MD1091Z - Process for micropropagation of Mentha gattefossei Maire plants in vitro - Google Patents
Process for micropropagation of Mentha gattefossei Maire plants in vitro Download PDFInfo
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- MD1091Z MD1091Z MDS20160032A MDS20160032A MD1091Z MD 1091 Z MD1091 Z MD 1091Z MD S20160032 A MDS20160032 A MD S20160032A MD S20160032 A MDS20160032 A MD S20160032A MD 1091 Z MD1091 Z MD 1091Z
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- 238000000034 method Methods 0.000 title claims abstract description 13
- 230000008569 process Effects 0.000 title claims abstract description 12
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- 238000000338 in vitro Methods 0.000 title claims abstract description 11
- 235000015097 nutrients Nutrition 0.000 claims description 10
- 238000005520 cutting process Methods 0.000 claims description 7
- 229910052500 inorganic mineral Inorganic materials 0.000 claims description 6
- 239000011707 mineral Substances 0.000 claims description 6
- 150000003839 salts Chemical class 0.000 claims description 6
- 230000006698 induction Effects 0.000 claims description 3
- 238000011081 inoculation Methods 0.000 claims description 3
- 238000005286 illumination Methods 0.000 claims description 2
- 241000196324 Embryophyta Species 0.000 description 15
- 235000010755 mineral Nutrition 0.000 description 5
- 235000016254 Mentha gattefossei Nutrition 0.000 description 3
- 241001479543 Mentha x piperita Species 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- 235000004357 Mentha x piperita Nutrition 0.000 description 2
- NWBJYWHLCVSVIJ-UHFFFAOYSA-N N-benzyladenine Chemical compound N=1C=NC=2NC=NC=2C=1NCC1=CC=CC=C1 NWBJYWHLCVSVIJ-UHFFFAOYSA-N 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 239000003630 growth substance Substances 0.000 description 2
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 2
- 238000012423 maintenance Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 2
- 239000000341 volatile oil Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 239000002028 Biomass Substances 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
- 229910021580 Cobalt(II) chloride Inorganic materials 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
- 239000007836 KH2PO4 Substances 0.000 description 1
- 241000207923 Lamiaceae Species 0.000 description 1
- 235000016257 Mentha pulegium Nutrition 0.000 description 1
- 229910004619 Na2MoO4 Inorganic materials 0.000 description 1
- 244000061176 Nicotiana tabacum Species 0.000 description 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
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- 230000003078 antioxidant effect Effects 0.000 description 1
- 238000004166 bioassay Methods 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
- 229910052927 chalcanthite Inorganic materials 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
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- 230000000694 effects Effects 0.000 description 1
- 229910052564 epsomite Inorganic materials 0.000 description 1
- 238000011066 ex-situ storage Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000001050 hortel pimenta 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
- 238000004519 manufacturing process Methods 0.000 description 1
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- 239000001771 mentha piperita Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 235000017807 phytochemicals Nutrition 0.000 description 1
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- 229930000223 plant secondary metabolite Natural products 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
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- 239000011684 sodium molybdate Substances 0.000 description 1
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- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
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- 238000012546 transfer Methods 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
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Invenţia se referă la biotehnologie, şi anume la un procedeu de micropropagare a plantelor de Mentha gattefossei Maire in vitro.Procedeul, conform invenţiei, include inocularea minibutaşilor şi cultivarea lăstarilor obţinuţi din mugurii axilari cu inducerea rizogenezei pe un mediu nutritiv, care conţine ½ săruri minerale faţă de mediul Murashige-Skoog, totodată cultivarea se efectuează la temperatura de 26°C, umiditatea relativă a aerului de 70%, cu o fotoperioadă de 16 ore lumină, intensitatea iluminării de 2000 lx şi pH-ul 5,8, timp de 4…5 săptămâni.The invention relates to biotechnology, namely to a process of micropropagation of Mentha gattefossei Maire plants in vitro. compared to the Murashige-Skoog environment, at the same time the cultivation is carried out at a temperature of 26 ° C, the relative humidity of the air of 70%, with a photoperiod of 16 hours light, the intensity of the lighting of 2000 lx and the pH 5.8, for 4 ... 5 weeks.
Description
Invenţia se referă la biotehnologie, şi anume la un procedeu de micropropagare a plantelor de Mentha gattefossei Maire in vitro. The invention relates to biotechnology, namely to a process for micropropagation of Mentha gattefossei Maire plants in vitro.
Mentha gattefossei Maire este o plantă medicinală rară din familia Lamiaceae, endemică din Maroc, cu un areal de răspândire foarte restrâns, estimat la 2000 km2. Mentha gattefossei Maire is a rare medicinal plant from the Lamiaceae family, endemic to Morocco, with a very limited distribution area, estimated at 2000 km2.
Partea aeriană a plantei sintetizează ulei volatil şi alte clase de compuşi biologic activi (polifenoli, flavonoide) cu efect antioxidant pronunţat. Conform studiului bibliografic de specialitate planta până în prezent rămâne a fi insuficient cercetată din punct de vedere fitochimic. The aerial part of the plant synthesizes volatile oil and other classes of biologically active compounds (polyphenols, flavonoids) with a pronounced antioxidant effect. According to the specialized bibliographic study, the plant remains insufficiently researched from a phytochemical point of view.
Specia este protejată la nivel internaţional, fiind inclusă în Lista Roşie a IUCN (International Union for the Conservation of Nature) la categoria plantelor Near Threatened. Efectivul populaţiilor naturale este în continuă descreştere ca rezultat al colectării intensive în scopuri medicinale şi ca sursă alimentară. Se exploatează, de asemenea, intensiv şi pentru extragerea uleiului volatil la nivel local şi internaţional. The species is internationally protected, being included in the IUCN (International Union for the Conservation of Nature) Red List as Near Threatened. The number of natural populations is constantly decreasing as a result of intensive collection for medicinal purposes and as a food source. It is also intensively exploited for the extraction of volatile oil locally and internationally.
În Republica Moldova se menţine pe terenul experimental al Grădinii Botanice (I) a AŞM, fiind obţinută prin schimbul internaţional de seminţe cu Grădina Botanică din Coimbra, Portugalia. Numărul redus de exemplare nu asigură materialul iniţial de înmulţire pentru o eventuală extindere în cultură pe suprafeţe mai mari, precum şi a materialului vegetal suficient pentru studii chimice ulterioare. Acest fapt implică necesitatea elaborării metodei de multiplicare rapidă în cultura in vitro a plantelor de Mentha gattefossei Maire, fiind în acelaşi timp o măsură eficientă de conservare ex situ a acestui taxon puternic periclitat la nivel mondial. In the Republic of Moldova it is maintained on the experimental field of the Botanical Garden (I) of the Academy of Sciences, having been obtained through the international exchange of seeds with the Botanical Garden of Coimbra, Portugal. The small number of specimens does not provide the initial propagation material for a possible expansion in culture on larger areas, as well as sufficient plant material for subsequent chemical studies. This fact implies the need to develop a method for rapid multiplication in in vitro culture of Mentha gattefossei Maire plants, being at the same time an effective measure for ex situ conservation of this highly endangered taxon worldwide.
Se cunoaşte procedeul de micropropagare a plantelor de Mentha piperita, ce constă în activarea mugurilor axilari, prezenţi la nivelul internodurilor, respectiv la subsuoara frunzelor. Pentru activarea şi creşterea intensivă a mugurilor axilari, explantele au fost cultivate pe mediul nutritiv de bază cu un conţinut de săruri minerale după Murashige-Skoog (Murashige T., Skoog F. A revised medium for rapid growth and bio-assays with tobaco tissue cultures. Physiology Plantarum, 1962, v.15, N95, p.473), suplinit cu 6-benzilaminopurină - 0,6 mg/l, agar - 0,6%; valoarea pH-lui a fost ajustată la 5,8 până la autoclavare [1]. The micropropagation process of Mentha piperita plants is known, which consists in activating the axillary buds, present at the level of the internodes, respectively at the leaf axils. For the activation and intensive growth of the axillary buds, the explants were cultivated on the basic nutrient medium with a content of mineral salts according to Murashige-Skoog (Murashige T., Skoog F. A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiology Plantarum, 1962, v.15, N95, p.473), supplemented with 6-benzylaminopurine - 0.6 mg/l, agar - 0.6%; the pH value was adjusted to 5.8 before autoclaving [1].
Însă acest procedeu are un dezavantaj, ce se manifestă prin perioada dublă de micropropagare. However, this process has a disadvantage, which is manifested by the double micropropagation period.
Dezavantajul menţionat poate fi înlăturat cu ajutorul procedeului revendicat. The aforementioned disadvantage can be eliminated by means of the claimed process.
Problema pe care o rezolvă invenţia constă în reducerea perioadei de micropropagare, sporirea coeficientului de multiplicare a plantelor şi, respectiv, diminuarea cheltuielilor pentru a obţine o plantă nouă de Mentha gattefossei Maire. The problem solved by the invention consists in reducing the micropropagation period, increasing the plant multiplication coefficient and, respectively, reducing the expenses to obtain a new Mentha gattefossei Maire plant.
Procedeul, conform invenţiei, include inocularea minibutaşilor şi cultivarea lăstarilor obţinuţi din mugurii axilari cu inducerea rizogenezei pe un mediu nutritiv, care conţine 1⁄2 săruri minerale faţă de mediul Murashige-Skoog, totodată cultivarea se efectuează la temperatura de 26°C, umiditatea relativă a aerului de 70%, cu o fotoperioadă de 16 ore lumină, intensitatea iluminării de 2000 lx şi pH-ul 5,8, timp de 4…5 săptămâni. The process, according to the invention, includes inoculating mini-cuttings and cultivating shoots obtained from axillary buds with rhizogenesis induction on a nutrient medium, which contains 1⁄2 mineral salts compared to the Murashige-Skoog medium, while cultivation is carried out at a temperature of 26°C, relative air humidity of 70%, with a photoperiod of 16 hours of light, illumination intensity of 2000 lx and pH 5.8, for 4...5 weeks.
Avantajele invenţiei sunt: The advantages of the invention are:
- posibilitatea de a obţine dintr-un explant (minibutaş) în decurs de 4…5 săptămâni, lăstari cu lungimea de 8…9 cm a câte 7…9 internoduri (invenţia), în comparaţie cu lăstari de 2…3 cm a câte 4…5 internoduri dintr-un explant în aceeaşi perioadă de timp (cea mai apropiată soluţie); - the possibility of obtaining from an explant (mini-cutting) within 4...5 weeks, shoots with a length of 8...9 cm and 7...9 internodes (the invention), compared to shoots of 2...3 cm and 4...5 internodes from an explant in the same period of time (the closest solution);
- sporirea coeficientului de micropropagare de la aproximativ 4000 plante în şase luni (cea mai apropiată soluţie) până la 150000 plante (invenţia) în aceeaşi perioadă de timp; - increasing the micropropagation coefficient from approximately 4000 plants in six months (the closest solution) to 150000 plants (the invention) in the same period of time;
- în comparaţie cu cea mai apropiată soluţie, procesul de micropropagare a plantelor de Mentha gattefossei Maire prin procedeul revendicat se caracterizează prin reducerea timpului de menţinere in vitro şi diminuarea numărului de explante necesare pentru multiplicare cu ≈50%. - compared to the closest solution, the process of micropropagation of Mentha gattefossei Maire plants by the claimed process is characterized by reducing the in vitro maintenance time and reducing the number of explants required for multiplication by ≈50%.
Rezultatul invenţiei constă în sporirea coeficientului de multiplicare de la aproximativ 4000 plante în şase luni (cea mai apropiată soluţie) până la 150000 plante (invenţia) de Mentha gattefossei Maire în aceeaşi perioadă de timp, datorită modificării conţinutului mediului nutritiv (1⁄2 săruri minerale după Murashige-Skoog). Aceste modificări induc creşterea lăstarilor cu majorarea numărului de internoduri (ceea ce sporeşte coeficientul potenţial de multiplicare). The result of the invention consists in increasing the multiplication coefficient from approximately 4000 plants in six months (the closest solution) to 150000 plants (the invention) of Mentha gattefossei Maire in the same period of time, due to the modification of the content of the nutrient medium (1⁄2 mineral salts according to Murashige-Skoog). These modifications induce the growth of shoots with an increase in the number of internodes (which increases the potential multiplication coefficient).
Exemplu de realizare concretă a invenţiei Example of concrete embodiment of the invention
Plantele de Mentha gattefossei Maire au fost obţinute din lăstarii plantelor cultivate din seminţe pe terenul experimental al Grădinii Botanice (I) a AŞM. Pentru a asigura sterilizarea, lăstarii de Mentha gattefossei Maire au fost trataţi cu preparatul comercial ACE (agent de înălbire pe bază de clor) de 20% pe parcursul a 10 minute şi spălaţi de trei ori cu apă sterilă. Inducerea lăstarilor şi creşterea plantulelor a fost realizată pe mediul nutritiv Murashige-Skoog (MS) cu pH-ul 5,8, la regimul de temperatură de 26°C, cu fotoperioada de 16 ore lumină (2000 lx) şi 8 ore întuneric. Mentha gattefossei Maire plants were obtained from the shoots of plants grown from seeds in the experimental field of the Botanical Garden (I) of the ASM. To ensure sterilization, the shoots of Mentha gattefossei Maire were treated with the commercial preparation ACE (chlorine bleach) 20% for 10 minutes and washed three times with sterile water. Shoot induction and seedling growth were performed on Murashige-Skoog (MS) nutrient medium with pH 5.8, at a temperature regime of 26°C, with a photoperiod of 16 hours of light (2000 lx) and 8 hours of darkness.
O parte din minibutaşi (controlul) a fost cultivată pe mediul nutritiv Murashige-Skoog, cu următorul conţinut de săruri minerale (mg/l): NH4NO3 - 1650, KNO3 - 1900, MgSO4·7H2O -370, KH2PO4 -170, CaCl2·2H2O - 440, H3BO3 - 6,2, MnSO4·4H2O - 22,3, CoCl2·6H2O-0,025, CuSO4 ·5H2O - 0,025, ZnSO4·7 H2O - 8,6, Na2MoO4·2H2O - 0,25, KI - 0,83, FeSO4·7H2O - 27,8, Na2EDTA - 37,3; agar - 0,6%. Altă parte a fost cultivată pe acelaşi mediu, dar suplimentat cu regulatorul de creştere 6-benzilaminopurină, valoarea pH-lui fiind ajustată înainte de autoclavare până la 5,8 (cea mai apropiată soluţie). A treia parte de minibutaşi a fost cultivată pe mediul nutritiv Murashige-Skoog ce conţinea 1⁄2 săruri minerale indicate în control, pH-ul fiind 5,8, fără fitohormoni (invenţia). În aşa fel mediile de cultivare se deosebeau după concentraţia şi conţinutul unor componenţi. Minibutaşii celor trei variante au fost cultivaţi la temperatura 26°C, cu fotoperioada de 16 ore lumină (2000 lx) şi 8 ore întuneric. Umiditatea relativă a aerului a fost de 70%. Part of the mini-cuttings (control) was cultivated on Murashige-Skoog nutrient medium, with the following content of mineral salts (mg/l): NH4NO3 - 1650, KNO3 - 1900, MgSO4·7H2O -370, KH2PO4 -170, CaCl2·2H2O - 440, H3BO3 - 6.2, MnSO4·4H2O - 22.3, CoCl2·6H2O-0.025, CuSO4·5H2O - 0.025, ZnSO4·7 H2O - 8.6, Na2MoO4·2H2O - 0.25, KI - 0.83, FeSO4·7H2O - 27.8, Na2EDTA - 37.3; agar - 0.6%. Another part was cultivated on the same medium, but supplemented with the growth regulator 6-benzylaminopurine, the pH value being adjusted before autoclaving to 5.8 (the closest solution). The third part of the mini-cuttings was cultivated on the Murashige-Skoog nutrient medium containing 1⁄2 of the mineral salts indicated in the control, the pH being 5.8, without phytohormones (the invention). In this way, the cultivation media differed in the concentration and content of some components. The mini-cuttings of the three variants were cultivated at a temperature of 26°C, with a photoperiod of 16 hours of light (2000 lx) and 8 hours of darkness. The relative humidity of the air was 70%.
Creşterea şi dezvoltarea lăstarilor de la baza mugurilor axilari şi a rădăcinilor, pe mediul nutritiv conform invenţiei, a fost observată după 3 zile de la momentul inoculării, iar după 4…5 săptămâni lăstarii au atins înălţimea de 7…9 cm. După 4…5 săptămâni de la momentul inoculării, fiecare plantă a fost secţionată în internoduri, care ulterior au fost inoculaţi pentru următorul ciclu de dezvoltare pe acelaşi mediu nutritiv. În aşa fel, pe acelaşi mediu nutritiv este posibilă iniţierea culturii in vitro şi cultivarea minibutaşilor pentru a obţine plantule, care sunt pregătite pentru cultivarea în condiţii ex vitro. În cazul celei mai apropiate soluţii, lăstarii cultivaţi după 4…5 săptămâni au atins înălţimea de numai 2,5…3 cm. The growth and development of shoots from the base of the axillary buds and roots, on the nutrient medium according to the invention, was observed after 3 days from the moment of inoculation, and after 4…5 weeks the shoots reached a height of 7…9 cm. After 4…5 weeks from the moment of inoculation, each plant was sectioned into internodes, which were subsequently inoculated for the next development cycle on the same nutrient medium. In this way, on the same nutrient medium it is possible to initiate in vitro culture and cultivate mini-cuttings to obtain seedlings, which are ready for cultivation under ex vitro conditions. In the case of the closest solution, the shoots cultivated after 4…5 weeks reached a height of only 2.5…3 cm.
Spre deosebire de cea mai apropiată soluţie, procedeul revendicat contribuie la micşorarea de două ori a cheltuielilor pentru reagenţii utilizaţi la cultivarea in vitro; diminuarea duratei de cultivare in vitro pentru obţinerea unei noi generaţii de plantule în decurs de 4…5 săptămâni; sporirea coeficientului de micropropagare de la 4 la 8. Pe parcursul a 3 cicluri s-au obţinut aproximativ 500 de plante (invenţia), faţă de cele 60 plante (cea mai apropiată soluţie), luând în consideraţie faptul că în conformitate cu invenţia propusă se obţin până la unsprezece cicluri pe parcursul unui an, faţă de cele cinci cicluri (cea mai apropiată soluţie), pe parcursul a jumătate de an se pot obţine cel puţin 150000 plante. Unlike the closest solution, the claimed process contributes to a twofold reduction in the costs of reagents used for in vitro cultivation; reducing the duration of in vitro cultivation to obtain a new generation of seedlings within 4...5 weeks; increasing the micropropagation coefficient from 4 to 8. During 3 cycles, approximately 500 plants were obtained (the invention), compared to 60 plants (the closest solution), taking into account the fact that according to the proposed invention, up to eleven cycles are obtained during a year, compared to five cycles (the closest solution), during half a year at least 150,000 plants can be obtained.
Prin urmare, procedeul revendicat permite de a diminua de două ori cheltuielile pentru reagenţi, durata menţinerii in vitro pentru a obţine o nouă generaţie pregătită pentru transferul ex vitro; de a spori de 35 ori numărul de plantule multiplicate pe parcursul unui an, utilizând aceeaşi suprafaţă a camerei de cultivare. Therefore, the claimed process allows to reduce by two the expenses for reagents, the duration of in vitro maintenance to obtain a new generation ready for ex vitro transfer; to increase by 35 times the number of seedlings multiplied during a year, using the same area of the cultivation chamber.
1. Santoro M.V., Nievas F., Zygadlo J., Giordano W., Banchio E. Effects of Growth Regulators on Biomass and the Production of Secondary Metabolites in Peppermint (Mentha piperita) Micropropagated in vitro. American Journal of Plant Sciences, 2013, 4, 49-55 <http://dx.doi.org/10.4236.2013.45A008> Mai, 2013 1. Santoro M.V., Nievas F., Zygadlo J., Giordano W., Banchio E. Effects of Growth Regulators on Biomass and the Production of Secondary Metabolites in Peppermint (Mentha piperita) Micropropagated in vitro. American Journal of Plant Sciences, 2013, 4, 49-55 <http://dx.doi.org/10.4236.2013.45A008> May, 2013
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| MD605Y (en) * | 2012-07-09 | 2013-03-31 | Inst De Genetica Si Fiziol A Plantelor Al Academiei De Stiinte A Moldovei | Process for microclonal propagation of Actinidia arguta plants in vitro |
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| MD605Y (en) * | 2012-07-09 | 2013-03-31 | Inst De Genetica Si Fiziol A Plantelor Al Academiei De Stiinte A Moldovei | Process for microclonal propagation of Actinidia arguta plants in vitro |
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| Title |
|---|
| Santoro M.V., Nievas F., Zygadlo J., Giordano W., Banchio E. Effects of Growth Regulators on Biomass and the Production of Secondary Metabolites in Peppermint (Mentha piperita) Micropropagated in vitro. American Journal of Plant Sciences, 2013, 4, 49-55 <http://dx.doi.org/10.4236.2013.45A008> Mai, 2013 * |
Also Published As
| Publication number | Publication date |
|---|---|
| MD1091Y (en) | 2016-11-30 |
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