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Molecular insight into ERF109 function in light regulated retrograde signaling
S164
Abstracts / New Biotechnology 33S (2016) S1–S213
P18-35 Content of oenothein B in clonally propagated Chamaenerion angustifolium (L.) Scop. genotypes ´ Bogna Opala, Mariola Dreger ∗ , Agnieszka Gryszczynska, Przemysław Łukasz Mikołajczak, Karolina Wielgus Institute of Natural Fibres & Medicinal Plants, Poland Rosebay willow herb (Chamaenerion angustifolium (L.) Scop. syn. Epilobium angustifolium L.) from Onagraceae family is a valuable medicinal plant that has been used in the treatment of urogenital disorders including BPH (benign prostatic hypertrophy). Oenothein B is a major constituent and has been regarded as one of main active compounds. The aim of this study was to obtain different genotypes of C. angustifolium in in vitro cultures and determine the content of oenothein B in clonally propagated plants. C. angustifolium seeds from Garden of Medicinal Plants (INF&MP) as well as seeds bought in Rieger-Hofmann GmbH were used for induction of in vitro cultures. Multi-shoots cultures were initiated from root’s explants according to Turker (Turker et al., 2008) in slight modification. Ten C. angustifolium genotypes originated from different individuals were obtained and propagated. Separated shoots were rooted on ½ MS medium with IAA (0.5 mg/L) and vitamin C. Rooted plants were harvested at the age of 5 weeks and dried at room temperature. HPLC method was applied for quantitative and qualitative determination of oenothein B. Content of oenothein oscillated between 3.2% and 1.68% (±0.1%) in dried weight. Results of the study confirmed that plants cultivated in vitro are able to synthesize oenothein B. According to our best knowledge oenothein B was found for the first time in C. angustifolium raw material derived from in vitro cultures. Acknowledgments: The research was financially supported by The National Centre for Research and Development, grant number PBS2/A8/23/2013. http://dx.doi.org/10.1016/j.nbt.2016.06.1287
P18-36 Molecular insight into ERF109 function in light regulated retrograde signaling 2 , Anna Czajkowska 1 , Marcin Anna Barczak 1,∗ , Barbara Karpinska ´ 1 Filipecki 1 2
Warsaw University of Life Sciences – SGGW, Poland University of Leeds, United Kingdom
Plants as a sessile organisms have to endure a wide variety of biotic and abiotic stresses. Fluctuations in light quality and intensity produce diverse signals that provide information to the nucleus enabling appropriate acclimation, development and growth of plants. Changes in redox state of photosynthetic electron transport components were detected allowing to regulate chloroplast and nuclear gene expression and provide coordinated and complex response. In order to discover the components of this regulation the role of ERF109 was analyzed. ERF109 is one of the candidates contributing in locally induced photo-oxidative stress in light-exposed leaves which results in systemic acquired acclimation (SAA) and transcriptional reprogramming. ERF109 is nuclear localized DNA-binding, highly JA-responsive, ethylene response factor which transcripts accumulate in systemic leaves as a result of SAA. Moreover, ERF109 binds directly to GCC-boxes in the promoters of genes participating in auxin biosynthesis to regulate lateral root formation. Despite
its evident involvement in SAA as well as JA and auxin crosstalk, protein partners of ERF109 are still unidentified. To discover ERF109-interacting proteins we performed GST-pull down experiment followed by mass spectrometry analysis, obtaining list of potential interactors. After analysis we chose the most promising 8 candidates which were tested in yeast two hybrid assay. Our results verify the role of ERF109 in light regulated retrograde signaling. http://dx.doi.org/10.1016/j.nbt.2016.06.1288
P18-37 Ploidy level evaluation of highbush blueberry parental genotypes and ther hybrids ∗ , Slanisław Pluta ´ Małgorzata Podwyszynska
Research Institute of Horticulture, Poland Highbush blueberry (Vaccinium corymbosum L.) is grown in Poland on commercial scale recently. With the level of blueberry production of 13.0 thousand metric tons in 2015, Poland currently ranks first in Europe and fifth in the world. Due to the high interest in new cultivars, the Research Institute of Horticulture in Skierniewice started the new blueberry breeding program in 2009. It is believed that the most of highbush blueberry cultivars are tetraploids, but pentaploids and diploids are also known. The ploidy level can affect fertility of parental genotypes as well as fruit and seed formation at hybridization. However, except of three cultivars which are known to be tetraploids, the ploidy level of other parental genotypes selected for our breeding program has been unknown. In our previous study we have observed low seed formation in some crosses. The aim of the present study was to elucidate whether the poor seed production could result different ploidy levels of parental cultivars. Twelve parental genotypes (used for the factorial crossing design) and 100 selected seedlings (hybrids) of V. corymbosum have been used for our studies. Assessment of the ploidy level was performed by the flow cytometry. Cytometric analysis has shown that all the parental cultivars are tetraploids. In case of their hybrids, except of six aneuploids, all other seedlings have been proven to be tetraploids. Since our results demonstrate that all the parental genotypes of V. corymbosum are tetraploids, other causative agent of poor seed production in some crosses should be considered. http://dx.doi.org/10.1016/j.nbt.2016.06.1289
P18-38 Modification of anther selection mode for the successful androgenesis induction in the cassava (Manihot esculenta) anther cultures Elzbieta Maria Golemiec International Center for Tropical Agriculture, Poland Decades of effort dedicated to work on cassava double haploid production protocol establishment, did not result in cassava plant regeneration as in many other species (Wedzony, 2009; Perrera, 2012). Existing protocols still need to be modified. Cassava flower buds size, ranging from 2.3 to 2.5 mm is considered as a most suitable for the androgenic cultures establishment. Performed experiments reviled variability in the tone of isolated anthers: transparent, white and yellowish. The aim of this work was to confirm if the difference in the androgenic answer in the culture depends on the type of plated
Abstracts / New Biotechnology 33S (2016) S1–S213
P18-35 Content of oenothein B in clonally propagated Chamaenerion angustifolium (L.) Scop. genotypes ´ Bogna Opala, Mariola Dreger ∗ , Agnieszka Gryszczynska, Przemysław Łukasz Mikołajczak, Karolina Wielgus Institute of Natural Fibres & Medicinal Plants, Poland Rosebay willow herb (Chamaenerion angustifolium (L.) Scop. syn. Epilobium angustifolium L.) from Onagraceae family is a valuable medicinal plant that has been used in the treatment of urogenital disorders including BPH (benign prostatic hypertrophy). Oenothein B is a major constituent and has been regarded as one of main active compounds. The aim of this study was to obtain different genotypes of C. angustifolium in in vitro cultures and determine the content of oenothein B in clonally propagated plants. C. angustifolium seeds from Garden of Medicinal Plants (INF&MP) as well as seeds bought in Rieger-Hofmann GmbH were used for induction of in vitro cultures. Multi-shoots cultures were initiated from root’s explants according to Turker (Turker et al., 2008) in slight modification. Ten C. angustifolium genotypes originated from different individuals were obtained and propagated. Separated shoots were rooted on ½ MS medium with IAA (0.5 mg/L) and vitamin C. Rooted plants were harvested at the age of 5 weeks and dried at room temperature. HPLC method was applied for quantitative and qualitative determination of oenothein B. Content of oenothein oscillated between 3.2% and 1.68% (±0.1%) in dried weight. Results of the study confirmed that plants cultivated in vitro are able to synthesize oenothein B. According to our best knowledge oenothein B was found for the first time in C. angustifolium raw material derived from in vitro cultures. Acknowledgments: The research was financially supported by The National Centre for Research and Development, grant number PBS2/A8/23/2013. http://dx.doi.org/10.1016/j.nbt.2016.06.1287
P18-36 Molecular insight into ERF109 function in light regulated retrograde signaling 2 , Anna Czajkowska 1 , Marcin Anna Barczak 1,∗ , Barbara Karpinska ´ 1 Filipecki 1 2
Warsaw University of Life Sciences – SGGW, Poland University of Leeds, United Kingdom
Plants as a sessile organisms have to endure a wide variety of biotic and abiotic stresses. Fluctuations in light quality and intensity produce diverse signals that provide information to the nucleus enabling appropriate acclimation, development and growth of plants. Changes in redox state of photosynthetic electron transport components were detected allowing to regulate chloroplast and nuclear gene expression and provide coordinated and complex response. In order to discover the components of this regulation the role of ERF109 was analyzed. ERF109 is one of the candidates contributing in locally induced photo-oxidative stress in light-exposed leaves which results in systemic acquired acclimation (SAA) and transcriptional reprogramming. ERF109 is nuclear localized DNA-binding, highly JA-responsive, ethylene response factor which transcripts accumulate in systemic leaves as a result of SAA. Moreover, ERF109 binds directly to GCC-boxes in the promoters of genes participating in auxin biosynthesis to regulate lateral root formation. Despite
its evident involvement in SAA as well as JA and auxin crosstalk, protein partners of ERF109 are still unidentified. To discover ERF109-interacting proteins we performed GST-pull down experiment followed by mass spectrometry analysis, obtaining list of potential interactors. After analysis we chose the most promising 8 candidates which were tested in yeast two hybrid assay. Our results verify the role of ERF109 in light regulated retrograde signaling. http://dx.doi.org/10.1016/j.nbt.2016.06.1288
P18-37 Ploidy level evaluation of highbush blueberry parental genotypes and ther hybrids ∗ , Slanisław Pluta ´ Małgorzata Podwyszynska
Research Institute of Horticulture, Poland Highbush blueberry (Vaccinium corymbosum L.) is grown in Poland on commercial scale recently. With the level of blueberry production of 13.0 thousand metric tons in 2015, Poland currently ranks first in Europe and fifth in the world. Due to the high interest in new cultivars, the Research Institute of Horticulture in Skierniewice started the new blueberry breeding program in 2009. It is believed that the most of highbush blueberry cultivars are tetraploids, but pentaploids and diploids are also known. The ploidy level can affect fertility of parental genotypes as well as fruit and seed formation at hybridization. However, except of three cultivars which are known to be tetraploids, the ploidy level of other parental genotypes selected for our breeding program has been unknown. In our previous study we have observed low seed formation in some crosses. The aim of the present study was to elucidate whether the poor seed production could result different ploidy levels of parental cultivars. Twelve parental genotypes (used for the factorial crossing design) and 100 selected seedlings (hybrids) of V. corymbosum have been used for our studies. Assessment of the ploidy level was performed by the flow cytometry. Cytometric analysis has shown that all the parental cultivars are tetraploids. In case of their hybrids, except of six aneuploids, all other seedlings have been proven to be tetraploids. Since our results demonstrate that all the parental genotypes of V. corymbosum are tetraploids, other causative agent of poor seed production in some crosses should be considered. http://dx.doi.org/10.1016/j.nbt.2016.06.1289
P18-38 Modification of anther selection mode for the successful androgenesis induction in the cassava (Manihot esculenta) anther cultures Elzbieta Maria Golemiec International Center for Tropical Agriculture, Poland Decades of effort dedicated to work on cassava double haploid production protocol establishment, did not result in cassava plant regeneration as in many other species (Wedzony, 2009; Perrera, 2012). Existing protocols still need to be modified. Cassava flower buds size, ranging from 2.3 to 2.5 mm is considered as a most suitable for the androgenic cultures establishment. Performed experiments reviled variability in the tone of isolated anthers: transparent, white and yellowish. The aim of this work was to confirm if the difference in the androgenic answer in the culture depends on the type of plated