CN105638480B - A kind of capsicum variety, which is cultivated, uses flower pesticide Fiber differentiation based formulas - Google Patents
A kind of capsicum variety, which is cultivated, uses flower pesticide Fiber differentiation based formulas Download PDFInfo
- Publication number
- CN105638480B CN105638480B CN201610086925.1A CN201610086925A CN105638480B CN 105638480 B CN105638480 B CN 105638480B CN 201610086925 A CN201610086925 A CN 201610086925A CN 105638480 B CN105638480 B CN 105638480B
- Authority
- CN
- China
- Prior art keywords
- culture
- flower pesticide
- vitamin
- capsicum
- culture medium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Developmental Biology & Embryology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention provides a kind of capsicum variety cultivation flower pesticide inducing culture, the formula of the culture medium is by a certain proportion of KNO3、NH4NO3、KH2PO4、MgSO4∙H2O、CaCl2∙2H2O、Na2‑EDTA、FeSO4∙7H2O、MnSO4∙H2O、ZnSO4∙7H2O、H3BO3、KI、CuSO4∙5H2O、CoCl∙6H2O、NaMoO4∙2H2O、AgNO3, inositol, vitamin B1, vitamin B6, nicotinic acid, glycine, folic acid, cyclic guanosine monophosphate, maltose, sucrose, plant gel, 2,4 D, kinetin, multiple phthalein nucleic acid, biotin, vitamin C, activated carbon and protein hydrolysate constituted.Culture medium of the present invention has the advantages that expeditiously induction pepper anther produces pollen callus, can significantly improve pepper anther culture efficiency, accelerate capsicum prevalent variety cultivation process.
Description
Technical field
The present invention relates to a kind of capsicum variety cultivation flower pesticide Fiber differentiation based formulas, and in particular to one kind significantly improves peppery
The Fiber differentiation based formulas of green pepper flower pesticide healing rate, belongs to technical field of agriculture science.
Background technology
Capsicum（Capsicum annuumL.）, also known as chilly, hot pepper, green pepper, green pepper etc. are solanaceous vegetable crop, one
Important industrial crops are planted, are also a kind of vegetable crop generally cultivated in the world.Containing abundant in chili peel, placental tissue
Carrotene and vitamin C, particularly Vc contents occupy the hat of vegetables in line green pepper, thus nutritive value is high.Contain in capsicum
Capsaicine has the effect whetting the appetite, help digest, strengthening resistance of human body, resisting a variety of diseases.Yield of hot pepper is high, supply
Season is long, and edible way is various, nutritious, is a kind of vegetables being loved by people.
Torrid areas of the capsicum originating from Central and South America and the band of Mexico one, is initially taken to western class by Columbus in 1493
Tooth, travels to Europe, then China is passed in the 17th century through India, is just planted extensively in China various regions afterwards.Although China capsicum
The cultivation history that plantation only has 300 years so far, but have developed rapidly, China's paprike cultivated area and annual production are continuous
Increase.In recent years, in addition to conventional outdoor cropping, facility cultivated area is also constantly expanding, and the whole nation has formed several paprike rule
Modelling production base.According to statistics, China's paprike cultivated area in 2005 reaches 1,300,000 hm2, account for national vegetable cultivation area
10%, various regions market paprike realizes year-round supply substantially.Present China has been pepper planting state maximum in the world, together
When, it is also the man of main producting and exporting country of chilli in the world, finds a good sale in Russia, Singapore, Malaysia, South Korea and Europe
Deng state and area.Capsicum has highly important status in China's agricultural production, carries out the genetic breeding research work of capsicum
It is significant.
As the improvement of people's living standards, the requirement to Quality of Capsicum is improved constantly；While Protectorate cultivation technology
Promote, the kind that seed selection is applied to Protectorate cultivation refer in face of numerous scientific workers；Because China not yet carries out greatly
Scale intensive manufacture, continuous cropping is serious, causes the large area such as virosis, anthracnose and epidemic disease to occur, it has also become influence capsicum life
The major obstacles of production.Therefore, seed selection is best in quality, be adapted to different condition cultivation and disease-resistant capsicum new varieties, it has also become vast
Breeder's urgent problem.
Breeding for heterosis being utilized conventional pepper breeding, i.e., coordinate force are high, Heterosis from two hybridization more
Obvious and character pure line cultivar with strong complementarity or self-mating system or sterile line carry out artificial pollination production hybrid seed.But capsicum
It is a kind of Constantly allogamous plant, natural hybrization rate is 4%~10%, and some kinds are purified up to 25% by selfing isolation and selection
And stable self-mating system, generally requiring the selection of the generation of carry out 5~6 could obtain, and not only the time is long, and exist between gene aobvious hidden
Sexual intercourse, it is difficult to select Comprehensive Traits it is excellent, with outstanding advantages and coordinate force it is strong, suitably as the product of strong advantage parent
System, and crop genetic diversity can be caused to decline the loss with desirable genes.With the development of plant tissue culture technique, plant
There is thing cell totipotency to be confirmed extensively, therefore, in vitro culture flower pesticide, the artificial development way for changing microspore
Footpath, makes the termination of its Development of Gametophytes approach, turns to sporinite development, occurs by embryo or adventitious organogenesis, is formed completely
Haplobiont, effective means is provided for artificial a large amount of production haplobionts.
Haploid breeding is as a kind of new breeding methods, by cultivating monoploid, and then monoploid material is added
Times, pure lines suitably as strong advantage parent just can be selected within a short period of time, simultaneously because in the absence of intergenic aobvious
Recessive relation, therefore some characters controlled by recessive gene just can be fully used, and so both substantially reduce breeding
The time limit, and the hereditary basis of selected excellent strain can be enriched, such strain is easily achieved the apolegamy of parent.Therefore, carry out peppery
The research of green pepper monoploid culture technique, undoubtedly has the parental line of outstanding advantages for seed selection, accelerates breeding process, cultivates strong
The combination of advantage capsicum is significant.
Pepper anther culture work beginning is more early, from the early 1970s, lot of domestic and foreign scholar trains to pepper anther
Support and studied.1973, Wang Yuying and George reports obtained capsicum haplobiont.Dumas in 1981 etc. is utilized
The technology of Anther Culture have successfully been obtained the colony of 3 different genetic origins, and the principal element for influenceing Anther Culture is carried out
More systematic research, it is proposed that a set of effective cultural method.Nineteen ninety Li Chunling etc., which is reported, successfully to be selected both at home and abroad
First with flower training method be bred as capsicum variety " seaflower No. three ", later but in succession be bred as Haifeng county 1, Haifeng county 2, Haifeng county 5,
6 paprike kinds or the cenospecies such as Haifeng county 12, Haifeng county 14, and the popularization and application in production, show obvious precocity
Property and yielding ability.Chen Xiaoshi etc. carries out Anther Culture using local varieties " Yi County pimento ", and being bred as colored training kind, " plug spends one
Number ".Anther Culture is widely used in pepper breeding work as emerging technology.
At present, though pepper anther culture research achieves greater advance, some problems are above still suffered from theoretical and application.
Changed on chilli microspore from Development of Gametophytes approach to sporinite approach, and then form monoploid or double haploid
Developmental regulation mechanism is still not clear.Medium component and condition of culture interphase interaction ambiguity Chu, embryoid or callus group
The inductivity and differentiation rate knitted are relatively low, have larger gap from application request.Therefore, accelerate capsicum Anther culture breeding mechanism to grind
Study carefully, the higher culture medium prescription of antherderived callus inductivity differentiation rate is developed, with important current demand.
Culture medium is the material base of Anther Culture, is directly connected to the growth and differentiation of culture, and nutrient media components is
The successful very important factor of influence flower training, a variety of capsicum training experiments show：The suitable amounts of culture medium each component,
And certain syntagmatic between them, Efficiency can be significantly affected, finds and optimum organization Anther Culture is organic additional
Material, can further improve Anther Culture Ability.Therefore, the optimization of medium component, growth regulator species, with when concentration
Regulation, the screening of Organic additives matter etc., it is the important research object for filtering out the higher flower training culture medium of culture efficiency.It is logical
Groping and put into practice after pepper anther culture for many years, we further optimize minimal medium formula, and continuously attempt to plus
Enter the organic additive of some raising pepper anther induction forces and combine the collocation of its species and concentration, finally found out a kind of peppery
Green pepper breed of variety flower pesticide Fiber differentiation based formulas.Industry Promotion of our investigative technique for promotion capsicum Anther culture breeding
Undoubtedly there is great realistic meaning.
The content of the invention
The invention provides a kind of capsicum variety cultivation flower pesticide inducing culture, it is characterised in that：The culture medium is matched somebody with somebody
Side is as follows：
KNO3900~1000mg/L, NH4NO3800~850mg/L, KH2PO4650~700mg/L, MgSO4∙H2O
1100~1300mg/L, CaCl2∙2H2O 200~250mg/L, Na2- EDTA 72~78mg/L, FeSO4∙7H253~57mg/ of O
L, MnSO4∙H2O 16~18mg/L, ZnSO4∙7H2O 17~19mg/L, H3BO35.5~6.5mg/L, KI 0.75~
0.85mg/L, CuSO4∙5H2O 0.02~0.03mg/L, CoCl 6H2O 0.025~0.035mg/L, NaMoO4∙2H2O 0.2~
0.3mg/L, AgNO34.5~5.5mg/L, 90~110mg/L of inositol, 0.45~0.55mg/L of vitamin B1, vitamin B6
0.45~0.55mg/L, 4.5~5.5mg/L of nicotinic acid, 1.8~2.2mg/L of glycine, 0.45~0.55mg/L of folic acid, ring phosphorus
Sour 0.25~0.35g/L of guanosine, 18~22g/L of maltose, 14~16g/L of sucrose, 4.5~5.5g/L of plant gel；
2,4-D 0.5~0.7mg/L, 0.4~0.6mg/L of kinetin, multiple phthalein 0.7~0.9mg/L of nucleic acid, biotin
0.04~0.06mg/L, vitamin C 40~50 μm of ol/L, 150~200mg/L of activated carbon, 0.35~0.45g/ of protein hydrolysate
L。
The optimum content of described culture medium prescription is：KNO3950mg/L, NH4NO3825mg/L, KH2PO4 675mg/
L, MgSO4∙H2O 1200mg/L, CaCl2∙2H2O 225mg/L, Na2- EDTA 75mg/L, FeSO4∙7H2O 55mg/L, MnSO4∙
H2O 17mg/L, ZnSO4∙7H2O 18mg/L, H3BO36.0mg/L, KI 0.8mg/L, CuSO4∙5H2O 0.025mg/L, CoCl
∙6H2O 0.03mg/L, NaMoO4∙2H2O 0.25mg/L, AgNO35.0mg/L, inositol 100mg/L, vitamin B1 0.5mg/
L, vitamin B6 0.5mg/L, nicotinic acid 5.0mg/L, glycine 2.0mg/L, folic acid 0.5mg/L, cyclic guanosine monophosphate 0.3g/
L, maltose 20g/L, sucrose 15g/L, plant gel 5.0g/L；
2,4-D 0.6mg/L, kinetin 0.5mg/L, multiple phthalein nucleic acid 0.8mg/L, biotin 0.05mg/L, vitamin C
45 μm of ol/L, activated carbon 175mg/L, protein hydrolysate 0.4g/L.
The secure ph of described culture medium prescription is：5.6~6.0, optimal secure ph is 5.8.
Culture medium of the present invention has the advantages that expeditiously induction pepper anther produces pollen callus, can be notable
Pepper anther culture efficiency is improved, accelerates capsicum prevalent variety cultivation process.The following examples and contrast experiment can be clearly
The characteristics of reflecting culture medium of the present invention.
Embodiment
Embodiment 1
It is formulated as follows the culture medium of formula：
KNO3950mg/L, NH4NO3825mg/L, KH2PO4675mg/L, MgSO4∙H2O 1200mg/L, CaCl2∙2H2O
225mg/L, Na2- EDTA 75mg/L, FeSO4∙7H2O 55mg/L, MnSO4∙H2O 17mg/L, ZnSO4∙7H2O 18mg/L,
H3BO36.0mg/L, KI 0.8mg/L, CuSO4∙5H2O 0.025mg/L, CoCl 6H2O 0.03mg/L, NaMoO4∙2H2O
0.25mg/L, AgNO35.0mg/L, inositol 100mg/L, vitamin B1 0.5mg/L, vitamin B6 0.5mg/L, nicotinic acid
5.0mg/L, glycine 2.0mg/L, folic acid 0.5mg/L, cyclic guanosine monophosphate 0.3g/L, maltose 20g/L, sucrose 15g/
L, plant gel 5.0g/L；
2,4-D 0.6mg/L, kinetin 0.5mg/L, multiple phthalein nucleic acid 0.8mg/L, biotin 0.05mg/L, vitamin C
45 μm of ol/L, activated carbon 175mg/L, protein hydrolysate 0.4g/L, pH 5.8.
Capsicum variety Xinfeng 5 and capsicum annum fasciculatum are selected as Anther Culture donor, fine degree/day is relatively low, sunshine is weaker
In morning, choose the bud endangered without disease pest for being grown on open country（The nearly isometric or petal of calyx and petal is slightly longer than calyx, microscopy this
When microspore be in monokaryon late period, for the best period of culture）, preserved immediately in 4 DEG C of refrigerators after taking.By calyx before inoculation
10-15min is rinsed with running water, 75% ethanol leaching 20s, 0.1% mercuric chloride sterilization 10min, then aseptic water washing 3-4 times is taken out
Flower pesticide is placed in standby in the culture dish for filling sterile blotting paper.Flower pesticide carefully is cut out from bud during inoculation, is inoculated with
In filling in the 60mm × 15mm culture dish of 10ml culture mediums, 25-30 pieces of flower pesticide is inoculated with per ware, is then sealed with sealed membrane.
Light culture 8 days or so under prior to 33 DEG C high temperature after flower pesticide inoculation, 25 DEG C of light cultures are to embryoid appearance afterwards, and it is rearmounted that flower pesticide goes out embryo
In being cultivated under 25 DEG C of light, callus induction rate is calculated, regeneration culture medium is then transferred to.
Callus induction rate（%）=（Callus produces number/inoculation flower pesticide sum）×100.
Embodiment 2
It is formulated as follows the culture medium of formula（AgNO3Add the preferred of concentration）：
KNO3950mg/L, NH4NO3825mg/L, KH2PO4675mg/L, MgSO4∙H2O 1200mg/L, CaCl2∙2H2O
225mg/L, Na2- EDTA 75mg/L, FeSO4∙7H2O 55mg/L, MnSO4∙H2O 17mg/L, ZnSO4∙7H2O 18mg/L,
H3BO36.0mg/L, KI 0.8mg/L, CuSO4∙5H2O 0.025mg/L, CoCl 6H2O 0.03mg/L, NaMoO4∙2H2O
0.25mg/L, inositol 100mg/L, vitamin B1 0.5mg/L, vitamin B6 0.5mg/L, nicotinic acid 5.0mg/L, glycine
2.0mg/L, folic acid 0.5mg/L, cyclic guanosine monophosphate 0.3g/L, maltose 20g/L, sucrose 15g/L, plant gel 5.0g/
L；
2,4-D 0.6mg/L, kinetin 0.5mg/L, multiple phthalein nucleic acid 0.8mg/L, biotin 0.05mg/L, vitamin C
45 μm of ol/L, activated carbon 175mg/L, protein hydrolysate 0.4g/L, pH 5.8.
AgNO3Addition concentration set following 8 kinds：0；1.0mg/L；2.0mg/L；3.0mg/L；4.0mg/L；6.0mg/L；
7.0mg/L；8.0mg/L.
Two capsicum varieties are equally chosen as flower pesticide donor, operating method, cultural method be the same as Example 1, calculating is cured
Injured tissue inductivity.
Embodiment 3
It is formulated as follows the culture medium of formula（Glycine adds the preferred of concentration）：
KNO3950mg/L, NH4NO3825mg/L, KH2PO4675mg/L, MgSO4∙H2O 1200mg/L, CaCl2∙2H2O
225mg/L, Na2- EDTA 75mg/L, FeSO4∙7H2O 55mg/L, MnSO4∙H2O 17mg/L, ZnSO4∙7H2O 18mg/L,
H3BO36.0mg/L, KI 0.8mg/L, CuSO4∙5H2O 0.025mg/L, CoCl 6H2O 0.03mg/L, NaMoO4∙2H2O
0.25mg/L, AgNO35.0mg/L, inositol 100mg/L, vitamin B1 0.5mg/L, vitamin B6 0.5mg/L, nicotinic acid
5.0mg/L, folic acid 0.5mg/L, cyclic guanosine monophosphate 0.3g/L, maltose 20g/L, sucrose 15g/L, plant gel 5.0g/
L；
2,4-D 0.6mg/L, kinetin 0.5mg/L, multiple phthalein nucleic acid 0.8mg/L, biotin 0.05mg/L, vitamin C
45 μm of ol/L, activated carbon 175mg/L, protein hydrolysate 0.4g/L, pH 5.8.
The addition concentration of glycine sets following 4 kinds：0；1.0mg/L；3.0mg/L；4.0mg/L.
Two capsicum varieties are equally chosen as flower pesticide donor, operating method, cultural method be the same as Example 1, calculating is cured
Injured tissue inductivity.
Embodiment 4
It is formulated as follows the culture medium of formula（Cyclic guanosine monophosphate adds the preferred of concentration）：
KNO3950mg/L, NH4NO3825mg/L, KH2PO4675mg/L, MgSO4∙H2O 1200mg/L, CaCl2∙2H2O
225mg/L, Na2- EDTA 75mg/L, FeSO4∙7H2O 55mg/L, MnSO4∙H2O 17mg/L, ZnSO4∙7H2O 18mg/L,
H3BO36.0mg/L, KI 0.8mg/L, CuSO4∙5H2O 0.025mg/L, CoCl 6H2O 0.03mg/L, NaMoO4∙2H2O
0.25mg/L, AgNO35.0mg/L, inositol 100mg/L, vitamin B1 0.5mg/L, vitamin B6 0.5mg/L, nicotinic acid
5.0mg/L, glycine 2.0mg/L, folic acid 0.5mg/L, maltose 20g/L, sucrose 15g/L, plant gel 5.0g/L；
2,4-D 0.6mg/L, kinetin 0.5mg/L, multiple phthalein nucleic acid 0.8mg/L, biotin 0.05mg/L, vitamin C
45 μm of ol/L, activated carbon 175mg/L, protein hydrolysate 0.4g/L, pH 5.8.
The addition concentration of cyclic guanosine monophosphate sets following 5 kinds：0；0.1g/L；0.2g/L；0.4g/L；0.5g/L.
Two capsicum varieties are equally chosen as flower pesticide donor, operating method, cultural method be the same as Example 1, calculating is cured
Injured tissue inductivity.
Embodiment 5
It is formulated as follows the culture medium of formula（Plant gel adds the preferred of concentration）：
KNO3950mg/L, NH4NO3825mg/L, KH2PO4675mg/L, MgSO4∙H2O 1200mg/L, CaCl2∙2H2O
225mg/L, Na2- EDTA 75mg/L, FeSO4∙7H2O 55mg/L, MnSO4∙H2O 17mg/L, ZnSO4∙7H2O 18mg/L,
H3BO36.0mg/L, KI 0.8mg/L, CuSO4∙5H2O 0.025mg/L, CoCl 6H2O 0.03mg/L, NaMoO4∙2H2O
0.25mg/L, AgNO35.0mg/L, inositol 100mg/L, vitamin B1 0.5mg/L, vitamin B6 0.5mg/L, nicotinic acid
5.0mg/L, glycine 2.0mg/L, folic acid 0.5mg/L, cyclic guanosine monophosphate 0.3g/L, maltose 20g/L, sucrose 15g/
L；
2,4-D 0.6mg/L, kinetin 0.5mg/L, multiple phthalein nucleic acid 0.8mg/L, biotin 0.05mg/L, vitamin C
45 μm of ol/L, activated carbon 175mg/L, protein hydrolysate 0.4g/L, pH 5.8.
The addition concentration of plant gel sets following 2 kinds：2.5g/L plant gel+2.5g/L agar；5.0g/ agar.
Two capsicum varieties are equally chosen as flower pesticide donor, operating method, cultural method be the same as Example 1, calculating is cured
Injured tissue inductivity.
Embodiment 6
It is formulated as follows the culture medium of formula（Vitamin C adds the preferred of concentration）：
KNO3950mg/L, NH4NO3825mg/L, KH2PO4675mg/L, MgSO4∙H2O 1200mg/L, CaCl2∙2H2O
225mg/L, Na2- EDTA 75mg/L, FeSO4∙7H2O 55mg/L, MnSO4∙H2O 17mg/L, ZnSO4∙7H2O 18mg/L,
H3BO36.0mg/L, KI 0.8mg/L, CuSO4∙5H2O 0.025mg/L, CoCl 6H2O 0.03mg/L, NaMoO4∙2H2O
0.25mg/L, AgNO35.0mg/L, inositol 100mg/L, vitamin B1 0.5mg/L, vitamin B6 0.5mg/L, nicotinic acid
5.0mg/L, glycine 2.0mg/L, folic acid 0.5mg/L, cyclic guanosine monophosphate 0.3g/L, maltose 20g/L, sucrose 15g/
L, plant gel 5.0g/L；
2,4-D 0.6mg/L, kinetin 0.5mg/L, multiple phthalein nucleic acid 0.8mg/L, biotin 0.05mg/L, activated carbon
175mg/L, protein hydrolysate 0.4g/L, pH 5.8.
Ascorbic addition concentration sets following 5 kinds：0；15μmol/L；30μmol/L；60μmol/L；75μmol/L.
Two capsicum varieties are equally chosen as flower pesticide donor, operating method, cultural method be the same as Example 1, calculating is cured
Injured tissue inductivity.
Embodiment 7
It is formulated as follows the culture medium of formula（Activated carbon adds the preferred of concentration）：
KNO3950mg/L, NH4NO3825mg/L, KH2PO4675mg/L, MgSO4∙H2O 1200mg/L, CaCl2∙2H2O
225mg/L, Na2- EDTA 75mg/L, FeSO4∙7H2O 55mg/L, MnSO4∙H2O 17mg/L, ZnSO4∙7H2O 18mg/L,
H3BO36.0mg/L, KI 0.8mg/L, CuSO4∙5H2O 0.025mg/L, CoCl 6H2O 0.03mg/L, NaMoO4∙2H2O
0.25mg/L, AgNO35.0mg/L, inositol 100mg/L, vitamin B1 0.5mg/L, vitamin B6 0.5mg/L, nicotinic acid
5.0mg/L, glycine 2.0mg/L, folic acid 0.5mg/L, cyclic guanosine monophosphate 0.3g/L, maltose 20g/L, sucrose 15g/
L, plant gel 5.0g/L；
2,4-D 0.6mg/L, kinetin 0.5mg/L, multiple phthalein nucleic acid 0.8mg/L, biotin 0.05mg/L, vitamin C
45 μm of ol/L, protein hydrolysate 0.4g/L, pH 5.8.
The addition concentration of activated carbon sets following 8 kinds：0；35mg/L；70mg/L；105mg/L；140mg/L；210mg/L；
245mg/L；280mg/L.
Two capsicum varieties are equally chosen as flower pesticide donor, operating method, cultural method be the same as Example 1, calculating is cured
Injured tissue inductivity.
Embodiment 8
It is formulated as follows the culture medium of formula（Protein hydrolysate adds the preferred of concentration）：
KNO3950mg/L, NH4NO3825mg/L, KH2PO4675mg/L, MgSO4∙H2O 1200mg/L, CaCl2∙2H2O
225mg/L, Na2- EDTA 75mg/L, FeSO4∙7H2O 55mg/L, MnSO4∙H2O 17mg/L, ZnSO4∙7H2O 18mg/L,
H3BO36.0mg/L, KI 0.8mg/L, CuSO4∙5H2O 0.025mg/L, CoCl 6H2O 0.03mg/L, NaMoO4∙2H2O
0.25mg/L, AgNO35.0mg/L, inositol 100mg/L, vitamin B1 0.5mg/L, vitamin B6 0.5mg/L, nicotinic acid
5.0mg/L, glycine 2.0mg/L, folic acid 0.5mg/L, cyclic guanosine monophosphate 0.3g/L, maltose 20g/L, sucrose 15g/
L, plant gel 5.0g/L；
2,4-D 0.6mg/L, kinetin 0.5mg/L, multiple phthalein nucleic acid 0.8mg/L, biotin 0.05mg/L, vitamin C
45 μm of ol/L, activated carbon 175mg/L, pH 5.8.
The addition concentration of protein hydrolysate sets following 6 kinds：0；0.1g/L；0.2g/L；0.3g/L；0.5g/L；0.6g/L.
Two capsicum varieties are equally chosen as flower pesticide donor, operating method, cultural method be the same as Example 1, calculating is cured
Injured tissue inductivity.
Culture medium contrast experiment
Embodiment 2-8 is contrasted with embodiment 1 respectively, the result of contrast is as shown in the table：
It can be seen that and induced using culture medium ratio provided by the present invention using other any contrast culture mediums from table 1-7
Pepper anther callus it is more efficient, show that the culture medium prescription that provides of the present invention is by a large amount of single-factors, multiple-factor
Screened optimum combination is tested, we have also made the change optimization of small range single-factor around the combination matching of the present invention for many years
Composite test, substantial amounts of experimental study also show that the component proportion of the present invention is optimal.Use culture provided by the present invention
Base is up to 18.4% and 20.2% to the callus induction rate of capsicum variety Xinfeng 5, capsicum annum fasciculatum, has absolutely proved that the present invention is carried
The culture medium of confession is a kind of excellent pepper anther inducing culture, has filled up the current pepper anther induction for having application value and has trained
The blank of base is supported, with higher industry promotional value and theoretical research value.
Embodiment 9
In order to which culture medium relatively more provided by the present invention induces pepper anther callus effect with the culture medium that forefathers use
Difference, our 4 pepper anther calli induction medias formulas from related literature search are configured to antherderived callus Fiber differentiation
Base, same to choose capsicum variety Xinfeng 5, capsicum annum fasciculatum as flower pesticide donor induction antherderived callus, operating method, cultural method are same
Embodiment 1, calculates callus induction rate.
Other 4 inducing cultures are：
MS1Culture medium：MS minimal medium+3g/L activated carbon+0.5mg/LNAA+1.0mg/LKT+0.1mg/L 6-
BA+5mg/L AgNO3, pH is 5.8（Documents come from Zhao Ji etc.《Shadow of the different culture media to pepper anther culture
Ring》）；
NTH1Culture medium：NTH minimal medium+3g/L activated carbons+0.5mg/LNAA+1.0mg/LKT+0.1mg/L
6-BA+5mg/L AgNO3, pH is 5.8（Documents come from Zhao Ji etc.《Shadow of the different culture media to pepper anther culture
Ring》）；
B5 medium：B5 minimal medium+30g/L sucrose+6g/L agar+3g/L activated carbon+0.5mg/L NAA+
1.0mg/L KT, pH are 5.8（Documents come from that to pay text graceful etc.《Different culture media and phase of picking flowers are to pepper anther culture
Influence》）；；
CP culture mediums：CP minimal medium+30g/L sucrose+6g/L agar+3g/L activated carbon+0.5mg/L NAA+
1.0mg/L KT, pH are 5.8（Documents come from that to pay text graceful etc.《Different culture media and phase of picking flowers are to pepper anther culture
Influence》）.
Culture medium contrast experiment
Embodiment 9 is contrasted with embodiment 1, the result of contrast is as shown in the table：
From the point of view of the inducing effect of different culture media, culture medium of the present invention is to the average inductivity of pepper anther callus
19.3%, and MS1Culture medium, NTH1Culture medium, B5 medium and CP culture mediums then only have 8.6%, 8.7%, 2.3% and 2.4% respectively,
It can be found that the present invention will be significantly higher than other 3 kinds of inducing cultures to the inductivity of pepper anther callus.
9 embodiments of the above can illustrate that the component proportion for the culture medium prescription that the present invention is provided is optimum combination, this hair
The pepper anther inducing culture that the culture medium of bright offer is found to the induction ratio forefathers of pepper anther callus has obvious
Advantage.Our result of study is for further genralrlization and application capsicum haploid breeding without being suspected to have larger realistic meaning and reality
With value.
Those skilled in the art can be according to present disclosure and the art technology grasped in the present invention
Appearance makes replacement or modification, but these are replaced or modification is all not regarded as a departure from present inventive concept, and these are replaced or modification
In claimed interest field.
Claims (1)
1. a kind of capsicum variety breeding method, the formula of wherein flower pesticide inducing culture is as follows：
KNO3950mg/L, NH4NO3825mg/L, KH2PO4675mg/L, MgSO4∙H2O 1200mg/L, CaCl2∙2H2O
225mg/L, Na2- EDTA 75mg/L, FeSO4∙7H2O 55mg/L, MnSO4∙H2O 17mg/L, ZnSO4∙7H2O 18mg/L,
H3BO36.0mg/L, KI 0.8mg/L, CuSO4∙5H2O 0.025mg/L, CoCl2∙6H2O 0.03mg/L, Na2MoO4∙2H2O
0.25mg/L, AgNO35.0mg/L, inositol 100mg/L, vitamin B1 0.5mg/L, vitamin B6 0.5mg/L, nicotinic acid
5.0mg/L, glycine 2.0mg/L, folic acid 0.5mg/L, cyclic guanosine monophosphate 0.3g/L, maltose 20g/L, sucrose 15g/L, plant
Gel 5.0g/L；2,4-D 0.6mg/L, kinetin 0.5mg/L, multiple phthalein nucleic acid 0.8mg/L, biotin 0.05mg/L, vitamin C
45 μm of ol/L, activated carbon 175mg/L, protein hydrolysate 0.4g/L；
The pH value of the flower pesticide Fiber differentiation based formulas is 5.8；
Breeding method comprises the following steps：
Capsicum variety Xinfeng 5 and capsicum annum fasciculatum are selected as Anther Culture donor, in the morning that fine degree/day is relatively low, sunshine is weaker
On, the bud endangered without disease pest for being grown on open country is chosen, is preserved immediately in 4 DEG C of refrigerators after taking；Calyx is used certainly before inoculation
Water rinses 10-15min, and flower pesticide is taken out in 75% ethanol leaching 20s, 0.1% mercuric chloride sterilization 10min, then aseptic water washing 3-4 times
It is placed in standby in the culture dish for filling sterile blotting paper；Flower pesticide carefully is cut out from bud during inoculation, Sheng is inoculated in
Have in the 60mm × 15mm culture dish of 10ml flower pesticide inducing cultures, 25-30 pieces of flower pesticide is inoculated with per ware, is then sealed with sealed membrane
Mouthful；Light culture 8 days under prior to 33 DEG C high temperature after flower pesticide inoculation, 25 DEG C of light cultures are to embryoid appearance afterwards, and it is rearmounted that flower pesticide goes out embryo
In being cultivated under 25 DEG C of light, callus induction rate is calculated, regeneration culture medium is then transferred to.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610086925.1A CN105638480B (en) | 2016-02-16 | 2016-02-16 | A kind of capsicum variety, which is cultivated, uses flower pesticide Fiber differentiation based formulas |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610086925.1A CN105638480B (en) | 2016-02-16 | 2016-02-16 | A kind of capsicum variety, which is cultivated, uses flower pesticide Fiber differentiation based formulas |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105638480A CN105638480A (en) | 2016-06-08 |
| CN105638480B true CN105638480B (en) | 2017-09-01 |
Family
ID=56488471
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610086925.1A Active CN105638480B (en) | 2016-02-16 | 2016-02-16 | A kind of capsicum variety, which is cultivated, uses flower pesticide Fiber differentiation based formulas |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105638480B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108401901B (en) * | 2018-03-06 | 2021-06-01 | 山东寿光蔬菜种业集团有限公司 | Method for cultivating disease-resistant homozygote of capsicum |
| CN116746489B (en) * | 2023-06-28 | 2024-08-16 | 漳州市农业科学研究所 | Sweet pepper tissue culture breeding medium and method based on two-step seedling method |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100526161B1 (en) * | 2004-03-04 | 2005-11-08 | 김문자 | A method for culturing isolated microspore of pepper |
| CN102047843B (en) * | 2010-10-15 | 2012-05-23 | 北京市农林科学院 | Method for obtaining dihaploid plants of sweet peppers |
| CN103444542B (en) * | 2013-09-13 | 2015-08-12 | 北京海花生物科技有限公司 | A kind of pepper anther directly obtains cultural method and the medium of plant |
| CN104041414B (en) * | 2014-06-18 | 2016-04-27 | 南京市蔬菜科学研究所 | A kind of pepper anther culture obtains the method for monoploid regeneration plant |
-
2016
- 2016-02-16 CN CN201610086925.1A patent/CN105638480B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN105638480A (en) | 2016-06-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111226792B (en) | High-throughput breeding method for leaf seedlings of cymbidium sinense | |
| CN105532480B (en) | A kind of potato flower pesticide inducing culture and Anther culture breeding method | |
| CN105284620A (en) | Method for rescuing grown-up seedlings of special precocious peach hybrid embryos | |
| CN101785431B (en) | Method for improving proliferation and differentiation of protocorm of Oncidium by utilizing concentrated coconut juice | |
| CN106434524A (en) | Chinese cabbage isolated microspore inducing medium formula | |
| CN112841037A (en) | Embryogenic callus induction culture medium for rubber tree, method for inducing somatic embryogenesis of rubber tree and application | |
| CN110506635B (en) | Marigold pollen induction culture medium and induction culture method | |
| CN102007867A (en) | Efficient rooting method for oil tea clone tissue culture seedlings | |
| CN102405830B (en) | Induction method of rosa chinensis receptacle callus tissues | |
| CN103749293A (en) | Method for obtaining regenerated plantlet through anther culture of asparagus officinalis and asparagus dauricus interspecific crossing F1 generation | |
| CN105638480B (en) | A kind of capsicum variety, which is cultivated, uses flower pesticide Fiber differentiation based formulas | |
| CN104996304B (en) | Culture medium and culture method for inducing callus differentiation through peony leaves | |
| CN102870683A (en) | Microbody propagation expanding method of aquilaria malaccensis | |
| CN108739396B (en) | Rapid seedling method for stem segments of red honeysuckle | |
| CN106258984A (en) | A kind of Chinese cabbage sporidiole differentiation culture based formulas | |
| CN114027182A (en) | Tissue culture propagation method for dolichos succulent plants in crassulaceae echeveria | |
| CN101584300A (en) | The method of isolated culturing hypericum regenerated plant | |
| CN103563747B (en) | The detoxifying fast breeding method of favour building Chinese yam | |
| CN105010141A (en) | Anthurium rapid propagation method | |
| CN112868527A (en) | Method for rapidly propagating flamingo pepper grass | |
| CN104082145A (en) | Method for rapidly propagating adiantum soboliferum | |
| CN102550404B (en) | Efficient induction method of plum blossom blade callus | |
| CN109220809B (en) | Koelreuteria paniculata somatic embryogenesis and plant regeneration culture method | |
| CN104823861B (en) | Oil tea radicle Induce aerosor obtains the method for regeneration plant | |
| CN105340752A (en) | Method for in vitro culture of hemerocallis middendorfii Trautv. et Mey. tissue |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information | ||
| CB03 | Change of inventor or designer information |
Inventor after: Zhang Xindian Inventor before: Zhang Ruiming |
|
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20170802 Address after: Ma Ling town Wang Zhuang street Xinyi city 221400 Jiangsu city of Xuzhou Province Applicant after: Xinyi best Trading Co., Ltd. Address before: 222000 Jiangsu, Lianyungang, Haizhou District, Haining Road, building 8-3, No. 4 shops Applicant before: JIANGSU QIANGNONG AGRICULTURE TECHNOLOGY SERVICE CO., LTD. |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |