CN101292629A - Flax free pollen culture medium and flax free pollen culture method - Google Patents
Flax free pollen culture medium and flax free pollen culture method Download PDFInfo
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- CN101292629A CN101292629A CNA2008100647514A CN200810064751A CN101292629A CN 101292629 A CN101292629 A CN 101292629A CN A2008100647514 A CNA2008100647514 A CN A2008100647514A CN 200810064751 A CN200810064751 A CN 200810064751A CN 101292629 A CN101292629 A CN 101292629A
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- 241000208202 Linaceae Species 0.000 title claims abstract description 67
- 235000004431 Linum usitatissimum Nutrition 0.000 title claims abstract description 67
- 239000001963 growth medium Substances 0.000 title abstract description 6
- 238000012136 culture method Methods 0.000 title abstract description 3
- 206010020649 Hyperkeratosis Diseases 0.000 claims abstract description 41
- 238000000034 method Methods 0.000 claims abstract description 18
- 230000004069 differentiation Effects 0.000 claims abstract description 16
- 230000001939 inductive effect Effects 0.000 claims abstract description 11
- 239000000725 suspension Substances 0.000 claims abstract description 6
- 238000001914 filtration Methods 0.000 claims abstract description 3
- 239000002609 medium Substances 0.000 claims description 28
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 claims description 18
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 12
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 12
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 claims description 12
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims description 12
- 238000005286 illumination Methods 0.000 claims description 10
- 239000006160 differential media Substances 0.000 claims description 8
- 229920001817 Agar Polymers 0.000 claims description 6
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 6
- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 claims description 6
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 claims description 6
- 241000196324 Embryophyta Species 0.000 claims description 6
- 239000004471 Glycine Substances 0.000 claims description 6
- 229930006000 Sucrose Natural products 0.000 claims description 6
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 6
- 239000008272 agar Substances 0.000 claims description 6
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 6
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 6
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 6
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 6
- 239000004327 boric acid Substances 0.000 claims description 6
- 239000001110 calcium chloride Substances 0.000 claims description 6
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 6
- 235000019301 disodium ethylene diamine tetraacetate Nutrition 0.000 claims description 6
- 239000011790 ferrous sulphate Substances 0.000 claims description 6
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 6
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 6
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 6
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 6
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 6
- 239000011702 manganese sulphate Substances 0.000 claims description 6
- 235000007079 manganese sulphate Nutrition 0.000 claims description 6
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 6
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 6
- 235000001968 nicotinic acid Nutrition 0.000 claims description 6
- 229960003512 nicotinic acid Drugs 0.000 claims description 6
- 239000011664 nicotinic acid Substances 0.000 claims description 6
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 6
- 239000004323 potassium nitrate Substances 0.000 claims description 6
- 235000010333 potassium nitrate Nutrition 0.000 claims description 6
- ZUFQODAHGAHPFQ-UHFFFAOYSA-N pyridoxine hydrochloride Chemical compound Cl.CC1=NC=C(CO)C(CO)=C1O ZUFQODAHGAHPFQ-UHFFFAOYSA-N 0.000 claims description 6
- 229960004172 pyridoxine hydrochloride Drugs 0.000 claims description 6
- 235000019171 pyridoxine hydrochloride Nutrition 0.000 claims description 6
- 239000011764 pyridoxine hydrochloride Substances 0.000 claims description 6
- 239000012882 rooting medium Substances 0.000 claims description 6
- 239000005720 sucrose Substances 0.000 claims description 6
- 229960000344 thiamine hydrochloride Drugs 0.000 claims description 6
- 235000019190 thiamine hydrochloride Nutrition 0.000 claims description 6
- 239000011747 thiamine hydrochloride Substances 0.000 claims description 6
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 6
- 239000011686 zinc sulphate Substances 0.000 claims description 6
- 235000009529 zinc sulphate Nutrition 0.000 claims description 6
- 239000008223 sterile water Substances 0.000 claims description 4
- 238000001467 acupuncture Methods 0.000 claims description 2
- 239000000575 pesticide Substances 0.000 claims description 2
- 230000001954 sterilising effect Effects 0.000 claims description 2
- 238000004659 sterilization and disinfection Methods 0.000 claims description 2
- 238000005406 washing Methods 0.000 claims description 2
- 230000006698 induction Effects 0.000 abstract description 3
- 238000012258 culturing Methods 0.000 abstract 1
- 239000012883 rooting culture medium Substances 0.000 abstract 1
- 239000005556 hormone Substances 0.000 description 4
- 229940088597 hormone Drugs 0.000 description 4
- 239000003375 plant hormone Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 238000009395 breeding Methods 0.000 description 1
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- 210000000349 chromosome Anatomy 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000012533 medium component Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
A flax free pollen culture medium and a flax free pollen culture method, belonging to the biotechnology field; its flax free pollen callus induction culture medium HF1Containing 1/2N61.5mg/L2.4D, 0.5 mg/L6-BA; flax free pollen callus differentiation culture medium HF2Containing 1/2N62.0 mg/L6-BA, 0.5 mg/LIAA; flax pollen green seedling rooting culture medium HF3Containing 1/2N60.2mg/LNAA, 0.2 mg/LIAA; the method comprises the following steps of preparing pollen suspension by filtering and centrifugally collecting pollen, inducing pollen callus, inducing pollen green seedling differentiation and rooting culture of the pollen green seedling to finish the culture of the flax free pollen; the invention screens flax free pollen culture medium HF1、HF2And HF3The method for culturing the free pollen of flax is established for the first time, the complex process of ploidy identification of flax is simplified, and the inductivity of the pollen callus, the differentiation rate of the pollen green seedlings and the rooting rate of the pollen green seedlings are obviously higher than those of the prior art.
Description
Technical field
The present invention relates to flax biological technical field, relate generally to flax free pollen culture technique.
Background technology
China's flax Research Work on Anther Culture starts from twentieth century seventies, and from flax anther culture, obtain pollen plant first by China, but the inductivity of flax anther callus and green seedling differentiation rate are very low, and there is a large amount of liploid plants from somatocyte development, bring very big difficulty to the monoploid evaluation, seriously restricted flax haploid breeding process.The cultivation of free pollen has its unique advantages, and at first, free pollen is single sexual cell doubly, the cultivation of free pollen not only can reduce the workload that ploidy is identified, and it also can be used as cell screening system efficiently, makes economical character stable rapidly, avoids offspring's separation.Secondly, it can be used as genetically modified acceptor, by chromosome doubling, makes foreign gene pure and mild fast.In flax free pollen incubation, cultural method and medium are to influence the main external cause condition of differentiation rate that pollen produces callus induction rate and the green seedling of pollen.Therefore, this area presses for and sets up flax free pollen cultural method and medium.
Summary of the invention
Purpose of the present invention just provides a kind of novel flax free pollen medium and flax free pollen cultural method, with reach set up flax free pollen culture systems, further improve the inductivity of flax pollen callus and the green seedling differentiation rate of pollen, accelerate the purpose that flax is cultivated the kind process.
The object of the present invention is achieved like this:
A kind of flax free pollen medium comprises flax free pollen callus inducing culture HF
1, flax free pollen callus differential medium HF
2With the green seedling rooting medium of flax pollen HF
31., flax free pollen callus inducing culture HF, it is characterized in that:
1Contain 1/2N
6, 1.5mg/12.4-D, 0.5mg/L6-BA; 1/2N
6Composition be: potassium nitrate 1415mg/L, ammonium sulfate 231mg/L, magnesium sulfate 92mg/L, calcium chloride 83mg/L, potassium dihydrogen phosphate 200mg/L, boric acid 0.8mg/L, manganese sulphate 2.2mg/L, potassium iodide 0.4mg/L, zinc sulphate 0.75mg/L, glycine 1.0mg/L, thiamine hydrochloride 1.0mg/L, pyridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, ferrous sulfate 27.8mg/L, disodium ethylene diamine tetraacetate 37.3mg/L, sucrose 30g/L, agar 8g/L; PH value is 5.5;
2., flax free pollen callus differential medium HF
2Contain 1/2N
6, 2.0mg/L6-BA, 0.5mg/LI AA; 1/2N
6Composition be: potassium nitrate 1415mg/L, ammonium sulfate 231mg/L, magnesium sulfate 92mg/L, calcium chloride 83mg/L, potassium dihydrogen phosphate 200mg/L, boric acid 0.8mg/L, manganese sulphate 2.2mg/L, potassium iodide 0.4mg/L, zinc sulphate 0.75mg/L, glycine 1.0mg/L, thiamine hydrochloride 1.0mg/L, pyridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, ferrous sulfate 27.8mg/L, disodium ethylene diamine tetraacetate 37.3mg/L, sucrose 30g/L, agar 8g/L; PH value is 5.5;
3., the green seedling rooting medium of flax pollen HF
3Contain 1/2N
6, 0.2mg/LNAA, 0.2mg/LIAA; 1/2N
6Composition be: potassium nitrate 1415mg/L, ammonium sulfate 231mg/L, magnesium sulfate 92mg/L, calcium chloride 83mg/L, potassium dihydrogen phosphate 200mg/L, boric acid 0.8mg/L, manganese sulphate 2.2mg/L, potassium iodide 0.4mg/L, zinc sulphate 0.75mg/L, glycine 1.0mg/L, thiamine hydrochloride 1.0mg/L, pyridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, ferrous sulfate 27.8mg/L, disodium ethylene diamine tetraacetate 37.3mg/L, sucrose 30g/L, agar 8g/L; PH value is 5.5;
A kind of flax free pollen cultural method that uses above-mentioned flax free pollen medium is characterized in that the method comprising the steps of and is:
1., get flax bud that the pollen microspore is in the monokaryon middle and later periods, put into 2% liquor natrii hypochloritis sterilization 15 minutes, use aseptic water washing again 3 times, take out flower pesticide and put into the culture dish that fills sterile water, with the acupuncture method for releasing pollen grain is discharged in the sterile water, again by filtering centrifugal collection pollen;
2., adjust pollen density 5.0 * 10
2Individual/ml, be prepared into pollen suspension, be inoculated into flax pollen callus inducing culture HF
1In, induce pollen callus, be to cultivate under 2000LX, 8 hours/day the condition of light application time at 18-25 ℃, intensity of illumination;
3., when flax pollen callus length to about the 3-5mm, change flax pollen callus differential medium HF over to
2In, induce the differentiation of the green seedling of pollen, be to cultivate under 2000LX, 12 hours/day the condition of light application time at 18-25 ℃, intensity of illumination;
4., the plant that 3. step is obtained changes the green seedling rooting medium of flax pollen HF over to
3In, be that 2000LX, light application time are to cultivate under 15 hours/day the condition at 18-25 ℃, intensity of illumination, can take root successively in general 7-15 days.
Key of the present invention is to have found the suitable culture method, filter out efficient culture medium, this invention is to cultivate on the basis that reaches multiple medium application study at flax pollen for many years, starts with from adjustment macroelement, trace element, plant hormone, filters out flax free pollen medium HF
1, HF
2And HF
3, and set up flax free pollen cultural method first, simplified the complex process that flax ploidy is identified, and with HF
1The pollen callus inductivity of medium, HF
2Green seedling differentiation rate of the pollen of medium and HF
3The green seedling rooting rate of the pollen of medium is all apparently higher than prior art.
Embodiment
Give an actual example below the present invention and implementation result are described in detail.
(1) with 1/2N
6Medium component is the basis, and the concentration by adjusting plant hormone 2.4-D (0.1-3.0mg/L), 6-BA (0.1-3.0mg/L) is also carried out proportioning test, filters out the best inducing culture HF of flax free pollen
1, its hormone combination concentration is 1.5mg/L2.4-D, 0.5mg/L6-BA; Concentration by adjusting plant hormone 6-BA (0.1-3.0mg/L), (0.1-2.0mg/L) I AA is also carried out proportioning test, filters out flax free pollen callus differential medium HF
2, its hormone combination concentration is 1.5mg/L6-BA, 0.5mg/LIAA; Carry out proportioning test by the concentration of adjusting plant hormone NAA (0-0.5mg/L), (0-0.5mg/L) I AA, filter out flax pollen plant root media HF
3, its hormone combination concentration is 0.2mg/LNAA, 0.2mg/LIAA;
(2) inducing culture HF
1Middle flax free pollen callus inductivity.
We inoculate flax pollen of 5 hybrid combinations.(pollen density is 5.0 * 10 to 5 each 2ml pollen suspensions of hybrid combination
2Individual/as ml) to be seeded in HF respectively
1On the medium, carry out inducing of pollen callus, at 18-25 ℃, intensity of illumination is 2000LX, and light application time 8 hours was cultivated 15-30 days, and experimental result shows medium HF
1The inductivity of pollen callus is 20.6-30.8%.
Table 2 medium HF
1Middle Different Cross Combinations is to the influence of pollen callus inductivity
| The hybrid combination code name | Inoculation pollen number (individual) | Callus (piece) | Callus induction rate (%) |
| 9601 | 1000 | 308 | 30.8 |
| 9605 | 1000 | 286 | 28.6 |
| 9712 | 1000 | 251 | 25.1 |
| 9715 | 1000 | 206 | 20.6 |
| 9718 | 1000 | 219 | 21.9 |
(3) HF
1With MS, B
5, N
6The medium contrast experiment.
The pollen of getting same hybrid combination is seeded in MS, B
5, N
6, HF
1On the medium, every kind of medium is all inoculated 4ml pollen suspension pollen suspension, and (pollen density is 5.0 * 10
2Individual/ml), hormone combination is 1.5mg/L2.4-D, 0.5mg/L6-BA in four kinds of medium.At 18-25 ℃, intensity of illumination is 2000LX, and light application time 8 hours was cultivated 15-30 days, and experimental result shows: HF
1, MS, B
5, N
6The inductivity of four kinds of medium pollen callus is respectively 29.7%, 6.2%, 8.6%, 18.5%.With HF
1Medium pollen callus inductivity is the highest.
The different medium of table 3 are to the influence of pollen callus inductivity
| Medium | Inoculation pollen number (individual) | Pollen callus (piece) | Pollen callus inductivity (%) |
| HF 1 | 2000 | 594 | 29.7 |
| MS | 2000 | 124 | 6.2 |
| B 5 | 2000 | 172 | 8.6 |
| N 6 | 2000 | 370 | 18.5 |
(4) the green seedling differentiation of flax pollen.When pollen callus length to about the 3-5mm, change flax pollen callus differential medium HF over to
2In, induce the differentiation of the green seedling of pollen.Flax pollen callus of having tested 5 hybrid combinations is at medium HF
2The differentiation situation of the middle green seedling of pollen.Condition of culture is temperature 18-25 ℃, and intensity of illumination is 2000LX, and light application time 12 hours was cultivated 20-30 days, and the differentiation rate of the green seedling of statistics pollen.Table 4 experimental result shows: HF
2The green seedling differentiation rate of the pollen of medium is 4.5-11.5%.
The callus of table 4 Different Cross Combinations is at HF
2The green seedling differentiation rate of pollen in the medium
| The hybrid combination code name | Callus (piece) | Green seedling number | The green seedling differentiation rate of pollen (%) |
| 9601 | 200 | 9 | 4.5 |
| 9605 | 200 | 15 | 7.5 |
| 9712 | 200 | 18 | 8.0 |
| 9715 | 200 | 13 | 6.6 |
| 9718 | 200 | 23 | 11.5 |
(5) flax pollen plant root media HF
3, when the green seedling length of pollen arrives the 5cm left and right sides, forward it to HF
3In the medium, at 18-25 ℃, intensity of illumination is 2000LX, and light application time was cultivated under the condition in 15 hours.Corotation moves the experiment of taking root of 568 green seedlings of pollen, and 541 seedling rootings are arranged, and rooting rate reaches 95.2%.
Claims (2)
1, a kind of flax free pollen medium comprises flax free pollen callus inducing culture HF
1, flax free pollen callus differential medium HF
2With the green seedling rooting medium of flax pollen HF
31., flax free pollen callus inducing culture HF, it is characterized in that:
1Contain 1/2N
6, 1.5mg/L2.4-D, 0.5mg/L6-BA; 1/2N
6Composition be: potassium nitrate 1415mg/L, ammonium sulfate 231mg/L, magnesium sulfate 92mg/L, calcium chloride 83mg/L, potassium dihydrogen phosphate 200mg/L, boric acid 0.8mg/L, manganese sulphate 2.2mg/L, potassium iodide 0.4mg/L, zinc sulphate 0.75mg/L, glycine 1.0mg/L, thiamine hydrochloride 1.0mg/L, pyridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, ferrous sulfate 27.8mg/L, disodium ethylene diamine tetraacetate 37.3mg/L, sucrose 30g/L, agar 8g/L;
2., flax free pollen callus differential medium HF
2Contain 1/2N
6, 2.0mg/L6-BA, 0.5mg/LIAA; 1/2N
6Composition be: potassium nitrate 1415mg/L, ammonium sulfate 231mg/L, magnesium sulfate 92mg/L, calcium chloride 83mg/L, potassium dihydrogen phosphate 200mg/L, boric acid 0.8mg/L, manganese sulphate 2.2mg/L, potassium iodide 0.4mg/L, zinc sulphate 0.75mg/L, glycine 1.0mg/L, thiamine hydrochloride 1.0mg/L, pyridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, ferrous sulfate 27.8mg/L, disodium ethylene diamine tetraacetate 37.3mg/L, sucrose 30g/L, agar 8g/L;
3., the green seedling rooting medium of flax pollen HF
3Contain 1/2N
6, 0.2mg/LNAA, 0.2mg/LIAA; 1/2N
6Composition be: potassium nitrate 1415mg/L, ammonium sulfate 231mg/L, magnesium sulfate 92mg/L, calcium chloride 83mg/L, potassium dihydrogen phosphate 200mg/L, boric acid 0.8mg/L, manganese sulphate 2.2mg/L, potassium iodide 0.4mg/L, zinc sulphate 0.75mg/L, glycine 1.0mg/L, thiamine hydrochloride 1.0mg/L, pyridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, ferrous sulfate 27.8mg/L, disodium ethylene diamine tetraacetate 37.3mg/L, sucrose 30g/L, agar 8g/L.
2, a kind of use flax free pollen cultural method of flax free pollen medium according to claim 1 is characterized in that the method comprising the steps of and is:
1., get flax bud that the pollen microspore is in the monokaryon middle and later periods, put into 2% liquor natrii hypochloritis sterilization 15 minutes, use aseptic water washing again 3 times, take out flower pesticide and put into the culture dish that fills sterile water, with the acupuncture method for releasing pollen grain is discharged in the sterile water, again by filtering centrifugal collection pollen;
2., adjust pollen density 5.0 * 10
2Individual/ml, be prepared into pollen suspension, be inoculated into flax pollen callus inducing culture HF
1In, induce pollen callus, be to cultivate under 2000LX, 8 hours/day the condition of light application time at 18-25 ℃, intensity of illumination;
3., when flax pollen callus length to about the 3-5mm, change flax pollen callus differential medium HF over to
2In, induce the differentiation of the green seedling of pollen, be to cultivate under 2000LX, 12 hours/day the condition of light application time at 18-25 ℃, intensity of illumination;
4., the plant that 3. step is obtained changes the green seedling rooting medium of flax pollen HF over to
3In, be that 2000LX, light application time are to cultivate under 15 hours/day the condition at 18-25 ℃, intensity of illumination.
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| CN2008100647514A CN101292629B (en) | 2008-06-18 | 2008-06-18 | Flax free farina culture medium and flax free farina culture method |
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|---|---|---|---|
| CN2008100647514A CN101292629B (en) | 2008-06-18 | 2008-06-18 | Flax free farina culture medium and flax free farina culture method |
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| CN101292629B CN101292629B (en) | 2011-01-05 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101531990B (en) * | 2009-04-15 | 2011-06-01 | 黑龙江省科学院大庆分院 | Flax monoploid cell suspension system culture medium |
| CN102119662A (en) * | 2010-12-24 | 2011-07-13 | 黑龙江八一农垦大学 | Polyploidy induction method for fiber linum usitatissimum under isolated culture condition |
| CN103396981A (en) * | 2013-07-01 | 2013-11-20 | 镇江瑞繁农艺有限公司 | In vitro culture and vitality determination method for water lily pollen |
| CN103421737A (en) * | 2013-06-04 | 2013-12-04 | 塔里木大学 | Culture medium for promoting germination of Amygdalus L. pollen, culture method and application thereof |
| CN107114245A (en) * | 2017-06-29 | 2017-09-01 | 成都苗夫现代苗木科技有限公司 | One grows flax tissue culture method |
-
2008
- 2008-06-18 CN CN2008100647514A patent/CN101292629B/en not_active Expired - Fee Related
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101531990B (en) * | 2009-04-15 | 2011-06-01 | 黑龙江省科学院大庆分院 | Flax monoploid cell suspension system culture medium |
| CN102119662A (en) * | 2010-12-24 | 2011-07-13 | 黑龙江八一农垦大学 | Polyploidy induction method for fiber linum usitatissimum under isolated culture condition |
| CN102119662B (en) * | 2010-12-24 | 2012-06-06 | 黑龙江八一农垦大学 | Polyploidy induction method for fiber linum usitatissimum under isolated culture condition |
| CN103421737A (en) * | 2013-06-04 | 2013-12-04 | 塔里木大学 | Culture medium for promoting germination of Amygdalus L. pollen, culture method and application thereof |
| CN103421737B (en) * | 2013-06-04 | 2015-04-15 | 塔里木大学 | Culture medium for promoting germination of Amygdalus L. pollen, culture method and application thereof |
| CN103396981A (en) * | 2013-07-01 | 2013-11-20 | 镇江瑞繁农艺有限公司 | In vitro culture and vitality determination method for water lily pollen |
| CN103396981B (en) * | 2013-07-01 | 2015-04-29 | 镇江瑞繁农艺有限公司 | In vitro culture and vitality determination method for water lily pollen |
| CN107114245A (en) * | 2017-06-29 | 2017-09-01 | 成都苗夫现代苗木科技有限公司 | One grows flax tissue culture method |
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