CN107637523A - A kind of method for culturing seedlings of horseshoe - Google Patents
A kind of method for culturing seedlings of horseshoe Download PDFInfo
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Abstract
The invention discloses a kind of method for culturing seedlings of horseshoe, belong to agricultural plantation technology field, the implantation methods comprise the following steps：The pretreatment of kind shepherd's purse, Fiber differentiation, Multiplying culture, culture of rootage, nursery prepare and transplanted；Wherein, during Fiber differentiation, Multiplying culture and culture of rootage, tissue cultures are carried out by MS culture mediums and various additives；In the set-up procedure of nursery, nursery fertilizer made from the following raw material is applied with to nursery：Well-rotted farmyard manure, humic acid, dregs of beans, edible fungi residue, microbial bacteria, Poria cocos, pseudo-ginseng, agaric, fruit of negundo, phoenix tree flower, calcium superphosphate part and lime part；The problem of present invention solves the tissue-cultured seedling growth vigor that traditional tissue cultures obtain and unobvious, and the seedling resistance against diseases finally obtained is weaker.
Description
Technical field
The invention belongs to agricultural plantation technology field, and in particular to a kind of method for culturing seedlings of horseshoe.
Background technology
Horseshoe, alias water chestnut, monocot genus Cyperaceae, eaten using underground bulb as fruits and vegetables for people.Horseshoe epidermis is black
It is bright, internal pure white, delicious flavour and the effect of change is eaten, because its nutritive value is known as " underground pyrus nivalis ".Horseshoe is rich in people
Several mineral materials, vitamin needed for body, amino acid, there is clearing heat and detoxicating, diuresis, decompression and other effects, be a kind of excellent medicine
Eat the fruits and vegetables group food of dual-purpose, and one of key farm products of China's export.At present, horseshoe is disappeared by vast deeply in international market
The person of expense welcomes, and its major product has water chestnut starch, peeled water chestnut can and quick-frozen horseshoe.With the raising of quality of the life, horseshoe
Market demand also increases year by year, has good plantation prospect.In existing planting technology, horseshoe mainly carries out nothing using bulb
Sexual reproduction, but cultivation easily causes bulb deterioration all the year round, causes bulb not rounding, and size is uneven；And because maternal plant carries disease germs seriously, lead
Cause newborn bulb quality decline, yield reduction and unstable.In addition, there is also male water chestnut in production, plant strain growth downgrade and
Not balling；Therefore, the method by the progress nursery of tissue cultures is more and more concerned, but the group that traditional tissue cultures obtain
Seedling growth vigor and unobvious are trained, and finally obtain the problem of seedling resistance against diseases is weaker.
The content of the invention
The goal of the invention of the present invention is to provide a kind of method for culturing seedlings of horseshoe, to solve the group that traditional tissue cultures obtain
Seedling growth vigor and unobvious are trained, and finally obtain the problem of seedling resistance against diseases is weaker.
To reach above-mentioned purpose, the technical solution adopted in the present invention is：
A kind of method for culturing seedlings of horseshoe, comprises the following steps：
(1) shepherd's purse pretreatment is planted：First, horseshoe plentiful and substantial, without disease pest is chosen as kind of a shepherd's purse；Then, described kind of shepherd's purse is carried out
Cleaning, sterilizing, vernalization processing；Then, the master of described kind of shepherd's purse, lateral bud are cut as explant；Finally, the explant is carried out
Clean, disinfect it is rear stand-by；
(2) Fiber differentiation：The explant sterilized in step (1) is seeded to inducing culture；The induction training
Base is supported using MS culture mediums as minimal medium, and is appended below the additive of mass concentration：6- benzyl aminoadenines 2-3mg/L,
Methyl α-naphthyl acetate 0.1-0.2mg/L, dichlorphenoxyacetic acid 2-4mg/L, white granulated sugar 60-65mg/L, activated carbon 20-25mg/L, vitamin
C18-24mg/L, mannitol 15-18mg/L, diethyl aminoethyl hexanoate 0.01-0.08mg/L and agar 5000-7000mg/L；The explant
After body culture 10-12 days, grow from bud；
(3) Multiplying culture：When described in step (2) is from bud length to 2-3cm, it is cut into simple bud and is transferred to Multiplying culture
Multiplying culture is carried out in base；The proliferated culture medium is appended below adding for mass concentration using MS culture mediums as minimal medium
Add agent：6- benzyl aminoadenines 2-3mg/L, methyl α-naphthyl acetate 0.1-0.2mg/L, dichlorphenoxyacetic acid 2-4mg/L, white granulated sugar 60-
65mg/L, activated carbon 20-25g/L, corn oligopeptide powder 15-20g/L, coconut palm breast 100-120mg/L and agar 5000-
7000mg/L；
(4) culture of rootage：Treat that the simple bud in step (3) grows up to 3-5cm seedling, the seedling is cut, and connects
Kind carries out culture of rootage into root media；The root media adds such as using 1/2MS culture mediums as minimal medium
The additive of lower mass concentration：Methyl α-naphthyl acetate 0.5-0.6mg/L, white granulated sugar 60-65mg/L, activated carbon 20-25mg/L, zeatin 4-
6mg/L and agar 5000-8000mg/L；Seedling culture 30-35 days, you can obtain tissue-cultured seedling；
(5) nursery prepares：Selecting flat, fertile, the convenient paddy field of draining, every mu applies nursery fertilizer 1000- as nursery
1200kg；The nursery fertilizer is made by the raw material of following parts by weight：Well-rotted farmyard manure 150-200 parts, humic acid 8-10 parts, dregs of beans
12-20 parts, edible fungi residue 4-6 parts, microbial bacteria 0.2-0.4 parts, Poria cocos 1-2 parts, pseudo-ginseng 1-2 parts, agaric 2-3 parts, five-leaved chaste tree
Sub- 2-3 parts, phoenix tree flower 1-2 parts, calcium superphosphate 10-15 parts and lime 20-30 parts；
(6) tissue culture transplantation of seedlings：The nursery that the tissue culture transplantation of seedlings obtained in step (4) is obtained to step (5),
Seedling is obtained after cultivating one month.
Preferably, in step (1), the method that the explant carries out disinfection comprises the following steps：First, by the explant
75% Ethanol Treatment 30-60s of body；Then, then with 0.1% mercuric chloride sterilize 10-12 minutes；Finally, by the explant nothing
Bacterium water cleans 5-6 times, and blots the sterilized water on the explant with aseptic paper.
Preferably, in step (5), the microbial bacteria includes：Nitrogen-fixing bacteria, phosphate solubilizing bacteria, potassium solubilizing bacteria, bacillus, unwrapping wire
Bacterium, saccharomycete.
Preferably, insect pest preventing and controlling are carried out in step (6), after tissue-cultured seedling transplant survival, the insect pest preventing and controlling include stalk blight,
Powdery mildew and the preventing and treating of snout moth's larva, specific prevention and controls are as follows：
For stalk blight, every mu sprays：Propiconazole 40mL+ Flusilazole 40mL+ Tebuconazoles 20mL；
For powdery mildew, every mu sprays：Propiconazole 30mL+ triazolone 60mL+ Difenoconazoles 20mL；
For snout moth's larva, every mu sprays：Chlopyrifos 20mL+ AVMs 30m.
Preferably, the PH of the inducing culture, the proliferated culture medium and the root media is 5.8-
6.0。
Preferably, during the Fiber differentiation, the Multiplying culture and the culture of rootage, cultivation temperature control
At 24-28 DEG C, intensity of illumination is controlled in 1200-1400Lux.
Due to using above-mentioned technical proposal, the invention has the advantages that：
1. the present invention applies tissue cultures in horseshoe is bred, it can be very good solve the problems, such as seedling degeneration, so as to
Obtain excellent seedling；The present invention carries out tissue training stage by stage by inducing culture, proliferated culture medium and root media
Support.Contain in inducing culture：MS culture mediums, 6- benzyls aminoadenine, methyl α-naphthyl acetate, dichlorphenoxyacetic acid, white granulated sugar, activity
Charcoal, vitamin C, mannitol, diethyl aminoethyl hexanoate and agar；Wherein, MS culture mediums provide abundant nutriment and absorbed for explant；
Dichlorphenoxyacetic acid induction explant forms callus, and when concentration is 2-4mg/L, inductivity highest, concentration it is too low or
Person's excessive concentration can cause inductivity to reduce；6- benzyl aminoadenines further promote calli induction, its concentration with
Dichlorphenoxyacetic acid concentration is suitable, and facilitation effect is optimal.Methyl α-naphthyl acetate can promote the cell division of explant, improve from bud
Formation speed.During white granulated sugar is Fiber differentiation, the indispensable carbon source material of explant, its concentration is too low to be resulted in
It is few from bud, excessive concentration to from the formation of bud play containment effect；Activated carbon, vitamin C and mannitol are supported except providing
Point, moreover it is possible to effectively suppress the activity of polyphenol oxidase, reduce the generation of noxious material, predominantly reduce and produced in cellular process
Raw tannin, phenolic compound are in the presence of polyphenol oxidase, the quinones substance that changes after being oxidized.Diethyl aminoethyl hexanoate can improve
Plant inner chlorophyll, protein, the content and photosynthetic rate of nucleic acid, and peroxidase and nitrate reductase can be improved
Activity, promote the carbon and nitrogen metabolism of plant, promote the division and elongation of plant cell, so as to strengthen absorption of the plant to nutrient；Amine
The addition of fresh ester, the quality of obtained tissue-cultured seedling can be effectively improved.Agar is mainly used in stablizing the form of inducing culture.Increase
Grow the main component of culture medium and the main component of inducing culture is substantially the same, it is fast that it contains MS culture mediums, 6- benzyl amino glands
Purine, methyl α-naphthyl acetate, dichlorphenoxyacetic acid, white granulated sugar, activated carbon, corn oligopeptide powder, coconut palm breast and agar；Wherein, corn peptide is made
The mixture of a variety of small peptides obtained for zein by enzyme degraded, except with can directly absorb, dissolubility by force, stably
Property the strong, characteristic such as security is high beyond, also have the function that it is antiviral, anti-oxidant, reduce aberration rate；Coconut palm breast is organic addition
Thing, can be to provide necessary physiological activator and micro constitutent from the growth of bud.Contain in root media：1/2MS is trained
Support base, methyl α-naphthyl acetate, white granulated sugar, activated carbon, zeatin and agar；Wherein, a kind of natural basic element of cell division of zeatin, its with
Methyl α-naphthyl acetate shares, and can promote the division of root cell, so as to improve seedling root absorption and the ability using nutrient.The present invention
The tissue-cultured seedling obtained, the tissue-cultured seedling that relatively conventional type of rearing obtains, its growth ability, plant recovery of nutrient and growth vigor
Become apparent from, therefore, the horseshoe yield obtained after plantation is higher, has good quality.
2. the nursery fertilizer that the present invention in the set-up procedure of nursery, uses, can not only promote tissue-cultured seedling to grow, can also carry
The immunity and resistance against diseases of high tissue-cultured seedling；Nursery fertilizer is made by the following raw material：Well-rotted farmyard manure, humic acid, dregs of beans, edible mushroom
Bacteria residue, microbial bacteria, Poria cocos, pseudo-ginseng, agaric, fruit of negundo, phoenix tree flower, calcium superphosphate part and lime；Wherein, decomposed farmers'
Fertilizer, dregs of beans and edible fungi residue, calcium superphosphate can not only provide abundant nutritional ingredient, and it is organic can also to update soil
Matter, promote microbial reproduction；Humic acid can have fertilizer to increase with element humic acid fertilizer with reference to made of such as nitrogen, phosphorus, potassium
Effect, improved soil, stimulate tissue-cultured seedling to grow, improve the functions such as horseshoe quality.Microbial bacteria is with well-rotted farmyard manure, dregs of beans and food
Be carrier with bacterium bacteria residue, and under the stimulation of humic acid, produce a variety of catalytic decomposition enzymes and CEC, UGF somatomedin,
The beneficial microbe colonies such as disease-resistant factor, soil activating cytokine are induced, in culture fertility, improve chemical fertilizer utilization ratio, improvement horse
Hoof quality, enhancing tissue-cultured seedling disease and insect resistance and drought-resistant ability, purification and rehabilitating soil etc. have preferable effect.In addition, Fu
Siberian cocklebur energy antibacterial, Perishing prevention disease, droop；The anti-inflammatory of pseudo-ginseng energy, anti-virus, logical cardinal vein, toxin expelling elemen, activating soil, can anti-yellowtop.
Agaric can reduce the persticide residue in soil, can control bacterium, toxin expelling, logical cardinal vein；Fruit of negundo can strengthen horseshoe rice shoot immunity,
Antibacterial and increase soil permeability, conductibility；Phoenix tree flower being capable of antifungal, bacterium；Above-mentioned all medicines are used in combination, and improve tissue-cultured seedling
Immunity and resistance against diseases.Quick lime has sterilization functions, can eliminate the harmful levels of pathogens in nursery to a certain extent,
Avoid ensuring the healthy growth of tissue-cultured seedling.
Embodiment
Technical scheme is further described in detail with reference to embodiment.
Embodiment 1
A kind of method for culturing seedlings of horseshoe, comprises the following steps：
(1) shepherd's purse pretreatment is planted：First, horseshoe plentiful and substantial, without disease pest is chosen as kind of a shepherd's purse；Then, described kind of shepherd's purse is carried out
Cleaning, sterilizing, vernalization processing；Then, the master of described kind of shepherd's purse, lateral bud are cut as explant；Finally, the explant is carried out
Clean, disinfect it is rear stand-by.The method that the explant carries out disinfection comprises the following steps：First, the explant is used
75% Ethanol Treatment 30s；Then, then with 0.1% mercuric chloride sterilize 10 minutes；Finally, by explant sterile water wash 5
Time, and blot the sterilized water on the explant with aseptic paper.
(2) Fiber differentiation：The explant sterilized in step (1) is seeded to inducing culture.The induction training
Base is supported using MS culture mediums as minimal medium, and is appended below the additive of mass concentration：6- benzyl aminoadenines 2mg/L, naphthalene
Acetic acid 0.1mg/L, dichlorphenoxyacetic acid 2mg/L, white granulated sugar 60mg/L, activated carbon 20mg/L, vitamin C 18mg/L, mannitol
15mg/L, diethyl aminoethyl hexanoate 0.01mg/L and agar 5000mg/L；The PH of the inducing culture is 5.8.During Fiber differentiation,
At 24 DEG C, intensity of illumination is controlled in 1200Lux for cultivation temperature control.After the explant culture 10 days, grow from bud.
(3) Multiplying culture：When described in step (2) is from bud length to 2cm, it is cut into simple bud and is transferred to proliferated culture medium
Middle carry out Multiplying culture.The proliferated culture medium is appended below the addition of mass concentration using MS culture mediums as minimal medium
Agent：6- benzyl aminoadenines 2mg/L, methyl α-naphthyl acetate 0.1mg/L, dichlorphenoxyacetic acid 2mg/L, white granulated sugar 60mg/L, activated carbon
20g/L, corn oligopeptide powder 15g/L, coconut palm breast 100mg/L and agar 5000mg/L.The PH of the proliferated culture medium is 5.8.
During Multiplying culture, at 24 DEG C, intensity of illumination is controlled in 1200Lux for cultivation temperature control.
(4) culture of rootage：Treat that the simple bud in step (3) grows up to 3cm seedling, the seedling is cut, and is inoculated with
Culture of rootage is carried out into root media.The root media is appended below using 1/2MS culture mediums as minimal medium
The additive of mass concentration：Methyl α-naphthyl acetate 0.5mg/L, white granulated sugar 60mg/L, activated carbon 20mg/L, zeatin 4mg/L and agar
5000mg/L.The PH of the root media is 5.8.In process of rooting culture, cultivation temperature control is at 24 DEG C, intensity of illumination control
System is in 1200Lux.The seedling culture 30 days, you can obtain tissue-cultured seedling.
(5) nursery prepares：Flat, fertile, the convenient paddy field of draining is selected as nursery, every mu applies nursery fertilizer
1000kg.The nursery fertilizer is made by the raw material of following parts by weight：150 parts of well-rotted farmyard manure, 8 parts of humic acid, 12 parts of dregs of beans, food
With 4 parts of bacterium bacteria residue, 0.2 part of microbial bacteria, 1 part of Poria cocos, 1 part of pseudo-ginseng, 2 parts of agaric, 2 parts of fruit of negundo, 1 part of phoenix tree flower, peroxophosphoric acid
20 parts of 10 parts of calcium and lime.The microbial bacteria includes：Nitrogen-fixing bacteria, phosphate solubilizing bacteria, potassium solubilizing bacteria, bacillus, actinomyces, yeast
Bacterium.
(6) tissue culture transplantation of seedlings：The nursery that the tissue culture transplantation of seedlings obtained in step (4) is obtained to step (5),
Seedling is obtained after cultivating one month.Wherein, after tissue-cultured seedling transplant survival, insect pest preventing and controlling are carried out.It is withered that the insect pest preventing and controlling include stalk
Disease, powdery mildew and the preventing and treating of snout moth's larva, specific prevention and controls are as follows：
For stalk blight, every mu sprays：Propiconazole 40mL+ Flusilazole 40mL+ Tebuconazoles 20mL；
For powdery mildew, every mu sprays：Propiconazole 30mL+ triazolone 60mL+ Difenoconazoles 20mL；
For snout moth's larva, every mu sprays：Chlopyrifos 20mL+ AVMs 30m.
Embodiment 2
A kind of method for culturing seedlings of horseshoe, comprises the following steps：
(1) shepherd's purse pretreatment is planted：First, horseshoe plentiful and substantial, without disease pest is chosen as kind of a shepherd's purse；Then, described kind of shepherd's purse is carried out
Cleaning, sterilizing, vernalization processing；Then, the master of described kind of shepherd's purse, lateral bud are cut as explant；Finally, the explant is carried out
Clean, disinfect it is rear stand-by.The method that the explant carries out disinfection comprises the following steps：First, the explant is used
75% Ethanol Treatment 60s；Then, then with 0.1% mercuric chloride sterilize 12 minutes；Finally, by explant sterile water wash 5-6
Time, and blot the sterilized water on the explant with aseptic paper.
(2) Fiber differentiation：The explant sterilized in step (1) is seeded to inducing culture.The induction training
Base is supported using MS culture mediums as minimal medium, and is appended below the additive of mass concentration：6- benzyl aminoadenines 3mg/L, naphthalene
Acetic acid 0.2mg/L, dichlorphenoxyacetic acid 4mg/L, white granulated sugar 65mg/L, activated carbon 25mg/L, Catergen 4mg/L, mannitol
18mg/L, diethyl aminoethyl hexanoate 0.08mg/L and agar 7000mg/L；The PH of the inducing culture is 6.0.During Fiber differentiation,
At 28 DEG C, intensity of illumination is controlled in 1400Lux for cultivation temperature control.After the explant culture 12 days, grow from bud.
(3) Multiplying culture：When described in step (2) is from bud length to 3cm, it is cut into simple bud and is transferred to proliferated culture medium
Middle carry out Multiplying culture.The proliferated culture medium is appended below the addition of mass concentration using MS culture mediums as minimal medium
Agent：6- benzyl aminoadenines 3mg/L, methyl α-naphthyl acetate 0.2mg/L, dichlorphenoxyacetic acid 4mg/L, white granulated sugar 65mg/L, activated carbon
25g/L, corn oligopeptide powder 20g/L, coconut palm breast 120mg/L and agar 7000mg/L.The PH of the proliferated culture medium is 6.0.
During Multiplying culture, at 28 DEG C, intensity of illumination is controlled in 1400Lux for cultivation temperature control.
(4) culture of rootage：Treat that the simple bud in step (3) grows up to 5cm seedling, the seedling is cut, and is inoculated with
Culture of rootage is carried out into root media.The root media is appended below using 1/2MS culture mediums as minimal medium
The additive of mass concentration：Methyl α-naphthyl acetate 0.6mg/L, white granulated sugar 65mg/L, activated carbon 25mg/L, zeatin 6mg/L and agar
8000mg/L.The PH of the root media is 6.0.In process of rooting culture, cultivation temperature control is at 28 DEG C, intensity of illumination control
System is in 1400Lux.The seedling culture 35 days, you can obtain tissue-cultured seedling.
(5) nursery prepares：Flat, fertile, the convenient paddy field of draining is selected as nursery, every mu applies nursery fertilizer
1200kg.The nursery fertilizer is made by the raw material of following parts by weight：200 parts of well-rotted farmyard manure, 10 parts of humic acid, 20 parts of dregs of beans,
6 parts of edible fungi residue, 0.4 part of microbial bacteria, 2 parts of Poria cocos, 2 parts of pseudo-ginseng, 3 parts of agaric, 3 parts of fruit of negundo, excessively 2 parts of phoenix tree flower, phosphorus
30 parts of 15 parts of sour calcium and lime.The microbial bacteria includes：Nitrogen-fixing bacteria, phosphate solubilizing bacteria, potassium solubilizing bacteria, bacillus, actinomyces, ferment
Female bacterium.
(6) tissue culture transplantation of seedlings：The nursery that the tissue culture transplantation of seedlings obtained in step (4) is obtained to step (5),
Seedling is obtained after cultivating one month.Wherein, after tissue-cultured seedling transplant survival, insect pest preventing and controlling are carried out.It is withered that the insect pest preventing and controlling include stalk
Disease, powdery mildew and the preventing and treating of snout moth's larva, specific prevention and controls are as follows：
For stalk blight, every mu sprays：Propiconazole 40mL+ Flusilazole 40mL+ Tebuconazoles 20mL；
For powdery mildew, every mu sprays：Propiconazole 30mL+ triazolone 60mL+ Difenoconazoles 20mL；
For snout moth's larva, every mu sprays：Chlopyrifos 20mL+ AVMs 30m.
Embodiment 3
A kind of method for culturing seedlings of horseshoe, comprises the following steps：
(1) shepherd's purse pretreatment is planted：First, horseshoe plentiful and substantial, without disease pest is chosen as kind of a shepherd's purse；Then, described kind of shepherd's purse is carried out
Cleaning, sterilizing, vernalization processing；Then, the master of described kind of shepherd's purse, lateral bud are cut as explant；Finally, the explant is carried out
Clean, disinfect it is rear stand-by.The method that the explant carries out disinfection comprises the following steps：First, the explant is used
75% Ethanol Treatment 45s；Then, then with 0.1% mercuric chloride sterilize 11 minutes；Finally, by explant sterile water wash 6
Time, and blot the sterilized water on the explant with aseptic paper.
(2) Fiber differentiation：The explant sterilized in step (1) is seeded to inducing culture.The induction training
Base is supported using MS culture mediums as minimal medium, and is appended below the additive of mass concentration：6- benzyl aminoadenines 2.5mg/L,
Methyl α-naphthyl acetate 0.15mg/L, dichlorphenoxyacetic acid 3mg/L, white granulated sugar 62mg/L, activated carbon 22mg/L, Catergen 1mg/L, sweet dew
Alcohol 16mg/L, diethyl aminoethyl hexanoate 0.05mg/L and agar 6000mg/L；The PH of the inducing culture is 5.9.Fiber differentiation process
In, at 26 DEG C, intensity of illumination is controlled in 1300Lux for cultivation temperature control.After the explant culture 11 days, grow from bud.
(3) Multiplying culture：When described in step (2) is from bud length to 2.5cm, it is cut into simple bud and is transferred to Multiplying culture
Multiplying culture is carried out in base.The proliferated culture medium is appended below adding for mass concentration using MS culture mediums as minimal medium
Add agent：6- benzyl aminoadenines 2.5mg/L, methyl α-naphthyl acetate 0.15mg/L, dichlorphenoxyacetic acid 3mg/L, white granulated sugar 62mg/L, activity
Charcoal 22g/L, corn oligopeptide powder 17g/L, coconut palm breast 110mg/L and agar 6000mg/L.The PH of the proliferated culture medium is
5.9.During Multiplying culture, at 26 DEG C, intensity of illumination is controlled in 1300Lux for cultivation temperature control.
(4) culture of rootage：Treat that the simple bud in step (3) grows up to 4cm seedling, the seedling is cut, and is inoculated with
Culture of rootage is carried out into root media.The root media is appended below using 1/2MS culture mediums as minimal medium
The additive of mass concentration：Methyl α-naphthyl acetate 0.55mg/L, white granulated sugar 62mg/L, activated carbon 22mg/L, zeatin 5mg/L and agar
6500mg/L.The PH of the root media is 5.9.In process of rooting culture, cultivation temperature control is at 26 DEG C, intensity of illumination control
System is in 1300Lux.The seedling culture 32 days, you can obtain tissue-cultured seedling.
(5) nursery prepares：Flat, fertile, the convenient paddy field of draining is selected as nursery, every mu applies nursery fertilizer
1100kg.The nursery fertilizer is made by the raw material of following parts by weight：180 parts of well-rotted farmyard manure, 9 parts of humic acid, 16 parts of dregs of beans, food
With 5 parts of bacterium bacteria residue, 0.3 part of microbial bacteria, 1.5 parts of Poria cocos, 1.5 parts of pseudo-ginseng, 2.5 parts of agaric, 2.5 parts of fruit of negundo, phoenix tree flower 1.5
25 parts of part, 12 parts of calcium superphosphate and lime.The microbial bacteria includes：Nitrogen-fixing bacteria, phosphate solubilizing bacteria, potassium solubilizing bacteria, bacillus, put
Line bacterium, saccharomycete.
(6) tissue culture transplantation of seedlings：The nursery that the tissue culture transplantation of seedlings obtained in step (4) is obtained to step (5),
Seedling is obtained after cultivating one month.Wherein, after tissue-cultured seedling transplant survival, insect pest preventing and controlling are carried out.It is withered that the insect pest preventing and controlling include stalk
Disease, powdery mildew and the preventing and treating of snout moth's larva, specific prevention and controls are as follows：
For stalk blight, every mu sprays：Propiconazole 40mL+ Flusilazole 40mL+ Tebuconazoles 20mL；
For powdery mildew, every mu sprays：Propiconazole 30mL+ triazolone 60mL+ Difenoconazoles 20mL；
For snout moth's larva, every mu sprays：Chlopyrifos 20mL+ AVMs 30m.
On the other hand, applicant have also been devised a kind of comparative example of the method for culturing seedlings of horseshoe of the present invention, it is specific as follows：
Comparative example 1
When performing step (2), inducing culture used in the present invention is changed into conventional inducing culture, i.e.,：MS is cultivated
Base+6- benzyls aminoadenine+white granulated sugar+agar；When performing step (3), proliferated culture medium used in the present invention is changed into often
Proliferated culture medium is advised, i.e.,：MS culture medium+6- benzyls aminoadenine, methyl α-naphthyl acetate+white granulated sugar+agar；When performing step (4), it incite somebody to action this
Root media used in invention changes conventional root media into, i.e.,：Methyl α-naphthyl acetate+white granulated sugar+activated carbon+agar.Its elsewhere
Reason is same as Example 3.
Comparative example 2
When performing step (5), nursery fertilizer used in the present invention is changed into conventional use of farm manure and composite fertilizer, other
Processing is same as Example 3.
The cultivation situation of the tissue-cultured seedling obtained to embodiment 1, embodiment 2, embodiment 3, comparative example 1 and comparative example 2 is entered
Row statistics, statistical result are as shown in table 1.
Table 1
As can be seen from Table 1, the indices in embodiment 3 are significantly better than that comparative example 1, illustrate used in the present invention
Inducing culture, increment culture medium and the relatively conventional tissue culture medium (TCM) of root media, it promotes the effect of tissue-cultured seedling growth
Fruit is more notable, can more obtain the obvious seedling of growth vigor." tissue-cultured seedling illness rate " and " tissue-cultured seedling average mark in embodiment 3
This two indexs of tiller number " are significantly better than that comparative example 2, illustrate the relatively conventional fertilizer of nursery fertilizer used in the present invention, its is right
The growth promoting function of tissue-cultured seedling is more notable, and can improve the immunity and resistance against diseases of tissue-cultured seedling, more strong so as to obtain
Strong seedling.The present invention is improved the yield and quality of seedling, made by the seedling management of science and the Fertilizer ratio of science
Planting household obtains good economic benefit, is worthy to be popularized.
Described above is the detailed description for the present invention preferably possible embodiments, but embodiment is not limited to this hair
Bright patent claim, the equal change completed or modification change under the technical spirit suggested by all present invention, all should belong to
Cover the scope of the claims in the present invention.
Claims (6)
1. a kind of method for culturing seedlings of horseshoe, it is characterised in that comprise the following steps：
(1) shepherd's purse pretreatment is planted：First, horseshoe plentiful and substantial, without disease pest is chosen as kind of a shepherd's purse；Then, described kind of shepherd's purse is cleaned,
Sterilizing, vernalization processing；Then, the master of described kind of shepherd's purse, lateral bud are cut as explant；Finally, the explant is cleaned,
Disinfect rear stand-by；
(2) Fiber differentiation：The explant sterilized in step (1) is seeded to inducing culture；The inducing culture
Using MS culture mediums as minimal medium, and it is appended below the additive of mass concentration：6- benzyl aminoadenines 2-3mg/L, naphthalene second
Sour 0.1-0.2mg/L, dichlorphenoxyacetic acid 2-4mg/L, white granulated sugar 60-65mg/L, activated carbon 20-25mg/L, vitamin C 18-
24mg/L, mannitol 15-18mg/L, diethyl aminoethyl hexanoate 0.01-0.08mg/L and agar 5000-7000mg/L；The explant training
After supporting 10-12 days, grow from bud；
(3) Multiplying culture：When described in step (2) is from bud length to 2-3cm, it is cut into simple bud and is transferred in proliferated culture medium
Carry out Multiplying culture；The proliferated culture medium is appended below the additive of mass concentration using MS culture mediums as minimal medium：
6- benzyl aminoadenines 2-3mg/L, methyl α-naphthyl acetate 0.1-0.2mg/L, dichlorphenoxyacetic acid 2-4mg/L, white granulated sugar 60-65mg/L,
Activated carbon 20-25g/L, corn oligopeptide powder 15-20g/L, coconut palm breast 100-120mg/L and agar 5000-7000mg/L；
(4) culture of rootage：Treat that the simple bud in step (3) grows up to 3-5cm seedling, the seedling is cut, and is seeded to
Culture of rootage is carried out in root media；The root media is appended below matter using 1/2MS culture mediums as minimal medium
Measure the additive of concentration：Methyl α-naphthyl acetate 0.5-0.6mg/L, white granulated sugar 60-65mg/L, activated carbon 20-25mg/L, zeatin 4-6mg/
L and agar 5000-8000mg/L；Seedling culture 30-35 days, you can obtain tissue-cultured seedling；
(5) nursery prepares：Selecting flat, fertile, the convenient paddy field of draining, every mu applies nursery fertilizer 1000- as nursery
1200kg；The nursery fertilizer is made by the raw material of following parts by weight：Well-rotted farmyard manure 150-200 parts, humic acid 8-10 parts, dregs of beans
12-20 parts, edible fungi residue 4-6 parts, microbial bacteria 0.2-0.4 parts, Poria cocos 1-2 parts, pseudo-ginseng 1-2 parts, agaric 2-3 parts, five-leaved chaste tree
Sub- 2-3 parts, phoenix tree flower 1-2 parts, calcium superphosphate 10-15 parts and lime 20-30 parts；
(6) tissue culture transplantation of seedlings：The nursery that the tissue culture transplantation of seedlings obtained in step (4) is obtained to step (5), cultivate
Seedling is obtained after one month.
2. the method for culturing seedlings of a kind of horseshoe according to claim 1, it is characterised in that in step (1), the explant enters
The method of row sterilization comprises the following steps：First, by the explant with 75% Ethanol Treatment 30-60s；Then, then with 0.1%
Mercuric chloride sterilizes 10-12 minutes；Finally, by explant sterile water wash 5-6 times, and the explant is blotted with aseptic paper
On sterilized water.
A kind of 3. method for culturing seedlings of horseshoe according to claim 1, it is characterised in that in step (5), the microbial bacteria
Including：Nitrogen-fixing bacteria, phosphate solubilizing bacteria, potassium solubilizing bacteria, bacillus, actinomyces, saccharomycete.
A kind of 4. method for culturing seedlings of horseshoe according to claim 1, it is characterised in that in step (6), tissue culture transplantation of seedlings into
Insect pest preventing and controlling are carried out after work；The insect pest preventing and controlling include stalk blight, powdery mildew and the preventing and treating of snout moth's larva, and specific prevention and controls are such as
Under：
For stalk blight, every mu sprays：Propiconazole 40mL+ Flusilazole 40mL+ Tebuconazoles 20mL；
For powdery mildew, every mu sprays：Propiconazole 30mL+ triazolone 60mL+ Difenoconazoles 20mL；
For snout moth's larva, every mu sprays：Chlopyrifos 20mL+ AVMs 30m.
5. the method for culturing seedlings of a kind of horseshoe according to claim 1, it is characterised in that the inducing culture, the increasing
The PH for growing culture medium and the root media is 5.8-6.0.
6. the method for culturing seedlings of a kind of horseshoe according to claim 5, it is characterised in that the Fiber differentiation, the propagation
During culture and the culture of rootage, at 24-28 DEG C, intensity of illumination is controlled in 1200- for cultivation temperature control
1400Lux。
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