg, glycine 2.5mg, thiamine hydrochloride (VB1) 0.2mg, hydrochloric acid pyrrole
Tremble alcohol (VB6) 0.8mg, niacin 0.8mg, 6- benzyl purine 5.0mg, methyl α-naphthyl acetate (NAA) 0.5mg, sucrose 30g, agar 7g, surplus
For water, pH 5.6 is adjusted, preparation method is to be uniformly mixed all the components by its content, adjusts pH value, sterilization.
(6) adventitious bud for high 3~5cm that step (5) obtains adventitious bud rooting seedling culture: is transferred to strong plantlets and rootage training
It supports in base, is carried out seedling culture 60 days under conditions of being 29 DEG C, illuminance 3000lx, illumination 12h/d in cultivation temperature, height of seedling can
Reach 5~8cm, radical 3~5, rooting rate is 100% or more.Seedling numbers corresponding its adventitious bud number.Described taking root is strong
Every liter of seedling culture medium containing spending No. 1 2g of treasured, spend treasured No. 2 2g, ferrous sulfate (FeSO4.7H2O) 40mg, disodium ethylene diamine tetraacetate
40mg, peptone 2g, mashed potatoes 60g, activated carbon 1000mg, inositol 120mg, glycine 2.5mg, thiamine hydrochloride (VB1)
10mg, puridoxine hydrochloride (VB6) 1.0mg, niacin 10mg, methyl α-naphthyl acetate (NAA) 1.0mg, sucrose 30g, agar 7g, surplus is water,
PH 5.6 is adjusted, preparation method is to be uniformly mixed all the components by its content, adjusts pH value, sterilization.
(7) at transplantation of seedlings and cultivation: when the seedling of adventitious bud length to 5~8cm of height of seedling, culture bottle being transferred to natural light
Lower refining seedling 20 days, then take out it from vial, clean the culture medium of root, move into blue stone: sawdust: bark volume ratio is
In the mixed-matrix of 1:1:1, appropriate ventilation and enough humidity are kept, the survival rate of transplanting is built up to 90% or more
Blue and Herba Renantherae coccineae generic cross kind.Plant after 30d or so is survived can generate new root system, may move into greenhouse at this time and carry out just
Ordinary water, fertilizer, pencil reason.
Embodiment 3
(1) material is made with the long seedling to 5~8cm of height of seedling of the adventitious bud of embodiment 1, cuts the tender shoots of seedling strain, into
(tender shoots → dried cotton wipes the cotton that 1 time → 75% ethyl alcohol impregnates and wipes the extra blade of 1 time → removal and cut for row disinfection
At segment, every section is sufficiently impregnated 3 times → 0.1%HgCl of 30s → sterile water wash comprising a bud point → 75% ethyl alcohol2Sufficiently leaching
Bract → 0.1%HgCl is peelled off after bubble 4~6min → aseptic water washing 3 times → drying excessive moisture2Abundant 1~2min → drying
Excessive moisture), the tender shoots after disinfection is placed in rhizomes induced medium GB1, the dark culturing 7d at 26 ± 2 DEG C, so
Afterwards in intensity of illumination 1000lx, light application time 10h/d, 26 ± 2 DEG C of cultivation temperature, cultivates 90 days, induce rhizomes, it is described
Every liter of GB1 of rhizomes induced medium containing spending No. 1 1g of treasured, spend treasured No. 2 1g, ferrous sulfate (FeSO4.7H2O) 20mg, ethylenediamine
Tetraacethyl disodium 20mg, peptone 0.5g, coconut milk 50mL, inositol 80mg, 1.5 mg of glycine, thiamine hydrochloride (VB1)
0.05mg, puridoxine hydrochloride (VB6) 0.4mg, niacin 0.4mg, 6- benzyl purine 0.5mg, methyl α-naphthyl acetate (NAA) 1.0mg, activity
Charcoal 0.5g, sucrose 15g and agar 6g, surplus are water, adjust pH 5.4, and preparation method is to mix all the components by its content
It is even, adjust pH value, sterilization.
(2) rhizomes be proliferated: by the single rhizomes that step (1) obtains be transferred in single rhizomes proliferated culture medium into
Row clonal expansion culture, subculture cycle are 55 days, and the separately culture of one, each rhizomes number carries out rhizomes proliferation, by 4
The rhizomes of~8 shoot proliferation cultures, each number can be proliferated out consistent shape of 1000~400000 inheritance stabilities
Stem group, described every liter of 180mg containing ammonium nitrate of rhizomes proliferated culture medium, potassium dihydrogen phosphate 90mg, 180 mg of potassium chloride, sulphur
Sour magnesium 90mg, calcium nitrate 90mg, zinc sulfate 2mg, boric acid 2mg, manganese sulfate 0.2mg, 0.04 mg of copper sulphate, nickel chloride 0.04mg,
Potassium iodide 0.03mg, ferrous sulfate 18mg, vitamin C 40mg, niacin 9mg, thiamine hydrochloride 4mg, puridoxine hydrochloride 0.4mg,
Glutamic acid 0.4mg, cobalamin 0.8mg, inositol 100mg, methyl α-naphthyl acetate 1mg, benzyl aminoadenine 2mg, active carbon 1.5g, banana
80g, sucrose 15g, agar 5g, surplus are water, and pH 5.7, condition of culture is 26 DEG C of cultivation temperature, illuminance 1800lx, light
According to 12h/d.
(3) rhizomes differentiation culture: the rhizomes group that step (2) obtains is transferred in differential medium, is being cultivated
Temperature carries out differentiation culture 45 days under conditions of being 28 DEG C, illuminance 2500lx, illumination 12h/d after, rhizomes group is being cultivated
The adventitious bud of high 3~5cm is differentiated to form on base, this adventitious bud material number corresponds to its rhizomes number.The differentiation culture
Every liter of base containing spending No. 1 1.5g of treasured, spend treasured No. 2 1.5g, ferrous sulfate (FeSO4.7H2O) 30mg, disodium ethylene diamine tetraacetate 30mg,
Peptone 1g, banana puree 65g, activated carbon 75mg, inositol 100mg, glycine 2mg, thiamine hydrochloride (VB1) 0.1mg, hydrochloric acid pyrrole
Tremble alcohol (VB6) 0.6mg, niacin 0.6mg, 6- benzyl purine 4.0mg, methyl α-naphthyl acetate (NAA) 0.45mg, sucrose 25g, agar 6.5g,
Surplus is water, adjusts pH 5.5, and preparation method is to be uniformly mixed all the components by its content, adjusts pH value, sterilization.
(4) adventitious bud for high 3~5cm that step (3) obtains adventitious bud rooting seedling culture: is transferred to strong plantlets and rootage training
It supports in base, is carried out seedling culture 60 days under conditions of being 28 DEG C, illuminance 2500lx, illumination 12h/d in cultivation temperature, height of seedling can
Reach 5~8cm, radical 3~5, rooting rate is 100% or more.Seedling numbers corresponding its adventitious bud number.Described taking root is strong
Every liter of seedling culture medium containing spending No. 1 1.5g of treasured, spend treasured No. 2 1.5g, ferrous sulfate (FeSO4.7H2O) 30mg, ethylenediamine tetra-acetic acid two
Sodium 30mg, peptone 1g, mashed potatoes 55g, activated carbon 800mg, inositol 100mg, glycine 2mg, thiamine hydrochloride (VB1)8mg、
Puridoxine hydrochloride (VB6) 0.8mg, niacin 8mg, methyl α-naphthyl acetate (NAA) 0.8mg, sucrose 20g, agar 6.5g, surplus is water, adjusts pH
5.5, preparation method is to be uniformly mixed all the components by its content, adjusts pH value, sterilization.
(5) at transplantation of seedlings and cultivation: when adventitious bud it is long to 5~8cm of height of seedling when, culture bottle is transferred to natural light lower refining seedling
20 days, then it is taken out from vial, clean the culture medium of root, move into blue stone: sawdust: bark volume ratio is 1:1:1
Mixed-matrix in, keep appropriate ventilation and enough humidity, the survival rate of transplanting up to 90% or more, obtains sword-leaved cymbidium and fire
Flame Cymbidium intermolecular hybrid kind.Plant after 30d or so is survived can generate new root system, may move at this time greenhouse carry out normal water,
Fertilizer, pencil reason.
Embodiment 4
(1) material is made with the long seedling to 5~8cm of height of seedling of adventitious bud, cuts the tender shoots of seedling strain, carries out disinfection (tender
Bud → dried cotton wipes the cotton that 1 time → 75% ethyl alcohol impregnates and wipes the extra blade of 1 time → removal and be cut into segment, often
Section sufficiently impregnates 3 times → 0.1%HgCl of 30s → sterile water wash comprising a bud point → 75% ethyl alcohol2Sufficiently impregnate 4~6min
Bract → 0.1%HgCl is peelled off after → aseptic water washing 3 times → drying excessive moisture2Abundant 1~2min → drying superfluous water
Point), the tender shoots after disinfection is placed in rhizomes induced medium GB1, the dark culturing 10d at 26 ± 2 DEG C, then in light
According to intensity 2000lx, light application time 10h/d, 26 ± 2 DEG C of cultivation temperature, cultivates 100 days, induce rhizomes, the root shape
Every liter of GB1 of stem induced medium containing spending No. 1 2g of treasured, spend treasured No. 2 2g, ferrous sulfate (FeSO4.7H2O) 40mg, ethylenediamine tetra-acetic acid
Disodium 40mg, peptone 2g, coconut milk 100mL, inositol 120mg, glycine 2.5mg, thiamine hydrochloride (VB1) 0.2mg, hydrochloric acid
Pyridoxol (VB6) 0.8mg, niacin 0.8mg, 6- benzyl purine 1.0mg, methyl α-naphthyl acetate (NAA) 2.0mg, active carbon 1.0g, sucrose
30g and agar 7g, surplus are water, adjust pH 5.6, and preparation method is to be uniformly mixed all the components by its content, adjust pH value,
Sterilization.
(2) rhizomes be proliferated: by the single rhizomes that step (1) obtains be transferred in single rhizomes proliferated culture medium into
Row clonal expansion culture, subculture cycle are 60 days, and the separately culture of one, each rhizomes number carries out rhizomes proliferation, by 4
The rhizomes of~8 shoot proliferation cultures, each number can be proliferated out consistent shape of 1000~400000 inheritance stabilities
Stem group, described every liter of 160mg containing ammonium nitrate of rhizomes proliferated culture medium, potassium dihydrogen phosphate 100mg, 160 mg of potassium chloride,
Magnesium sulfate 80mg, calcium nitrate 100mg, zinc sulfate 3mg, boric acid 3mg, manganese sulfate 0.4mg, 0.04 mg of copper sulphate, nickel chloride
0.03mg, potassium iodide 0.03mg, ferrous sulfate 18mg, vitamin C 40mg, niacin 9mg, thiamine hydrochloride 4mg, hydrochloric acid pyrrole are trembled
Alcohol 0.4mg, glutamic acid 0.3mg, cobalamin 0.8mg, inositol 90mg, methyl α-naphthyl acetate 1.5mg, benzyl aminoadenine 1.5mg, active carbon
1g, banana 90g, sucrose 15g, agar 5g, surplus are water, and pH 5.7, condition of culture is 26 DEG C of cultivation temperature, illuminance
1500lx, illumination 12h/d.
(3) rhizomes differentiation culture: the rhizomes group that step (2) obtains is transferred in differential medium, is being cultivated
Temperature carries out differentiation culture 45 days under conditions of being 29 DEG C, illuminance 3000lx, illumination 12h/d after, rhizomes group is being cultivated
The adventitious bud of high 3~5cm is differentiated to form on base, this adventitious bud material number corresponds to its rhizomes number.The differentiation culture
Every liter of base containing spending No. 1 1.5g of treasured, spend treasured No. 2 1.5g, ferrous sulfate (FeSO4.7H2O) 40mg, disodium ethylene diamine tetraacetate 30mg,
Peptone 1.5g, banana puree 70g, activated carbon 90mg, inositol 100mg, glycine 2.5mg, thiamine hydrochloride (VB1) 0.2mg, salt
Sour pyridoxol (VB6) 0.8mg, niacin 0.5mg, 6- benzyl purine 4.0mg, methyl α-naphthyl acetate (NAA) 0.5mg, sucrose 30g, agar 7g,
Surplus is water, adjusts pH 5.6, and preparation method is to be uniformly mixed all the components by its content, adjusts pH value, sterilization.
(4) adventitious bud for high 3~5cm that step (3) obtains adventitious bud rooting seedling culture: is transferred to strong plantlets and rootage training
It supports in base, is carried out seedling culture 60 days under conditions of being 29 DEG C, illuminance 3000lx, illumination 12h/d in cultivation temperature, height of seedling can
Reach 5~8cm, radical 3~5, rooting rate is 100% or more.Seedling numbers corresponding its adventitious bud number.Described taking root is strong
Every liter of seedling culture medium containing spending No. 1 2g of treasured, spend treasured No. 2 1g, ferrous sulfate (FeSO4.7H2O) 30mg, disodium ethylene diamine tetraacetate
30mg, peptone 1.5g, mashed potatoes 60g, activated carbon 700mg, inositol 90mg, glycine 2.5mg, thiamine hydrochloride (VB1)
8mg, puridoxine hydrochloride (VB6) 1.0mg, niacin 8mg, methyl α-naphthyl acetate (NAA) 0.5mg, sucrose 20g, agar 7g, surplus is water, is adjusted
PH 5.6, preparation method are to be uniformly mixed all the components by its content, adjust pH value, sterilization.
(5) at transplantation of seedlings and cultivation: when adventitious bud it is long to 5~8cm of height of seedling when, culture bottle is transferred to natural light lower refining seedling
20 days, then it is taken out from vial, clean the culture medium of root, move into blue stone: sawdust: bark volume ratio is 1:1:1
Mixed-matrix in, keep appropriate ventilation and enough humidity, the survival rate of transplanting up to 90% or more, obtains sword-leaved cymbidium and fire
Flame Cymbidium intermolecular hybrid kind.Plant after 30d or so is survived can generate new root system, may move at this time greenhouse carry out normal water,
Fertilizer, pencil reason.
The above is only the preferred embodiment of the present invention, it is noted that above-mentioned preferred embodiment is not construed as pair
Limitation of the invention, protection scope of the present invention should be defined by the scope defined by the claims..For the art
For those of ordinary skill, without departing from the spirit and scope of the present invention, several improvements and modifications can also be made, these change
It also should be regarded as protection scope of the present invention into retouching.
Claims (10)
1. a kind of method that sword-leaved cymbidium and Herba Renantherae coccineae generic cross rapidly and efficiently cultivate orchid new varieties, which is characterized in that including with
Lower step:
(1) pollen collecting and preservation: when male parent Herba Renantherae coccineae single flower petal is just unfolded, orchid is picked out using the picking tool of sterilizing
Anther cap, then orchid massula chosen into the container of cleaning sterile be sealed with the picking tool of sterilizing, be placed in 4~6 DEG C and keep away
Light saves；
(2) artificial pollination and hybridization fruit are cultivated: when maternal sword-leaved cymbidium blooms, the massula that step (1) is saved is in room temperature item
Thaw 20~30min under part, then adds sterile water and carries out rewetting 20~30min of processing, obtains having great-hearted pollen；It will
Has great-hearted pollen pollination in the cultivation on the flower column cap of maternal sword-leaved cymbidium, carrying out hybridization capsule；
(3) Hybrids embryo culture development is rhizomes: when hybridization capsule is developed to 180~200 days, picking hybridization fruit is sterilized
Hybridization fruit is cut afterwards, and powdered hybrid embryo is seeded to progress sterile culture in embryo rescue culture medium and obtains rhizomes, it is described
Embryo save every liter of culture medium contain 160~200mg of ammonium nitrate, 80~100mg of potassium dihydrogen phosphate, 160~200mg of potassium chloride, sulphur
Sour 80~100mg of magnesium, 80~100mg of calcium nitrate, 1~3mg of zinc sulfate, 1~3mg of boric acid, 0.1~0.5mg of manganese sulfate, copper sulphate
0.03~0.05mg, 0.03~0.05mg of nickel chloride, 0.01~0.05mg of potassium iodide, 16~20mg of ferrous sulfate, vitamin C
30~50mg, 8~10mg of niacin, 3~5mg of thiamine hydrochloride, 0.3~0.5mg of puridoxine hydrochloride, 0.3~0.5mg of glutamic acid,
0.5~1mg of cobalamin, 0.05~0.1mg of 6- benzyl purine, 0.2~0.5mg of methyl α-naphthyl acetate, 50~100mL of coconut milk, banana puree
50~80g, 15~30g of sucrose, 6~7g of agar, surplus are water, pH 5.4~5.6；
(4) rhizomes is proliferated: the single rhizomes that step (3) obtains is transferred to progress clonal expansion culture in proliferated culture medium
The consistent rhizomes group of inheritance stability is obtained, described every liter of proliferated culture medium contains 160~200mg of ammonium nitrate, biphosphate
80~100mg of potassium, 160~200mg of potassium chloride, 80~100mg of magnesium sulfate, 80~100mg of calcium nitrate, 1~3mg of zinc sulfate, boron
1~3mg of acid, 0.1~0.5mg of manganese sulfate, 0.03~0.05mg of copper sulphate, 0.03~0.05mg of nickel chloride, potassium iodide 0.01~
0.05mg, 16~20mg of ferrous sulfate, 30~50mg of vitamin C, 8~10mg of niacin, 3~5mg of thiamine hydrochloride, hydrochloric acid pyrrole
Tremble 0.3~0.5mg of alcohol, 0.3~0.5mg of glutamic acid, 0.5~1mg of cobalamin, 80~120mg of inositol, methyl α-naphthyl acetate 0.2~
2.0mg, 0.5~3.0mg of benzyl aminoadenine, 1~2.0g of active carbon, 50~150g of banana, 10~20g of sucrose, agar 5~
6g, surplus are water, pH 5.5~5.7；
(5) rhizomes differentiation culture: the rhizomes group that step (4) obtains is transferred in differential medium and carries out differentiation training
Support, until rhizomes Population Differentiation forms the adventitious bud of high 3~5cm, described every liter of differential medium containing spend No. 1 1~2g of treasured,
Spend No. 2 1~2g of treasured, 20~40mg of ferrous sulfate, 20~40mg of disodium ethylene diamine tetraacetate, 0.5~2g of peptone, banana puree 50
~80g, 50~100mg of activated carbon, 80~120mg of inositol, 1.5~2.5mg of glycine, 0.05~0.2mg of thiamine hydrochloride, salt
Sour 0.4~0.8mg of pyridoxol, 0.4~0.8mg of niacin, 3.0~5.0mg of 6- benzyl purine, 0.4~0.5mg of methyl α-naphthyl acetate, sucrose
20~30g, 6~7g of agar, surplus are water, pH 5.4~5.6；
(6) adventitious bud rooting seedling culture: the adventitious bud that step (5) obtains is transferred in Rooting and hardening-off culture base and carries out seedling
Culture, described every liter of Rooting and hardening-off culture base containing spending No. 1 1~2g of treasured, spend treasured No. 2 1~2g, 20~40mg of ferrous sulfate, second
20~40mg of edetate disodium, 0.5~2g of peptone, 50~60g of mashed potatoes, 500~1000mg of activated carbon, inositol 80~
120mg, 1.5~2.5mg of glycine, 5~10mg of thiamine hydrochloride, 0.5~1.0mg of puridoxine hydrochloride, 5~10mg of niacin, naphthalene
0.5~1.0mg of acetic acid, 15~30g of sucrose, 6~7g of agar, surplus are water, pH 5.4~5.6；
(7) at transplantation of seedlings and cultivation: when the seedling of the long supreme 5~8cm of adventitious bud, being transferred to natural light lower refining seedling 10~20
It, then moves into Lan Shi: sawdust: cultivating in the mixed-matrix that bark volume ratio is 1:1:1, obtains sword-leaved cymbidium and be mixed with flame Cymbidium
Hand over kind.
2. the method that sword-leaved cymbidium according to claim 1 and Herba Renantherae coccineae generic cross rapidly and efficiently cultivate orchid new varieties,
It is characterized in that, the step (3) carries out sterile culture, condition of culture in embryo rescue culture medium are as follows: cultivation temperature 24~
28 DEG C, 1500~2000lx of illuminance, illumination 12h/d.
3. the method that sword-leaved cymbidium according to claim 1 and Herba Renantherae coccineae generic cross rapidly and efficiently cultivate orchid new varieties,
It is characterized in that, the step (4) carries out Multiplying culture, condition of culture in proliferated culture medium are as follows: cultivation temperature 24~
28 DEG C, 1500~2000lx of illuminance, illumination 12h/d.
4. the method that sword-leaved cymbidium according to claim 1 and Herba Renantherae coccineae generic cross rapidly and efficiently cultivate orchid new varieties,
It is characterized in that, the step (5) carries out differentiation culture, condition of culture in differential medium are as follows: cultivation temperature 25
~29 DEG C, 2000~3000lx of illuminance, illumination 12h/d.
5. the method that sword-leaved cymbidium according to claim 1 and Herba Renantherae coccineae generic cross rapidly and efficiently cultivate orchid new varieties,
It is characterized in that, the step (6) carries out seedling culture, condition of culture are as follows: cultivation temperature in Rooting and hardening-off culture base
It is 25~29 DEG C, 2000~3000lx of illuminance, illumination 12h/d.
6. the asexual reproduction method of a kind of sword-leaved cymbidium and Herba Renantherae coccineae intergeneric conjugal transfer, which comprises the following steps:
(1) rhizomes induces: the tender shoots after cutting the disinfection of the seedling of the step (7) in claim 1 is inoculated in rhizomes and lures
It leads in culture medium GB1, is first placed in 7~10d of dark culturing at 26 ± 2 DEG C, then illumination cultivation, intensity of illumination 1000~
2000lx, light application time 10h/d, 26 ± 2 DEG C of cultivation temperature, culture induces rhizomes；The rhizomes induced medium
GB1: every liter containing spend No. 1 1~2g of treasured, spend No. 2 1~2g of treasured, 20~40mg of ferrous sulfate, disodium ethylene diamine tetraacetate 20~
40mg, 0.5~2g of peptone, 50~100mL of coconut milk, 80~120mg of inositol, 1.5~2.5mg of glycine, thiamine hydrochloride
0.05~0.2mg, 0.4~0.8mg of puridoxine hydrochloride, 0.4~0.8mg of niacin, 0.5~1.0mg of 6- benzyl purine, methyl α-naphthyl acetate
1.0~2.0mg, 0.5~1.0g of active carbon, 15~30g of sucrose, 6~7g of agar, surplus are water, pH 5.4~5.6；
(2) rhizomes is proliferated: rhizomes is transferred to progress Multiplying culture in proliferated culture medium and obtains rhizomes group, it is described
Every liter of proliferated culture medium contains 160~200mg of ammonium nitrate, 80~100mg of potassium dihydrogen phosphate, 160~200mg of potassium chloride, magnesium sulfate
80~100mg, 80~100mg of calcium nitrate, 1~3mg of zinc sulfate, 1~3mg of boric acid, 0.1~0.5mg of manganese sulfate, copper sulphate 0.03
~0.05mg, 0.03~0.05mg of nickel chloride, 0.01~0.05mg of potassium iodide, 16~20mg of ferrous sulfate, vitamin C 30~
50mg, 8~10mg of niacin, 3~5mg of thiamine hydrochloride, 0.3~0.5mg of puridoxine hydrochloride, 0.3~0.5mg of glutamic acid, cobalt amine
0.5~1mg of element, inositol 80-120mg, methyl α-naphthyl acetate 0.2-2.0mg, benzyl aminoadenine 0.5-3.0mg, active carbon 1-2.0g, perfume (or spice)
Any of several broadleaf plants 50-150g, sucrose 10-20g, agar 5-6g, surplus are water, pH 5.5~5.7；
(3) rhizomes differentiation culture: the rhizomes group that step (2) obtains is transferred in differential medium and carries out differentiation training
It supports, until protocorms are differentiated to form the adventitious bud of high 3~5cm, described every liter of differential medium containing spending precious No. 1 1~2g, flower
No. 2 1~2g of treasured, 20~40mg of ferrous sulfate, 20~40mg of disodium ethylene diamine tetraacetate, 0.5~2g of peptone, banana puree 50~
80g, 50~100mg of activated carbon, 80~120mg of inositol, 1.5~2.5mg of glycine, 0.05~0.2mg of thiamine hydrochloride, hydrochloric acid
0.4~0.8mg of pyridoxol, 0.4~0.8mg of niacin, 3.0~5.0mg of 6- benzyl purine, 0.4~0.5mg of methyl α-naphthyl acetate, sucrose 20
~30g, 6~7g of agar, surplus are water, pH 5.4~5.6；
(4) adventitious bud rooting seedling culture: the adventitious bud that step (3) obtains is transferred in Rooting and hardening-off culture base and carries out seedling
Culture, described every liter of Rooting and hardening-off culture base containing spending No. 1 1~2g of treasured, spend treasured No. 2 1~2g, 20~40mg of ferrous sulfate, second
20~40mg of edetate disodium, 0.5~2g of peptone, 50~60g of mashed potatoes, 500~1000mg of activated carbon, inositol 80~
120mg, 1.5~2.5mg of glycine, 5~10mg of thiamine hydrochloride, 0.5~1.0mg of puridoxine hydrochloride, 5~10mg of niacin, naphthalene
0.5~1.0mg of acetic acid, 15~30g of sucrose, 6~7g of agar, surplus are water, pH 5.4~5.6；
(5) at transplantation of seedlings and cultivation: when the seedling of adventitious bud length to 5~8cm of height of seedling, being transferred to natural light lower refining seedling 10~20
It, then moves into Lan Shi: sawdust: cultivating in the mixed-matrix that bark volume ratio is 1:1:1, obtains sword-leaved cymbidium and be mixed with flame Cymbidium
Hand over kind.
7. asexual reproduction method according to claim 6, which is characterized in that inducing in rhizomes for the step (1) is trained
It supports and is cultivated in base GB1, condition of culture are as follows: 24~28 DEG C of cultivation temperature, 1500~2000lx of illuminance, illumination 12h/d.
8. asexual reproduction method according to claim 6, which is characterized in that the step (2) in proliferated culture medium
Middle carry out Multiplying culture, condition of culture are as follows: 24~28 DEG C of cultivation temperature, 1500~2000lx of illuminance, illumination 12h/d.
9. asexual reproduction method according to claim 6, which is characterized in that the step (3) in differential medium
In carry out differentiation culture, condition of culture are as follows: cultivation temperature be 25~29 DEG C, 2000~3000lx of illuminance, illumination 12h/d.
10. asexual reproduction method according to claim 6, which is characterized in that the step (4) is trained in strong plantlets and rootage
It supports and carries out seedling culture, condition of culture in base are as follows: cultivation temperature is 25~29 DEG C, 2000~3000lx of illuminance, illumination
12h/d。
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