Pub Date : 2022-01-01DOI: 10.56304/s0234275822060084
В. Ю. Кислицин, А. М. Чулкин, И. Н. Зоров, И.Г. Синельников, А. П. Синицын, А. М. Рожкова
{"title":"Функция транскрипционного фактора XlnR у мицелиального гриба\u0000 \u0000 Penicillium verruculosum\u0000","authors":"В. Ю. Кислицин, А. М. Чулкин, И. Н. Зоров, И.Г. Синельников, А. П. Синицын, А. М. Рожкова","doi":"10.56304/s0234275822060084","DOIUrl":"https://doi.org/10.56304/s0234275822060084","url":null,"abstract":"","PeriodicalId":8850,"journal":{"name":"Biotekhnologiya","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70856449","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.56304/s0234275822060096
Ю. Ю. Миллер, Т. Ф. Киселева
{"title":"Биотехнологический подход к интенсификации производства соевого солода","authors":"Ю. Ю. Миллер, Т. Ф. Киселева","doi":"10.56304/s0234275822060096","DOIUrl":"https://doi.org/10.56304/s0234275822060096","url":null,"abstract":"","PeriodicalId":8850,"journal":{"name":"Biotekhnologiya","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70856459","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.21519/0234-2758-2021-37-3-20-28
Ya. A. Delegan, T. Antipova, M. Vainshtein
The methods for increasing the isolation of carotenoids from yeast producer cultures have been compared. It was shown that the optimal procedure is microwave pretreatment (2450 GHz, 600 W, 40 s) with subsequent extraction of the target products with methanol. Additives of sodium persulfate or ammonium persulfate to aerated culture medium provided additional oxygen sources for the yeast and increased the medium redox potential. The introduction of persulfate led to a 1.3-fold increase in the content of colored carotenoids in the R. dubiovatum VKPM Y-305 biomass as compared to the control without persulfate. In the R. sphaerocarpum VKPM Y-1559 biomass, persulfate at concentrations of 0.15 g/L and 0.50 g/L increased the content of colored carotenoids by 1.3 and 1.4 times, respectively. The optimal persulfate concentent depends on the yeast growth phase and time of exposure to the compound. Higher persulfate concentrations resulted in increased redox potential and discoloration of carotenoids, though without yeast cell death. yeasts, carotenoids, medium redox potential, persulfate
{"title":"Effect of Persulfates on Production of Carotenoids in the Rhodosporidium sphaerocarpum, R. diobovatum and Rhodotorula glutinis Yeast Cultures","authors":"Ya. A. Delegan, T. Antipova, M. Vainshtein","doi":"10.21519/0234-2758-2021-37-3-20-28","DOIUrl":"https://doi.org/10.21519/0234-2758-2021-37-3-20-28","url":null,"abstract":"The methods for increasing the isolation of carotenoids from yeast producer cultures have been compared. It was shown that the optimal procedure is microwave pretreatment (2450 GHz, 600 W, 40 s) with subsequent extraction of the target products with methanol. Additives of sodium persulfate or ammonium persulfate to aerated culture medium provided additional oxygen sources for the yeast and increased the medium redox potential. The introduction of persulfate led to a 1.3-fold increase in the content of colored carotenoids in the R. dubiovatum VKPM Y-305 biomass as compared to the control without persulfate. In the R. sphaerocarpum VKPM Y-1559 biomass, persulfate at concentrations of 0.15 g/L and 0.50 g/L increased the content of colored carotenoids by 1.3 and 1.4 times, respectively. The optimal persulfate concentent depends on the yeast growth phase and time of exposure to the compound. Higher persulfate concentrations resulted in increased redox potential and discoloration of carotenoids, though without yeast cell death. yeasts, carotenoids, medium redox potential, persulfate","PeriodicalId":8850,"journal":{"name":"Biotekhnologiya","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67999046","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.21519/0234-2758-2021-37-4-32-42
M. Mollaev, A. Zabolotskii, D. A. Mazalev, N. Gorokhovets, M. Sokol, M. R. Mollaeva, M. Fomicheva, A. Pshenichnikova, E. D. Nikolskaya
Previous studies have demonstrated the applicability of alpha-fetoprotein or its receptor-binding domain fused to the 6-histidine tag at the C-terminus (rAFP3D-His6) as vectors for targeted delivery of antitumor agents. This tag is undesirable for further preclinical trials. Therefore, we designed a recombinant protein rAFP3D without any affine tags, and accessed its functional activity. The protein was produced as inclusion bodies in Escherichia coli BL21 (DE3) cells. Optimal conditions for washing inclusion bodies and refolding the target protein were selected. We used a saturated solution of (NH4)2SO4 as the primary purification step to precipitate the main fraction of protein admixtures. The second purification step included hydrophobic chromatography using butyl-cellulose. The identity of the protein sequence was confirmed by tandem mass spectrometry. Circular dichroism demonstrated the authenticity of the secondary structure. Fluorescently labeled rAFP3D actively penetrated into MCF-7 tumor cells. These results indicate that rAFP3D can be used for targeted drug delivery. Key words: alpha-fetoprotein, receptor-binding domain, recombinant protein, MALDI-MS, HPLC, drug delivery system Funding - The project was supported with Russian Foundation for Basic Research (project no. 18-29-09022/20).
{"title":"Obtaining and Purification of Recombinant Domain III of Human Alpha-Fetoprotein","authors":"M. Mollaev, A. Zabolotskii, D. A. Mazalev, N. Gorokhovets, M. Sokol, M. R. Mollaeva, M. Fomicheva, A. Pshenichnikova, E. D. Nikolskaya","doi":"10.21519/0234-2758-2021-37-4-32-42","DOIUrl":"https://doi.org/10.21519/0234-2758-2021-37-4-32-42","url":null,"abstract":"Previous studies have demonstrated the applicability of alpha-fetoprotein or its receptor-binding domain fused to the 6-histidine tag at the C-terminus (rAFP3D-His6) as vectors for targeted delivery of antitumor agents. This tag is undesirable for further preclinical trials. Therefore, we designed a recombinant protein rAFP3D without any affine tags, and accessed its functional activity. The protein was produced as inclusion bodies in Escherichia coli BL21 (DE3) cells. Optimal conditions for washing inclusion bodies and refolding the target protein were selected. We used a saturated solution of (NH4)2SO4 as the primary purification step to precipitate the main fraction of protein admixtures. The second purification step included hydrophobic chromatography using butyl-cellulose. The identity of the protein sequence was confirmed by tandem mass spectrometry. Circular dichroism demonstrated the authenticity of the secondary structure. Fluorescently labeled rAFP3D actively penetrated into MCF-7 tumor cells. These results indicate that rAFP3D can be used for targeted drug delivery. Key words: alpha-fetoprotein, receptor-binding domain, recombinant protein, MALDI-MS, HPLC, drug delivery system Funding - The project was supported with Russian Foundation for Basic Research (project no. 18-29-09022/20).","PeriodicalId":8850,"journal":{"name":"Biotekhnologiya","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68000667","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.21519/0234-2758-2021-37-5-80-87
A. Tkachev, M. Osetrova, D. Smirnov, O. Efimova, E.E. Khrameev
Abstract-Lipids make up more than half of the dry matter of the human brain and play a key role in its functioning. However, the lipid composition of the brain anatomical structures remains poorly understood. The first such studies were carried out more than 50 years ago, but since then, a small number of works have been published describing the concentration of lipids in only a few areas of the human brain. A fundamentally new step towards understanding the molecular organization of the brain and identifying the molecular basis of human cognitive abilities should be a detailed large-scale study of the brain lipidome. However, there is no description in the literature of methods optimized for studying the lipid composition of the human brain. In this work, we develop and present methods for lipid extraction and mass spectrometry analysis, which ensure simultaneous detection of the maximum amount of different lipid classes and individual substances in the human brain, as well as the approaches to bioinformatics analysis of the obtained data. Their use makes it possible to create a comprehensive picture of the molecular organization of the human brain, which has no analogues in the world in terms of its completeness. Key words: lipidome, brain, mass spectrometry, high-performance liquid chromatography, bioinformatics The reported study was funded by the Russian Foundation for Basic Research, project no. 20-34-70077.
{"title":"Application of Mass Spectrometry Methods to the Analysis of Lipid Composition of the Human Brain","authors":"A. Tkachev, M. Osetrova, D. Smirnov, O. Efimova, E.E. Khrameev","doi":"10.21519/0234-2758-2021-37-5-80-87","DOIUrl":"https://doi.org/10.21519/0234-2758-2021-37-5-80-87","url":null,"abstract":"Abstract-Lipids make up more than half of the dry matter of the human brain and play a key role in its functioning. However, the lipid composition of the brain anatomical structures remains poorly understood. The first such studies were carried out more than 50 years ago, but since then, a small number of works have been published describing the concentration of lipids in only a few areas of the human brain. A fundamentally new step towards understanding the molecular organization of the brain and identifying the molecular basis of human cognitive abilities should be a detailed large-scale study of the brain lipidome. However, there is no description in the literature of methods optimized for studying the lipid composition of the human brain. In this work, we develop and present methods for lipid extraction and mass spectrometry analysis, which ensure simultaneous detection of the maximum amount of different lipid classes and individual substances in the human brain, as well as the approaches to bioinformatics analysis of the obtained data. Their use makes it possible to create a comprehensive picture of the molecular organization of the human brain, which has no analogues in the world in terms of its completeness. Key words: lipidome, brain, mass spectrometry, high-performance liquid chromatography, bioinformatics The reported study was funded by the Russian Foundation for Basic Research, project no. 20-34-70077.","PeriodicalId":8850,"journal":{"name":"Biotekhnologiya","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68003851","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.21519/0234-2758-2021-37-5-108-116
E. Dubrovskaya, V. Grinev, O. Turkovskaya
Abstract-A model system for the successful mycoremediation of soil contaminated with real oil pollution has been developed on the basis of Pleurotus ostreatus Florida grown on a natural substrate (husk, straw). It was shown that under these conditions, mycoremediation was observed during 2 weeks at 24° C with oil contamination of up to 22 g/kg and inoculum/soil ratio of 1:10. The solid-phase P. ostreatus inoculums grown on straw induced the production of highly active ligninolytic enzymes, laccases and hybrid peroxidases. This allows us to recommend this inoculum for the development of technologies for mycoremediation of soils contaminated with mixtures of pollutants with a high content of aromatic fractions. Based on the solid-phase inoculum of Schizophyllum commune grown on a natural substrate (pine sawdust), a semi-submerged model system was developed, which can be repeatedly used for the degradation of surfactants in the soil. In this case, sawdust can be considered not only as a source of carbon and energy for the fungus, but also as a kind of natural carrier for its immobilization. This model system based on S. commune grown on a natural substrate can also be recommended for the development of ecological biotechnologies on its basis. Key words: mycoremediation, basidiomycetes, oil, surfactants, ligninolytic enzymes This work was carried out and funded within the framework of theme No. 121031700141-7 (IBPPM RAS) with partial support of the Russian Science Foundation (project no. 16-14-00081 "Fungi for Environmentally Significant Biotechnologies") (degradation of surfactants) and of the Russian Foundation for Basic Research (project no. 18-29-05062 "Development of Scientific Foundations of a New Integrated Biotechnology for Oil Spill Response in the Arid Climate Zone") (mycoremediation of oil-contaminated soil).
{"title":"Prospects for the Use of Xylotrophic Fungi Pleurotus ostreatus Florida and Schizophyllum commune for Mycoremediation of Soils Contaminated with Oil Hydrocarbons and Surfactants","authors":"E. Dubrovskaya, V. Grinev, O. Turkovskaya","doi":"10.21519/0234-2758-2021-37-5-108-116","DOIUrl":"https://doi.org/10.21519/0234-2758-2021-37-5-108-116","url":null,"abstract":"Abstract-A model system for the successful mycoremediation of soil contaminated with real oil pollution has been developed on the basis of Pleurotus ostreatus Florida grown on a natural substrate (husk, straw). It was shown that under these conditions, mycoremediation was observed during 2 weeks at 24° C with oil contamination of up to 22 g/kg and inoculum/soil ratio of 1:10. The solid-phase P. ostreatus inoculums grown on straw induced the production of highly active ligninolytic enzymes, laccases and hybrid peroxidases. This allows us to recommend this inoculum for the development of technologies for mycoremediation of soils contaminated with mixtures of pollutants with a high content of aromatic fractions. Based on the solid-phase inoculum of Schizophyllum commune grown on a natural substrate (pine sawdust), a semi-submerged model system was developed, which can be repeatedly used for the degradation of surfactants in the soil. In this case, sawdust can be considered not only as a source of carbon and energy for the fungus, but also as a kind of natural carrier for its immobilization. This model system based on S. commune grown on a natural substrate can also be recommended for the development of ecological biotechnologies on its basis. Key words: mycoremediation, basidiomycetes, oil, surfactants, ligninolytic enzymes This work was carried out and funded within the framework of theme No. 121031700141-7 (IBPPM RAS) with partial support of the Russian Science Foundation (project no. 16-14-00081 \"Fungi for Environmentally Significant Biotechnologies\") (degradation of surfactants) and of the Russian Foundation for Basic Research (project no. 18-29-05062 \"Development of Scientific Foundations of a New Integrated Biotechnology for Oil Spill Response in the Arid Climate Zone\") (mycoremediation of oil-contaminated soil).","PeriodicalId":8850,"journal":{"name":"Biotekhnologiya","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68001923","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.21519/0234-2758-2021-37-5-72-79
L. Yakupova, А.А. Skuredina, Еlena V Kudryashova
Abstract-Moxifloxacin encapsulation in polymer cyclodextrin-based particles with an average hydrodynamic diameter of 150--200 nm has led to the formation of potential delivery systems with a degree of moxifloxacin inclusion of more than 80%. Cross-linking of the moxifloxacin-cyclodextrin complexes caused a pronounced slowdown in the release of the drug molecules in acidic media to less than 10% per day. In the presence of trypsin, the drug release was accelerated by 15--20% within 90 min. It was shown by Fourier-transform infrared spectroscopy that this acceleration was due to the partial enzymatic degradation of the urethane bonds of the polymer matrix near the surface of the particles. The results obtained are important for the development of highly effective oral dosage forms of prolonged action. Key words: fluoroquinolones, cyclodextrins, FTIR spectroscopy, trypsin
{"title":"Trypsinolysis of Polyurethane Particles Based on Сyclodextrin with Encapsulated Moxifloxacin","authors":"L. Yakupova, А.А. Skuredina, Еlena V Kudryashova","doi":"10.21519/0234-2758-2021-37-5-72-79","DOIUrl":"https://doi.org/10.21519/0234-2758-2021-37-5-72-79","url":null,"abstract":"Abstract-Moxifloxacin encapsulation in polymer cyclodextrin-based particles with an average hydrodynamic diameter of 150--200 nm has led to the formation of potential delivery systems with a degree of moxifloxacin inclusion of more than 80%. Cross-linking of the moxifloxacin-cyclodextrin complexes caused a pronounced slowdown in the release of the drug molecules in acidic media to less than 10% per day. In the presence of trypsin, the drug release was accelerated by 15--20% within 90 min. It was shown by Fourier-transform infrared spectroscopy that this acceleration was due to the partial enzymatic degradation of the urethane bonds of the polymer matrix near the surface of the particles. The results obtained are important for the development of highly effective oral dosage forms of prolonged action. Key words: fluoroquinolones, cyclodextrins, FTIR spectroscopy, trypsin","PeriodicalId":8850,"journal":{"name":"Biotekhnologiya","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68002896","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.21519/0234-2758-2021-37-3-11-19
E.V. Viazau, R. Goncharik, R. Vasilov, A. Selishcheva
The effect of the natural carotenoid astaxanthin (AST) and its esters as inhibitors of chlorophyll (Chl) a photodegradation in the Haematococcus pluvialis unicellular alga has been studied. It was shown that the presence of AST or its esters in low concentrations ((0.67 μM)) suppressed by 15-17% the Chl a destruction as a result of irradiation of a Chl a solution in acetone with white light (250 μmol photons∙m-2∙s-1 for 2 h 20 min). When Chl a solutions were irradiated with blue light (500 μmol∙photons∙m-2∙s-1 for 5 h) in the presence of AST or its esters at high concentrations (AST : Chl a = 5 : 1 mol/mol), the Chl a degradation was suppressed by 40-50%. In a highly polar medium (methanol), the rate of Chl a photodegradation was much higher than in an apolar medium (hexane : acetone = 94 : 6). Esterified AST forms are better protectors of Chl a than unsubstituted AST, which is consistent with the general idea of the predominance of AST esters in the microalga H. pluvialis cells in vivo. The results indicate a possible protective role of AST against Chl a photooxidation in H. pluvialis cells. astaxanthin, chlorophyll, singlet oxygen, photodegradation, Haematococcus pluvialis This work was supported by the Belarusian Republican Foundation for Basic Research (project no. B19РМ-010) and Russian Foundation for Basic Research (project no. 19-54-04003).
{"title":"Protective Properties of Astaxanthin and its Esters Isolated from the Haematococcus pluvialis Alga during the Photodegradation of Chlorophyll a","authors":"E.V. Viazau, R. Goncharik, R. Vasilov, A. Selishcheva","doi":"10.21519/0234-2758-2021-37-3-11-19","DOIUrl":"https://doi.org/10.21519/0234-2758-2021-37-3-11-19","url":null,"abstract":"The effect of the natural carotenoid astaxanthin (AST) and its esters as inhibitors of chlorophyll (Chl) a photodegradation in the Haematococcus pluvialis unicellular alga has been studied. It was shown that the presence of AST or its esters in low concentrations ((0.67 μM)) suppressed by 15-17% the Chl a destruction as a result of irradiation of a Chl a solution in acetone with white light (250 μmol photons∙m-2∙s-1 for 2 h 20 min). When Chl a solutions were irradiated with blue light (500 μmol∙photons∙m-2∙s-1 for 5 h) in the presence of AST or its esters at high concentrations (AST : Chl a = 5 : 1 mol/mol), the Chl a degradation was suppressed by 40-50%. In a highly polar medium (methanol), the rate of Chl a photodegradation was much higher than in an apolar medium (hexane : acetone = 94 : 6). Esterified AST forms are better protectors of Chl a than unsubstituted AST, which is consistent with the general idea of the predominance of AST esters in the microalga H. pluvialis cells in vivo. The results indicate a possible protective role of AST against Chl a photooxidation in H. pluvialis cells. astaxanthin, chlorophyll, singlet oxygen, photodegradation, Haematococcus pluvialis This work was supported by the Belarusian Republican Foundation for Basic Research (project no. B19РМ-010) and Russian Foundation for Basic Research (project no. 19-54-04003).","PeriodicalId":8850,"journal":{"name":"Biotekhnologiya","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67998983","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.21519/0234-2758-2021-37-4-14-19
N. R. Mikheeva, A. Nalbandyan, N. Cherkasova
The MTP4 (Metal Tolerance Protein 4) gene, which controls the resistance of sugar beet plants to heavy metals and is located on the 3rd chromosome has been studied. Micro clones grown in vitro with lethal Cd doses were used as explants. For PCR, specific primers MTP4A F/R and MTP4B F/R, covering the 2nd, 3rd and 4th exons and introns between them in the MTP4 gene, were used. Samples with resistance to heavy metals (in particular, Cd) contained some nucleotide changes in the 4th exon resulting in amino acid substitutions in the polypeptide chain. Based on the results of the bioinformatics analysis, it can be assumed with a high probability that the resistance of the studied breeding materials to heavy metals arose due to the presence of certain polymorphisms in the analyzed sugar beet gene. Geneious Prime software as a tool for the MTP4 gene sequence analysis will probably allow the in vitro selection of resistant forms for the subsequent use in the breeding scheme. Key words: sugar beet, МТР genes, genetic polymorphism, PCR, molecular genetic markers, breeding, DNA
研究了位于甜菜第3染色体上控制甜菜对重金属抗性的MTP4 (Metal Tolerance Protein 4)基因。以致死剂量Cd培养的微无性系为外植体。PCR采用特异性引物MTP4A F/R和MTP4B F/R，分别覆盖MTP4基因的第2、3、4外显子和内含子。对重金属(特别是Cd)具有抗性的样品在第4外显子上含有一些核苷酸变化，导致多肽链上的氨基酸取代。根据生物信息学分析的结果，我们可以很有可能认为所研究的育种材料对重金属的抗性是由于所分析的甜菜基因中存在一定的多态性。genous Prime软件作为MTP4基因序列分析的工具，可能允许在体外选择抗性形式，以便随后在育种方案中使用。关键词:甜菜，МТР基因，遗传多态性，PCR，分子遗传标记，育种，DNA
{"title":"Screening of Sugar Beet Regenerants for the Heavy Metal Resistance MTP4 Gene","authors":"N. R. Mikheeva, A. Nalbandyan, N. Cherkasova","doi":"10.21519/0234-2758-2021-37-4-14-19","DOIUrl":"https://doi.org/10.21519/0234-2758-2021-37-4-14-19","url":null,"abstract":"The MTP4 (Metal Tolerance Protein 4) gene, which controls the resistance of sugar beet plants to heavy metals and is located on the 3rd chromosome has been studied. Micro clones grown in vitro with lethal Cd doses were used as explants. For PCR, specific primers MTP4A F/R and MTP4B F/R, covering the 2nd, 3rd and 4th exons and introns between them in the MTP4 gene, were used. Samples with resistance to heavy metals (in particular, Cd) contained some nucleotide changes in the 4th exon resulting in amino acid substitutions in the polypeptide chain. Based on the results of the bioinformatics analysis, it can be assumed with a high probability that the resistance of the studied breeding materials to heavy metals arose due to the presence of certain polymorphisms in the analyzed sugar beet gene. Geneious Prime software as a tool for the MTP4 gene sequence analysis will probably allow the in vitro selection of resistant forms for the subsequent use in the breeding scheme. Key words: sugar beet, МТР genes, genetic polymorphism, PCR, molecular genetic markers, breeding, DNA","PeriodicalId":8850,"journal":{"name":"Biotekhnologiya","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68000995","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.21519/0234-2758-2021-37-6-14-24
N. Gessler, E. Isakova, Y. Deryabina
Using the extremophilic yeast of Yarrowia lipolytica, a new model has been proposed to study the protective properties of stilbene polyphenols, namely resveratrol and pinosylvin, under heat shock. It was shown that a short-term thermal exposure of yeast cells (55 C, 25 min) led to a 40% decrease in the colony-forming ability of the population, a fivefold decrease in the respiration rate, and a growth of cyanide resistance and catalase activity, which indicated the adaptive yeast response to heat stress. Under these conditions, natural biologically active stilbenes, resveratrol and pinosylvin, at a concentration of 10 μM each increased yeast survival by 28% and 13%, respectively. In heat shock, resveratrol additionally raised catalase activity, while pinosylvin increased the cell respiration rate and decreased cyanide resistance and catalase activity. The results obtained indicate that resveratrol acts as a mild pro-oxidant inducing antioxidant protection during the adaptive response of the yeast to heat shock. Unlike resveratrol, pinosylvin increases cell survival stabilizing mitochondrial function and preserving the ATP-generating component of respiration. Yarrowia lipolytica yeast, polyphenols, stilbenoids, resveratrol, pinosylvin, cellular respiratory activity, heat shock, superoxide dismutase, catalase
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