The phenyl glycoside myconoside, extracted from Balkan endemic Haberlea rhodopensis, has a positi... more The phenyl glycoside myconoside, extracted from Balkan endemic Haberlea rhodopensis, has a positive effect on human health, but the exact molecular mechanism of its action is still unknown. The cell membrane and its associated junctional complex are the first targets of exogenous compound action. We aimed to study the effect of myconoside on membrane organization and cytoskeleton components involved in the maintenance of cell polarity in the MDCKII cell line. By fluorescent spectroscopy and microscopy, we found that at low concentrations, myconoside increases the cell viability by enhancing membrane lipid order and adherent junctions. The opposite effect is observed in high myconoside doses. We hypothesized that the cell morphological and physicochemical changes of the analyzed cell compartments are directly related to cell viability and cell apical-basal polarity. Our finding contributes to a better understanding of the beneficial application of phytochemical myconoside in pharmaco...
Studies have been carried out on the effects of the phenyl glycoside myconoside, extracted from t... more Studies have been carried out on the effects of the phenyl glycoside myconoside, extracted from the relict, Balkan endemic resurrection plant Haberlea rhodopensis on the plasma membrane structural organization and the actin cytoskeleton. Because the plasma membrane is the first target of exogenous bioactive compounds, we focused our attention on the influence of myconoside on the membrane lipid order and actin cytoskeleton in human lung adenocarcinoma A549 cells, using fluorescent spectroscopy and microscopy techniques. We found that low myconoside concentration (5 μg/ml) did not change cell viability but was able to increase plasma membrane lipid order of the treated cells. Higher myconoside concentration (20 μg/ml) inhibited cell viability by decreasing plasma membrane lipid order and impairing actin cytoskeleton. We hypothesize that the observed changes in the plasma membrane structural organization and the actin cytoskeleton are functionally connected to cell viability. Biomimetic membranes were used to demonstrate that myconoside is able to reorganize the membrane lipids by changing the fraction of sphingomyelin-cholesterol enriched domains. Thus, we propose a putative mechanism of action of myconoside on A549 cells plasma membrane lipids as well as on actin filaments in order to explain its cytotoxic effect at high myconoside concentration.
Atanas G. Atanasov and Dongdong Wang acknowledge the support by the Polish KNOW (Leading National... more Atanas G. Atanasov and Dongdong Wang acknowledge the support by the Polish KNOW (Leading National Research Centre) Scientific Consortium “Healthy Animal - Safe Food,” decision of Ministry of Science and Higher Education No. 05-1/KNOW2/2015 and the European Union under the European Regional Development Fund (Homing/2017-4/41). Antoni Sureda was supported by the Institute of Health Carlos III (Project CIBEROBN CB12/03/30038).
The research team of Kobe University’s Graduate School of Agriculture Science, led by Professor D... more The research team of Kobe University’s Graduate School of Agriculture Science, led by Professor Dr Roumiana Tsenkova and a research group from Agrobioinstitute in Sofia, Bulgaria led by Professor Dr Dimitar Djilianov, recently made a significant step forward in understanding the “drying without dying” phenomenon in resurrection plants – a small group of plant species which are able to survive long periods without water. Using aquaphotomics and near infrared spectroscopy, the entire process of desiccation and subsequent rehydration in one such plant – Haberlea rhodopensis was monitored non-destructively and compared with botanically similar, non-resurrection species Deinostigma eberhardtii. The research found that during drying, resurrection plant performs controlled, organized restructuring of water molecular network in its leaves as a preparation for full desiccation which is characterized by accumulation of water molecular dimers and water molecules with four hydrogen bonds, while...
Plant stress responses are complex phenomena encompassing an array of changes at molecular and ce... more Plant stress responses are complex phenomena encompassing an array of changes at molecular and cellular level. They leave imprint on the molecular structure of water in the plant tissues which can be measured in vivo, in a completely non-destructive and reagent-free way, using aquaphotomics near infrared spectroscopy. Analysis of the near infrared spectrum regions, where water absorbs, provides indirect insight into the dynamics and functionality of the whole biological system.In our efforts to perform Responsible Research and Innovations (RRI) practices, we display openly the details of our approach showing the advantages and the eventual difficulties. The protocol described here is developed for non-destructive monitoring of plant response to desiccation stress, particularly focused on the extreme desiccation tolerance of resurrection plants. With alterations in plant material and experimental conditions, the methods described allow detection, monitoring and analysis of plant resp...
Breast cancer is among the most problematic diseases and a leading cause of death in women. The m... more Breast cancer is among the most problematic diseases and a leading cause of death in women. The methods of therapy widely used, so far, are often with many side effects, seriously hampering patients’ quality of life. To overcome these constraints, new cancer treatment alternatives are constantly tested, including bioactive compounds of plant origin. Our aim was to study the effects of Haberlea rhodopensis methanol extract fractions on cell viability and proliferation of two model breast cancer cell lines with different characteristics. In addition to the strong reduction in cell viability, two of the fractions showed significant influence on the proliferation rate of the hormone receptor expressing MCF7 and the triple negative MDA-MB231 breast cancer cell lines. No significant effects on the benign MCF10A cell line were observed. We applied a large scale non-targeted approach to purify and identify highly abundant compounds from the active fractions of H. rhodopensis extracts. By th...
PETROVA, G. and D. DJILIANOV, 2013. Agrobacterium – mediated genetic transformation of the resurr... more PETROVA, G. and D. DJILIANOV, 2013. Agrobacterium – mediated genetic transformation of the resurrection plant Haberlea rhodopensis Friv. Bulg. J. Agric. Sci., Supplement 2, 19: 10–14 Resurrection plants are widely used models for desiccation tolerance studies. Several genes have been successfully isolated from these species. In an attempt to study their role, these genes have been successfully transferred in other model plants. Since resurrection plants are as a rule, polyploids, they are pure targets for mutational studies. In this respect, the establishment of effi cient and repeatable transformation system will contribute signifi cantly for the elucidation of stress tolerance. In this study we describe for the fi rst time, a procedure for Agrobacterium tumefaciens – mediated genetic transformation of the resurrection plant Haberlea rhodopensis Friv. For in vitro regeneration of Haberlea, we used liquid WPM media. It enables us to achieve direct regeneration and transformation sys...
Contemporary agriculture faces the great challenges to meet the increasing demands of the society... more Contemporary agriculture faces the great challenges to meet the increasing demands of the society for food and row materials and the progressing global warming. To solve these problems we need to develop new crop varieties with high yield and highly tolerant to abitoic stress and at the same time to ensure sustainable development of the society and environment. <br> In this respect it is expected that modern biotechnologies will help plant breeding. The development of genetically modified crops with increased abiotic stress tolerance is a priority of many labs in the world. Despite the fact that the presence of such GM forms on the market is negligible at the moment, it is of major importance to develop a specific procedure for risk assessment. Such procedure should cover all stages of GM forms development, release in the environment and participation in food chains.
Haberlea rhodopensis is a Balkan endemic plant that belongs to the Gesneriaceae family, and is be... more Haberlea rhodopensis is a Balkan endemic plant that belongs to the Gesneriaceae family, and is believed to have medicinal use and health-promoting properties. This study aimed to (i) prepare aqueous (HAE) and ethanolic (HEE) extracts from the leaves of H. rhodopensis from in vitro propagated plants, (ii) screen for their potential antiproliferative and antimigratory activities, and (iii) chemically characterize both HAE and HEE by identifying compounds which may contribute to their observed bioactivity thereby further supporting their potential use in biomedical applications. The antiproliferative activity of both extracts was assessed against six human cancer cell lines by employing the sulforhodamine-B (SRB) assay. HEE was found to be more potent in inhibiting cancer cell growth as compared to HAE. Therefore, HEE’s antimigratory effects were further studied in hepatocellular carcinoma (HepG2) and non-small cell lung adenocarcinoma (A459) cell lines as they were among the most sens...
Figure S2. Confirmation of RNA-seq data using qRT-PCR and the KEGG analysis of up- and down-regul... more Figure S2. Confirmation of RNA-seq data using qRT-PCR and the KEGG analysis of up- and down-regulated transcripts. (a) Validation of RNA-seq data using qPCR. (b) KEGG analysis of up- and down-regulated unigenes of each stage. (TIF 721â kb)
Table S3. Expression of genes that were known as dehydration inducible genes (ELIP, GS, beta-amyl... more Table S3. Expression of genes that were known as dehydration inducible genes (ELIP, GS, beta-amylase, catalase, LEA etc.). (XLSX 22â kb)
The phenyl glycoside myconoside, extracted from Balkan endemic Haberlea rhodopensis, has a positi... more The phenyl glycoside myconoside, extracted from Balkan endemic Haberlea rhodopensis, has a positive effect on human health, but the exact molecular mechanism of its action is still unknown. The cell membrane and its associated junctional complex are the first targets of exogenous compound action. We aimed to study the effect of myconoside on membrane organization and cytoskeleton components involved in the maintenance of cell polarity in the MDCKII cell line. By fluorescent spectroscopy and microscopy, we found that at low concentrations, myconoside increases the cell viability by enhancing membrane lipid order and adherent junctions. The opposite effect is observed in high myconoside doses. We hypothesized that the cell morphological and physicochemical changes of the analyzed cell compartments are directly related to cell viability and cell apical-basal polarity. Our finding contributes to a better understanding of the beneficial application of phytochemical myconoside in pharmaco...
Studies have been carried out on the effects of the phenyl glycoside myconoside, extracted from t... more Studies have been carried out on the effects of the phenyl glycoside myconoside, extracted from the relict, Balkan endemic resurrection plant Haberlea rhodopensis on the plasma membrane structural organization and the actin cytoskeleton. Because the plasma membrane is the first target of exogenous bioactive compounds, we focused our attention on the influence of myconoside on the membrane lipid order and actin cytoskeleton in human lung adenocarcinoma A549 cells, using fluorescent spectroscopy and microscopy techniques. We found that low myconoside concentration (5 μg/ml) did not change cell viability but was able to increase plasma membrane lipid order of the treated cells. Higher myconoside concentration (20 μg/ml) inhibited cell viability by decreasing plasma membrane lipid order and impairing actin cytoskeleton. We hypothesize that the observed changes in the plasma membrane structural organization and the actin cytoskeleton are functionally connected to cell viability. Biomimetic membranes were used to demonstrate that myconoside is able to reorganize the membrane lipids by changing the fraction of sphingomyelin-cholesterol enriched domains. Thus, we propose a putative mechanism of action of myconoside on A549 cells plasma membrane lipids as well as on actin filaments in order to explain its cytotoxic effect at high myconoside concentration.
Atanas G. Atanasov and Dongdong Wang acknowledge the support by the Polish KNOW (Leading National... more Atanas G. Atanasov and Dongdong Wang acknowledge the support by the Polish KNOW (Leading National Research Centre) Scientific Consortium “Healthy Animal - Safe Food,” decision of Ministry of Science and Higher Education No. 05-1/KNOW2/2015 and the European Union under the European Regional Development Fund (Homing/2017-4/41). Antoni Sureda was supported by the Institute of Health Carlos III (Project CIBEROBN CB12/03/30038).
The research team of Kobe University’s Graduate School of Agriculture Science, led by Professor D... more The research team of Kobe University’s Graduate School of Agriculture Science, led by Professor Dr Roumiana Tsenkova and a research group from Agrobioinstitute in Sofia, Bulgaria led by Professor Dr Dimitar Djilianov, recently made a significant step forward in understanding the “drying without dying” phenomenon in resurrection plants – a small group of plant species which are able to survive long periods without water. Using aquaphotomics and near infrared spectroscopy, the entire process of desiccation and subsequent rehydration in one such plant – Haberlea rhodopensis was monitored non-destructively and compared with botanically similar, non-resurrection species Deinostigma eberhardtii. The research found that during drying, resurrection plant performs controlled, organized restructuring of water molecular network in its leaves as a preparation for full desiccation which is characterized by accumulation of water molecular dimers and water molecules with four hydrogen bonds, while...
Plant stress responses are complex phenomena encompassing an array of changes at molecular and ce... more Plant stress responses are complex phenomena encompassing an array of changes at molecular and cellular level. They leave imprint on the molecular structure of water in the plant tissues which can be measured in vivo, in a completely non-destructive and reagent-free way, using aquaphotomics near infrared spectroscopy. Analysis of the near infrared spectrum regions, where water absorbs, provides indirect insight into the dynamics and functionality of the whole biological system.In our efforts to perform Responsible Research and Innovations (RRI) practices, we display openly the details of our approach showing the advantages and the eventual difficulties. The protocol described here is developed for non-destructive monitoring of plant response to desiccation stress, particularly focused on the extreme desiccation tolerance of resurrection plants. With alterations in plant material and experimental conditions, the methods described allow detection, monitoring and analysis of plant resp...
Breast cancer is among the most problematic diseases and a leading cause of death in women. The m... more Breast cancer is among the most problematic diseases and a leading cause of death in women. The methods of therapy widely used, so far, are often with many side effects, seriously hampering patients’ quality of life. To overcome these constraints, new cancer treatment alternatives are constantly tested, including bioactive compounds of plant origin. Our aim was to study the effects of Haberlea rhodopensis methanol extract fractions on cell viability and proliferation of two model breast cancer cell lines with different characteristics. In addition to the strong reduction in cell viability, two of the fractions showed significant influence on the proliferation rate of the hormone receptor expressing MCF7 and the triple negative MDA-MB231 breast cancer cell lines. No significant effects on the benign MCF10A cell line were observed. We applied a large scale non-targeted approach to purify and identify highly abundant compounds from the active fractions of H. rhodopensis extracts. By th...
PETROVA, G. and D. DJILIANOV, 2013. Agrobacterium – mediated genetic transformation of the resurr... more PETROVA, G. and D. DJILIANOV, 2013. Agrobacterium – mediated genetic transformation of the resurrection plant Haberlea rhodopensis Friv. Bulg. J. Agric. Sci., Supplement 2, 19: 10–14 Resurrection plants are widely used models for desiccation tolerance studies. Several genes have been successfully isolated from these species. In an attempt to study their role, these genes have been successfully transferred in other model plants. Since resurrection plants are as a rule, polyploids, they are pure targets for mutational studies. In this respect, the establishment of effi cient and repeatable transformation system will contribute signifi cantly for the elucidation of stress tolerance. In this study we describe for the fi rst time, a procedure for Agrobacterium tumefaciens – mediated genetic transformation of the resurrection plant Haberlea rhodopensis Friv. For in vitro regeneration of Haberlea, we used liquid WPM media. It enables us to achieve direct regeneration and transformation sys...
Contemporary agriculture faces the great challenges to meet the increasing demands of the society... more Contemporary agriculture faces the great challenges to meet the increasing demands of the society for food and row materials and the progressing global warming. To solve these problems we need to develop new crop varieties with high yield and highly tolerant to abitoic stress and at the same time to ensure sustainable development of the society and environment. <br> In this respect it is expected that modern biotechnologies will help plant breeding. The development of genetically modified crops with increased abiotic stress tolerance is a priority of many labs in the world. Despite the fact that the presence of such GM forms on the market is negligible at the moment, it is of major importance to develop a specific procedure for risk assessment. Such procedure should cover all stages of GM forms development, release in the environment and participation in food chains.
Haberlea rhodopensis is a Balkan endemic plant that belongs to the Gesneriaceae family, and is be... more Haberlea rhodopensis is a Balkan endemic plant that belongs to the Gesneriaceae family, and is believed to have medicinal use and health-promoting properties. This study aimed to (i) prepare aqueous (HAE) and ethanolic (HEE) extracts from the leaves of H. rhodopensis from in vitro propagated plants, (ii) screen for their potential antiproliferative and antimigratory activities, and (iii) chemically characterize both HAE and HEE by identifying compounds which may contribute to their observed bioactivity thereby further supporting their potential use in biomedical applications. The antiproliferative activity of both extracts was assessed against six human cancer cell lines by employing the sulforhodamine-B (SRB) assay. HEE was found to be more potent in inhibiting cancer cell growth as compared to HAE. Therefore, HEE’s antimigratory effects were further studied in hepatocellular carcinoma (HepG2) and non-small cell lung adenocarcinoma (A459) cell lines as they were among the most sens...
Figure S2. Confirmation of RNA-seq data using qRT-PCR and the KEGG analysis of up- and down-regul... more Figure S2. Confirmation of RNA-seq data using qRT-PCR and the KEGG analysis of up- and down-regulated transcripts. (a) Validation of RNA-seq data using qPCR. (b) KEGG analysis of up- and down-regulated unigenes of each stage. (TIF 721â kb)
Table S3. Expression of genes that were known as dehydration inducible genes (ELIP, GS, beta-amyl... more Table S3. Expression of genes that were known as dehydration inducible genes (ELIP, GS, beta-amylase, catalase, LEA etc.). (XLSX 22â kb)
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