All Russia Research Institute of Agricultural Biotechnology

Moscow, Russia

All Russia Research Institute of Agricultural Biotechnology

Moscow, Russia
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Miroshnichenko D.,RAS Institute of Basic Biological Problems | Chaban I.,All Russia Research Institute of Agricultural Biotechnology | Chernobrovkina M.,All Russia Research Institute of Agricultural Biotechnology | Dolgov S.,RAS Institute of Chemistry
PLoS ONE | Year: 2017

Einkorn (Triticum monococcum L.) is A-genome diploid wheat that has a potential to become a useful model for understanding the biology and genomics in Triticeae. Unfortunately, the application of modern technologies such as genetic engineering, RNAi-based gene silencing and genome editing is not available for einkorn as there is no efficient in vitro tissue culture and plant regeneration system. In the present study an efficient and simple protocol for plant regeneration via direct or indirect somatic embryogenesis and organogenesis has been developed. Various auxins used as sole inductors in einkorn displayed low effect for morphogenesis (0±8%) and plant regeneration (1±2 shoots per explant). The addition of Daminozide, the inhibitor of biosynthesis of gibberellins, together with auxin significantly improved the formation of morphogenic structures, especially when Dicamba (51.4%) and Picloram (56.6%) were used for combination; furthermore, the simultaneous addition of cytokinin into induction medium significantly promoted in vitro performance. Among the tested cytokinins, the urea-type substances, such as TDZ and CPPU were more effective than the adenine type ones, BA and Zeatin, for the regulation of morphogenesis; especially, TDZ was more effective than CPPU for shoot formation (11.73 vs. 7.04 per regenerating callus). The highest morphogenic response of 90.2% with the production of more than 10 shoots per initial explant was observed when 3.0 mg/L Dicamba, 50.0 mg/L Daminozide and 0.25 mg/L TDZ were combined together. Along with the identification of appropriate induction medium, the optimal developmental stage for einkorn was found as partially transparent immature embryo in size of around 1.0 mm. Although in the present study the critical balance between plant growth regulators was established for einkorn only, we assume that further the proposed strategy could be successfully applied to other recalcitrant cereal species and genotypes. © 2017 Miroshnichenko et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Fedorov A.A.,Russian Academy of Sciences | Berdnikov A.S.,Russian Academy of Sciences | Sochivko D.G.,JSC Syntol | Varlamov D.A.,All Russia Research Institute of Agricultural Biotechnology | And 2 more authors.
Doklady Biochemistry and Biophysics | Year: 2016

Macroscopic kinetic models describing the process of polymerase chain reaction (PCR) are currently solved only by numerical methods, which hampers the development of effective software algorithms for processing the results of the reaction. This paper considers the application of the homotopy perturbation method for obtaining approximate analytical solution of the simplest system of enzymatic kinetic equations describing the synthesis of nucleic acid molecules during PCR. The resulting approximate analytic solution with high accuracy reproduces the results of a numerical solution of the system in a wide range of ratios of enzyme and substrate concentrations both for the case of a large excess of the substrate over the enzyme and vice versa. © 2016, Pleiades Publishing, Ltd.


Khvatkov P.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Chernobrovkina M.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Okuneva A.,All Russia Research Institute of Agricultural Biotechnology | Shvedova A.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | And 2 more authors.
Plant Cell, Tissue and Organ Culture (PCTOC) | Year: 2014

The development of tissue-culture systems in duckweed has been limited to species of the genus Lemna. Here we report on a tissue-culture system (callus induction, callus growth and plant regeneration) for the rootless duckweed Wolffia arrhiza. We developed a two-step procedure for callus induction in Wolffia using Schenk & Hildebrandt (SH) medium containing glucose, mannitol and sorbitol. In the first stage, explants were precultivated in the presence of 5.0 mg l-1 2,4-dichlorophenoxyacetic acid and 0.5 mg l-1 N6-benzyladenine (BA) for 16 weeks. In the second stage, BA was replaced with 12.5 mg l-1 Picloram (PCL) for 4 weeks. The resulting callus could be maintained in vitro for a long time (about 1 year) with a relatively low concentration of PCL (4.0 mg l-1), or used to regenerate whole plants by transferring it to growth regulator-free SH medium. The protocols created for callus induction and regeneration were highly efficient at each stage (with the efficiency of 97 % for callus formation and 78 % for regeneration). © 2014 Springer Science+Business Media Dordrecht.


Miroshnichenko D.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Chernobrovkina M.,All Russia Research Institute of Agricultural Biotechnology | Dolgov S.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry
Plant Cell, Tissue and Organ Culture | Year: 2016

The efficiency of immature embryo-derived in vitro culture of G genome wheats is significantly influenced by various auxins and sugars which are used for induction of embryogenic response, and by regeneration media composition for promotion of plant development from subcultured embryogenic calli. The embryogenic calli of Triticum timopheevii has demonstrated the highest regeneration ability when the initial explants were cultured on the media supplemented with 4 mg l−1 of Picloram (29.0 %), 4 mg l−1 of Dicamba (28.7 %) or 3 mg l−1 of 2,4-D (29.1 %). The media supplemented with 5–6 mg l−1 of Picloram were considered to be the most effective for promotion of embryogenic/regenerable callus production in Triticumkiharae cultures (73.7–75.0 %). Both T. timopheevii and T. kiharae embryogenic structures were characterized by the formation of green and albino plantlets. Generally the medium that was initially supplemented with Picloram promoted the formation of lower albino plants fraction rather than 2,4-D and Dicamba. As it was measured by the total green plant production per initial explant, the overall efficiency has been reduced when sucrose was substituted by glucose or maltose. The regeneration medium supplemented with 0.25 mg l−1 TDZ significantly enhanced the regeneration capacity of embryogenic callus in T. kiharae. In culture of T. timopheevii the difference between the medium lack of growth regulators and the medium supplemented with TDZ was not prominent, though both of the media have demonstrated the greater efficacy as compared to those supplemented with BA and Zeatin. © 2016 Springer Science+Business Media Dordrecht


Sochivko D.G.,JSC Syntol | Fedorov A.A.,Russian Academy of Sciences | Varlamov D.A.,All Russia Research Institute of Agricultural Biotechnology | Kurochkin V.E.,Russian Academy of Sciences | Petrov R.V.,Federal Biomedical Agency
Doklady Biochemistry and Biophysics | Year: 2016

The paper reviews different approaches to the mathematical analysis of polymerase chain reaction (PCR) kinetic curves. The basic principles of PCR mathematical analysis are presented. Approximation of PCR kinetic curves and PCR efficiency curves by various functions is described. Several PCR models based on chemical kinetics equations are suggested. Decision criteria for an optimal function to describe PCR efficiency are proposed. © 2016, Pleiades Publishing, Ltd.


Komakhin R.A.,All Russia Research Institute of Agricultural Biotechnology | Vysotskii D.A.,All Russia Research Institute of Agricultural Biotechnology | Shukurov R.R.,IBC Generium LLC | Voblikova V.D.,All Russia Research Institute of Agricultural Biotechnology | And 4 more authors.
BMC Biotechnology | Year: 2016

Background: In a previous study we found that in chickweed the expression level of the pro-SmAMP2 gene was comparable or even higher to that of the β-actin gene. This high level of the gene expression has attracted our attention as an opportunity for the identification of novel strong promoters of plant origin, which could find its application in plant biotechnology. Therefore, in the present study we focused on the nucleotide sequence identification and the functional characteristics of the pro-SmAMP2 promoter in transgenic plants. Results: In chickweed (Stellaria media), a 2120 bp promoter region of the pro-SmAMP2 gene encoding antifungal peptides was sequenced. Six 5'-deletion variants -2120, -1504, -1149, -822, -455, and -290 bp of pro-SmAMP2 gene promoter were fused with the coding region of the reporter gene gusA in the plant expression vector pCambia1381Z. Independent transgenic plants of tobacco Nicotiana tabacum were obtained with each genetic structure. GUS protein activity assay in extracts from transgenic plants showed that all deletion variants of the promoter, except -290 bp, expressed the gusA gene. In most transgenic plants, the GUS activity level was comparable or higher than in plants with the viral promoter CaMV 35S. GUS activity remains high in progenies and its level correlates positively with the amount of gusA gene mRNA in T3 homozygous plants. The activity of the pro-SmAMP2 promoter was detected in all organs of the transgenic plants studied, during meiosis and in pollen as well. Conclusion: Our results show that the pro-SmAMP2 promoter can be used for target genes expression control in transgenic plants. © 2016 Komakhin et al.


PubMed | Russian Academy of Sciences, JSC Syntol, Federal Biomedical Agency and All Russia Research Institute of Agricultural Biotechnology
Type: | Journal: Doklady. Biochemistry and biophysics | Year: 2016

The paper reviews different approaches to the mathematical analysis of polymerase chain reaction (PCR) kinetic curves. The basic principles of PCR mathematical analysis are presented. Approximation of PCR kinetic curves and PCR efficiency curves by various functions is described. Several PCR models based on chemical kinetics equations are suggested. Decision criteria for an optimal function to describe PCR efficiency are proposed.


PubMed | Russian Academy of Sciences, JSC Syntol, Federal Biomedical Agency and All Russia Research Institute of Agricultural Biotechnology
Type: Journal Article | Journal: Doklady. Biochemistry and biophysics | Year: 2016

Macroscopic kinetic models describing the process of polymerase chain reaction (PCR) are currently solved only by numerical methods, which hampers the development of effective software algorithms for processing the results of the reaction. This paper considers the application of the homotopy perturbation method for obtaining approximate analytical solution of the simplest system of enzymatic kinetic equations describing the synthesis of nucleic acid molecules during PCR. The resulting approximate analytic solution with high accuracy reproduces the results of a numerical solution of the system in a wide range of ratios of enzyme and substrate concentrations both for the case of a large excess of the substrate over the enzyme and vice versa.


PubMed | Russian Academy of Sciences, JSC Syntol, Federal Biomedical Agency and All Russia Research Institute of Agricultural Biotechnology
Type: Journal Article | Journal: Doklady. Biochemistry and biophysics | Year: 2017

Development of methods for obtaining approximate analytical solutions of nonlinear differential equations and their systems is a rapidly developing field of mathematical physics. Earlier, an approximate solution of the simplest system of kinetic enzymatic equations for calculating dynamics of complementary strands of nucleic acids was obtained. In this study, we consider an alternative approach to selecting the basic linear approximation of the used method, which makes it possible to obtain more accurate analytical solutions of the set problem.


PubMed | All Russia Research Institute of Agricultural Biotechnology and IBC Generium LLC
Type: Journal Article | Journal: BMC biotechnology | Year: 2016

In a previous study we found that in chickweed the expression level of the pro-SmAMP2 gene was comparable or even higher to that of the -actin gene. This high level of the gene expression has attracted our attention as an opportunity for the identification of novel strong promoters of plant origin, which could find its application in plant biotechnology. Therefore, in the present study we focused on the nucleotide sequence identification and the functional characteristics of the pro-SmAMP2 promoter in transgenic plants.In chickweed (Stellaria media), a 2120bp promoter region of the pro-SmAMP2 gene encoding antifungal peptides was sequenced. Six 5-deletion variants -2120, -1504, -1149, -822, -455, and -290bp of pro-SmAMP2 gene promoter were fused with the coding region of the reporter gene gusA in the plant expression vector pCambia1381Z. Independent transgenic plants of tobacco Nicotiana tabacum were obtained with each genetic structure. GUS protein activity assay in extracts from transgenic plants showed that all deletion variants of the promoter, except -290bp, expressed the gusA gene. In most transgenic plants, the GUS activity level was comparable or higher than in plants with the viral promoter CaMV 35S. GUS activity remains high in progenies and its level correlates positively with the amount of gusA gene mRNA in T3 homozygous plants. The activity of the ro-SmAMP2 promoter was detected in all organs of the transgenic plants studied, during meiosis and in pollen as well.Our results show that the ro-SmAMP2 promoter can be used for target genes expression control in transgenic plants.

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