CN101805721B - Method for culturing corn haploid coleoptile section tissue and specific culture medium thereof - Google Patents
Method for culturing corn haploid coleoptile section tissue and specific culture medium thereof Download PDFInfo
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Abstract
The invention discloses a method for culturing corn haploid coleoptile section tissue and a specific culture medium thereof. The differentiation culture medium provided by the invention is the culture medium used in the differentiation and shoot formation of corn haploid coleoptile section induced callus, and is prepared by adding the following substances into MS basic culture medium: proline, casein hydrolyzate, carbon and gel, wherein the final concentration of the proline is 0.7g/L, and the final concentration of the casein hydrolyzat is 0.5g/L. The method of the invention adopts corn coleoptile section as the explant induced callus, and then differentiates the callus into a haploid strain to establish a stable and high-efficiency tissue culture system. The invention can be used for differentiation rate genetic research, material selection for tissue culture, and the like.
Description
Technical field
The present invention relates to plant tissue culture and crop breeding field, particularly corn haploid bud scale joint tissue culture and doubling monoploids and special culture media.
Background technology
Corn (Zea mays L.) also claim Zea mays, maize, maize, ear of maize; Guangdong language is called maize, and the south of Fujian Province language is called a kind wheat, is the annual gramineae herbage, also is the highest food crop of whole world ultimate production.Class of trade is mainly divided according to the quality of seed, is divided into horse tooth kind, hard kind, powder seed, explosion kind and Glutinous Semen Maydis, sweet corn etc.Seed top depression causes because of the ridity of hard starch of seed and soft starch is unequal.It is few that flint corn contains soft starch, dry back top depression.Flour corn mainly contains soft starch, and opaque is prone to pulverize.Sweet corn is sent out wrinkle, and is transparent, and sugar is not converted into starch.Pop corn is the extreme form of jadeite rice, and seed is little and hard, does not contain soft starch, and ICW expands during heating, the seed explosion.Can improve the corn type with good hybridization between selfed lines.
Corn is as feed, food and industrial raw material, at the main food of many areas conduct.Except that edible, corn also is the main raw material of industrial spirit and liquor.Other part purposes of milpa is also quite extensive: cornstalk is used for papermaking and system wallboard; Foreskin can be done packing material and the braiding of grass skill; Corn cob can be made fuel, also is used for preparing industrial solvent, and cauline leaf is except that being used as animal feed, or the methane-generating pit good raw material.
Utilize haploid breeding can overcome hybrid and separate, shortening the breeding cycle, improve selection rate.Through obtaining haplobiont, pass through chromosome doubling again, can obtain the diploid of isozygotying very soon, like this general shortening the breeding cycle greatly.
Corn haploid bud scale joint tissue culture obtains the technology of haplobiont, and not seeing at present has report.
Summary of the invention
The object of the present invention is to provide a kind of special culture media that is used for corn haploid bud scale joint callus differentiation culture.
Division culture medium provided by the invention is in the MS basic culture solution, to add the solid medium that following material obtains: proline(Pro), casein hydrolysate, carbon source and gelifying agent;
The solvent of above-mentioned MS basic culture solution is that water, solute are as shown in table 1;
The final concentration of proline(Pro) is that the final concentration of 0.6-0.8g/L, casein hydrolysate is 0.4-0.6g/L in the above-mentioned division culture medium.
The solute of table 1.MS basic culture solution
| Macroelement | Concentration (gL in the substratum -1) |
| NH 4NO 3 | 1.65 |
| KNO 3 | 1.9 |
| KH 2PO 4 | 0.17 |
| MgSO 4.7H 2O | 0.37 |
| CaCl 2 | 0.44 |
| Trace element | Concentration (mgL in the substratum -1) |
| FeSO 4.7H 2O | 27.8 |
| Na 2EDTA | 37.3 |
| MnSO 4.4H 2O | 22.3 |
| ZnSO 4.4H 2O | 8.6 |
| H 3BO 3 | 6.2 |
| KI | 0.83 |
| Na 2MoO 4.2H 2O | 0.25 |
| CuSO 4.5H 2O | 0.025 |
| CoCl 2.6H 2O | 0.025 |
| Organic composition | Concentration (mgL in the substratum -1) |
| Glycocoll | 2.0 |
| Vitamin | 0.4 |
| Pyridoxine hydrochloride | 0.5 |
| Nicotinic acid | 0.5 |
| Inositol | 100 |
Above-mentioned gelifying agent is tissue culture such as agar powder, carrageenin or Gelrite solidifying agent commonly used; Gelrite preferably; The consumption of Gelrite (gelatin) is the basis with the substratum hardness that can reach, and can suitably adjust consumption, and the concentration of Gelrite in division culture medium specifically can be 3.0g/L.
Above-mentioned carbon source is glucose, SANMALT-S or sucrose, is preferably sucrose, and the concentration of sucrose in division culture medium specifically can be 30g/L.
Further, in the above-mentioned division culture medium final concentration of proline(Pro) and casein hydrolysate preferably proline(Pro) be that 0.7g/L, casein hydrolysate are 0.5g/L.
The present invention also aims to provide a kind of method of producing the corn haplobiont.
The present invention provides the method for producing corn monoploid regeneration plant, may further comprise the steps:
1) mature embryo with the monoploid corn kernel places the MSVS34 substratum to cultivate, and sprouts to obtain plumule, chooses colourless monoploid plumule;
2) with above-mentioned steps 1) in the monoploid plumule that obtains cut the coleoptile joint and be inoculated in the MSW57 callus inducing medium and cultivate, obtain the monoploid callus;
3) with above-mentioned steps 2) in the monoploid callus that obtains place above-mentioned division culture medium to cultivate, obtain the monoploid regeneration plant.
Above-mentioned MSVS34 substratum and MSW57 callus inducing medium all are substratum commonly used in the plant tissue culture.
The method of production corn haplobiont of the present invention is in said step 2) and step 3) between also can comprise succeeding transfer culture; Said succeeding transfer culture is with said step 2) in the monoploid callus that obtains place N6 subculture medium multiplication by culture, the monoploid callus that obtains breeding.
Above-mentioned N6 subculture medium is in the N6 basic culture solution, to add casein hydrolysate, proline(Pro), 2, the solid medium that 4-D, carbon source and gelifying agent obtain; Said carbon source is preferably sucrose, and gelifying agent is preferably Gelrite.The final concentration of said sucrose in said N6 subculture medium is 30.0g/L, and the final concentration of said Gelrite in said N6 subculture medium is 2.7g/L
The solvent of said N6 basic culture solution is that water, solute are as shown in table 2; Wherein casein hydrolysate, proline(Pro) and 2, the final concentration of 4-D is respectively following 1) or 2):
1) casein hydrolysate is 0.5g/L, and proline(Pro) is 0.7g/L, 2, and 4-D is 1.5-2.5mg/L;
2) casein hydrolysate is 0.5g/L, and proline(Pro) is 0.7g/L, 2, and 4-D is 2mg/L.
Casein hydrolysate, proline(Pro) and 2 in the optimum N6 subculture medium, the final concentration of 4-D is above-mentioned 2) said.
The solute of table 2.N6 basic culture solution
| Macroelement | Concentration (gL in the substratum -1) |
| KNO 3 | 2.83 |
| (NH 4) 2SO 4 | 0.46 |
| KH 2PO 4 | 0.4 |
| MgSO 4.7H 2O | 0.185 |
| CaCl 2.2H 2O | 0.166 |
| Trace element | Concentration (mgL in the substratum -1) |
| FeSO 4.7H 2O | 27.8 |
| Na 2EDTA | 37.3 |
| MnSO 4.4H 2O | 4.4 |
| ZnSO 4.7H 2O | 1.5 |
| H 3BO 3 | 1.6 |
| KI | 0.8 |
| Organic composition | Concentration (mgL in the substratum -1) |
| Glycocoll | 2.0 |
| Vitamin | 1.0 |
| Pyridoxine hydrochloride | 0.5 |
| Nicotinic acid | 0.5 |
The method of above-mentioned production corn haplobiont also can comprise: the haplobiont that obtains in the step 3) is inoculated into carries out root culture, the haplobiont that obtains taking root in the root media.
Above-mentioned root media is in the MS basic culture solution, to add the solid medium that NAA, carbon source and gelifying agent obtain;
The solvent of said MS basic culture solution is that water, solute are as shown in table 1; Said carbon source is preferably sucrose, and gelifying agent is preferably Gelrite.The final concentration of said sucrose in said root media is 30g/L, and the final concentration of said Gelrite in said root media is 1.2g/L.Wherein the final concentration of NAA is 0.25-0.7mg/L, preferably 0.5mg/L.
Division culture medium of the present invention can be divided into plant with maize bud scale joint inductive callus effectively, and the differentiation adventitious buds rate reaches 36%, and after cultivating through root media, rooting rate reaches 88.9%.
Method of the present invention is to utilize the coleoptile joint of corn haploid bud differentiation obtains haplobiont as the explant induction callus; The tissue culturing system that the present invention sets up can be used for haploid breeding research effectively; For keeping good monoploid material, and good monoploid material is doubled as zygoid good material is provided.
Description of drawings
Fig. 1 is a corn monoploid seed.
The monoploid plumule that Fig. 2 sprouts for mature embryo of the present invention.
Fig. 3 is the present invention's haploid embryo bud scale joint to be inoculated.
Fig. 4 induces the monoploid callus that obtains for haploid embryo bud scale joint.
Fig. 5 is the monoploid callus of subculture.
The monoploid regeneration plant that Fig. 6 obtains for the callus induction differentiation.
Fig. 7 is that rooting and transplant becomes haplobiont alive.
Embodiment
Below in conjunction with specific embodiment the present invention is described further, but the present invention is not limited to following examples.
Among the following embodiment,, be ordinary method like no specified otherwise.
The preparation of embodiment 1, corn haplobiont and statistical observation
One, the acquisition of monoploid seed
, hybridize as male parent with corn haploid inducing line Stock6 (U.S. profound scholar gets company, Garst Seed Co) with agricultural university 108 (buying the greatly healthy scientific and technological development of middle peasant ltd) in Beijing.After seed maturity, the drying, select the seed of the white embryo in purple top, subsequent use.
Two, the acquisition of monoploid plumule and statistical observation
1, the pre-treatment of monoploid seed
The purple that from step 1, obtains pushes up in the corn kernel (being corn monoploid seed, as shown in Figure 1) of white embryo and selects 100, the envelope of packing into, and opening is put into moisture eliminator with envelope.Use the beaker of 250mL, contain 200mL 5.25% (percent by volume) chlorine bleach liquor, in beaker, add the 2mL concentrated hydrochloric acid again, put into moisture eliminator central authorities, encapsulation process 8-15 hour.Seed is taken out, put into beaker, in Bechtop, add 70% (percent by volume) Ethanol Treatment 1min; Remove 70% ethanol, add 2% (percent by volume) chlorine bleach liquor, soak 20min; With aseptic washing 4-5 time, use the sterilized water soaked overnight at last.
2, obtain the monoploid plumule
On super clean bench, the seed of soaked overnight is peeled off, obtain mature embryo; Be inoculated in the MSVS34 substratum, place in the illumination cultivation chamber and cultivated 7-10 days, culture condition is in the illumination cultivation chamber: place 25 ℃ of illumination cultivation; Intensity of illumination 2000lx, illumination every day 16h cultivates 7-10d.Plumule after obtaining sprouting is rejected the plumule material that radicle, embryo or plumule are purple, chooses acquisition radicle, embryo and plumule and is colourless monoploid plumule (as shown in Figure 2) material.
Above-mentioned MSVS34 substratum is in the MS basic culture solution, to add the substratum that following material obtains: casein hydrolysate, MES (2-morpholinoethanesulfonic acid), magnesium chloride, Stimulina, xitix, picloram (picloram), BAP (benzyladenine), carbon source and gelifying agent; The solvent of MS basic culture solution is that water, solute are as shown in table 1; Wherein the final concentration of casein hydrolysate is 0.1g/L, and the final concentration of MES is 1.95g/L, and the final concentration of magnesium chloride is 0.75g/L; The final concentration of Stimulina is 0.5g/L; The final concentration of xitix is 0.1g/L, and the final concentration of picloram is 10mg/L, and the final concentration of BAP is 3mg/L; Above-mentioned carbon source is a SANMALT-S, and gelifying agent is an agar; The final concentration of SANMALT-S is 40g/L, and the final concentration of agar is 8g/L, can suitably adjust its consumption, and the pH of MSVS34 substratum is 5.8.
3, statistical observation
Experiment repetition 3 times, the MSVS34 substratum is induced the result of mature embryo sprouting and the yield of monoploid plumule in the statistical observation above-mentioned steps 2.Experimental result sees the following form 3.Can find out that from table 3 after mature embryo was sprouted, rejecting radicle, plumule were the material of purple, keeping radicle and plumule is the monoploid material of non-purple.
Table 3.MSVS34 substratum is induced mature embryo sprouting situation statistics
Three, the acquisition of callus and statistical observation
1, the acquisition of callus
When the monoploid plumule of treating in the above-mentioned steps two grows to about 5-6cm monoploid seedling, cut the coleoptile joint (upper and lower each 0.5cm of stipes, as shown in Figure 3) of about 1em; And use the scalpel five equilibrium, tangent plane is close to substratum, is inoculated in the MSW57 callus inducing medium; 25 ℃ of illumination cultivation, intensity of illumination 2000lx, illumination every day 16h; Illumination cultivation 21d obtains monoploid callus (as shown in Figure 4).
Above-mentioned MSW57 callus inducing medium is in the MS basic culture solution, to add vitamin, casamino acids, proline(Pro), Silver Nitrate, 2, the substratum that 4-D, picloram, carbon source and gelifying agent obtain; The solvent of MS basic culture solution is that water, solute are as shown in table 1; Wherein the final concentration of vitamin is 0.9mg/L, and the final concentration of casamino acids is 0.5g/L, and the final concentration of proline(Pro) is 1.38g/L, and the final concentration of Silver Nitrate is 3.4mg/L, 2, and the final concentration of 4-D is 0.5mg/L, the final concentration of picloram is 2.2mg/L; Above-mentioned carbon source is a sucrose, and gelifying agent is an agar; The final concentration of sucrose is 30g/L, and the final concentration of agar is 8g/L, and the pH of substratum is 5.8.
2, statistical observation
Experiment repetition 3 times, in the statistical observation above-mentioned steps 1 callus induce situation, experimental result sees the following form 4, the inductivity of callus can reach more than 36.9%.
Table 4. callus induction rate
Four, the succeeding transfer culture of callus and statistical observation
1, succeeding transfer culture
The callus that obtains in the step 3 is transferred in the N6 subculture medium, is under 25 ℃ the condition in temperature, the dark cultivation, and per 21 days subcultures once obtain the monoploid callus (as shown in Figure 5) of required propagation quantity.
Above-mentioned N6 subculture medium is in the N6 basic culture solution, to add the substratum that following material obtains: casein hydrolysate, proline(Pro), 2,4-D, carbon source and gelifying agent; Wherein carbon source is a sucrose, and the final concentration of sucrose in substratum is 30g/L, and gelifying agent is Gelrite, and the final concentration of Gelrite is 2.7g/L, and the final concentration of casein hydrolysate is 0.5g/L, and the final concentration of proline(Pro) is 0.7g/L, 2, and the final concentration of 4-D is 2mg/L; The pH of substratum is 5.8, and the solute of N6 basic culture solution is as shown in table 2, and its solvent is a water.
Five, the acquisition of monoploid regeneration plant and statistical observation
1, obtains the monoploid regeneration plant
The monoploid callus that succeeding transfer culture obtains in monoploid callus that above-mentioned steps three is obtained or the step 4 is cut into small pieces and changes in the division culture medium; Illumination cultivation, culture condition are that temperature is 25 ℃; Be placed under the intensity of illumination of 2000lx and cultivate; Every day, light was cultivated 16 hours, secretly cultivated 8 hours, and differentiation obtains monoploid regeneration plant (as shown in Figure 6).
Above-mentioned division culture medium is in the MS basic culture solution, to add the substratum that proline(Pro), casein hydrolysate, carbon source and gelifying agent obtain, and the solvent of MS basic culture solution is that water, solute are as shown in table 1; Wherein carbon source is that final concentration is the sucrose of 30g/L, and gelifying agent is Gelrite, and the final concentration of Gelrite is 3.0g/L, and the final concentration of proline(Pro) is 0.7g/L, and the final concentration of casein hydrolysate is 0.5g/L, and the pH of substratum is 5.8.
2, statistical observation
Experiment repetition 3 times, the situation of regeneration differentiation in the statistical observation above-mentioned steps 1, experimental result sees the following form 5, can find out from table 5, and the differentiation of calli rate can reach more than 54%, and the differentiation rate of indefinite bud reaches more than 22%.
The differentiation rate of table 5. regeneration differentiation culture
Six, root culture and statistical observation
1, the monoploid acquisition of plant of taking root
When the haplobiont that obtains in the above-mentioned steps five is grown to 3-5cm, change over to and carry out root culture in the root media, culture condition is 25 ℃ of illumination cultivation; Intensity of illumination 2000lx; Illumination every day 16h cultivated about 21 days, the good monoploid that obtains the taking root plant that takes root.
Above-mentioned root media is in the MS basic culture solution, to add the substratum that NAA, carbon source and gelifying agent obtain, and the solvent of MS basic culture solution is that water, solute are as shown in table 1; Wherein carbon source is that final concentration is the sucrose of 30g/L, and gelifying agent is Gelrite, and the final concentration of Gelrite is 1.2g/L, and the final concentration of NAA is 0.5mg/L, and the pH of substratum is 5.8.
2, statistical observation
Experiment repetition 3 times, the root culture situation of statistical observation above-mentioned steps 1, experimental result sees the following form 6, and rooting rate reaches more than 81.7%, and every strain haplobiont is on average taken root 2.1.
The rooting rate of table 6. root culture
Seven, acclimatization and transplants and check and analysis
1, acclimatization and transplants
The good monoploid that takes root in the above-mentioned steps six is taken root behind the plant refining seedling, transplant to flowerpot, growth is 7-10 days in the greenhouse, and the plant of transplant survival is planted in the greenhouse, obtains the haplobiont (being the regenerated plant, as shown in Figure 7) of transplant survival.The surviving rate of transplanting is 75%.
2, check and analysis
When regeneration plant in the above-mentioned steps 1 (be monoploid take root plant) is transplanted flowerpot, cut 1-2 the tip of a root, with Kano stationary liquid (absolute ethyl alcohol: Glacial acetic acid min. 99.5=3: 1) fixedly more than the 2h.Use ddH
2After the O cleaning many times, add an amount of cellulase and polygalacturonase mixture enzymolysis 2h.Compressing tablet is examined under a microscope chromosome number, and the result finds that chromosome number is 10 all, explains that the regenerated plant all is a haplobiont.
Claims (9)
1. a division culture medium is in the MS basic culture solution, to add the solid medium that following material obtains: proline(Pro), casein hydrolysate, carbon source and gelifying agent;
The solvent of said MS basic culture solution is that water, solute are as shown in table 1;
The final concentration of proline(Pro) is that the final concentration of 0.6-0.8g/L, casein hydrolysate is 0.4-0.6g/L in the said division culture medium.
2. division culture medium according to claim 1 is characterized in that: said gelifying agent is agar powder, carrageenin or Gelrite, and wherein the final concentration of Gelrite in said division culture medium is 3.0g/L; Said carbon source is glucose, SANMALT-S or sucrose, and wherein the final concentration of sucrose in said division culture medium is 30g/L.
3. division culture medium according to claim 1 and 2 is characterized in that: said gelifying agent is Gelrite; Said carbon source is a sucrose.
4. division culture medium according to claim 3 is characterized in that: the final concentration of proline(Pro) and casein hydrolysate is respectively in the said division culture medium: proline(Pro) is that 0.7g/L, casein hydrolysate are 0.5g/L.
5. method of producing corn monoploid regeneration plant may further comprise the steps:
1) mature embryo with the monoploid corn kernel places the MSVS34 substratum to cultivate, and sprouts to obtain plumule, chooses colourless monoploid plumule;
Said monoploid corn kernel be with corn haploid inducing line Stock6 as male parent, hybridize with agricultural university 108 and to obtain;
Said MSVS34 substratum is in the MS basic culture solution, to add the substratum that following material obtains: casein hydrolysate, 2-morpholinoethanesulfonic acid, magnesium chloride, Stimulina, xitix, picloram, benzyladenine, carbon source and gelifying agent; Wherein the final concentration of casein hydrolysate is 0.1g/L, and the final concentration of 2-morpholinoethanesulfonic acid is 1.95g/L, and the final concentration of magnesium chloride is 0.75g/L; The final concentration of Stimulina is 0.5g/L; The final concentration of xitix is 0.1g/L, and the final concentration of picloram is 10mg/L, and the final concentration of benzyladenine is 3mg/L; Said carbon source is a SANMALT-S, and said gelifying agent is an agar; The final concentration of said SANMALT-S is 40g/L, and the final concentration of said agar is 8g/L;
2) with above-mentioned steps 1) in the monoploid plumule that obtains cut the coleoptile joint and be inoculated in the MSW57 callus inducing medium and cultivate, obtain the monoploid callus;
Said cultivation is 25 ℃ of illumination cultivation, intensity of illumination 2000lx, illumination every day 16h, illumination cultivation 21d;
Said MSW57 callus inducing medium is in the MS basic culture solution, to add vitamin, casamino acids, proline(Pro), Silver Nitrate, 2, the substratum that 4-D, picloram, carbon source and gelifying agent obtain; Wherein the final concentration of vitamin is 0.9mg/L, and the final concentration of casamino acids is 0.5g/L, and the final concentration of proline(Pro) is 1.38g/L, and the final concentration of Silver Nitrate is 3.4mg/L, 2, and the final concentration of 4-D is 0.5mg/L, the final concentration of picloram is 2.2mg/L; Said carbon source is a sucrose, and said gelifying agent is an agar; The final concentration of said sucrose is 30g/L, and the final concentration of said agar is 8g/L;
3) with above-mentioned steps 2) in the monoploid callus that obtains place the arbitrary described division culture medium of claim 1-4 to cultivate, obtain the monoploid regeneration plant;
Said culture condition is that temperature is 25 ℃, is placed under the intensity of illumination of 2000lx to cultivate, and every day, light was cultivated 16 hours, secretly cultivated 8 hours.
6. method according to claim 5; It is characterized in that: in said step 2) and step 3) between also comprise succeeding transfer culture; Said succeeding transfer culture is with said step 2) in the monoploid callus that obtains place N6 subculture medium multiplication by culture, the monoploid callus that obtains breeding;
Said N6 subculture medium is in the N6 basic culture solution, to add casein hydrolysate, proline(Pro), 2, the solid medium that 4-D, carbon source and gelifying agent obtain;
The solvent of said N6 basic culture solution is that water, solute are as shown in table 2;
Wherein casein hydrolysate, proline(Pro) and 2, the final concentration of 4-D is respectively as follows: casein hydrolysate is 0.5g/L, proline(Pro) is 0.7g/L, 2,4-D is 1.5-2.5mg/L.
7. method according to claim 6 is characterized in that:
Said N6 subculture medium, wherein casein hydrolysate, proline(Pro) and 2, the final concentration of 4-D is as follows: casein hydrolysate is 0.5g/L, proline(Pro) is 0.7g/L, 2,4-D is 2mg/L;
Said carbon source is a sucrose, and said gelifying agent is Gelrite;
The final concentration of said sucrose in said N6 subculture medium is 30.0g/L, and the final concentration of said Gelrite in said N6 subculture medium is 2.7g/L.
8. according to arbitrary described method among the claim 5-7; It is characterized in that: the method for said production corn monoploid regeneration plant also comprises: the haplobiont that obtains in the step 3) is inoculated into carries out root culture, the haplobiont that obtains taking root in the root media;
Said root media is in the MS basic culture solution, to add the solid medium that NAA, carbon source and gelifying agent obtain;
The solvent of said MS basic culture solution is that water, solute are as shown in table 1;
Wherein the final concentration of NAA is 0.25-0.7mg/L;
Carbon source in the said root media is a sucrose, and gelifying agent is Gelrite; The final concentration of said sucrose in said root media is 30g/L, and the final concentration of said Gelrite in said root media is 1.2g/L.
9. method according to claim 8 is characterized in that: the final concentration of the NAA in the said root media is 0.5mg/L.
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| CN103190346A (en) * | 2013-04-22 | 2013-07-10 | 北京金冠丰生物技术有限公司 | Method for constructing corn reproduction system taking coleoptile section as explant |
| CN103173487A (en) * | 2013-04-23 | 2013-06-26 | 北京金冠丰生物技术有限公司 | Anniversary large-scale maize transformation method |
| CN104026017B (en) * | 2014-06-20 | 2017-03-29 | 四川农业大学 | The breeding method of Semen Maydiss haplobiont |
| CA3000133A1 (en) * | 2015-10-02 | 2017-04-06 | Keygene N.V. | Method for the production of haploid and subsequent doubled haploid plants |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101011031A (en) * | 2007-02-07 | 2007-08-08 | 华中农业大学 | Callus induction and plant regeneration method by corn mature embryo approach |
| US20080274547A1 (en) * | 2003-12-19 | 2008-11-06 | Monsanto Technology Llc | Use of nitric oxide modulators in agrobacterium-mediated plant transformation |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US20080274547A1 (en) * | 2003-12-19 | 2008-11-06 | Monsanto Technology Llc | Use of nitric oxide modulators in agrobacterium-mediated plant transformation |
| CN101011031A (en) * | 2007-02-07 | 2007-08-08 | 华中农业大学 | Callus induction and plant regeneration method by corn mature embryo approach |
Non-Patent Citations (1)
| Title |
|---|
| Vladimir Sidorov et al.Agrobacterium-mediated transformation of seedling-derived maize callus.《Plant Cell Rep》.2005,第25卷(第4期),320-328. * |
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