MD1059Z - Process for treating melon seeds - Google Patents
Process for treating melon seeds Download PDFInfo
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- MD1059Z MD1059Z MDS20160029A MDS20160029A MD1059Z MD 1059 Z MD1059 Z MD 1059Z MD S20160029 A MDS20160029 A MD S20160029A MD S20160029 A MDS20160029 A MD S20160029A MD 1059 Z MD1059 Z MD 1059Z
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- pvp
- fe3o4
- nanoparticles
- seeds
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- 238000000034 method Methods 0.000 title claims abstract description 18
- 241000353135 Psenopsis anomala Species 0.000 title claims abstract description 4
- SZVJSHCCFOBDDC-UHFFFAOYSA-N ferrosoferric oxide Chemical compound O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 claims abstract description 88
- 229920000191 poly(N-vinyl pyrrolidone) Polymers 0.000 claims abstract description 69
- WHNWPMSKXPGLAX-UHFFFAOYSA-N N-Vinyl-2-pyrrolidone Chemical compound C=CN1CCCC1=O WHNWPMSKXPGLAX-UHFFFAOYSA-N 0.000 claims abstract description 61
- 239000002069 magnetite nanoparticle Substances 0.000 claims abstract description 11
- 238000002791 soaking Methods 0.000 claims abstract description 3
- 238000009331 sowing Methods 0.000 claims abstract description 3
- 239000002105 nanoparticle Substances 0.000 description 34
- 239000000243 solution Substances 0.000 description 34
- 230000035784 germination Effects 0.000 description 20
- 241000196324 Embryophyta Species 0.000 description 12
- 230000000877 morphologic effect Effects 0.000 description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 12
- 244000241257 Cucumis melo Species 0.000 description 11
- 230000015572 biosynthetic process Effects 0.000 description 11
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 11
- 238000003786 synthesis reaction Methods 0.000 description 10
- UQSXHKLRYXJYBZ-UHFFFAOYSA-N Iron oxide Chemical compound [Fe]=O UQSXHKLRYXJYBZ-UHFFFAOYSA-N 0.000 description 8
- 244000052616 bacterial pathogen Species 0.000 description 8
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 7
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 7
- 239000000203 mixture Substances 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 6
- 239000007864 aqueous solution Substances 0.000 description 6
- 229910001868 water Inorganic materials 0.000 description 6
- 235000009842 Cucumis melo Nutrition 0.000 description 5
- 244000098338 Triticum aestivum Species 0.000 description 5
- 239000012153 distilled water Substances 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 4
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 4
- 235000011114 ammonium hydroxide Nutrition 0.000 description 4
- 239000012300 argon atmosphere Substances 0.000 description 4
- 238000000975 co-precipitation Methods 0.000 description 4
- 239000013068 control sample Substances 0.000 description 4
- 235000015510 Cucumis melo subsp melo Nutrition 0.000 description 3
- 240000008042 Zea mays Species 0.000 description 3
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 description 3
- 239000000908 ammonium hydroxide Substances 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- 230000007226 seed germination Effects 0.000 description 3
- 239000002689 soil Substances 0.000 description 3
- 239000003381 stabilizer Substances 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 239000011550 stock solution Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical group N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 238000001157 Fourier transform infrared spectrum Methods 0.000 description 2
- 235000021307 Triticum Nutrition 0.000 description 2
- 238000002441 X-ray diffraction Methods 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- 239000011554 ferrofluid Substances 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 230000002363 herbicidal effect Effects 0.000 description 2
- 239000004009 herbicide Substances 0.000 description 2
- 239000002122 magnetic nanoparticle Substances 0.000 description 2
- 238000000386 microscopy Methods 0.000 description 2
- 239000011858 nanopowder Substances 0.000 description 2
- 230000035515 penetration Effects 0.000 description 2
- 239000000575 pesticide Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonium chloride Substances [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 1
- 235000000832 Ayote Nutrition 0.000 description 1
- 235000002566 Capsicum Nutrition 0.000 description 1
- 235000008534 Capsicum annuum var annuum Nutrition 0.000 description 1
- 240000008384 Capsicum annuum var. annuum Species 0.000 description 1
- 240000007092 Cucurbita argyrosperma Species 0.000 description 1
- 235000003949 Cucurbita mixta Nutrition 0.000 description 1
- 244000020551 Helianthus annuus Species 0.000 description 1
- 235000003222 Helianthus annuus Nutrition 0.000 description 1
- 238000004566 IR spectroscopy Methods 0.000 description 1
- 240000004296 Lolium perenne Species 0.000 description 1
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 1
- 239000006002 Pepper Substances 0.000 description 1
- 235000016761 Piper aduncum Nutrition 0.000 description 1
- 235000017804 Piper guineense Nutrition 0.000 description 1
- 244000203593 Piper nigrum Species 0.000 description 1
- 235000008184 Piper nigrum Nutrition 0.000 description 1
- 240000003768 Solanum lycopersicum Species 0.000 description 1
- 235000007244 Zea mays Nutrition 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 229920000249 biocompatible polymer Polymers 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000009614 chemical analysis method Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000005294 ferromagnetic effect Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000003324 growth hormone secretagogue Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000003607 modifier Substances 0.000 description 1
- 230000003562 morphometric effect Effects 0.000 description 1
- 238000013425 morphometry Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 239000001301 oxygen Chemical group 0.000 description 1
- 229910052760 oxygen Chemical group 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- RWPGFSMJFRPDDP-UHFFFAOYSA-L potassium metabisulfite Chemical compound [K+].[K+].[O-]S(=O)S([O-])(=O)=O RWPGFSMJFRPDDP-UHFFFAOYSA-L 0.000 description 1
- 229940043349 potassium metabisulfite Drugs 0.000 description 1
- 235000010263 potassium metabisulphite Nutrition 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000001878 scanning electron micrograph Methods 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000000021 stimulant Substances 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- ZSDSQXJSNMTJDA-UHFFFAOYSA-N trifluralin Chemical compound CCCN(CCC)C1=C([N+]([O-])=O)C=C(C(F)(F)F)C=C1[N+]([O-])=O ZSDSQXJSNMTJDA-UHFFFAOYSA-N 0.000 description 1
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- Pretreatment Of Seeds And Plants (AREA)
Abstract
Description
Invenţia se referă la utilizarea nanotehnologiilor în agricultură, în particular la tratarea seminţelor şi creşterea răsadurilor. The invention relates to the use of nanotechnologies in agriculture, in particular to seed treatment and seedling growth.
Este cunoscut un procedeu de tratare cu lichid feromagnetic a culturii Zea mays (porumb) într-un mediu cultural. Concentraţia nanoparticulelor magnetice este de 4,4%, iar dimensiunea medie a acestora este de 10 nm. La utilizarea acestei concentraţii de ferofluid se observă un efect de stimulare [1]. A ferromagnetic fluid treatment process is known for Zea mays (corn) culture in a culture medium. The concentration of magnetic nanoparticles is 4.4%, and their average size is 10 nm. When using this concentration of ferrofluid, a stimulating effect is observed [1].
Dezavantajul acestui procedeu constă în utilizarea concentraţiilor înalte de ferofluid, care este scump, de asemenea, pe parcursul sintezei nu se utilizează un agent stabilizator, ceea ce împiedică pătrunderea substanţelor active prin peretele celular al seminţelor. The disadvantage of this process is the use of high concentrations of ferrofluid, which is expensive, and also, during the synthesis, a stabilizing agent is not used, which prevents the penetration of active substances through the cell wall of the seeds.
Este cunoscută compoziţia pentru reţinerea umidităţii în sol, care contribuie la îmbunătăţirea creşterii plantelor în sol uscat. Compoziţia lichidului cultural conţine un amestec din particule de oxid de fier cu dimensiunile de 50 µm şi metabisulfit de potasiu. Compoziţia dată nu inhibă germinarea seminţelor de ardei, roşii, floarea-soarelui şi grâu, iar germinarea seminţelor de porumb şi de ardei verde a fost parţial stimulată. În comparaţie cu plantele de control, probele tratate cu soluţia de oxid de fier au dezvoltat un sistem radicular puternic [2]. The composition for retaining moisture in the soil, which contributes to improving plant growth in dry soil, is known. The composition of the culture liquid contains a mixture of iron oxide particles with a size of 50 µm and potassium metabisulfite. This composition does not inhibit the germination of pepper, tomato, sunflower and wheat seeds, and the germination of corn and green pepper seeds was partially stimulated. Compared with the control plants, the samples treated with the iron oxide solution developed a strong root system [2].
Dezavantajul acestei compoziţii constă în utilizarea oxidului de fier de dimensiuni micronice, ceea ce cauzează obţinerea unor parametri morfologici insuficienţi. The disadvantage of this composition is the use of micron-sized iron oxide, which causes insufficient morphological parameters to be obtained.
Este cunoscut procedeul de tratare a seminţelor de Lolium perenne (secară) şi Cucurbita mixta cu nanoparticule de Fe3O4, acoperite cu un înveliş de poli-N-vinilpirolidonă (PVP). Seminţele plantelor au fost tratate cu suspensie de Fe3O4 /PVP şi lăsate pentru germinare în amestecul cultural cu concentraţia de 30 şi 100 mg/L. Lungimea rădăcinilor şi lungimea germenilor de plante nu au variat semnificativ faţă de plantele de control [3]. The process of treating Lolium perenne (rye) and Cucurbita mixta seeds with Fe3O4 nanoparticles, coated with a poly-N-vinylpyrrolidone (PVP) coating is known. The seeds of the plants were treated with Fe3O4 /PVP suspension and left for germination in the cultural mixture with a concentration of 30 and 100 mg/L. The length of the roots and the length of the plant sprouts did not vary significantly compared to the control plants [3].
Dezavantajul acestui procedeu constă în utilizarea PVP în concentraţie mare, ceea ce aparent blochează pătrunderea nanoparticulelor în celulele vegetale ale seminţei. The disadvantage of this process is the use of high concentration of PVP, which apparently blocks the penetration of nanoparticles into the plant cells of the seed.
Este cunoscut procedeul de tratare a seminţelor de grâu de primăvară Triticum aestivum cu diferite forme de fier. La tratarea seminţelor cu soluţia apoasă coloidală de nanoparticule de magnetit, stabilizate cu poli-N-vinilpirolidonă în concentraţie de 0,125 g/L, germinarea seminţelor a atins randamentul de 95%. Nanoparticulele de magnetit au stimulat dezvoltarea sistemului radicular şi a lăstarilor de Triticum aestivum, fiind observată o creştere maximă a valorilor parametrilor morfometrici în cazul utilizării nanoparticulelor de magnetit în concentraţie de 0,25 g/L (creşterea lungimii lăstarilor cu 23,3%) şi de 0,125 g/L (creşterea lungimii rădăcinilor cu 68,6% şi a numărului de rădăcini cu 9,5%) [4]. The process of treating spring wheat seeds Triticum aestivum with different forms of iron is known. When treating seeds with colloidal aqueous solution of magnetite nanoparticles, stabilized with poly-N-vinylpyrrolidone in concentration of 0.125 g/L, seed germination reached a yield of 95%. Magnetite nanoparticles stimulated the development of the root system and shoots of Triticum aestivum, a maximum increase in the values of morphometric parameters was observed in the case of using magnetite nanoparticles in concentration of 0.25 g/L (increase in shoot length by 23.3%) and 0.125 g/L (increase in root length by 68.6% and in the number of roots by 9.5%) [4].
Dezavantajul acestui procedeu constă în concentraţia mare a soluţiei de magnetit. The disadvantage of this process is the high concentration of the magnetite solution.
Cea mai apropiată soluţie este stimulatorul de creştere şi destructorul de pesticide în formă de nanoparticule de Fe3O4 (magnetit) învelite cu PVP, care se utilizează în calitate de stabilizator, cu dimensiunile nanoparticulelor de 55…200 nm, în funcţie de tehnologia aplicată. Efectul maxim morfologic este atins la utilizarea soluţiei de Fe3O4/PVP în concentraţie de 50 mg/L, lungimea rădăcinii plantei crescând cu cca 40%, iar lungimea hipocotililor - cu 180% în raport cu probele de control. S-a demonstrat şi eficienţa acestui preparat la descompunerea erbicidului trifluralin - un erbicid din grupa III de toxicitate [5]. The closest solution is the growth stimulator and pesticide destroyer in the form of Fe3O4 (magnetite) nanoparticles coated with PVP, which are used as a stabilizer, with nanoparticle sizes of 55…200 nm, depending on the applied technology. The maximum morphological effect is achieved when using the Fe3O4/PVP solution in a concentration of 50 mg/L, the length of the plant root increasing by about 40%, and the length of the hypocotyls - by 180% compared to the control samples. The effectiveness of this preparation in the decomposition of the herbicide trifluralin - a herbicide of toxicity group III [5].
Dezavantajul acestei soluţii constă în diametrul mare al nanoparticulelor, care împiedică pătrunderea acestora prin membrana celulară a seminţelor. The disadvantage of this solution lies in the large diameter of the nanoparticles, which prevents them from penetrating the seed cell membrane.
Problema pe care o rezolvă invenţia constă în mărirea permeabilităţii celulare a seminţelor culturii Cucumis melo (pepene galben), îmbunătăţirea parametrilor morfologici ai răsadurilor şi sporirea energiei germinative şi a germinaţiei seminţelor. The problem solved by the invention consists in increasing the cellular permeability of the seeds of the Cucumis melo crop (melon), improving the morphological parameters of the seedlings and increasing the germinative energy and germination of the seeds.
Procedeul, conform invenţiei, înlătură dezavantajul menţionat mai sus prin aceea că include înmuierea seminţelor înainte de semănat timp de 2…3 ore într-o soluţie apoasă coloidală de nanoparticule de magnetit stabilizate cu poli-N-vinilpirolidonă Fe3O4/PVP în concentraţie de 20…100 mg/L şi cu dimensiunile de 10…20 nm, concentraţia poli-N-vinilpirolidonei utilizate fiind de 0,005…0,01 mol/L. The process, according to the invention, eliminates the above-mentioned disadvantage by including soaking the seeds before sowing for 2…3 hours in an aqueous colloidal solution of magnetite nanoparticles stabilized with poly-N-vinylpyrrolidone Fe3O4/PVP in a concentration of 20…100 mg/L and with dimensions of 10…20 nm, the concentration of poly-N-vinylpyrrolidone used being 0.005…0.01 mol/L.
Rezultatul tehnic al invenţiei constă în îmbunătăţirea parametrilor morfologici ai răsadurilor şi sporirea energiei germinative şi a germinaţiei seminţelor. Efectul maximal morfologic a fost demonstrat prin creşterea lungimii plantelor cu 43...50% şi a lungimii hipocotilului cu 179…267%. The technical result of the invention consists in improving the morphological parameters of seedlings and increasing the germination energy and seed germination. The maximum morphological effect was demonstrated by increasing the length of plants by 43...50% and the length of the hypocotyl by 179...267%.
Invenţia se explică prin desenele din fig. 1-5, care reprezintă: The invention is explained by the drawings in Fig. 1-5, which represent:
- fig. 1, imaginile SEM ale nanoparticulelor de Fe3O4/PVP; - Fig. 1, SEM images of Fe3O4/PVP nanoparticles;
- fig. 2, difractograma XRD a nanoparticulelor de Fe3O4/PVP; - fig. 2, XRD diffractogram of Fe3O4/PVP nanoparticles;
- fig. 3, spectrul FTIR al PVP; - Fig. 3, FTIR spectrum of PVP;
- fig. 4, spectrul FTIR al nanoparticulelor de Fe3O4/PVP; - Fig. 4, FTIR spectrum of Fe3O4/PVP nanoparticles;
- fig. 5, mostrele culturii Cucumis melo, prelucrate cu soluţie de nanoparticule de Fe3O4/PVP (S.12) la a treia (а) şi a şaptea zi de germinare (b), în care: - Fig. 5, samples of Cucumis melo culture, processed with Fe3O4/PVP nanoparticle solution (S.12) on the third (а) and seventh day of germination (b), in which:
1 - mostra de control tratată cu H2O; 1 - control sample treated with H2O;
2 - mostra tratată cu soluţie apoasă coloidală de nanoparticule de Fe3O4/PVP în concentraţie de 20 mg/L; 2 - sample treated with colloidal aqueous solution of Fe3O4/PVP nanoparticles at a concentration of 20 mg/L;
3 - mostra tratată cu soluţie apoasă coloidală de nanoparticule de Fe3O4/PVP în concentraţie de 50 mg/L; 3 - sample treated with colloidal aqueous solution of Fe3O4/PVP nanoparticles at a concentration of 50 mg/L;
4 - mostra tratată cu soluţie apoasă coloidală de nanoparticule de Fe3O4/PVP în concentraţie de 100 mg/L. 4 - sample treated with colloidal aqueous solution of Fe3O4/PVP nanoparticles at a concentration of 100 mg/L.
Soluţia apoasă coloidală de nanoparticule de magnetit, stabilizate cu polimerul biocompatibil poli-N-vinilpirolidonă Fe3O4/PVP, introdus în timpul sintezei, se utilizează în calitate de stimulator. Culoarea soluţiei devine cafenie în timpul sintezei. The aqueous colloidal solution of magnetite nanoparticles, stabilized with the biocompatible polymer poly-N-vinylpyrrolidone Fe3O4/PVP, introduced during the synthesis, is used as a stimulator. The color of the solution becomes brown during the synthesis.
Procedeul de producere a soluţiei apoase coloidale de nanoaparticule de Fe3O4/PVP se realizează în modul următor. The process for producing the aqueous colloidal solution of Fe3O4/PVP nanoparticles is carried out in the following manner.
Procedeul de sintetizare. The synthesis process.
Nanoparticulele de Fe3O4/PVP au fost obţinute prin metoda coprecipitării, folosind soluţiile apoase ale sulfatului de fier (II), clorurii de fier (III), în prezenţa PVP în raport de Fe2+: Fe3+: PVP=1:2:0,1. Raportul PVP către componentele iniţiale a variat în intervalul de 1…3. Amestecul a fost supus hidrolizei în prezenţa soluţiei de amoniac, în atmosferă de argon, în decurs de 5 ore. Variind cantitatea polimerului şi masa moleculară a acestuia de la 8 000 până la 40 000, s-a sedimentat faza solidă în formă de nanopulbere. Astfel, s-a obţinut soluţia coloidală a oxidului de fier stabilizat cu PVP. Fe3O4/PVP nanoparticles were obtained by the coprecipitation method, using aqueous solutions of iron (II) sulfate, iron (III) chloride, in the presence of PVP in the ratio of Fe2+: Fe3+: PVP=1:2:0.1. The ratio of PVP to the initial components varied in the range of 1…3. The mixture was subjected to hydrolysis in the presence of ammonia solution, in an argon atmosphere, within 5 hours. Varying the amount of polymer and its molecular weight from 8,000 to 40,000, the solid phase was sedimented in the form of nanopowder. Thus, the colloidal solution of iron oxide stabilized with PVP was obtained.
Soluţia coloidală se obţine prin omogenizare în baie cu ultrasunet a suspensiei de nanopulbere de magnetit, în concentraţie de 50 mg/L şi apă distilată. Apoi prin diluare s-au obţinut soluţiile cu concentraţiile necesare. În acest mod, utilizarea PVP în calitate de modificator (stabilizator) al suprafeţei permite formarea nanoparticulelor solubile în apă, cu obţinerea unor coloizi stabili. The colloidal solution is obtained by homogenizing in an ultrasonic bath the suspension of magnetite nanopowder, in a concentration of 50 mg/L and distilled water. Then, by dilution, the solutions with the required concentrations were obtained. In this way, the use of PVP as a surface modifier (stabilizer) allows the formation of water-soluble nanoparticles, obtaining stable colloids.
a) Sinteza nanoparticulelor de Fe3O4/PVP cu dimensiunile de 10 nm. a) Synthesis of Fe3O4/PVP nanoparticles with dimensions of 10 nm.
Nanoparticulele de Fe3O4/PVP au fost pregătite conform metodei coprecipitării chimice, folosind sulfatul de fier (II) şi clorura de fier (III) în prezenţa PVP. În acest scop, 0,560 g de sulfat de fier (II) au fost dizolvate în 35 mL de apă distilată, 3 mL de soluţie de PVP cu concentraţia molară 0,01 mol/L şi 3 mL de soluţie saturată de clorură de fier (III) au fost adăugaţi picătură cu picătură în soluţia stoc. Peste 30 min de la începutul reacţiei au fost adăugaţi 15 mL de hidroxid de amoniu. Sinteza a durat 1,5 ore, la temperatura de 55°C, în atmosferă de argon, la agitare. Pulberea neagră de magnetit a fost separată de soluţie prin spălare cu etanol şi uscată la 100°C. Fe3O4/PVP nanoparticles were prepared according to the chemical coprecipitation method, using iron (II) sulfate and iron (III) chloride in the presence of PVP. For this purpose, 0.560 g of iron (II) sulfate was dissolved in 35 mL of distilled water, 3 mL of PVP solution with a molar concentration of 0.01 mol/L and 3 mL of saturated iron (III) chloride solution were added dropwise to the stock solution. After 30 min from the start of the reaction, 15 mL of ammonium hydroxide was added. The synthesis lasted 1.5 hours, at a temperature of 55°C, in an argon atmosphere, with stirring. The black magnetite powder was separated from the solution by washing with ethanol and dried at 100°C.
A fost determinat, %: Fe 71,73; O 28,05; C 0,19; H 0,024. It was determined, %: Fe 71.73; O 28.05; C 0.19; H 0.024.
A fost calculat, %: Fe 69,04; O 30,64; C 0,21; H 0,11. It was calculated, %: Fe 69.04; O 30.64; C 0.21; H 0.11.
b) Sinteza nanoparticulelor de Fe3O4/PVP cu dimensiunile de 20 nm. b) Synthesis of Fe3O4/PVP nanoparticles with dimensions of 20 nm.
Nanoparticulele de Fe3O4 au fost pregătite conform metodei coprecipitării chimice, folosind sulfat de fier (II) şi clorură de fier (III) în prezenţa PVP. În acest scop, 0,560 g de sulfat de fier (II) au fost dizolvate în 35 mL de apă distilată, 3 mL de soluţie de PVP cu concentraţia molară 0,01 mol/L şi 3 mL de soluţie saturată de clorură de fier (III) au fost adăugaţi picătură cu picătură în soluţia stoc. Peste 30 min de la începutul reacţiei au fost adăugaţi 15 mL de hidroxid de amoniu. Sinteza a durat 5 ore, la temperatura de 75°C, în atmosferă de argon, la agitare. Pulberea neagră de magnetit a fost separată de soluţie prin spălare cu etanol şi uscată la 100°C. Fe3O4 nanoparticles were prepared according to the chemical coprecipitation method, using iron (II) sulfate and iron (III) chloride in the presence of PVP. For this purpose, 0.560 g of iron (II) sulfate was dissolved in 35 mL of distilled water, 3 mL of PVP solution with a molar concentration of 0.01 mol/L and 3 mL of saturated iron (III) chloride solution were added dropwise to the stock solution. After 30 min from the start of the reaction, 15 mL of ammonium hydroxide was added. The synthesis lasted 5 hours, at a temperature of 75°C, in an argon atmosphere, with stirring. The black magnetite powder was separated from the solution by washing with ethanol and dried at 100°C.
A fost determinat, %: Fe 71,73; O 28,05; C 0,19; H 0,024. It was determined, %: Fe 71.73; O 28.05; C 0.19; H 0.024.
A fost calculat, %: Fe 70,01; O 29,15; C 0,16; H 0,68. It was calculated, %: Fe 70.01; O 29.15; C 0.16; H 0.68.
c) Sinteza nanoparticulelor de Fe3O4/PVP cu diferite concentraţii de PVP. c) Synthesis of Fe3O4/PVP nanoparticles with different PVP concentrations.
Nanoparticulele de Fe3O4 au fost pregătite conform metodei coprecipitării chimice, folosind sulfatul de fier (II) şi clorura de fier (III) în prezenţa PVP. În acest scop, 0,560 g de sulfat de fier (II) au fost dizolvate în 35 mL de apă distilată, 3 mL de soluţie de PVP cu concentraţia molară de 0,01…0,005 mol/L şi 3 mL de soluţie saturată de clorură de fier (III) au fost adăugaţi picătură cu picătură în soluţia stoc. Peste 30 min de la începutul reacţiei au fost adăugaţi 15 mL de hidroxid de amoniu. Sinteza a durat 1,5 ore, la temperatura de 55°C, în atmosferă de argon, la agitare. Pulberea neagră de magnetit a fost separată de soluţie prin spălare cu etanol şi uscată la 100°C. Fe3O4 nanoparticles were prepared according to the chemical coprecipitation method, using iron (II) sulfate and iron (III) chloride in the presence of PVP. For this purpose, 0.560 g of iron (II) sulfate was dissolved in 35 mL of distilled water, 3 mL of PVP solution with a molar concentration of 0.01…0.005 mol/L and 3 mL of saturated iron (III) chloride solution were added dropwise to the stock solution. 30 min after the start of the reaction, 15 mL of ammonium hydroxide was added. The synthesis lasted 1.5 hours, at a temperature of 55°C, in an argon atmosphere, with stirring. The black magnetite powder was separated from the solution by washing with ethanol and dried at 100°C.
d) Obţinerea soluţiilor apoase coloidale de nanoparticule de magnetit cu concentraţii diferite. d) Obtaining colloidal aqueous solutions of magnetite nanoparticles with different concentrations.
Soluţiile coloidale ale nanoparticulelor de Fe3O4/PVP au fost obţinute din pulbere neagră sintetizată conform metodei (a). Pulberea a fost dizolvată în apă distilată obţinând soluţii cu concentraţii de 20, 50 şi 100 mg/L. Colloidal solutions of Fe3O4/PVP nanoparticles were obtained from black powder synthesized according to method (a). The powder was dissolved in distilled water obtaining solutions with concentrations of 20, 50 and 100 mg/L.
Identificarea produselor obţinute a fost efectuată prin metodele analizei chimice, SEM-microscopie (fig. 1), XRD (fig. 2), IR-spectroscopie (fig. 3 şi 4). The identification of the obtained products was carried out by chemical analysis methods, SEM-microscopy (fig. 1), XRD (fig. 2), IR-spectroscopy (fig. 3 and 4).
Din rezultatele SEM-microscopiei este evident că nanoparticulele sintetizate sunt aglomerate în entităţi mari cu dimensiunile de 25…35 nm. Dimensiunile cristalitelor nanoparticulelor de magnetit a fost calculată în corespundere cu lungimea de undă a difracţiei razelor X, fiind de d=(15±1) nm. Din comparaţia spectrelor nanoparticulelor de Fe3O4/PVP şi PVP rezultă că are loc coordonarea PVP la nanoparticulele de magnetit prin legăturile de azot şi oxigen din PVP. From the SEM-microscopy results it is evident that the synthesized nanoparticles are agglomerated in large entities with dimensions of 25…35 nm. The crystallite dimensions of magnetite nanoparticles were calculated in accordance with the wavelength of X-ray diffraction, being d=(15±1) nm. From the comparison of the spectra of Fe3O4/PVP and PVP nanoparticles it results that the coordination of PVP to magnetite nanoparticles occurs through the nitrogen and oxygen bonds in PVP.
Experimentul cu seminţele culturii Cucumis melo (pepene galben) a fost efectuat în felul următor. The experiment with Cucumis melo (melon) seeds was carried out as follows.
Seminţele de pepene galben au fost germinate în ceşti Petri, pe substrat de hârtie de filtru timp de 7 zile, în corespundere cu GOST 12038-84 Seminţele culturilor agricole, metode de determinare a nivelului de germinare. Germinarea şi energia de germinare (GOST 12038-66) este procentul seminţelor care au germinat normal, preluate pentru analiză. Au fost numărate 4 loturi de seminţe, a câte 50 de seminţe în lot. Seminţele au fost plasate în ceştile Petri pe substratul din hârtie de filtru umezită. În fiecare ceaşcă s-au turnat câte 2 mL de soluţie apoasă coloidală de nanoparticule de Fe3O4/PVP. În seria ceştilor de control s-au adăugat suplimentar 2 mL de apă. După adăugarea soluţiei în fiecare dintre ceştile Petri, acestea au fost acoperite cu hârtie de filtru umezită şi cu capac. Ceştile Petri au fost plasate într-un termostat cu temperatura constantă de 25°C. Germinarea a avut loc la întuneric. Zilnic era verificată temperatura şi umiditatea, menţinându-le constante ca în starea iniţială, suplimentar se adăuga apă deionizată. Seminţele care au germinat normal au fost numărate de două ori: prima dată a fost determinată energia de germinare (a treia zi), a doua oară - germenii (a şaptea zi). Melon seeds were germinated in Petri dishes on a filter paper substrate for 7 days, in accordance with GOST 12038-84 Seeds of agricultural crops, methods for determining the level of germination. Germination and germination energy (GOST 12038-66) is the percentage of seeds that germinated normally, taken for analysis. 4 batches of seeds were counted, 50 seeds per batch. The seeds were placed in Petri dishes on a substrate of moistened filter paper. 2 mL of an aqueous colloidal solution of Fe3O4/PVP nanoparticles was poured into each cup. An additional 2 mL of water was added to the series of control cups. After adding the solution to each of the Petri dishes, they were covered with moistened filter paper and a lid. The Petri dishes were placed in a thermostat with a constant temperature of 25°C. Germination took place in the dark. Temperature and humidity were checked daily, keeping them constant as in the initial state, additionally deionized water was added. Seeds that germinated normally were counted twice: the first time the germination energy was determined (third day), the second time - the germs (seventh day).
Pentru evaluarea germinării seminţelor, a fost calculat numărul total de seminţe normal germinate, luând în consideraţie energia germinării şi numărul de germeni obţinuţi, exprimat în procente ca media aritmetică a 4 repetări. Au fost calculate energia de germinare, procentul de germinare şi parametrii morfologici ai plantelor, precum lungimea rădăcinilor şi hipocotilului. To evaluate seed germination, the total number of normally germinated seeds was calculated, taking into account the germination energy and the number of germs obtained, expressed in percentage as the arithmetic mean of 4 repetitions. Germination energy, germination percentage and plant morphological parameters, such as root and hypocotyl length, were calculated.
A fost calculat intervalul de încredere. Calculele au fost efectuate, folosind nivelul de încredere de 95%. The confidence interval was calculated. The calculations were performed using the 95% confidence level.
Exemple de realizare a invenţiei Examples of embodiments of the invention
Exemplul 1 Example 1
Efectul de stimulare a creşterii realizat prin tratarea seminţelor culturii Cucumis melo L. cu soluţie apoasă coloidală de nanoparticule de magnetit stabilizate cu poli-N-vinilpirolidonă (Fe3O4/PVP) de diferite dimensiuni. Growth-stimulating effect achieved by treating Cucumis melo L. seeds with aqueous colloidal solution of magnetite nanoparticles stabilized with poly-N-vinylpyrrolidone (Fe3O4/PVP) of different sizes.
Influenţa soluţiilor de nanoparticule de Fe3O4/PVP de diferite dimensiuni asupra energiei de germinare, germenilor şi a parametrilor morfologici ai pepenilor speciei Cucumis melo L. este prezentată în tabelul 1. The influence of Fe3O4/PVP nanoparticle solutions of different sizes on germination energy, germs and morphological parameters of Cucumis melo L. melons is presented in Table 1.
Tabelul 1 Table 1
Varianta d (Fe3O4/PVP), nm C (Fe3O4/PVP), mg/L C (PVP), mol/L Energia de germinare, % Germeni, % Lungimea rădăcinilor, mm Lungimea hipocotilului, mm M ±m % de la control M ±m % de la control Controlul (H2O) 0 0 0 90 95 40,57 ± 1,27 100,00 7,28 ±1,29 100,00 1 10 50 0,01 97 98,6 61,21± 1,02 150,87 27,43± 0,17 367,78 2 20 50 0,01 95 97,5 58,04 ± 1,07 143,06 20,32 ± 5,71 279,12Variant d (Fe3O4/PVP), nm C (Fe3O4/PVP), mg/L C (PVP), mol/L Germination energy, % Germs, % Root length, mm Hypocotyl length, mm M ±m % of control M ±m % of control Control (H2O) 0 0 0 90 95 40.57 ± 1.27 100.00 7.28 ±1.29 100.00 1 10 50 0.01 97 98.6 61.21± 1.02 150.87 27.43± 0.17 367.78 2 20 50 0.01 95 97.5 58.04 ± 1.07 143.06 20.32 ± 5.71 279.12
Efectul maximal morfologic a fost atins la utilizarea soluţiei de nanoparticule de Fe3O4/PVP cu dimensiunile de 10 nm. Lungimea plantelor a crescut cu 50%, iar lungimea hipocotilului cu 267% comparativ cu mostra de control. The maximum morphological effect was achieved when using the Fe3O4/PVP nanoparticles solution with dimensions of 10 nm. The length of the plants increased by 50%, and the length of the hypocotyl by 267% compared to the control sample.
Exemplul 2 Example 2
Influenţa soluţiilor de nanoparticule de Fe3O4/PVP cu diferită concentraţie de PVP asupra energiei de germinare, germenilor şi a parametrilor morfologici ai pepenilor speciei Cucumis melo L. este prezentată în tabelul 2. The influence of Fe3O4/PVP nanoparticle solutions with different PVP concentrations on germination energy, germs and morphological parameters of Cucumis melo L. melons is presented in Table 2.
Tabelul 2 Table 2
Varianta d (Fe3O4/PVP), nm C (Fe3O4/PVP), mg/L C (PVP), mol/L Energia de germinare, % Germeni, % Lungimea rădăcinilor, mm Lungimea hipocotilului, mm M ±m % de la control M ±m % de la control Controlul (H2O) 0 0 0 90 95 40,57 ± 1,27 100,00 7,28 ±1,29 100,00 1 10 50 0,01 95 97,5 58,04 ± 1,07 143,06 20,32 ± 5,71 279,12 2 10 50 0,005 98 98,1 59,15± 2,05 145,79 22,17± 3,14 304,53Variant d (Fe3O4/PVP), nm C (Fe3O4/PVP), mg/L C (PVP), mol/L Germination energy, % Germs, % Root length, mm Hypocotyl length, mm M ±m % of control M ±m % of control Control (H2O) 0 0 0 90 95 40.57 ± 1.27 100.00 7.28 ±1.29 100.00 1 10 50 0.01 95 97.5 58.04 ± 1.07 143.06 20.32 ± 5.71 279.12 2 10 50 0.005 98 98.1 59.15± 2.05 145.79 22.17± 3.14 304.53
Efectul maximal morfologic a fost atins la utilizarea soluţiei de nanoparticule de Fe3O4/PVP cu concentraţia PVP de 0,005%. Lungimea plantelor a crescut cu 45%, iar lungimea hipocotilului cu 204% comparativ cu mostra de control. The maximum morphological effect was achieved when using the Fe3O4/PVP nanoparticle solution with a PVP concentration of 0.005%. The length of the plants increased by 45%, and the length of the hypocotyl by 204% compared to the control sample.
Exemplul 3 Example 3
Influenţa soluţiilor de nanoparticule de Fe3O4/PVP cu diferită concentraţie asupra energiei de germinare, germenilor şi a parametrilor morfologici ai pepenilor speciei Cucumis melo L. este prezentată în tabelul 3. The influence of Fe3O4/PVP nanoparticle solutions with different concentrations on germination energy, germs and morphological parameters of Cucumis melo L. melons is presented in Table 3.
Tabelul 3 Table 3
Varianta d (Fe3O4/PVP), nm C (Fe3O4/PVP), mg/L C (PVP), mol/L Energia de germinare, % Germeni, % Lungimea rădăcinilor, mm Lungimea hipocotilului, mm M ±m % de la control M ±m % de la control Controlul (H2O) 0 0 0 90 95 40,57 ± 1,27 100,00 7,28 ±1,29 100,00 1 10 20 0,01 90 90 30,44 ± 5,06 75,03 7,63 ± 2,45 104,81 2 10 50 0,01 95 97,5 58,04 ± 1,07 143,06 20,32 ± 5,71 279,12 3 10 100 0,01 92,5 95 36.21 ± 8,43 89,25 6,77 ± 6,55 92,99 Prototip (Triticum aestivum L.)[4] - 125 - - 95 - 68,6 - -Variant d (Fe3O4/PVP), nm C (Fe3O4/PVP), mg/L C (PVP), mol/L Germination energy, % Germs, % Root length, mm Hypocotyl length, mm M ±m % of control M ±m % of control Control (H2O) 0 0 0 90 95 40.57 ± 1.27 100.00 7.28 ±1.29 100.00 1 10 20 0.01 90 90 30.44 ± 5.06 75.03 7.63 ± 2.45 104.81 2 10 50 0.01 95 97.5 58.04 ± 1.07 143.06 20.32 ± 5.71 279.12 3 10 100 0.01 92.5 95 36.21 ± 8.43 89.25 6.77 ± 6.55 92.99 Prototype (Triticum aestivum L.)[4] - 125 - - 95 - 68.6 - -
Efectul maximal morfologic a fost atins la utilizarea soluţiei de nanoparticule de Fe3O4/PVP cu concentraţia de 50 mg/L. Lungimea plantelor a crescut cu 43%, iar lungimea hipocotilului cu 179% comparativ cu mostra de control. La concentraţia de 20 şi 100 mg/L, influenţa nanoparticulelor nu a fost considerabilă, ceea ce poate fi explicat prin stresul oxidativ, specific acestei specii de plantă agricolă. The maximum morphological effect was achieved when using the Fe3O4/PVP nanoparticle solution with a concentration of 50 mg/L. The length of the plants increased by 43%, and the length of the hypocotyl by 179% compared to the control sample. At the concentration of 20 and 100 mg/L, the influence of the nanoparticles was not considerable, which can be explained by the oxidative stress, specific to this agricultural plant species.
Exemplul 4 Example 4
Seminţele culturii Cucumis melo (pepene galben), în cantitate de 10 g, au fost înmuiate în apă timp de 24 ore. Ulterior, seminţele au fost tratate cu soluţie apoasă coloidală de nanoparticule de Fe3O4/PVP în concentraţie de 50 mg/L timp de 2…3 ore, după care au fost însămânţate în sol. Cucumis melo (melon) seeds, in an amount of 10 g, were soaked in water for 24 hours. Subsequently, the seeds were treated with an aqueous colloidal solution of Fe3O4/PVP nanoparticles at a concentration of 50 mg/L for 2…3 hours, after which they were sown in soil.
1. Răcuciu M., Creangă D. TMA-OH coated magnetic nanoparticles internalized in vegetal tissue. Romanian Journal of Physics, V. 52, 2007, p. 395-402 [online], [regăsit la 2016.06.16]. Regăsit din <http://www.ifin.ro/rjp/2007_52_3-4/0395_0403.pdf> 1. Răcuciu M., Creangă D. TMA-OH coated magnetic nanoparticles internalized in vegetal tissue. Romanian Journal of Physics, V. 52, 2007, p. 395-402 [online], [retrieved 2016.06.16]. Retrieved from <http://www.ifin.ro/rjp/2007_52_3-4/0395_0403.pdf>
2. WO 2015097486 A1 2015.07.02 2. WO 2015097486 A1 2015.07.02
3. Huanhua Wang, Xiaoming Kou, Zhiguo Pei, John Q. Xiao. Physiological effects of magnetite (Fe3O4) nanoparticles on perennial ryegrass (Loliumperenne L.) and pumpkin (Cucurbita mixta) plants. Nanotoxiology, nr. 5, 2011, p. 30-42[online], [regăsit la 2016.06.16]. Regăsit din <https://www.researchgate.net/publication/50591380_Physiological_effects_of_magnetite_Fe3O4_nanoparticles_on_perennial_ryegrass_Lolium_perenne_L_and_pumpkin_Cucurbita_mixta_plants> 3. Huanhua Wang, Xiaoming Kou, Zhiguo Pei, John Q. Xiao. Physiological effects of magnetite (Fe3O4) nanoparticles on perennial ryegrass (Loliumperenne L.) and pumpkin (Cucurbita mixta) plants. Nanotoxicology, no. 5, 2011, p. 30-42[online], [retrieved on 2016.06.16]. Retrieved from <https://www.researchgate.net/publication/50591380_Physiological_effects_of_magnetite_Fe3O4_nanoparticles_on_perennial_ryegrass_Lolium_perenne_L_and_pumpkin_Cucurbita_mixta_plants>
4. Кудрявцева Е. А., Анилова Л. В., Кузьмин С. Н., Шарыгина М.В. Влияние различных форм железа на прорастание семян Triticum aestivum L. Вестник ОГУ, № 6 (155), 2013, p. 46-48 [online], [regăsit la 2016.06.16]. Regăsit din < http://vestnik.osu.ru/2013_6/12.pdf> 4. Kudryavtseva Е. A., Anilova L. V., Kuzmin S. N., Sharygina M.V. The effect of different types of iron on the germination of seeds of Triticum aestivum L. Vestnik ОГУ, № 6 (155), 2013, p. 46-48 [online], [retrieved on 2016.06.16]. Retrieved from < http://vestnik.osu.ru/2013_6/12.pdf>
5. Cele mai valoroase realizări ştiinţifice. Nanoparticule de Fe3O4/PVP: stimulatori de creştere şi destructori de pesticide [online], [regăsit la 2016.06.16]. Regăsit din <http://www.asm.md/galerie/Realizari%20valoroase%20SSIT-2015.pdf> 5. The most valuable scientific achievements. Fe3O4/PVP nanoparticles: growth stimulants and pesticide destroyers [online], [retrieved on 2016.06.16]. Retrieved from <http://www.asm.md/galerie/Realizari%20valoroase%20SSIT-2015.pdf>
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| MD572F1 (en) * | 1995-02-21 | 1996-07-31 | Univ De Stat Din Moldova | Method for cocumber seeds treatment before sowing |
| MD450F1 (en) * | 1995-12-19 | 1996-08-30 | Univ De Stat Din Moldova | Method for beans treatment before sowing |
| MD3495F1 (en) * | 2007-06-07 | 2008-02-29 | Institutul De Genetica Si Fiziologie A Plantelor Al Academiei De Stiinte A Moldovei | Process for cultivation of vegetable crops |
| WO2015097486A1 (en) * | 2013-12-23 | 2015-07-02 | Invention Center Kft. | Iron (iii) oxide containing soil-binding composition |
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| MD572F1 (en) * | 1995-02-21 | 1996-07-31 | Univ De Stat Din Moldova | Method for cocumber seeds treatment before sowing |
| MD450F1 (en) * | 1995-12-19 | 1996-08-30 | Univ De Stat Din Moldova | Method for beans treatment before sowing |
| MD3495F1 (en) * | 2007-06-07 | 2008-02-29 | Institutul De Genetica Si Fiziologie A Plantelor Al Academiei De Stiinte A Moldovei | Process for cultivation of vegetable crops |
| WO2015097486A1 (en) * | 2013-12-23 | 2015-07-02 | Invention Center Kft. | Iron (iii) oxide containing soil-binding composition |
Non-Patent Citations (4)
| Title |
|---|
| Cele mai valoroase realizări ştiinţifice. Nanoparticule de Fe3O4/PVP: stimulatori de creştere şi destructori de pesticide [online], [regăsit la 2016.06.16]. Regăsit din <http://www.asm.md/galerie/Realizari%20valoroase%20SSIT-2015.pdf> * |
| Huanhua Wang, Xiaoming Kou, Zhiguo Pei, John Q. Xiao. Physiological effects of magnetite (Fe3O4) nanoparticles on perennial ryegrass (Loliumperenne L.) and pumpkin (Cucurbita mixta) plants. Nanotoxiology, nr. 5, 2011, p. 30-42[online], [regăsit la 2016.06.16]. Regăsit din <https://www.researchgate.net/publication/50591380_Physiological_effects_of_magnetite_Fe3O4_nanoparticles_on_perennial_ryegrass_Lolium_perenne_L_and_pumpkin_Cucurbita_mixta_plants> * |
| Răcuciu M., Creangă D. TMA-OH coated magnetic nanoparticles internalized in vegetal tissue. Romanian Journal of Physics, V. 52, 2007, p. 395-402 [online], [regăsit la 2016.06.16]. Regăsit din <http://www.ifin.ro/rjp/2007_52_3-4/0395_0403.pdf> * |
| Кудрявцева Е. А., Анилова Л. В., Кузьмин С. Н., Шарыгина М.В. Влияние различных форм железа на прорастание семян Triticum aestivum L. Вестник ОГУ, № 6 (155), 2013, p. 46-48 [online], [regăsit la 2016.06.16]. Regăsit din < http://vestnik.osu.ru/2013_6/12.pdf> * |
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