CN101250497A - Culture media for plant tissue culture - Google Patents
Culture media for plant tissue culture Download PDFInfo
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- CN101250497A CN101250497A CN 200810015674 CN200810015674A CN101250497A CN 101250497 A CN101250497 A CN 101250497A CN 200810015674 CN200810015674 CN 200810015674 CN 200810015674 A CN200810015674 A CN 200810015674A CN 101250497 A CN101250497 A CN 101250497A
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Abstract
The invention relates to culture medium which is used to culture plant tissues and is characterized in that the culture medium is prepared through adding rare earth compound on the basis of existing plant tissue culture medium. Since rare earth elements are added into the culture medium of the invention, when the culture medium is in use, the growth and the development of a plant test tube seedling root system can be promoted, the root system activity is increased, the root system differentiation and the metabolic activity are promoted, the photosynthesis is intensified, the plant quality is improved, and the capacity of plants to absorb nutrient elements is increased. The hardening-seedling survival rate can be increased by 10%-30% through using plant test tube seedlings which are cultured by the invention and the using effect is good.
Description
Affiliated technical field
The invention belongs to field of plant tissue culture technique.
Background technology
Plant tissue culture is meant the part with the cell of plant materials, tissue or organ, under aseptic condition, be inoculated on the specific substratum, in certain container, cultivate to obtain new individual method, break away from the parent cultivation because of belonging to, so be also referred to as isolated culture.At present, plant tissue culture technique is seized of critical role in biotechnology, and to garden crop, commodity trees etc., part wherein or most of nursery stock are provided by plant tissue culture technique from ornamental plant, asexually propagated crop.In plant tissue culture technique, the moiety of substratum is an important ring.
Through the research of over half a century, the prescription of existing some kinds of substratum comes out, main at present adopt MS, WHITE, B arranged
5, N
6, GS etc.Its main component comprises inorganic elements and organic element, and in the inorganic elements, its concentration is macroelement greater than 0.5mmol/L's, and less than 0.5mmol/L is trace element; The material that contains organic element comprises carbohydrate, VITAMIN, inositol, natural complex, amino acid etc.The difference of different substratum mainly is the content difference of certain or certain several element, to be adapted to the cultivation of different plants.For example, the substratum of high salt component comprises MS, LS, ER etc., is characterized in that the content of sylvite, ammonium salt and nitrate is all higher, is used for the cultivation of organ, flower pesticide, cell and the protoplastis of plant, wherein being most widely used with MS; The substratum that potassium nitrate content is higher mainly contains B
5, N
6, GS etc., be particularly suitable for the cultivation of grape, araucaria, pulse family and cress, wherein be most widely used with GS again.Yet above-mentioned substratum still has defective.When carrying out plant tissue culture with these cultivation machines, its test-tube plantlet hardening surviving rate is low, and result of use is poor.With GS is example, using GS to carry out in the tissue culture procedures of grape, because the root system absorptive function of test-tube plantlet is poor, a little less than the test-tube plantlet growing way, cause test-tube plantlet hardening surviving rate low, generally be not higher than 70%, not only influence commercial effect, and many research work are caused damage, seriously limited further developing and using of this technology.
Summary of the invention
The purpose of this invention is to provide the good culture media for plant tissue culture of a kind of result of use.
For achieving the above object, the feature of technical solution of the present invention is to contain macroelement compound, trace compound, organic element compound and rare-earth compound in the aqueous solution of this substratum.
A kind of compound that the said rare-earth compound of the present invention is made up of a kind of rare earth element; The also compound of two or more that can form by two or more rare earth element; Preferred NdCl
3And/or LaCl
3, by the milligram quantities of contained material in the aqueous solution of every liter of substratum, NdCl
3And/or LaCl
3Amount is 2.0-10.If use two or more rare-earth compound simultaneously, different rare-earth compounds can be an arbitrary proportion.
The said macroelement compound of the present invention is meant the inorganics of the concentration of effective element in the aqueous solution of substratum greater than 0.5mmol/L; Said trace compound is meant the inorganics of the concentration of effective element in the aqueous solution of substratum less than 0.5mmol/L.
The present invention can be made up of GS and rare-earth compound, also can be made up of MS and rare-earth compound.
A kind of technical scheme of the present invention be by contained material in the aqueous solution of every liter of substratum the milligram number contain following material:
The macroelement compound:
KNO
3?1000-1500；(NH
4)
2SO
4?53.6-80.4；Na
2HPO
4?140-210；MgSO
4·7H
2O100-150；FeSO
4·7H
2O?11.12-16.68；Na
2-EDTA?14.92-22.38；CaCl
2·2H
2O120-180；
Trace compound:
MnSO
4·H
2O?4-6；ZnSO
4·7H
2O?0.8-1.2；CoCl
2·6H
2O?0.010-0.015；CuSO
4·5H
2O0.010-0.015；H
3BO
3?1.2-1.8；KI?0.30-0.45；
Organic element compound:
Nicotinic acid (Vpp) 0.8-1.2; Pyridoxine hydrochloride (VB
6) 0.8-1.2; Vitamin (VB
1) 8-12; Inositol 20-30;
Rare-earth compound:
NdCl
3?2.0-10。
Another technical scheme of the present invention, by contained material in the aqueous solution of every liter of substratum the milligram number contain following material:
The macroelement compound:
NH
4NO
3?1320-1980；KNO
3?1520-2280；CaCl
2·2H
2O?352-528；MgSO
4·7H
2O296-444；KH
2PO
4?136-204；FeSO
4·7H
2O?22.96-34.44；Na
2-EDTA?29.84-44.76；
Trace compound:
MnSO
4·H
2O?17.84-26.76；ZnSO
4·7H
2O?6.88-10.32；CoCl
2·6H
2O?0.02-0.03；CuSO
4·5H
2O?0.02-0.03；H
3BO
3?4.96-7.44；Na
2MO
4·2H
2O?0.2-0.3；KI0.664-0.996；
Organic element compound:
Nicotinic acid (Vpp) 0.4-0.6; Pyridoxine hydrochloride (VB
6) 0.4-0.6; Vitamin (VB
1) 0.08-0.12; Inositol 80-120; Glycine 1.6-2.4;
Rare-earth compound:
NdCl
3?2.0-10。
Principal feature of the present invention is to have added rare earth element in existing plant tissue culture media.Rare earth element is 15 kinds of very close lanthanon of character in the periodic table of chemical element, is also referred to as transition element.The ionic configuration of these elements is similar, and institute is electrically charged identical, and ionic radius is also very nearly the same, so these+character of 3 valency elements is very similar.Rare earth element can react with elements such as nitrogen, hydrogen, carbon, phosphorus, is soluble in hydrochloric acid, sulfuric acid and the nitric acid.After adding rare earth element in the substratum, can be by rare earth ion to Ca
2+, Mg
2+, Fe
2+Deng the displacement of metal ion and competition and influence the physiological function of some biological enzymes, biomacromolecule.The present invention is owing to added rare earth element, in use, can promote growing of root system of plant, improve the root system vigor, promote root system differentiation and Metabolic activity, strengthen photosynthesis, improve plant quality, improve the receptivity of plant to nutritive element, can be widely used in the tissue culture of multiple draft such as grape, petunia, raspberry, the living poplar of speed, the tuber of pinellia, muskmelon, hemerocailis middendorffi, xylophyta, more existing substratum is compared, and its hardening surviving rate of plant of using the present invention to cultivate can improve 10%-30%, and result of use is good.Because China is the big country of rare earth resources, also be large agricultural country, so the present invention has application promise in clinical practice.
Embodiment
Embodiments of the invention 1-6 sees Table 1.Wherein the amount of each component is the amount of contained solute in the aqueous solution of every liter of substratum, and unit is mg/L.
The compound method of embodiment 1 is as follows:
1, the preparation of rare-earth compound solution: the content according to Nd among the embodiment 1 takes by weighing Nd
2O
3498mg is that hydrochloric acid and its reaction of 1N is NdCl with 8ml concentration approximately
3, fixed molten with distilled water to 1000ml, be mixed with the NdCl that concentration is 1mg/ml
3Solution.
2, the preparation of mother liquor: according to the amount of listed each component among the embodiment 1, weighing macroelement, trace element, organic element and calcium chloride reagent are also fixed molten, add the NdCl that has prepared
3Solution.
3, culture medium preparation: measure 500ml distilled water earlier, add 20g sucrose, 5g agar, above-mentioned mother liquor and IAA 0.2mg successively.Fixed molten with distilled water to 1L, with NaOH, HCl the ph value of solution is adjusted to 5.8-6.0.
4, medium sterilization: the above-mentioned medium liquid branch that has prepared of 1L is filled in 20-22 the 150ml Erlenmeyer flask, lie against in the high-pressure sterilizing pot after sealing, heat to pressure be steam bleeding behind the 0.05Pa, continue to heat to pressure be 0.12-0.15Pa, timing 15-20 minute, keep flat after the taking-up, cool off and get final product.
Through the aforesaid method prepared culture medium, can carry out the material inoculation.
The preparation of embodiment 2-6: take by weighing reagent by listed amount of substance in the table 1 respectively, prepare 1 liter of culture medium solution respectively by above-mentioned steps.
Table 1, embodiment 1-6
Press the prepared substratum of embodiment 1-6, but called after GSX.Learn that through test more existing GS compares, GSX has outstanding beneficial effect.Be that example gives comparative descriptions now with growth effect to test-tube grape seedling.
Choose 30 days test-tube grape seedling of growth, remove stem apex and basal part of stem, the stem section is cut into one section of a bud in the middle of getting, and is inoculated in respectively on GS and the GSX substratum, cultivate indexs such as measuring its radical, root length, plant height, the number of blade and chlorophyll content after 25 days respectively, and calculate mean number.
1. to the influence of test-tube grape seedling rooting, leave, plant height situation: see Table 2.
Table 2, press the prepared substratum of embodiment 1-6 (GSX) and former substratum (GS) relatively to the influence of test-tube grape seedling rooting, leave, plant height situation:
As can be seen, with the test-tube grape seedling that the test-tube grape seedling ratio of GSX cultivation is cultivated with GS, its number of on average taking root exceeds the 2.0-3.4 bar, the average root 0.7-1.2cm that grows tall out, average leave number exceeds the 0.5-1.2 sheet, and average plant height exceeds 0.8-1.5cm, and this illustrates that advantage of the present invention is apparent.
2. to the influence of test-tube grape seedling chlorophyll content: see Table 3.
Table 3, press the prepared substratum of embodiment 1-6 (GSX) and former substratum (GS) relatively to the influence of test-tube grape seedling chlorophyll content situation:
As can be seen, with the test-tube plantlet that the test-tube plantlet ratio of GSX cultivation is cultivated with GS, its test-tube plantlet blade chlorophyll content exceeds 15.6%-27.9%.Chlorophyllous raising has significant promoter action to the plant test-tube plantlet blade in this explanation the present invention.Chlorophyll content is the basis of photosynthesis of plant, and chlorophyllous raising illustrates that test-tube plantlet photosynthesis is strong, and the organic synthesis ability of test-tube plantlet is strengthened, and can guarantee that test-tube plantlet has vigorous growth potential.
Embodiment 7-12 sees Table 4.
Embodiment 7-12 preparation method:
Take by weighing reagent by the listed amount of substance of table 4 respectively, prepare 1 liter of culture medium solution respectively by the preparation steps of embodiment 1.
Table 4, embodiment 7-12
Press the prepared substratum of embodiment 7-12, but called after MSX.Learn that through test more existing MS compares, MSX has outstanding beneficial effect.Be that example gives comparative descriptions now with growth effect to the petunia test-tube plantlet.
Choose 30 days petunia test-tube plantlet of growth, remove stem apex and basal part of stem, the stem section is cut into one section of a bud in the middle of getting, and is inoculated in respectively on MS and the MSX substratum, cultivates indexs such as measuring its radical, root length, plant height, the number of blade after 25 days respectively, and calculates mean number.
The situation that influences to petunia rooting of vitro seedling, leave, plant height situation sees Table 5.
Table 5, press the prepared substratum of embodiment 7-12 (MSX) and former substratum (MS) relatively to the influence of petunia rooting of vitro seedling, leave, plant height situation:
As can be seen, the test-tube grape seedling of cultivating with MSX compares the petunia test-tube plantlet of cultivating with MS, its number of on average taking root exceeds the 2.6-3.6 bar, the average root 0.7-1.1cm that grows tall out, average leave number exceeds the 1-2 sheet, average plant height exceeds 1.6-2.2cm, illustrates that the present invention is apparent to the advantage that promotes plant-growth.
By above data as can be seen, GSX of the present invention, MSX substratum are compared with former substratum GS, MS, and the root system of grape, petunia is had remarkably influenced, and the number of on average taking root, average root is long all is significantly improved.Because the root growth of test-tube plantlet is improved, and has promoted the receptivity of test-tube plantlet, the cauline leaf growth of test-tube plantlet is also got help.In addition, also effectively improved the chlorophyll content of test-tube plantlet blade, thereby the stalwartness growth of test-tube plantlet is guaranteed.Test is learnt, after using the present invention, the hardening surviving rate that can make test-tube grape seedling is increased to more than 95% by original about 70%, the hardening surviving rate that makes petunia is increased to 100% by original about 90%, improve the efficient of plant tissue culture greatly, saved the reproductive-cost of test-tube plantlet.Except test-tube grape seedling and petunia test-tube plantlet, the present invention all has unusual effect to the various plants tissue culture, has the accelerating effect of broad-spectrum, is analyzed no longer one by one.
Claims (10)
1, a kind of culture media for plant tissue culture is characterized in that containing macroelement compound, trace compound, organic element compound and rare-earth compound in the aqueous solution of this substratum.
2,, it is characterized in that a kind of compound that said rare-earth compound is made up of a kind of rare earth element according to the described substratum of claim 1.
3, according to the described substratum of claim 1, the compound of two or more that it is characterized in that said rare-earth compound is made up of two or more rare earth element.
4,, it is characterized in that said rare-earth compound is NdCl according to the described substratum of claim 1
3And/or LaCl
3
5,, it is characterized in that the amount of the rare-earth compound that contained in the aqueous solution of every liter of substratum is 2.0-10mg according to claim 1,2,3 or 4 described substratum.
6,, it is characterized in that said macroelement compound is meant the inorganics of the concentration of effective element in the aqueous solution of substratum greater than 0.5mmol/L according to claim 1,2,3 or 4 described substratum.
7,, it is characterized in that said trace compound is meant the inorganics of the concentration of effective element in the aqueous solution of substratum less than 0.5mmol/L according to claim 1,2,3 or 4 described substratum.
8, according to claim 1,2,3 or 4 described substratum, it is characterized in that this substratum is made up of GS and rare-earth compound, also can form by MS and rare-earth compound.
9,, it is characterized in that containing following material by the milligram number of contained material in the aqueous solution of every liter of substratum according to claim 1,2,3 or 4 described substratum:
The macroelement compound:
KNO
3?1000-1500；(NH
4)
2SO
4?53.6-80.4；Na
2HPO
4?140-210；MgSO
4·7H
2O100-150；FeSO
4·7H
2O?11.12-16.68；Na
2-EDTA?14.92-22.38；CaCl
2·2H
2O120-180；
Trace compound:
MnSO
4·H
2O?4-6；ZnSO
4·7H
2O?0.8-1.2；CoCl·6H
2O?0.01-0.015；CuSO
4·5H
2O?0.01-0.015；H
3BO
3?1.2-1.8；KI?0.30-0.45；
Organic element compound:
Nicotinic acid (Vpp) 0.8-1.2; Pyridoxine hydrochloride (VB
6) 0.8-1.2; Vitamin (VB
1) 8-12; Inositol 20-30;
Rare-earth compound:
NdCl
3?2.0-10。
10,, it is characterized in that containing following material by the milligram number of contained material in the aqueous solution of every liter of substratum according to claim 1,2,3 or 4 described substratum:
The macroelement compound:
NH
4NO
3?1320-1980；KNO
3?1520-2280；CaCl
2·2H
2O?352-528；MgSO
4·7H
2O296-444；KH
2PO
4?136-204；FeSO
4·7H
2O?22.96-34.44；Na
2-EDTA?29.84-44.76；
Trace compound:
MnSO
4·H
2O?17.84-26.76；ZnSO
4·7H
2O?6.88-10.32；CoCl
2·6H
2O0.02-0.03；CuSO
4·5H
2O?0.02-0.03；H
3BO
3?4.96-7.44；Na
2MO
4·2H
2O?0.2-0.3；KI?0.664-0.996；
Organic element compound:
Nicotinic acid (Vpp) 0.4-0.6; Pyridoxine hydrochloride (VB
6) 0.4-0.6; Vitamin (VB
1) 0.08-0.12; Inositol 80-120; Glycine 1.6-2.4;
Rare-earth compound:
NdCl
3?2.0-10。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200810015674 CN101250497B (en) | 2008-04-14 | 2008-04-14 | Culture media for plant tissue culture |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200810015674 CN101250497B (en) | 2008-04-14 | 2008-04-14 | Culture media for plant tissue culture |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101250497A true CN101250497A (en) | 2008-08-27 |
| CN101250497B CN101250497B (en) | 2010-09-08 |
Family
ID=39954097
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200810015674 Expired - Fee Related CN101250497B (en) | 2008-04-14 | 2008-04-14 | Culture media for plant tissue culture |
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|---|---|
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102577970A (en) * | 2012-03-08 | 2012-07-18 | 重庆文理学院 | Tissue medium for Lonicera macranthoides Hand. Mazz Yulei No.1 sprouts |
| CN102845747A (en) * | 2012-09-01 | 2013-01-02 | 山东博然螺旋藻生物股份有限公司 | Preparation method of selenium-rich quick-frozen spiral algae |
| CZ306000B6 (en) * | 2014-06-24 | 2016-06-15 | Mikrobiologický ústav AV ČR, v. v. i. | Nutrient solution for culturing photosynthesizing microorganisms, process of its preparation and use |
| CN106212286A (en) * | 2016-08-10 | 2016-12-14 | 江苏绿洲园艺绿化有限公司 | A kind of laurustinus special culture media |
-
2008
- 2008-04-14 CN CN 200810015674 patent/CN101250497B/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102577970A (en) * | 2012-03-08 | 2012-07-18 | 重庆文理学院 | Tissue medium for Lonicera macranthoides Hand. Mazz Yulei No.1 sprouts |
| CN102845747A (en) * | 2012-09-01 | 2013-01-02 | 山东博然螺旋藻生物股份有限公司 | Preparation method of selenium-rich quick-frozen spiral algae |
| CN102845747B (en) * | 2012-09-01 | 2013-08-14 | 山东博然螺旋藻生物股份有限公司 | Preparation method of selenium-rich quick-frozen spiral algae |
| CZ306000B6 (en) * | 2014-06-24 | 2016-06-15 | Mikrobiologický ústav AV ČR, v. v. i. | Nutrient solution for culturing photosynthesizing microorganisms, process of its preparation and use |
| CN106212286A (en) * | 2016-08-10 | 2016-12-14 | 江苏绿洲园艺绿化有限公司 | A kind of laurustinus special culture media |
| CN106212286B (en) * | 2016-08-10 | 2019-06-25 | 江苏绿洲园艺绿化有限公司 | A kind of laurustinus special culture media |
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| Publication number | Publication date |
|---|---|
| CN101250497B (en) | 2010-09-08 |
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