CN104170743A - Culture medium for preserving tissue culture seedlings of stevia rebaudiana - Google Patents
Culture medium for preserving tissue culture seedlings of stevia rebaudiana Download PDFInfo
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- CN104170743A CN104170743A CN201410453016.8A CN201410453016A CN104170743A CN 104170743 A CN104170743 A CN 104170743A CN 201410453016 A CN201410453016 A CN 201410453016A CN 104170743 A CN104170743 A CN 104170743A
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- stevia rebaudiana
- tissue culture
- stevia
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Abstract
The invention discloses a culture medium for preserving tissue culture seedlings of stevia rebaudiana. The culture medium is characterized by using a peat and perlite mixed matrix instead of a traditional agar matrix and adding a nutrient solution containing various nutrient substances and plant hormones to the peat and perlite mixed matrix. Growth of stevia rebaudiana can be greatly slowed by culturing stevia rebaudiana on the culture medium, thus achieving the aim of preserving the tissue culture seedlings of stevia rebaudiana for a long time. Compared with the traditional plant tissue culture seedling preservation technology, the culture medium has the advantages that the culture medium is used for culturing stevia rebaudiana by adopting normal temperature conditions for stevia rebaudiana tissue culture, is low in expense and cost and has good effects.
Description
Technical field
a Storaged media for stevia group training seedling, relates to stevia tissue culture technique field, says more specifically a kind of stevia group training seedling is carried out to the long-term medium of preserving.
Background technology
it is the important means of carrying out the productions such as stevia Fast-propagation, genetic mutagenesis and research work that the tissue of stevia is cultivated.But because fast numerous production and scientific research have certain stage, inevitably have the lag phase of certain hour therebetween.For example production unit has completed according to client's needs the seedling amount that order needs, and new order also cannot be expected; Or R&D institution completed or come to an end the research of stevia aspect, within the regular hour, needn't breed again chrysanthemum seedling.Now the culture units of organizing of stevia will face the problem that group training seedling is preserved: how to allow the chrysanthemum seedling of group training grow lentamente, by growing slowly, the growth time of prolongation group training seedling in bottle, reduces the operation of subculture, has both saved and has manually also saved material.Because the group of stevia is trained the fast especially of seedling growth, the seedling of generally taking root just can cover with one bottle about 30 days, if do not carry out subculture cultivation, seedling body will be aging gradually, finally dead, made group training material loss.Now, if there is market demand, from explant, transform cultivation possibility the aid is too slow to be helpful temporarily, misses market good opportunity.If but chrysanthemum seedling is carried out to succeeding preservation, and will increase the consumption of human and material resources, cause waste.In order to address the above problem, the method that someone adopts low temperature to cultivate is carried out the preservation of stevia group training seedling, and the group training seedling of stevia is cultivated at 10 ℃, by the environment of low temperature, delays the growth of chrysanthemum seedling, can make the growth cycle of stevia group training seedling extend.But cultivating, low temperature needs certain facility condition, the consumption that can increase electric power.So invent, a kind of can to organize the technology of preserving stevia tissue culturing seedling under cultivation temperature condition in normal stevia be that industry is expected.
Summary of the invention
the object of the present invention is to provide a kind of medium that carries out the preservation of stevia group training seedling under cultivation temperature of organizing in normal stevia, to reduce stevia group training seedling, preserve the running cost of cultivating.
the present invention is achieved in that
1. stevia group is trained a Storaged media for seedling, and its formula is:
(1) matrix: matrix adopts the mixture of the peat composed of rotten mosses and perlite 1:1;
(2) nutrient solution: in nutrient solution, the concentration of macroelement, trace element and molysite is all according to the requirement preparation of 1/2MS medium, and the concentration of organic principle is 2 times of MS medium, separately adds 0.4mg/L NAA and 30g/L sucrose;
(3) consumption: by the tissue culture flasks of 250ml, every bottle adds mixed-matrix 50ml, nutrient solution 40ml.
operating procedure:
(1) preparation of matrix: the peat composed of rotten mosses is put together and mixed according to the weight portion of 1:1 with perlite;
(2) preparation nutrient solution;
(3) bottling: add matrix and nutrient solution in tissue culture bottle;
(4) sterilizing: carry out sterilizing with autoclave sterilizer;
(5) inoculation: treat after sterilizing that tissue culture bottle is cooling, inoculate in matrix, every bottle graft enters 1-5 single stem segment (being preferably 1) and adds a cover;
(6) cultivate: tissue culture bottle is put into group training chamber and according to normal stevia conditions of tissue culture, cultivate: 25 ± 1 ℃ of temperature, the about 1500lx(light source of intensity of illumination is 40 W fluorescent lamps), light application time 12 h/d.
beneficial effect of the present invention:
adopt Storaged media of the present invention to preserve the group training seedling of stevia, can organize under cultivation temperature in normal stevia, the holding time of the training of prolongation group greatly seedling, labor and material saving, reduces expenses.When there is the fast numerous demand of stevia, can from Storaged media, take out immediately chrysanthemum seedling, get back to and in agar medium, carry out fast numerous cultivation; If there is no fast numerous demand, unit can utilize medium of the present invention to carry out the long-term preservation of material, and growth cycle is long, and subculture is long interval time.The application of basal culture medium can improve the strategic maneuver of relevant enterprise and R&D institution, reduces expenditure and increases income.
embodiment:
1. the formula of the Storaged media of stevia group training seedling:
(1) matrix: matrix adopts the mixture of the peat composed of rotten mosses and perlite 1:1;
(2) nutrient solution: in nutrient solution, the concentration of macroelement, trace element and molysite is all according to the requirement preparation of 1/2MS medium, and the concentration of organic principle is 2 times of MS medium, separately adds 0.4mg/L NAA and 30g/L sucrose;
(3) consumption: by the tissue culture flasks of 250ml, every bottle adds mixed-matrix 50ml, nutrient solution 40ml.
operating procedure:
(1) preparation of matrix: the peat composed of rotten mosses is put together and mixed according to the weight portion of 1:1 with perlite;
(2) preparation nutrient solution;
(3) bottling: add matrix and nutrient solution in tissue culture bottle;
(4) sterilizing: carry out sterilizing with autoclave sterilizer;
(5) inoculation: treat after sterilizing that tissue culture bottle is cooling, inoculate in matrix, every bottle graft enters 1-5 single stem segment (being preferably 1) and adds a cover;
(6) cultivate: tissue culture bottle is put into group training chamber and according to normal stevia conditions of tissue culture, cultivate: 25 ± 1 ℃ of temperature, the about 1500lx(light source of intensity of illumination is 40 W fluorescent lamps), light application time 12 h/d.
test situation:
test in October 9 in 2013 Anhui Science and Technology College Plant Tissue Breeding laboratory carry out.According to single stem segment of above-mentioned formula and operating procedure inoculation stevia group training seedling, the agar medium medium of 1/2MS of take is contrast; 20 bottles of every processing inoculations, every bottle graft kind 5 strains.
growth investigation: as seen from Table 1, in Storaged media of the present invention, the tissue culturing seedling growth of stevia is obviously slack-off, from be inoculated into cover with bottle and from time of being inoculated into the death of half seedling all obviously contrast greatly increase, effectively extended the holding time of stevia group training seedling.
two kinds of medium stevia group training seedling growing state tables of table 1.
Claims (1)
1. a Storaged media for stevia group training seedling, is characterized in that, its formula is as follows:
(1) matrix: matrix is the mixture of the peat composed of rotten mosses and perlite 1:1;
(2) nutrient solution: in nutrient solution, the concentration of macroelement, trace element and molysite is all according to the requirement preparation of 1/2MS medium, and the concentration of organic principle is 2 times of MS medium, separately adds 0.4mg/L NAA and 30g/L sucrose;
(3) consumption: by the tissue culture flasks of 250ml, every bottle adds mixed-matrix 50ml, nutrient solution 40ml.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410453016.8A CN104170743A (en) | 2014-09-09 | 2014-09-09 | Culture medium for preserving tissue culture seedlings of stevia rebaudiana |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201410453016.8A CN104170743A (en) | 2014-09-09 | 2014-09-09 | Culture medium for preserving tissue culture seedlings of stevia rebaudiana |
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| CN104170743A true CN104170743A (en) | 2014-12-03 |
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| CN201410453016.8A Pending CN104170743A (en) | 2014-09-09 | 2014-09-09 | Culture medium for preserving tissue culture seedlings of stevia rebaudiana |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106613957A (en) * | 2016-11-03 | 2017-05-10 | 明光市大全甜叶菊专业合作社 | Stevia rebaudiana tissue culture seedling medium |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1178067A (en) * | 1997-10-10 | 1998-04-08 | 中国科学院昆明植物研究所 | Technology for germ plasma preservation and quick breeding by group culture of pyrethrum |
| CN101002541A (en) * | 2007-01-17 | 2007-07-25 | 南京农业大学 | Method for dislocation storage of germ resource of chrysanthemum |
| CN101006774A (en) * | 2007-01-17 | 2007-08-01 | 南京农业大学 | In vitro conservation method of churysanthemum by reducing macroelements in the medium |
| KR20110019552A (en) * | 2009-08-20 | 2011-02-28 | 박철우 | A making method of a composite for promotion sugar content growth of plant |
-
2014
- 2014-09-09 CN CN201410453016.8A patent/CN104170743A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1178067A (en) * | 1997-10-10 | 1998-04-08 | 中国科学院昆明植物研究所 | Technology for germ plasma preservation and quick breeding by group culture of pyrethrum |
| CN101002541A (en) * | 2007-01-17 | 2007-07-25 | 南京农业大学 | Method for dislocation storage of germ resource of chrysanthemum |
| CN101006774A (en) * | 2007-01-17 | 2007-08-01 | 南京农业大学 | In vitro conservation method of churysanthemum by reducing macroelements in the medium |
| KR20110019552A (en) * | 2009-08-20 | 2011-02-28 | 박철우 | A making method of a composite for promotion sugar content growth of plant |
Non-Patent Citations (5)
| Title |
|---|
| J.M. LECROY ET AL.: "Propagation of Stevia rebaudiana Stem Cuttings Infuenced by Media and Cutting Types", 《HORTSCIENCE》 * |
| SHAHID AKBAR KHALIL ET AL.: "Selection of suitable propagation method for consistent plantlets production in Stevia rebaudiana (Bertoni)", 《SAUDI JOURNAL OF BIOLOGICAL SCIENCES》 * |
| 吴仕超: "42份菊科花卉的离体保存及其遗传多样性的RAPD分析", 《福建农林大学硕士学位论文》 * |
| 崔广荣等: "甜叶菊的组织培养", 《安徽科技学院学报》 * |
| 董振红: "甜叶菊品系"1096"离体培养及其UGT76G1酶基因的克隆", 《东北农业大学硕士学位论文》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106613957A (en) * | 2016-11-03 | 2017-05-10 | 明光市大全甜叶菊专业合作社 | Stevia rebaudiana tissue culture seedling medium |
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Application publication date: 20141203 |