MD31Y - Micropropagation process of Echinacea purpurea L. Moench in vitro - Google Patents
Micropropagation process of Echinacea purpurea L. Moench in vitro Download PDFInfo
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- MD31Y MD31Y MDS20090037A MDS20090037A MD31Y MD 31 Y MD31 Y MD 31Y MD S20090037 A MDS20090037 A MD S20090037A MD S20090037 A MDS20090037 A MD S20090037A MD 31 Y MD31 Y MD 31Y
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- 235000014134 echinacea Nutrition 0.000 title claims abstract description 8
- 238000000034 method Methods 0.000 title claims abstract description 8
- 240000004530 Echinacea purpurea Species 0.000 title claims abstract description 7
- 238000000338 in vitro Methods 0.000 title claims abstract description 5
- 235000015097 nutrients Nutrition 0.000 claims abstract description 8
- 244000061458 Solanum melongena Species 0.000 claims abstract description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 5
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims abstract description 4
- 238000009835 boiling Methods 0.000 claims abstract description 4
- 238000000605 extraction Methods 0.000 claims abstract description 4
- 229930002534 steroid glycoside Natural products 0.000 claims abstract description 4
- 230000035784 germination Effects 0.000 claims abstract description 3
- 238000005286 illumination Methods 0.000 claims abstract description 3
- 235000002597 Solanum melongena Nutrition 0.000 claims description 5
- 150000008143 steroidal glycosides Chemical class 0.000 claims description 2
- 241000196324 Embryophyta Species 0.000 abstract description 9
- 230000008929 regeneration Effects 0.000 abstract description 3
- 238000011069 regeneration method Methods 0.000 abstract description 3
- 230000030118 somatic embryogenesis Effects 0.000 abstract description 3
- 230000011218 segmentation Effects 0.000 abstract 1
- 239000002609 medium Substances 0.000 description 11
- 230000012010 growth Effects 0.000 description 9
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 6
- 206010020649 Hyperkeratosis Diseases 0.000 description 5
- 230000018109 developmental process Effects 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 230000000977 initiatory effect Effects 0.000 description 2
- 229910052500 inorganic mineral Inorganic materials 0.000 description 2
- 230000000442 meristematic effect Effects 0.000 description 2
- 239000011707 mineral Substances 0.000 description 2
- 235000010755 mineral Nutrition 0.000 description 2
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 230000007226 seed germination Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 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 description 1
- 244000133098 Echinacea angustifolia Species 0.000 description 1
- 241000206672 Gelidium Species 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 229910004619 Na2MoO4 Inorganic materials 0.000 description 1
- 244000010815 Phlomis lychnitis Species 0.000 description 1
- YGYAWVDWMABLBF-UHFFFAOYSA-N Phosgene Chemical compound ClC(Cl)=O YGYAWVDWMABLBF-UHFFFAOYSA-N 0.000 description 1
- 244000042430 Rhodiola rosea Species 0.000 description 1
- 235000003713 Rhodiola rosea Nutrition 0.000 description 1
- 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 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 235000010419 agar Nutrition 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000007844 bleaching agent Substances 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 235000011148 calcium chloride Nutrition 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 229910000366 copper(II) sulfate Inorganic materials 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- FSBUXLDOLNLABB-ISAKITKMSA-N echinacoside Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](OC(=O)\C=C\C=2C=C(O)C(O)=CC=2)[C@@H](CO[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)O[C@@H](OCCC=2C=C(O)C(O)=CC=2)[C@@H]1O FSBUXLDOLNLABB-ISAKITKMSA-N 0.000 description 1
- NJYVDFDTLLZVMG-UHFFFAOYSA-N echinacoside Natural products CC1OC(OC2C(O)C(OCCc3ccc(O)c(O)c3)OC(COC4OC(CO)C(O)C(O)C4O)C2OC(=O)C=Cc5cc(O)cc(O)c5)C(O)C(O)C1O NJYVDFDTLLZVMG-UHFFFAOYSA-N 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 1
- 230000002439 hemostatic effect Effects 0.000 description 1
- 230000003308 immunostimulating effect Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 230000001338 necrotic effect Effects 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 235000010333 potassium nitrate Nutrition 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 239000011684 sodium molybdate Substances 0.000 description 1
- 235000015393 sodium molybdate Nutrition 0.000 description 1
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 239000012808 vapor phase Substances 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 1
- 229910000368 zinc sulfate Inorganic materials 0.000 description 1
- 239000011686 zinc sulphate Substances 0.000 description 1
- 235000009529 zinc sulphate Nutrition 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
Invenţia se referă la biotehnologie, în particular la un procedeu de micropropagare a plantelor de Echinacea purpurea L. Moench in vitro. The invention relates to biotechnology, in particular to a process for micropropagation of Echinacea purpurea L. Moench plants in vitro.
Echinacea purpurea L. Moench este o plantă decorativă şi medicinală preţioasă, multianuală. Acumulează şi sintetizează substanţele biologic active, este mult apreciată pentru calităţile sale terapeutice, posedă proprietăţi imunostimulente şi antivirale. În componenţa rădăcinilor se conţine substanţa anginoli. Preparatul din flori posedă proprietăţi hemostatice şi de vindecare a rănilor. Toate părţile componente ale plantei conţin glicozida steroidică echinacozidă şi ulei volatil eteric până la 1%. Echinacea purpurea L. Moench is a valuable, perennial ornamental and medicinal plant. It accumulates and synthesizes biologically active substances, is highly valued for its therapeutic qualities, possesses immunostimulating and antiviral properties. The roots contain the substance anginol. The flower preparation has hemostatic and wound-healing properties. All parts of the plant contain the steroid glycoside echinacoside and volatile essential oil up to 1%.
Este cunoscut procedeul de micropropagare a plantelor de Rhodiola rosea L., ce constă în obţinerea şi activarea meristemoizilor de la care pornesc lăstarii. Pentru activarea şi creşterea intensivă a plantulelor, explantele primare prelevate din plantule obţinute prin germinarea seminţelor au fost cultivate pe mediul nutritiv de bază ce conţine săruri minerale după Murashige-Skoog (MS) (Murashige T., Skoog F. Revised medium for rapid growth and bio-assays with tabaco tissue cultures. Physiology Plantarum, 1962, v. 15, N. 95, p. 473, suplimentat cu cărbune activat în cantitate de 1200 mg/l, valoarea pH-ului fiind ajustată la 6,5 până la autoclavare [1]. The micropropagation process of Rhodiola rosea L. plants is known, which consists in obtaining and activating the meristemoids from which the shoots start. For the activation and intensive growth of the seedlings, the primary explants taken from the seedlings obtained by seed germination were cultivated on the basic nutrient medium containing mineral salts according to Murashige-Skoog (MS) (Murashige T., Skoog F. Revised medium for rapid growth and bio-assays with tabaco tissue cultures. Physiology Plantarum, 1962, v. 15, N. 95, p. 473, supplemented with activated charcoal in a quantity of 1200 mg/l, the pH value being adjusted to 6.5 until autoclaving [1].
Dezavantajele acestui procedeu sunt: coeficientul de micropropagare scăzut, faptul că nu toate explantele asigură iniţierea creşterii meristemoizilor de la baza cărora pornesc lăstarii. The disadvantages of this process are: the low micropropagation coefficient, the fact that not all explants ensure the initiation of meristem growth from which shoots start.
Problema pe care o rezolvă invenţia constă în sporirea coeficientului de multiplicare a plantelor de Echinacea purpurea L. Moench pe calea modificării conţinutului mediului nutritiv utilizat pentru micropropagare. The problem solved by the invention consists in increasing the multiplication coefficient of Echinacea purpurea L. Moench plants by modifying the content of the nutrient medium used for micropropagation.
Esenţa invenţiei constă în aceea că procedeul include germinarea aseptică a seminţelor, obţinerea explantelor, segmentarea şi cultivarea lor pe mediul nutritiv Murashige-Skoog, ce conţine suplimentar glicozide steroidice obţinute din rădăcini de pătlăgele vinete Solanum melongena L. prin extracţie cu apă la fierbere, prelucrarea extractului obţinut cu butanol-1 şi concentrare, în concentraţie de (3,0…3,5)·10-3%, totodată cultivarea se efectuează la o temperatură de 22…26°C, umiditatea relativă a aerului de 70%, cu o fotoperioadă de 16 şi 8 ore noapte/zi şi intensitatea iluminării de 3000 lx. The essence of the invention is that the process includes aseptic germination of seeds, obtaining explants, segmenting and cultivating them on Murashige-Skoog nutrient medium, which additionally contains steroidal glycosides obtained from eggplant roots Solanum melongena L. by extraction with boiling water, processing the obtained extract with butanol-1 and concentration, in a concentration of (3.0…3.5)·10-3%, at the same time the cultivation is carried out at a temperature of 22…26°C, relative air humidity of 70%, with a photoperiod of 16 and 8 hours night/day and illumination intensity of 3000 lx.
Preparatul Somelangozidă a fost obţinut din rădăcini de pătlăgele vinete prin extracţie cu apă la fierbere, prelucrarea extractului obţinut cu butanol-1 şi conşentrare (SU 1473317 A 1988.12.15). The preparation Somelangoside was obtained from eggplant roots by extraction with boiling water, processing the obtained extract with butanol-1 and concentration (SU 1473317 A 1988.12.15).
Rezultatul invenţiei constă în sporirea embriogenezei somatice şi regenerării plantelor. The result of the invention consists in enhancing somatic embryogenesis and plant regeneration.
Exemplu de realizare a invenţiei Example of embodiment of the invention
Plantulele de Echinacea purpurea L. Moench au fost obţinute prin germinarea aseptică a seminţelor. Pentru a asigura sterilitatea, seminţele s-au tratat cu agent de înălbire pe bază de clor de 15%, timp de 20 min, spălându-se de 4...5 ori cu apă distilată sterilă. În continuare s-au inoculat pe mediul nutritiv de bază MS. Plantulele obţinute ajungând la mărimea de 7...10 cm, s-au segmentat şi cultivat în condiţii aseptice la hota cu flux de aer laminar steril. Echinacea purpurea L. Moench seedlings were obtained by aseptic seed germination. To ensure sterility, the seeds were treated with 15% chlorine bleach for 20 min, washed 4...5 times with sterile distilled water. They were then inoculated onto MS basic nutrient medium. The seedlings obtained, reaching a size of 7...10 cm, were segmented and cultivated under aseptic conditions in a sterile laminar air flow hood.
În calitate de material iniţial pentru iniţierea micropropagării de plantule au servit segmente de hipocotil şi frunze prelevate din pantulele germinate aseptic din seminţe. În vasele de sticlă de 50...100 ml s-au repartizat câte 15...20 ml de mediu nutritiv. S-au sterilizat prin autoclavare, în faza de vapori 10 min aerisire şi 15 min sterilizare în condiţiile de presiune (p=1 atm) şi temperatură (t=121°C, pH-ul fiind ajustat la 5,8...5,9). As initial material for the initiation of micropropagation of seedlings, hypocotyl segments and leaves taken from aseptically germinated seed pods served. 15...20 ml of nutrient medium were distributed in 50...100 ml glass vessels. They were sterilized by autoclaving, in the vapor phase 10 min of aeration and 15 min of sterilization under pressure (p=1 atm) and temperature (t=121°C, pH adjusted to 5.8...5.9).
Toate operaţiile au fost efectuate în condiţii strict aseptice la hota cu flux de aer laminar steril. În camera de creştere s-a menţinut un regim controlat de creştere constant, caracterizat prin temperatura de 24±2°C, umiditatea relativă a aerului de 70%, durata perioadei de iluminare 16 ore/zi şi 8 ore/noapte, intensitatea luminii 3000 lx. All operations were performed under strictly aseptic conditions in a sterile laminar flow hood. A constant controlled growth regime was maintained in the growth chamber, characterized by a temperature of 24±2°C, relative air humidity of 70%, duration of the lighting period of 16 hours/day and 8 hours/night, light intensity of 3000 lx.
O parte de explante (varianta control) s-au inoculat pe mediul de cultură de bază Murashige-Skoog, ce conţinea macroelemente (săruri minerale, mg/l): NH4NO3 - 1650, KNO3 - 1900, MgSO4 · 7H2O - 370, KH2PO4 - 170, CaCl2 · 7H2O - 440; microelemente, mg/l: H3BO3 - 6,2, MnSO4 · 4H2O - 22,3, COCl2 · 6H2O - 0,025, CuSO4 · 5H2O - 0,025, ZnSO4 · 7H2O - 8,6, Na2MoO4 · 2H2O - 0,25, KJ - 0,83, FeSO4 · 7H2O - 27,8, Na2EDTA - 37,3, Inozitol - 100, solidificat cu agar-agar 0,7%, zaharoză 3%. A part of the explants (control variant) were inoculated on the Murashige-Skoog basic culture medium, which contained macroelements (mineral salts, mg/l): NH4NO3 - 1650, KNO3 - 1900, MgSO4 · 7H2O - 370, KH2PO4 - 170, CaCl2 · 7H2O - 440; microelements, mg/l: H3BO3 - 6.2, MnSO4 · 4H2O - 22.3, COCl2 · 6H2O - 0.025, CuSO4 · 5H2O - 0.025, ZnSO4 · 7H2O - 8.6, Na2MoO4 · 2H2O - 0.25, KJ - 0.83, FeSO4 · 7H2O - 27.8, Na2EDTA - 37.3, Inositol - 100, solidified with agar-agar 0.7%, sucrose 3%.
Altă parte de explante (cea mai apropiată soluţie), s-a utilizat pe mediul de bază Murashige-Skoog, suplimentat cu cărbune activat în cantitate de 1200 mg/l, valoarea pH-ului fiind ajustată la 6,5. Another part of the explants (the closest solution) was used on Murashige-Skoog basic medium, supplemented with activated carbon in an amount of 1200 mg/l, the pH value being adjusted to 6.5.
A treia parte de explante au fost utilizate pe mediul de cultură de bază Murashige-Skoog, suplimentat cu Somelangozidă în concentraţie de 3,0...3,5·10-3. The third part of explants were used on Murashige-Skoog basic culture medium, supplemented with Somelangoside in a concentration of 3.0...3.5·10-3.
Proliferarea explantelor în varianta martor s-a remarcat foarte slab, creşterea şi dezvoltarea calusului fără zone meristematice, ca rezultat explantele inoculate pe acest mediu s-au necrotizat. The proliferation of explants in the control variant was very weak, the growth and development of callus without meristematic areas, as a result the explants inoculated on this medium became necrotic.
În al doilea caz la utilizarea mediului Murashige-Skoog cărbune activat - cea mai apropiată soluţie, proliferarea explantelor s-a realizat după 10 zile de la inoculare, inducerea, creşterea şi dezvoltarea calusului s-au observat după 18 zile. Apariţia plantulelor - după 25 zile. Creşterea şi dezvoltarea plantulelor s-au observat după o lună de zile. În acest caz micropropagarea plantulelor a constituit 3·104 plante anual. In the second case, when using the Murashige-Skoog activated carbon medium - the closest solution, the proliferation of explants was achieved after 10 days from inoculation, the induction, growth and development of callus were observed after 18 days. The emergence of seedlings - after 25 days. The growth and development of seedlings were observed after a month. In this case, the micropropagation of seedlings amounted to 3·104 plants per year.
Pe al treilea mediu, conform invenţiei, proliferarea explantelor a evoluat la 5 zile de la cultivare, la 8 zile formarea şi dezvoltarea calusului embrionar cu zone meristematice de culoare roz. Apariţia plantulelor regenerate s-a observat la 14 zile. Creşterea şi dezvoltarea plantulelor s-a observat la 20 zile de la momentul cultivării explantelor pe mediul de cultură Murashige - Skoog, suplimentat cu substanţe furostanolice Somelangozidă în concentraţie de 3,0·10-3. În această variantă raportul optimal al ingredienţilor prezenţi în mediul nutritiv de cultură artificial sporeşte considerabil coeficientul de micropropagare a plantulelor de Echinacea - cu 4·105 plante anual. Valoarea medie a masei celulare a calusului s-a caracterizat cu valori de 64,23...75,1%. Capacitatea embriogenezei somatice a constituit 77,5...83,6%, iar capacitatea de regenerare a evoluat între 74,2...76,4%. În general calusul embrionar a fost multiplicat în scopul variabilităţii genetice de micropropagare a plantulelor obţinute in vitro. On the third medium, according to the invention, the proliferation of explants evolved 5 days after cultivation, at 8 days the formation and development of embryonic callus with pink meristematic areas. The appearance of regenerated seedlings was observed at 14 days. The growth and development of seedlings was observed 20 days after the explants were cultivated on the Murashige - Skoog culture medium, supplemented with furostanol substances Somelangoside in a concentration of 3.0·10-3. In this variant, the optimal ratio of the ingredients present in the artificial culture nutrient medium considerably increases the micropropagation coefficient of Echinacea seedlings - by 4·105 plants annually. The average value of the callus cell mass was characterized by values of 64.23...75.1%. The capacity of somatic embryogenesis was 77.5...83.6%, and the regeneration capacity evolved between 74.2...76.4%. In general, the embryonic callus was multiplied for the purpose of genetic variability of micropropagation of the plantlets obtained in vitro.
1. MD 3375 G2 2007.08.31 1. MD 3375 G2 2007.08.31
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| MD605Z (en) * | 2012-07-09 | 2013-10-31 | Институт Генетики И Физиологии Растений Академии Наук Молдовы | Process for microclonal propagation of Actinidia arguta plants in vitro |
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| US4338745A (en) * | 1980-03-05 | 1982-07-13 | Kyowa Hakko Kogyo Kabushiki Kaisha | Process for mass propagation of plantlets |
| JPS6356284A (en) * | 1986-04-28 | 1988-03-10 | Noyaku Bio Technol Kaihatsu Gijutsu Kenkyu Kumiai | Cultivation of large amount of phryma leptostachya |
| RU2039427C1 (en) * | 1991-04-25 | 1995-07-20 | Всероссийский селекционно-технологический институт садоводства и питомниководства | Method for reproduction of currants in vitro |
| RU2080780C1 (en) * | 1994-05-11 | 1997-06-10 | Всероссийский научно-исследовательский институт сельскохозяйственной биотехнологии РАСХН | Clonal plant micro propagation method |
| RU2123256C1 (en) * | 1996-08-08 | 1998-12-20 | Всероссийский научно-исследовательский институт цветоводства и субтропических культур | Method of preparing tulip microbulbs from isolated embryos under in vitro conditions |
| MD1262G2 (en) * | 1997-03-03 | 2000-12-31 | Институт Генетики И Физиологии Растений Академии Наук Молдовы | Process for obtaining Stevia rebaudiana Bertoni seedlings |
| RU2180165C2 (en) * | 2000-01-31 | 2002-03-10 | Абукамель Ахмед Абдельсамад | Method for microclonal reproduction of gladiolus |
| RU2222933C2 (en) * | 2002-04-26 | 2004-02-10 | Государственное научное учреждение Всероссийский селекционно-технологический институт садоводства и питомниководства | Method for multiplying of in-vitro grown garden plants |
| RU2273987C2 (en) * | 2004-02-24 | 2006-04-20 | ГНУ Всероссийский научно-исследовательский институт цветоводства и субтропических культур РАСХН | Method for preparing tulip valuable plant-regenerates by culturing seed bud in vitro |
| RU2279212C2 (en) * | 2004-07-13 | 2006-07-10 | ФЕДЕРАЛЬНОЕ ГОСУДАРСТВЕННОЕ ОБРАЗОВАТЕЛЬНОЕ УЧРЕЖДЕНИЕ ВЫСШЕГО ПРОФЕССИОНАЛЬНОГО ОБРАЗОВАНИЯ "РОССИЙСКИЙ ГОСУДАРСТВЕННЫЙ АГРАРНЫЙ УНИВЕРСИТЕТ - МСХА имени К.А.ТИМИРЯЗЕВА" | METHOD FOR in-vitro MULTIPLICATION OF PLANTS |
| RU2286053C2 (en) * | 2005-02-17 | 2006-10-27 | Государственное образовательное учреждение высшего профессионального образования "Мордовский государственный университет им. Н.П. Огарева" | Method for in vitro multiplication of gladiolus |
| RU2324338C1 (en) * | 2007-01-25 | 2008-05-20 | Государственное образовательное учреждение высшего профессионального образования "Алтайский государственный технический университет им. И.И. Ползунова" (АлтГТУ) | Way of reception of biomass in vitro |
| MD3375G2 (en) * | 2007-03-23 | 2008-03-31 | Институт Генетики, Физиологии И Защиты Растений Академии Наук Молдовы | Process for micropropagation in vitro of Rhodiola rosea L. plants |
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2009
- 2009-02-23 MD MDS20090037A patent/MD31Z/en not_active IP Right Cessation
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| Publication number | Publication date |
|---|---|
| MD31Z (en) | 2010-01-31 |
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