CN102258010A - Slow refrigerating preservation method for paris polyphylla rootstock - Google Patents
Slow refrigerating preservation method for paris polyphylla rootstock Download PDFInfo
- Publication number
- CN102258010A CN102258010A CN2011102591816A CN201110259181A CN102258010A CN 102258010 A CN102258010 A CN 102258010A CN 2011102591816 A CN2011102591816 A CN 2011102591816A CN 201110259181 A CN201110259181 A CN 201110259181A CN 102258010 A CN102258010 A CN 102258010A
- Authority
- CN
- China
- Prior art keywords
- place
- rhizome
- paris rhizome
- magnificent
- rhizoma paris
- 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.)
- Granted
Links
- 238000000034 method Methods 0.000 title claims abstract description 28
- 241000244987 Daiswa polyphylla Species 0.000 title abstract description 12
- 238000004321 preservation Methods 0.000 title abstract description 11
- 239000000843 powder Substances 0.000 claims abstract description 9
- 235000020183 skimmed milk Nutrition 0.000 claims abstract description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 22
- 238000007710 freezing Methods 0.000 claims description 18
- 230000008014 freezing Effects 0.000 claims description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 239000007788 liquid Substances 0.000 claims description 11
- 229910052757 nitrogen Inorganic materials 0.000 claims description 11
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 claims description 10
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims description 10
- 229930006000 Sucrose Natural products 0.000 claims description 10
- 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 10
- 239000005720 sucrose Substances 0.000 claims description 10
- 238000005406 washing Methods 0.000 claims description 9
- 238000010257 thawing Methods 0.000 claims description 6
- 239000012531 culture fluid Substances 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 5
- 238000011010 flushing procedure Methods 0.000 claims description 5
- 238000002791 soaking Methods 0.000 claims description 5
- 239000000463 material Substances 0.000 abstract description 23
- 238000001816 cooling Methods 0.000 abstract description 10
- 230000004083 survival effect Effects 0.000 abstract description 10
- 230000000694 effects Effects 0.000 abstract description 5
- 231100000331 toxic Toxicity 0.000 abstract 1
- 230000002588 toxic effect Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 15
- 239000000243 solution Substances 0.000 description 14
- PKDBCJSWQUOKDO-UHFFFAOYSA-M 2,3,5-triphenyltetrazolium chloride Chemical compound [Cl-].C1=CC=CC=C1C(N=[N+]1C=2C=CC=CC=2)=NN1C1=CC=CC=C1 PKDBCJSWQUOKDO-UHFFFAOYSA-M 0.000 description 10
- 230000031700 light absorption Effects 0.000 description 6
- 239000003814 drug Substances 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 238000005138 cryopreservation Methods 0.000 description 4
- 238000004043 dyeing Methods 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
- 239000008055 phosphate buffer solution Substances 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 230000006378 damage Effects 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 239000013078 crystal Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 206010010904 Convulsion Diseases 0.000 description 1
- 241000243575 Daiswa yunnanensis Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000243542 Paris polyphylla var. chinensis Species 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 230000000202 analgesic effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000036461 convulsion Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 210000002977 intracellular fluid Anatomy 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000010297 mechanical methods and process Methods 0.000 description 1
- 230000005226 mechanical processes and functions Effects 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a slow refrigerating preservation method for a paris polyphylla rootstock. In the method, skimmed milk powder with no toxic or side effect is taken as a protectant, a normal refrigerator can be used for finishing cooling operation in the refrigerating preservation process instead of an expensive procedural cooling instrument, the preservation cost and instrument dependence of the material are greatly lowered, and the operation process of the slow refrigerating preservation method is effectively simplified. Due to the adoption of the method, the survival rate of a preserved paris polyphylla rootstock can be about 75-85 percent, and environmentally-friendly and efficient preservation of the paris polyphylla rootstock material is realized.
Description
Technical field
The present invention relates to a kind of cryopreservation method of plant germplasm resource, particularly relate to a kind of slow freezing store method of magnificent rhizoma paris rhizome.
Background technology
The Chinese medicine Paris polyphylla of Chinese Pharmacopoeia regulation is the subterranean stem of Yunnan Rhizoma Paridis (Paris polyphylla var.yunnanensis) and magnificent Paris polyphylla (Paris polyphylla var.chinensis), and the former main product is in Yunnan, and latter's main product is in Sichuan.The flea that Shennong's Herbal is put down in writing the earliest not is magnificent Paris polyphylla.That Paris polyphylla has is clearing heat and detoxicating, anti-inflammatory analgetic, activate blood circulation and disperse blood clots, the effect of cool liver arresting convulsion, be important magistral medicines.
Paris polyphylla heightens because of it has antiviral, antitumor action demand in recent years, and price raises up fast.The Paris polyphylla traditional Chinese medicine always provides medicinal with wild resource, because wild resource ground is excessively excavated, causes wild Paris polyphylla resource to suffer destructive destruction in recent years, and is now endangered.For better protection wild Paris chinensis Franch matter resource, the present invention has carried out the rhizomatous ultralow temperature of Paris polyphylla to China and has preserved Study on Technology.
The slow freezing method is long, preservation effect of an a kind of application time cryopreservation method preferably, and it is to be the freezing and storing method that theoretical foundation is set up with the protectiveness dehydration; This method is by finishing in procedural cooling instrument the progressively cooling of freezing preservation material to be handled, the intracellular moisture of material is fully deviate from, in refrigerating process, caused preserving the harm of material because of large quantity of moisture in the cell produces ice crystal thereby avoid preserving material effectively.Yet the cooling step by step of preserving material is handled because this method must use expensive procedural cooling instrument just can finish in actual mechanical process, concrete operations need be finished in procedural cooling instrument and will preserve material with per minute 1-2 ℃ cooling rate and drop to gradually about-100 ℃ from 0 ℃, material are dropped in the liquid nitrogen at last again and preserve.Therefore, conventional slow freezing store method must use expensive procedural cooling instrument on the one hand, and whole freezing preservation process need spend the long time on the other hand; And the conventional employed cryoprotector of slow freezing method mostly is organic reagent greatly, also is unfavorable for the preservation of traditional Chinese medicine germplasm materials.
Summary of the invention
The present invention is directed to the problem that has the store method existence now and the slow freezing store method that a kind of magnificent rhizoma paris rhizome is provided; this method adopts skim milk powder without any side effects as protectant, and has replaced the expensive program cooling instrument that must use originally with general refrigerator.
For achieving the above object, the solution that the present invention adopts comprises the following steps:
(1) preliminary treatment: get magnificent rhizoma paris rhizome, under 4~8 ℃ of conditions, placed 15~20 days earlier, be cut into the bulk of 5~20mm3 then, place pre-culture fluid again: sucrose 4~7%+ proline 0.2~0.5% pre-cultivation 20~24 hours;
(2) freezing processing: the magnificent rhizoma paris rhizome after will cultivating is in advance put into protectant cryovial is housed, place 2 ℃ of refrigerators to place 30~40min earlier, and then dislocation is in-16 ℃ refrigerator and begin to lower the temperature gradually, treat to place 30~60min after temperature is reduced to-24 ℃, drop in the liquid nitrogen rapidly immediately and preserve, described protectant is: skim milk powder 30~40%+ proline 1.0~1.5%;
(3) thaw and wash: magnificent rhizoma paris rhizome preserved at least 1 week in liquid nitrogen after, when needs use rhizome, cryovial is taken out, put into constant incubator rapidly and carry out quick-thawing, thaw finish after, sucrose solution with 0.4~0.5mol/L carries out washing by soaking 3~4 times, and each 10~15min is clean with distilled water flushing at last.
The temperature of constant incubator is 40~60 ℃ in the above-mentioned steps (3).
The survival rate of the rhizoma paris rhizome cell after thawing can detect by the following method: the magnificent rhizoma paris rhizome pH after will washing is 7.0 0.4%TTC (2,3, the 5-triphenyltetrazolium chloride) solution is in 25 ℃ of dark place dyeing 18h, remove TTC solution then, adding absolute ethyl alcohol, to carry out the extraction of TTF in 40 ℃ of thermostat water baths colourless until material, get supernatant and measure light absorption value in the 485nm place, and back material cell relative survival rate is frozen in calculating, cell relative survival rate %=handles the light absorption value X 100% of the light absorption value/material that is untreated of back material, the TTC solution that is adopted is to be formulated in 7.0 the phosphate buffer solution with the TTC material at pH, and compound concentration is 0.4%.
The present invention has adopted skim milk powder without any side effects as protectant, has realized that the green of rare traditional Chinese medicine is preserved; Though skim milk powder can not enter in the cell as the impermeability protectant; but it is dissolvable in water in the tissue juice between the cell to increase the concentration of tissue juice; thereby make and form permeable pressure head between intracellular fluid and tissue juice; can impel deviating from of ICW effectively; with the content of free water in the minimizing cell, thereby can avoid material in process of cryopreservation, to cause the injury of pair cell owing to the formation of ice crystal in the cell.Simultaneously, the present invention has realized finishing smoothly the cooling operation of process of cryopreservation in general refrigerator, really broken away from the dependence of conventional slow freezing store method to the expensive program cooling instrument, has reduced the time and the cost of the freezing preservation of material widely.
Below in conjunction with embodiment the present invention is described in further details, but content of the present invention is not limited in this.
Embodiment
Embodiment 1
(1) preliminary treatment: choose and grow more than 5 years and the magnificent rhizoma paris rhizome of healthy growth, under 4 ℃ of conditions, placed 20 days earlier, be cut into 15mm then
3Bulk, place pre-culture fluid again: sucrose 7%+ proline 0.5% is pre-to be cultivated 24 hours;
(2) freezing processing: the magnificent rhizoma paris rhizome after will cultivating is in advance put into protectant cryovial is housed, place 2 ℃ of refrigerators to place 30min earlier, and then dislocation is in-16 ℃ refrigerator and begin to lower the temperature gradually, treat to place 40min after temperature is reduced to-24 ℃, drop in the liquid nitrogen rapidly immediately and preserve, described protectant is: skim milk powder 40%+ proline 1.5%;
(3) thaw and wash: magnificent rhizoma paris rhizome preserved for 2 weeks in liquid nitrogen after, when needs use rhizome, cryovial is taken out, put into 55 ℃ of constant incubators rapidly and carry out quick-thawing, thaw finish after, sucrose solution with 0.5mol/L carries out washing by soaking 3 times, and each 10min is clean with distilled water flushing at last;
(4) freeze the detection of material cell survival rate afterwards: the magnificent rhizoma paris rhizome after will washing is put into beaker, add that an amount of to regulate pH with phosphate buffer solution be that 7.0 0.4%TTC solution is in 25 ℃ of constant temperature dark places dyeing 18h, inhale then and remove TTC solution, add absolute ethyl alcohol and in 40 ℃ of thermostat water baths, carry out the extraction of TF until colourless, get supernatant and measure light absorption value at the 485nm place on spectrophotometer, it is 85.4% that back material cell relative survival rate is frozen in calculating.
Embodiment 2
(1) preliminary treatment: choose and grow more than 5 years and the magnificent rhizoma paris rhizome of healthy growth, under 4 ℃ of conditions, placed 20 days earlier, be cut into 15mm then
3Bulk, place pre-culture fluid again: sucrose 4%+ proline 0.2% is pre-to be cultivated 24 hours;
(2) freezing processing: the magnificent rhizoma paris rhizome after will cultivating is in advance put into protectant cryovial is housed, place 2 ℃ of refrigerators to place 30min earlier, and then dislocation is in-16 ℃ refrigerator and begin to lower the temperature gradually, treat to place 40min after temperature is reduced to-24 ℃, drop in the liquid nitrogen rapidly immediately and preserve, described protectant is: skim milk powder 30%+ proline 1%;
(3) thaw and wash: magnificent rhizoma paris rhizome preserved for 2 weeks in liquid nitrogen after, when needs use rhizome, cryovial is taken out, put into 40 ℃ of constant incubators rapidly and carry out quick-thawing, thaw finish after, sucrose solution with 0.5mol/L carries out washing by soaking 3 times, and each 10min is clean with distilled water flushing at last;
(4) freeze the detection of material cell survival rate afterwards: the magnificent rhizoma paris rhizome after will washing is put into beaker, add that an amount of to regulate pH with phosphate buffer solution be that 7.0 0.4%TTC solution is in 25 ℃ of constant temperature dark places dyeing 18h, inhale then and remove TTC solution, add absolute ethyl alcohol and in 40 ℃ of thermostat water baths, carry out the extraction of TF until colourless, get supernatant and measure light absorption value at the 485nm place on spectrophotometer, it is 77.4% that back material cell relative survival rate is frozen in calculating.
Embodiment 3
(1) preliminary treatment: choose and grow more than 5 years and the magnificent rhizoma paris rhizome of healthy growth, under 4 ℃ of conditions, placed 15 days earlier, be cut into 5mm then
3Bulk, place pre-culture fluid again: sucrose 7%+ proline 0.5% is pre-to be cultivated 20 hours;
(2) freezing processing: the magnificent rhizoma paris rhizome after will cultivating is in advance put into protectant cryovial is housed, place 2 ℃ of refrigerators to place 30min earlier, and then dislocation is in-16 ℃ refrigerator and begin to lower the temperature gradually, treat to place 40min after temperature is reduced to-24 ℃, drop in the liquid nitrogen rapidly immediately and preserve, described protectant is: skim milk powder 40%+ proline 1.5%;
(3) thaw and wash: magnificent rhizoma paris rhizome preserved for 2 weeks in liquid nitrogen after, when needs use rhizome, cryovial is taken out, put into 55 ℃ of constant incubators rapidly and carry out quick-thawing, thaw finish after, sucrose solution with 0.4mol/L carries out washing by soaking 3 times, and each 10min is clean with distilled water flushing at last;
(4) freeze the detection of material cell survival rate afterwards: the magnificent rhizoma paris rhizome after will washing is put into beaker, add that an amount of to regulate pH with phosphate buffer solution be that 7.0 0.4%TTC solution is in 25 ℃ of constant temperature dark places dyeing 18h, inhale then and remove TTC solution, add absolute ethyl alcohol and in 40 ℃ of thermostat water baths, carry out the extraction of TF until colourless, get supernatant and measure light absorption value at the 485nm place on spectrophotometer, it is 80.4% that back material cell relative survival rate is frozen in calculating.
Claims (2)
1. the slow freezing store method of a magnificent rhizoma paris rhizome is characterized in that comprising the steps:
(1) preliminary treatment: get magnificent rhizoma paris rhizome, under 4~8 ℃ of conditions, placed 15~20 days earlier, be cut into 5~20mm then
3Bulk, place pre-culture fluid again: sucrose 4~7%+ proline 0.2~0.5% is pre-to be cultivated 20~24 hours;
(2) freezing processing: the magnificent rhizoma paris rhizome after will cultivating is in advance put into protectant cryovial is housed, place 2 ℃ of refrigerators to place 30~40min earlier, and then dislocation is in-16 ℃ refrigerator and begin to lower the temperature gradually, treat to place 30~60min after temperature is reduced to-24 ℃, drop in the liquid nitrogen rapidly immediately and preserve, described protectant is: skim milk powder 30~40%+ proline 1.0~1.5%;
(3) thaw and wash: magnificent rhizoma paris rhizome preserved at least 1 week in liquid nitrogen after, when needs use rhizome, cryovial is taken out, put into constant incubator rapidly and carry out quick-thawing, thaw finish after, sucrose solution with 0.4~0.5mol/L carries out washing by soaking 3~4 times, and each 10~15min is clean with distilled water flushing at last.
2. a kind of magnificent rhizoma paris rhizome slow freezing store method according to claim 1 is characterized in that: the temperature of constant incubator is 40~60 ℃ in the step (3).
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN 201110259181 CN102258010B (en) | 2011-09-02 | 2011-09-02 | Slow refrigerating preservation method for paris polyphylla rootstock |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN 201110259181 CN102258010B (en) | 2011-09-02 | 2011-09-02 | Slow refrigerating preservation method for paris polyphylla rootstock |
Publications (2)
Publication Number | Publication Date |
---|---|
CN102258010A true CN102258010A (en) | 2011-11-30 |
CN102258010B CN102258010B (en) | 2013-12-18 |
Family
ID=45005010
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN 201110259181 Expired - Fee Related CN102258010B (en) | 2011-09-02 | 2011-09-02 | Slow refrigerating preservation method for paris polyphylla rootstock |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN102258010B (en) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105123253A (en) * | 2015-10-15 | 2015-12-09 | 王婧 | Seedling method for paris polyphylla |
CN108684657A (en) * | 2018-03-27 | 2018-10-23 | 中国医学科学院药用植物研究所海南分所 | A kind of dalbergia wood seed cryopreservation method |
CN108719280A (en) * | 2018-07-14 | 2018-11-02 | 沿河后花园农业观光旅游综合开发有限公司 | A kind of store method of lily ball tissue |
CN111693475A (en) * | 2020-07-28 | 2020-09-22 | 扬州大学 | Improved method for measuring activity of plant root system in sections |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
SU1477325A1 (en) * | 1986-01-27 | 1989-05-07 | Крымское Научно-Производственное Объединение Винодельческой Промышленности | Method of preparing grapevine grafts for winter grafting |
JPH1056821A (en) * | 1996-08-22 | 1998-03-03 | Koichi Naito | Greening foundation member |
CN1532279A (en) * | 2003-03-26 | 2004-09-29 | ����������Ʒ�о��� | Freeze-drying and cryopreservation methods of ginseng callus cells |
CN101884321A (en) * | 2010-07-13 | 2010-11-17 | 大连普瑞康生物技术有限公司 | Cryopreservation method for Sinkiang saussurea involucrata cell line |
-
2011
- 2011-09-02 CN CN 201110259181 patent/CN102258010B/en not_active Expired - Fee Related
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
SU1477325A1 (en) * | 1986-01-27 | 1989-05-07 | Крымское Научно-Производственное Объединение Винодельческой Промышленности | Method of preparing grapevine grafts for winter grafting |
JPH1056821A (en) * | 1996-08-22 | 1998-03-03 | Koichi Naito | Greening foundation member |
CN1532279A (en) * | 2003-03-26 | 2004-09-29 | ����������Ʒ�о��� | Freeze-drying and cryopreservation methods of ginseng callus cells |
CN101884321A (en) * | 2010-07-13 | 2010-11-17 | 大连普瑞康生物技术有限公司 | Cryopreservation method for Sinkiang saussurea involucrata cell line |
Non-Patent Citations (2)
Title |
---|
王越 等: "观赏植物种质资源的超低温保存", 《植物生理学通讯》, vol. 42, no. 3, 30 June 2006 (2006-06-30), pages 562 - 2 * |
郭玉琼: "龙眼、荔枝胚性培养物超低温保存研究", 《福建农林大学博士学位论文》, no. 6, 15 December 2007 (2007-12-15), pages 22 - 2 * |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105123253A (en) * | 2015-10-15 | 2015-12-09 | 王婧 | Seedling method for paris polyphylla |
CN105123253B (en) * | 2015-10-15 | 2017-10-27 | 向君民 | A kind of method for culturing seedlings of paris polyphylla |
CN108684657A (en) * | 2018-03-27 | 2018-10-23 | 中国医学科学院药用植物研究所海南分所 | A kind of dalbergia wood seed cryopreservation method |
CN108684657B (en) * | 2018-03-27 | 2021-02-12 | 中国医学科学院药用植物研究所海南分所 | Ultralow-temperature preservation method for dalbergia wood seeds |
CN108719280A (en) * | 2018-07-14 | 2018-11-02 | 沿河后花园农业观光旅游综合开发有限公司 | A kind of store method of lily ball tissue |
CN111693475A (en) * | 2020-07-28 | 2020-09-22 | 扬州大学 | Improved method for measuring activity of plant root system in sections |
Also Published As
Publication number | Publication date |
---|---|
CN102258010B (en) | 2013-12-18 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN102302057B (en) | A kind of method of fresh-keeping lychee with plant essential oil | |
CN102258008B (en) | Vitrification ultra-low temperature preserving method for effectively preserving bitter buckwheat callus | |
CN102258010A (en) | Slow refrigerating preservation method for paris polyphylla rootstock | |
CN103975852B (en) | A kind of method that Saussurea involucrata Callus ultralow temperature is preserved | |
CN103548993A (en) | Fresh-keeping method for prolonging storage period of kiwi fruits | |
CN101965860A (en) | Method for processing quick-frozen fresh lotus | |
CN104127708A (en) | Rhizome gastrodiae freshness keeping method | |
CN104161110A (en) | Cress leaf processing process | |
CN102492622A (en) | Preservation method for stock spawn of straw mushrooms | |
CN102265818B (en) | Preservation method for seed nutgall of Chinese nutgall | |
CN103283717B (en) | Monkshood product and fresh-keeping method thereof | |
CN101884321A (en) | Cryopreservation method for Sinkiang saussurea involucrata cell line | |
CN1072917C (en) | Fresh keeping technology for fresh medicinal rhizomes | |
CN104542923A (en) | Quick-frozen asparagus lettuce processing technology | |
CN103518436A (en) | Method for preventing soil water erosion by means of juice of maize straw | |
CN103314758B (en) | Method for artificially cultivating saussurea involucrata | |
CN105145108B (en) | A kind of cultural methods of the raising golden flower without aspidistra survival rate | |
CN103893220A (en) | Preparation method of Cordyceps sinensis superfine powder | |
CN103039519B (en) | Special comprehensive anti-freezing, insecticiding, egg-killing and sterilization emulsion for yangtao trees, and preparation method of emulsion | |
CN107810949A (en) | A kind of Seed of Platycodon Grandiflorum cryopreservation method | |
CN105557397A (en) | Fruit tree anti-freezing method | |
CN107853293A (en) | A kind of radix ampelopsis seed cryopreservation method | |
CN107864953A (en) | A kind of rhizoma arisaematis seed cryopreservation method | |
CN106578020A (en) | Method for keeping freshness of litchi fruits by using Rhizoma Alpiniae Officinarum essential oil | |
de Kock | Quality management of plums following heat waves during the harvesting season |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
CF01 | Termination of patent right due to non-payment of annual fee | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20131218 Termination date: 20160902 |