Katarzyna Hnatuszko-Konka | University of Łódz (original) (raw)

Papers by Katarzyna Hnatuszko-Konka

Additional file 1: Fig. S1. A circular plasmid map of the modified pCAMBIA 1305.2 vector. Fig. S2... more Additional file 1: Fig. S1. A circular plasmid map of the modified pCAMBIA 1305.2 vector. Fig. S2. The regeneration from the Pionier cauliflower hypocotyls after R. radiobacter-mediated transformation approach: left side (a, c, e, g) after EHA 105 treatment, right side (b, d, f, h) after LBA 4404 treatment. a, b – growth of the hypocotyls on the selection medium I°; c, d – 10-day growth of the hypocotyls on the selection medium II° (shoot induction); e, f – 2-month shoot regeneration on the selection medium II° (all the regenerated shoots are gathered on one Petri dish per given strain); g, h – the effects of the GUS assay on the Pionier cauliflower hypocotyls after R. radiobacter-mediated transformation approach (EHA 105 and LBA 4404 strain respectively) .Table S1. The characteristic of the in vitro response of the Pionier hypocotyl explants to different variants of regeneration media. Table S2. The characteristics of the in vitro response of the Pionier cotyledon explants to diffe...

and two types of explant (hypocotyl and cotyledon) were tested for their potential to regenerate ... more and two types of explant (hypocotyl and cotyledon) were tested for their potential to regenerate under in vitro con-ditions. Hypocotyl and cotyledon explants from 10-d-old seedlings were subcultured onto different callus induction media based on Murashige and Skoog (MS) basal medium supplemented with 1 % sucrose and different concentra-tions and combinations of plant growth regulators. Hypo-cotyl explants were found to be more suitable for callus induction and organogenesis than cotyledon explants for all cultivars tested. In terms of regeneration, the cv. ‘Amager ’ was significantly more responsive than the other cultivars tested and produced the highest number of shoots/ buds per explant. Moreover, among five types of media tested, MS+8.88 μM 6-benzyloaminopurine (BAP)+ 0.53 μM α-naphthylacetic acid (NAA) was most effective for shoot regeneration. Rooting was achieved within 10– 15 d on all the rooting media, but MS medium containing 5.37 μM NAA produced the maximum number of stro...

plasminogen activator of bacterial origin, in potato (Solanum tuberosum L.) plants

Recombinant proteins can be expressed in transformed cell cultures of bacteria, yeasts, molds, ma... more Recombinant proteins can be expressed in transformed cell cultures of bacteria, yeasts, molds, mammals, plants, insects, or via transgenic plants and animals. Numerous factors influence quality, functionality, yield and protein production rate, so the choice of appropriate expression system is of primary importance. During last few years, plants have

International Journal of Molecular Sciences

The aim of this work was to determine the effect of stress conditions caused by different light s... more The aim of this work was to determine the effect of stress conditions caused by different light sources, i.e., blue LED (λ = 430 nm), red LED (λ = 670 nm), blue and red LED (70%:30%) and white LED (430–670 nm) on the growth and morphology of cultivated in vitro Dracocephalum forrestii shoot culture. It also examines the effects on bioactive phenolic compound production and photosynthetic pigment content, as well as on antioxidant enzyme activity (CAT, SOD, POD) and antioxidant properties. The most beneficial proliferation effect was observed under white LEDs (7.1 ± 2.1 shoots per explant). The white and blue lights stimulated the highest fresh weight gain, while red light induced the highest dry weight gain. The total phenolic acid content ranged from 13.824 ± 1.181 to 20.018 ± 801 mg g DW−1 depending on light conditions. The highest content of rosmarinic acid was found in the control shoots (cultivated under fluorescent lamps), followed by culture grown under red light. All LED tre...

Biotechnologia. Journal of Biotechnology, Computational Biology and Bionanotechnology, 2013

... Katarzyna HNATUSZKO-KONKA1, Piotr ŁUCHNIAK1, Aneta WIKTOREK-SMAGUR2, Aneta GERSZBERG1, Tomasz... more ... Katarzyna HNATUSZKO-KONKA1, Piotr ŁUCHNIAK1, Aneta WIKTOREK-SMAGUR2, Aneta GERSZBERG1, Tomasz KOWALCZYK1, Andrzej K. KONONOWICZ1 ... [31] RAKOCZY-TROJANOWSKA M. Alternative methods of plant transformation – a short review. ...

Genes

The ability to restore or replace injured tissues can be undoubtedly named among the most spectac... more The ability to restore or replace injured tissues can be undoubtedly named among the most spectacular achievements of plant organisms. One of such regeneration pathways is organogenesis, the formation of individual organs from nonmeristematic tissue sections. The process can be triggered in vitro by incubation on medium supplemented with phytohormones. Cytokinins are a class of phytohormones demonstrating pleiotropic effects and a powerful network of molecular interactions. The present study reviews existing knowledge on the possible sequence of molecular and genetic events behind de novo shoot organogenesis initiated by cytokinins. Overall, the review aims to collect reactions encompassed by cytokinin primary responses, starting from phytohormone perception by the dedicated receptors, to transcriptional reprogramming of cell fate by the last module of multistep-phosphorelays. It also includes a brief reminder of other control mechanisms, such as epigenetic reprogramming.

Biomolecules

Salvia bulleyana is a rare Chinese medicinal plant that due to the presence of polyphenols lowers... more Salvia bulleyana is a rare Chinese medicinal plant that due to the presence of polyphenols lowers the risk of some chronic diseases especially those related to the cardiovascular system. The present study examines the organogenic competence of various combinations and concentrations of plant growth regulators to develop an efficient protocol for in vitro regeneration of S. bulleyana via leaf explants, maintaining the high production of active constituents. The purpose of the study was also to assess the possibilities of using a cytokinin-based regeneration to effectively produce therapeutic compounds. The adventitious shoot formation was observed through direct organogenesis on media with purine derivatives (meta-topolin, mT and benzylaminopurine, BAP), and through indirect organogenesis on media with urea derivatives (tidiazuron, TDZ and forchlorfenuron, CPPU). The highest regeneration frequency (95%) with 5.2 shoots per explant was obtained on leaves cultured on Murashige and Skoo...

Scientific Reports

the tissue culture of Phaseolus vulgaris has always been considered difficult. Its regenerative c... more the tissue culture of Phaseolus vulgaris has always been considered difficult. Its regenerative capacity and response to culture conditions are highly genotype-dependent and hamper the application of genetic engineering. The objective of this study was to develop a repeatable technique for organogenic bud induction from selected explants of the common bean. Epicotyls and hypocotyls of six cultivars were investigated to determine the effect of the genotype, and four variants of two basal media (Murashige-Skoog and Gamborg) were tested. The composition of these medium variants was based on the published data suggesting the most universal medium compounds that show the advantage of being applicable to different cultivars. As a result, the common bean epicotyls showed undisputed regeneration superiority over the hypocotyls. Moreover, a well-known observation was confirmed, namely that common bean regeneration is cultivar-specific or at least specific to the cluster of related cultivars. However, efficient regeneration was achieved most often when the epicotyls were incubated on the MS or B5 media amended with AgNo 3 and BAP. Additionally, the positive synergistic influence of activated charcoal and silver nitrate on bud formation was demonstrated. The highest values of the epicotyl in vitro response for the common bean cultivars could be presented as follows: Czerwona (70.00%) > Goldpantera (58.89%) and Ibiza (58.89%) > Plus (55.56%) > Laponia (50.56%) > Złota Saxa (46.11%). The common bean is the most widely cultivated of Phaseolus species worldwide, since it is a very important source of protein and calories. This warm-season legume also plays an important role as a natural system of nitrogen-fixation. However, a number of biotic and abiotic stresses severely affect the yield of this crop. In vitro regeneration of the common bean is frequently considered to be difficult, which significantly hampers progress in the transformation. Phaseolus vulgaris displays extremely high diversity regarding the response to tissue culture conditions, hence a cultivar/genotype-specific protocol of regeneration is suggested 1. It was demonstrated, for example, in the contrasting results reported by Delgado-Sanchez et al. 2 and Gatica Arias et al. 3 regarding adenine usage. Of course, this genotype-dependent response is also determined by the type or state of the explant, cell or tissue specialization and by the cultivation conditions 4. Nevertheless, the common bean was regenerated via organogenic 5-7 and embryogenic pathways 8-10 from different plant explants: intact seedlings, embryonic axes, cotyledons, cotyledonary nodes, leaf petioles, internodes, auxiliary shoots, hypocotyls or leaves 3,11-13. However, plant regeneration employing the published protocols remains difficult to reproduce. The objective of the reported study was to develop a repeatable technique for organogenic bud induction from selected explants of young and mature plants of the common bean. However, the assumption was made that different published protocols could be retested to optimize them and check their usefulness regarding chosen cultivars and explants. The plant material included six varieties: Czerwona, Plus, Laponia, Złota Saxa, Ibiza and Goldpantera (the last four are listed in The Common Catalogue of Varieties of Vegetable Species, 35th complete edition, 2016) 14. The studies were conducted on randomly chosen cultivars as a starting point, following Malmberg's 15 suggestion, assuming that the wide screening of genetic lines might become a base of knowledge for plant regeneration 1,4. It should be noted here that different parts of young and mature plants were tested in

Biomolecules

The present study demonstrates hormonal control of Salvia viridis growth and development using fo... more The present study demonstrates hormonal control of Salvia viridis growth and development using four different purine-type cytokinins at different concentrations. The addition of cytokinins significantly increased biomass of cultures, proliferation rate, and, interestingly, secondary metabolite production. The best response in terms of multiplication ratio was recorded on Murashige and Skoog medium supplemented with 0.5 mg/L BPA (N-benzylotetrahydropyranyl adenine), while the greatest biomass accumulation was achieved when supplemented with 1 mg/L m-T (meta-topoline). Quantitative UPLC-DAD analysis of the hydromethanolic extract from S. viridis culture revealed the presence of 12 polyphenols: seven phenolic acids and five phenylethanoids. The highest total content of polyphenolic compounds was found in shoots cultivated on medium with 2 mg/L BPA (18.66 mg/g DW): almost twice that of control shoots. The medium was also the most optimal for the biosynthesis of rosmarinic acid, the pred...

Current Pharmaceutical Biotechnology

BACKGROUND This review is intended to draw the attention of pharmaceutical and biotechnological c... more BACKGROUND This review is intended to draw the attention of pharmaceutical and biotechnological communities to the untapped potential of the Scutellaria genus. Skullcaps, as they are more widely known, are found in one of the oldest materia medica in the world, that of ancient Chinese pharmacology, and their numerous wide range of medicinal bioactivities have been studied both in vivo and in vitro. For thousands of years, chemical compounds from the Scutellaria species have been safely used as antitumor, antibacterial, antiviral, anti-inflammatory, antioxidant or hepatoprotective factors. OBJECTIVE As these effects are well known, reflected in the presence of Scutellaria plants in national pharmacopoeias, it is clear that the plant has yet enormous unexploited potential. The European pharmacological market has turned to the resources of Scutellaria only in the last two decades, and although the construction and clinical processing of a new drug is a long process, the general impression is that very few medical products in pharmacies have been inspired by the phytochemistry of skullcaps. CONCLUSION This paper presents the current state of knowledge on the wealth of Scutellaria chemical compounds with treatment applications, its tissue culture and biotechnological achievements, especially in the context of the production of secondary metabolites.

AMB Express, Jan 6, 2018

Brassica oleracea var. botrytis, a very popular crop grown for its edible inflorescence, is bred ... more Brassica oleracea var. botrytis, a very popular crop grown for its edible inflorescence, is bred only as a mutated annual cultivar and does not naturally occur in environment. Since cauliflower is still described as the most troublesome of all the B. oleracea vegetables regarding transformation processes, it is fully justified to focus on the improvement of tools for its genetic modifications. Here, we present a successful protocol for genetic transformation of cauliflower employing the process of agroinfection. The primary analysis of in vitro response of five cultivars allowed us to have chosen Pionier as the most promising cultivar; in consequence the Pionier was transformed via Rhizobium-mediated techniques in order to evaluate both, R. radiobacter (EHA 105, LBA 4404) and R. rhizogenes (ATCC 18534, A4) species. However, the latter system turned out to be more effective and, the A4 strain, in particular (72% transformation efficiency, 55% confirmed by GUS assay). That shows a pro...

Plant Cell, Tissue and Organ Culture (PCTOC), 2016

Plant biotechnology is a dynamically developing science, which comprises many fields of knowledge... more Plant biotechnology is a dynamically developing science, which comprises many fields of knowledge. Novel plant genetic engineering findings highly influence the improvement of industrial production. These findings mostly concern cis-regulatory elements, which are sequences controlling gene expression at all developmental stages. They comprise of promoters, enhancers, insulators and silencers, which are used to construct synthetic expression cassettes. Examples of most important cis-regulatory elements are reviewed in the present paper. Variability among core promoters content and distal promoter regions impedes evaluation of interactions between them during the artificial promoters construction. Synthetic promoters and artificial expression cassettes trigger a significant increase in gene expression level, better properties and quality of a product. Accumulating knowledge about gene promoters, cis sequences and their cooperating factors allows uniform expression systems and highly predictable results.

World Journal of Microbiology and Biotechnology, 2016

Here, we present the application of microbiology and biotechnology for the production of recombin... more Here, we present the application of microbiology and biotechnology for the production of recombinant pharmaceutical proteins in plant cells. To the best of our knowledge and belief it is one of few examples of the expression of the prokaryotic staphylokinase (SAK) in the eukaryotic system. Despite the tremendous progress made in the plant biotechnology, most of the heterologous proteins still accumulate to low concentrations in plant tissues. Therefore, the composition of expression cassettes to assure economically feasible level of protein production in plants remains crucial. The aim of our research was obtaining a high concentration of the bacterial anticoagulant factor-staphylokinase, in Arabidopsis thaliana seeds. The coding sequence of staphylokinase was placed under control of the b-phaseolin promoter and cloned between the signal sequence of the seed storage protein 2S2 and the carboxy-terminal KDEL signal sequence. The engineered binary vector pATAG-sak was introduced into Arabidopsis thaliana plants via Agrobacterium tumefaciens-mediated transformation. Analysis of the subsequent generations of Arabidopsis seeds revealed both presence of the sak and nptII transgenes, and the SAK protein. Moreover, a plasminogen activator activity of staphylokinase was observed in the protein extracts from seeds, while such a reaction was not observed in the leaf extracts showing seed-specific activity of the b-phaseolin promoter.

Arabidopsis has been known to naturalists under various names since the sixteenth century, and it... more Arabidopsis has been known to naturalists under various names since the sixteenth century, and its "laboratory career" is more than 100 years long. Arabidopsis is the most thoroughly studied model organism in plant biology. It has become a major model system for investigating molecular, genetic and biochemical processes in plants. Genetic transformation of Arabidopsis by Agrobacterium caused many difficulties, even though the plant is clearly appropriate for any genetic manipulation. An important breakthrough in the methodology of genetic plant transformation has demonstrated that Arabidopsis is able to produce transgenic plants with no need to use in vitro cultures. This method is called in planta transformation. For Arabidopsis this transformation method is now routinely employed using the relatively low-tech approach of floral dipping. Floral dip method is very simple to perform. It requires minimal labor and no plant tissue culture and is easy, cheap and has a quick pr...

Plant Cell, Tissue and Organ Culture (PCTOC), 2014

Originating in the Andes, the tomato (Solanum lycopersicum L.) was imported to Europe in the 16th... more Originating in the Andes, the tomato (Solanum lycopersicum L.) was imported to Europe in the 16th century. At present, it is an important crop plant cultivated all over the world, and its production and consumption continue to increase. This popular vegetable is known as a major source of important nutrients including lycopene, bcarotene, flavonoids and vitamin C as well as hydroxycinnamic acid derivatives. Since the discovery that lycopene has anti-oxidative, anti-cancer properties, interest in tomatoes has grown rapidly. The development of genetic engineering tools and plant biotechnology has opened great opportunities for engineering tomato plants. This review presents examples of successful tissue culture and genetically modified tomatoes which resistance to a range of environmental stresses improved, along with fruit quality. Additionally, a successful molecular farming model was established.

Biotechnology Advances, 2014

Since the ability to genetically engineer plants was established, researchers have modified a gre... more Since the ability to genetically engineer plants was established, researchers have modified a great number of plant species to satisfy agricultural, horticultural, industrial, medicinal or veterinary requirements. Almost thirty years after the first approaches to the genetic modification of pulse crops, it is possible to transform many grain legumes. However, one of the most important species for human nutrition, Phaseolus vulgaris, still lacks some practical tools for genomic research, such as routine genetic transformation. Its recalcitrance towards in vitro regeneration and rooting significantly hampers the possibilities of improvement of the common bean that suffers from many biotic and abiotic constraints. Thus, an efficient and reproducible system for regeneration of a whole plant is desired. Although noticeable progress has been made, the rate of recovery of transgenic lines is still low. Here, the current status of tissue culture and recent progress in transformation methodology are presented. Some major challenges and obstacles are discussed and some examples of their solutions are presented.

Additional file 1: Fig. S1. A circular plasmid map of the modified pCAMBIA 1305.2 vector. Fig. S2... more Additional file 1: Fig. S1. A circular plasmid map of the modified pCAMBIA 1305.2 vector. Fig. S2. The regeneration from the Pionier cauliflower hypocotyls after R. radiobacter-mediated transformation approach: left side (a, c, e, g) after EHA 105 treatment, right side (b, d, f, h) after LBA 4404 treatment. a, b – growth of the hypocotyls on the selection medium I°; c, d – 10-day growth of the hypocotyls on the selection medium II° (shoot induction); e, f – 2-month shoot regeneration on the selection medium II° (all the regenerated shoots are gathered on one Petri dish per given strain); g, h – the effects of the GUS assay on the Pionier cauliflower hypocotyls after R. radiobacter-mediated transformation approach (EHA 105 and LBA 4404 strain respectively) .Table S1. The characteristic of the in vitro response of the Pionier hypocotyl explants to different variants of regeneration media. Table S2. The characteristics of the in vitro response of the Pionier cotyledon explants to diffe...

and two types of explant (hypocotyl and cotyledon) were tested for their potential to regenerate ... more and two types of explant (hypocotyl and cotyledon) were tested for their potential to regenerate under in vitro con-ditions. Hypocotyl and cotyledon explants from 10-d-old seedlings were subcultured onto different callus induction media based on Murashige and Skoog (MS) basal medium supplemented with 1 % sucrose and different concentra-tions and combinations of plant growth regulators. Hypo-cotyl explants were found to be more suitable for callus induction and organogenesis than cotyledon explants for all cultivars tested. In terms of regeneration, the cv. ‘Amager ’ was significantly more responsive than the other cultivars tested and produced the highest number of shoots/ buds per explant. Moreover, among five types of media tested, MS+8.88 μM 6-benzyloaminopurine (BAP)+ 0.53 μM α-naphthylacetic acid (NAA) was most effective for shoot regeneration. Rooting was achieved within 10– 15 d on all the rooting media, but MS medium containing 5.37 μM NAA produced the maximum number of stro...

plasminogen activator of bacterial origin, in potato (Solanum tuberosum L.) plants

Recombinant proteins can be expressed in transformed cell cultures of bacteria, yeasts, molds, ma... more Recombinant proteins can be expressed in transformed cell cultures of bacteria, yeasts, molds, mammals, plants, insects, or via transgenic plants and animals. Numerous factors influence quality, functionality, yield and protein production rate, so the choice of appropriate expression system is of primary importance. During last few years, plants have

International Journal of Molecular Sciences

The aim of this work was to determine the effect of stress conditions caused by different light s... more The aim of this work was to determine the effect of stress conditions caused by different light sources, i.e., blue LED (λ = 430 nm), red LED (λ = 670 nm), blue and red LED (70%:30%) and white LED (430–670 nm) on the growth and morphology of cultivated in vitro Dracocephalum forrestii shoot culture. It also examines the effects on bioactive phenolic compound production and photosynthetic pigment content, as well as on antioxidant enzyme activity (CAT, SOD, POD) and antioxidant properties. The most beneficial proliferation effect was observed under white LEDs (7.1 ± 2.1 shoots per explant). The white and blue lights stimulated the highest fresh weight gain, while red light induced the highest dry weight gain. The total phenolic acid content ranged from 13.824 ± 1.181 to 20.018 ± 801 mg g DW−1 depending on light conditions. The highest content of rosmarinic acid was found in the control shoots (cultivated under fluorescent lamps), followed by culture grown under red light. All LED tre...

Biotechnologia. Journal of Biotechnology, Computational Biology and Bionanotechnology, 2013

... Katarzyna HNATUSZKO-KONKA1, Piotr ŁUCHNIAK1, Aneta WIKTOREK-SMAGUR2, Aneta GERSZBERG1, Tomasz... more ... Katarzyna HNATUSZKO-KONKA1, Piotr ŁUCHNIAK1, Aneta WIKTOREK-SMAGUR2, Aneta GERSZBERG1, Tomasz KOWALCZYK1, Andrzej K. KONONOWICZ1 ... [31] RAKOCZY-TROJANOWSKA M. Alternative methods of plant transformation – a short review. ...

Genes

The ability to restore or replace injured tissues can be undoubtedly named among the most spectac... more The ability to restore or replace injured tissues can be undoubtedly named among the most spectacular achievements of plant organisms. One of such regeneration pathways is organogenesis, the formation of individual organs from nonmeristematic tissue sections. The process can be triggered in vitro by incubation on medium supplemented with phytohormones. Cytokinins are a class of phytohormones demonstrating pleiotropic effects and a powerful network of molecular interactions. The present study reviews existing knowledge on the possible sequence of molecular and genetic events behind de novo shoot organogenesis initiated by cytokinins. Overall, the review aims to collect reactions encompassed by cytokinin primary responses, starting from phytohormone perception by the dedicated receptors, to transcriptional reprogramming of cell fate by the last module of multistep-phosphorelays. It also includes a brief reminder of other control mechanisms, such as epigenetic reprogramming.

Biomolecules

Salvia bulleyana is a rare Chinese medicinal plant that due to the presence of polyphenols lowers... more Salvia bulleyana is a rare Chinese medicinal plant that due to the presence of polyphenols lowers the risk of some chronic diseases especially those related to the cardiovascular system. The present study examines the organogenic competence of various combinations and concentrations of plant growth regulators to develop an efficient protocol for in vitro regeneration of S. bulleyana via leaf explants, maintaining the high production of active constituents. The purpose of the study was also to assess the possibilities of using a cytokinin-based regeneration to effectively produce therapeutic compounds. The adventitious shoot formation was observed through direct organogenesis on media with purine derivatives (meta-topolin, mT and benzylaminopurine, BAP), and through indirect organogenesis on media with urea derivatives (tidiazuron, TDZ and forchlorfenuron, CPPU). The highest regeneration frequency (95%) with 5.2 shoots per explant was obtained on leaves cultured on Murashige and Skoo...

Scientific Reports

the tissue culture of Phaseolus vulgaris has always been considered difficult. Its regenerative c... more the tissue culture of Phaseolus vulgaris has always been considered difficult. Its regenerative capacity and response to culture conditions are highly genotype-dependent and hamper the application of genetic engineering. The objective of this study was to develop a repeatable technique for organogenic bud induction from selected explants of the common bean. Epicotyls and hypocotyls of six cultivars were investigated to determine the effect of the genotype, and four variants of two basal media (Murashige-Skoog and Gamborg) were tested. The composition of these medium variants was based on the published data suggesting the most universal medium compounds that show the advantage of being applicable to different cultivars. As a result, the common bean epicotyls showed undisputed regeneration superiority over the hypocotyls. Moreover, a well-known observation was confirmed, namely that common bean regeneration is cultivar-specific or at least specific to the cluster of related cultivars. However, efficient regeneration was achieved most often when the epicotyls were incubated on the MS or B5 media amended with AgNo 3 and BAP. Additionally, the positive synergistic influence of activated charcoal and silver nitrate on bud formation was demonstrated. The highest values of the epicotyl in vitro response for the common bean cultivars could be presented as follows: Czerwona (70.00%) > Goldpantera (58.89%) and Ibiza (58.89%) > Plus (55.56%) > Laponia (50.56%) > Złota Saxa (46.11%). The common bean is the most widely cultivated of Phaseolus species worldwide, since it is a very important source of protein and calories. This warm-season legume also plays an important role as a natural system of nitrogen-fixation. However, a number of biotic and abiotic stresses severely affect the yield of this crop. In vitro regeneration of the common bean is frequently considered to be difficult, which significantly hampers progress in the transformation. Phaseolus vulgaris displays extremely high diversity regarding the response to tissue culture conditions, hence a cultivar/genotype-specific protocol of regeneration is suggested 1. It was demonstrated, for example, in the contrasting results reported by Delgado-Sanchez et al. 2 and Gatica Arias et al. 3 regarding adenine usage. Of course, this genotype-dependent response is also determined by the type or state of the explant, cell or tissue specialization and by the cultivation conditions 4. Nevertheless, the common bean was regenerated via organogenic 5-7 and embryogenic pathways 8-10 from different plant explants: intact seedlings, embryonic axes, cotyledons, cotyledonary nodes, leaf petioles, internodes, auxiliary shoots, hypocotyls or leaves 3,11-13. However, plant regeneration employing the published protocols remains difficult to reproduce. The objective of the reported study was to develop a repeatable technique for organogenic bud induction from selected explants of young and mature plants of the common bean. However, the assumption was made that different published protocols could be retested to optimize them and check their usefulness regarding chosen cultivars and explants. The plant material included six varieties: Czerwona, Plus, Laponia, Złota Saxa, Ibiza and Goldpantera (the last four are listed in The Common Catalogue of Varieties of Vegetable Species, 35th complete edition, 2016) 14. The studies were conducted on randomly chosen cultivars as a starting point, following Malmberg's 15 suggestion, assuming that the wide screening of genetic lines might become a base of knowledge for plant regeneration 1,4. It should be noted here that different parts of young and mature plants were tested in

Biomolecules

The present study demonstrates hormonal control of Salvia viridis growth and development using fo... more The present study demonstrates hormonal control of Salvia viridis growth and development using four different purine-type cytokinins at different concentrations. The addition of cytokinins significantly increased biomass of cultures, proliferation rate, and, interestingly, secondary metabolite production. The best response in terms of multiplication ratio was recorded on Murashige and Skoog medium supplemented with 0.5 mg/L BPA (N-benzylotetrahydropyranyl adenine), while the greatest biomass accumulation was achieved when supplemented with 1 mg/L m-T (meta-topoline). Quantitative UPLC-DAD analysis of the hydromethanolic extract from S. viridis culture revealed the presence of 12 polyphenols: seven phenolic acids and five phenylethanoids. The highest total content of polyphenolic compounds was found in shoots cultivated on medium with 2 mg/L BPA (18.66 mg/g DW): almost twice that of control shoots. The medium was also the most optimal for the biosynthesis of rosmarinic acid, the pred...

Current Pharmaceutical Biotechnology

BACKGROUND This review is intended to draw the attention of pharmaceutical and biotechnological c... more BACKGROUND This review is intended to draw the attention of pharmaceutical and biotechnological communities to the untapped potential of the Scutellaria genus. Skullcaps, as they are more widely known, are found in one of the oldest materia medica in the world, that of ancient Chinese pharmacology, and their numerous wide range of medicinal bioactivities have been studied both in vivo and in vitro. For thousands of years, chemical compounds from the Scutellaria species have been safely used as antitumor, antibacterial, antiviral, anti-inflammatory, antioxidant or hepatoprotective factors. OBJECTIVE As these effects are well known, reflected in the presence of Scutellaria plants in national pharmacopoeias, it is clear that the plant has yet enormous unexploited potential. The European pharmacological market has turned to the resources of Scutellaria only in the last two decades, and although the construction and clinical processing of a new drug is a long process, the general impression is that very few medical products in pharmacies have been inspired by the phytochemistry of skullcaps. CONCLUSION This paper presents the current state of knowledge on the wealth of Scutellaria chemical compounds with treatment applications, its tissue culture and biotechnological achievements, especially in the context of the production of secondary metabolites.

AMB Express, Jan 6, 2018

Brassica oleracea var. botrytis, a very popular crop grown for its edible inflorescence, is bred ... more Brassica oleracea var. botrytis, a very popular crop grown for its edible inflorescence, is bred only as a mutated annual cultivar and does not naturally occur in environment. Since cauliflower is still described as the most troublesome of all the B. oleracea vegetables regarding transformation processes, it is fully justified to focus on the improvement of tools for its genetic modifications. Here, we present a successful protocol for genetic transformation of cauliflower employing the process of agroinfection. The primary analysis of in vitro response of five cultivars allowed us to have chosen Pionier as the most promising cultivar; in consequence the Pionier was transformed via Rhizobium-mediated techniques in order to evaluate both, R. radiobacter (EHA 105, LBA 4404) and R. rhizogenes (ATCC 18534, A4) species. However, the latter system turned out to be more effective and, the A4 strain, in particular (72% transformation efficiency, 55% confirmed by GUS assay). That shows a pro...

Plant Cell, Tissue and Organ Culture (PCTOC), 2016

Plant biotechnology is a dynamically developing science, which comprises many fields of knowledge... more Plant biotechnology is a dynamically developing science, which comprises many fields of knowledge. Novel plant genetic engineering findings highly influence the improvement of industrial production. These findings mostly concern cis-regulatory elements, which are sequences controlling gene expression at all developmental stages. They comprise of promoters, enhancers, insulators and silencers, which are used to construct synthetic expression cassettes. Examples of most important cis-regulatory elements are reviewed in the present paper. Variability among core promoters content and distal promoter regions impedes evaluation of interactions between them during the artificial promoters construction. Synthetic promoters and artificial expression cassettes trigger a significant increase in gene expression level, better properties and quality of a product. Accumulating knowledge about gene promoters, cis sequences and their cooperating factors allows uniform expression systems and highly predictable results.

World Journal of Microbiology and Biotechnology, 2016

Here, we present the application of microbiology and biotechnology for the production of recombin... more Here, we present the application of microbiology and biotechnology for the production of recombinant pharmaceutical proteins in plant cells. To the best of our knowledge and belief it is one of few examples of the expression of the prokaryotic staphylokinase (SAK) in the eukaryotic system. Despite the tremendous progress made in the plant biotechnology, most of the heterologous proteins still accumulate to low concentrations in plant tissues. Therefore, the composition of expression cassettes to assure economically feasible level of protein production in plants remains crucial. The aim of our research was obtaining a high concentration of the bacterial anticoagulant factor-staphylokinase, in Arabidopsis thaliana seeds. The coding sequence of staphylokinase was placed under control of the b-phaseolin promoter and cloned between the signal sequence of the seed storage protein 2S2 and the carboxy-terminal KDEL signal sequence. The engineered binary vector pATAG-sak was introduced into Arabidopsis thaliana plants via Agrobacterium tumefaciens-mediated transformation. Analysis of the subsequent generations of Arabidopsis seeds revealed both presence of the sak and nptII transgenes, and the SAK protein. Moreover, a plasminogen activator activity of staphylokinase was observed in the protein extracts from seeds, while such a reaction was not observed in the leaf extracts showing seed-specific activity of the b-phaseolin promoter.

Arabidopsis has been known to naturalists under various names since the sixteenth century, and it... more Arabidopsis has been known to naturalists under various names since the sixteenth century, and its "laboratory career" is more than 100 years long. Arabidopsis is the most thoroughly studied model organism in plant biology. It has become a major model system for investigating molecular, genetic and biochemical processes in plants. Genetic transformation of Arabidopsis by Agrobacterium caused many difficulties, even though the plant is clearly appropriate for any genetic manipulation. An important breakthrough in the methodology of genetic plant transformation has demonstrated that Arabidopsis is able to produce transgenic plants with no need to use in vitro cultures. This method is called in planta transformation. For Arabidopsis this transformation method is now routinely employed using the relatively low-tech approach of floral dipping. Floral dip method is very simple to perform. It requires minimal labor and no plant tissue culture and is easy, cheap and has a quick pr...

Plant Cell, Tissue and Organ Culture (PCTOC), 2014

Originating in the Andes, the tomato (Solanum lycopersicum L.) was imported to Europe in the 16th... more Originating in the Andes, the tomato (Solanum lycopersicum L.) was imported to Europe in the 16th century. At present, it is an important crop plant cultivated all over the world, and its production and consumption continue to increase. This popular vegetable is known as a major source of important nutrients including lycopene, bcarotene, flavonoids and vitamin C as well as hydroxycinnamic acid derivatives. Since the discovery that lycopene has anti-oxidative, anti-cancer properties, interest in tomatoes has grown rapidly. The development of genetic engineering tools and plant biotechnology has opened great opportunities for engineering tomato plants. This review presents examples of successful tissue culture and genetically modified tomatoes which resistance to a range of environmental stresses improved, along with fruit quality. Additionally, a successful molecular farming model was established.

Biotechnology Advances, 2014

Since the ability to genetically engineer plants was established, researchers have modified a gre... more Since the ability to genetically engineer plants was established, researchers have modified a great number of plant species to satisfy agricultural, horticultural, industrial, medicinal or veterinary requirements. Almost thirty years after the first approaches to the genetic modification of pulse crops, it is possible to transform many grain legumes. However, one of the most important species for human nutrition, Phaseolus vulgaris, still lacks some practical tools for genomic research, such as routine genetic transformation. Its recalcitrance towards in vitro regeneration and rooting significantly hampers the possibilities of improvement of the common bean that suffers from many biotic and abiotic constraints. Thus, an efficient and reproducible system for regeneration of a whole plant is desired. Although noticeable progress has been made, the rate of recovery of transgenic lines is still low. Here, the current status of tissue culture and recent progress in transformation methodology are presented. Some major challenges and obstacles are discussed and some examples of their solutions are presented.