- Email: [email protected]
Efficient in vitro regeneration of common daisy, a medicinal plant
New Biotechnology · Volume 29S · September 2012
tsuruhatensis with a GenBank accession number of AB075017. The cadmium-resistant isolate was shown to be resistant to other heavy metals like Al3+ , Li2+ , Ba2+ , Mn2+ , Pb2+ , Ag2+ , Sn2+ , Ni2+ , Zn2+ and Sr2+ . Resistance to the antibiotics ampicillin, netilmicin, oxacillin, penicillin and rifampin was also noted. Transformation and curing results revealed that the cadmium resistance ability of Delftia tsuruhatensis was chromosome-encoded. Keywords: Cadmium-resistant; 16S rDNA sequencing; Delftia tsuruhatensis; Heavy metal resistance http://dx.doi.org/10.1016/j.nbt.2012.08.496 Poster 5.0.57 Evaluation some seed quality characters of (Cupressus sempervirens var. horizantalis) species using accelerated aging test Fatemeh Ahmadloo1,∗ , Masoud Tabari2 1 2
Faculty of Natural Resources, Tarbiat Modares University, Nour, Iran College of Natural Resources, Tarbiat Modares University, Nour, Iran
The accelerated aging test is used for estimating a vigor index of seed lots and also for estimating their storage potential. Current research carried out for considering the effect accelerated ageing on some of the physiological characteristics of Cupressus sempervirens var. horizantalis seeds including germination percent, mean germination time, germination speed, germination energy and vigor index. The design was set up as a Completely Randomized Design (CRD) with five treatment and four replications 100 per petridish. The seed samples were exposed to 40◦ C and relative humidity of 100 percent for zero, 24, 48, 72, 96 and 120 h. Then seeds were kept for 37 days in germinator (16 h photoperiod at 1000 lux and 20◦ C). For measuring Electrical Conductivity (EC) test and Tetrazolium test (TZ), seeds treated exposed to 25◦ C in incubator for 24 h. The results indicated that increasing aging period significantly reduced most of seed physiological characteristics. The highest germination percent 81.1% and speed of germination 8.4 n/d was observed in control treatment and the lowest for 120 h (11.55% and 1.1 n/d respectively). Seed vigor strongly decreased and it was more affecting accelerated ageing. The EC values were the least and highest in control sample and all of accelerated ageing treatments, respectively. The viability chart illustrates the relationship between initial germination, seed deterioration rate and storage condition over time. Therefore, proper storage conditions of C. sempervirens seeds are important before sowing of the seeds next year. Keywords: Accelerated aging test; Electrical conductivity; Germination; Seed vigor http://dx.doi.org/10.1016/j.nbt.2012.08.497
S178
www.elsevier.com/locate/nbt
Poster 5.0.58 Inhibition of Escherichia coli by essential oil extracted from shiitake mushroom Fatma Ozen1,∗ , F. Yesim Ekinci1,2 , May Korachi1 1 2
Yeditepe University, Institute of Biotechnology, Istanbul, Turkey Yeditepe University, Department of Food Engineering, Istanbul, Turkey
Shiitake (Lentinula edodes) is an edible mushroom which is a popular food source in Asia, USA and Europe. The aim of this study was to investigate the antimicrobial activity of Shiitake essential oil extracts against Enterococcus faecalis ATCC 19433, Escherichia coli ATCC 25922, Listeria monocytogenes ATCC 15313, Shigella flexneri ATCC 12022, Staphylococcus aureus ATCC 25923, Yersinia enterocolitica ATCC 23715. Essential oil extracts were analysed by gas chromatography–mass spectrometry (GC–MS) for its qualitative and quantitative composition. Antimicrobial activity and minimum inhibitory concentrations (MIC) were performed (concentrations 0.24 mg/l to 30.7 mg/l). Shiitake essential oil demonstrated antimicrobial activity against E. coli (>15 ± 5.6 mm), L. monocytogenes (>9.5 ± 2.1 mm) and Y. enterocolitica (>9.5 ± 2.1 mm). The MIC value of essential oil for E. coli, Y. enterocolitica and L. monocytogenes were found at concentration of 1.9 mg/l. Keywords: Shiitake; Essential oil; Disc diffusion assay; MIC; Food-borne pathogens http://dx.doi.org/10.1016/j.nbt.2012.08.498 Poster 5.0.59 Efficient in vitro regeneration of common daisy, a medicinal plant F. Pehlivan Karakas∗ , A. Ucar Turker Department of Biology, Faculty of Arts and Sciences, Abant Izzet Baysal University, Bolu 14280, Turkey Bellis perennis L. (common daisy) is a medicinal plant that has been used to treat common cold, wounds, stomachache, eye diseases, eczema, gastritis, cancer, diarrhea, rheumatism, inflammation and infections of the upper respiratory tract. A highly efficient and rapid regeneration system via multiple shoot formation was developed for common daisy. Explants (leaf, pedicel, petiole and root segments) excised from sterile field-grown plants were cultured on medium supplemented with different concentrations and combinations of various plant growth regulators. The pedicel explants formed more shoots than other explants. The root explants did not form any shoot regeneration on any media. The best shoot proliferation was obtained from pedicel explants cultured on media with 0.5 mg/l TDZ and 0.5 mg/l IAA. Regenerated shoots were transferred to rooting media containing different concentrations of IAA, IBA, NAA or 2,4-D. Most shoots developed roots on medium with 1.0 mg/l IAA. Rooted explants were transferred to vermiculate in Magenta containers for acclimatization and after 3 weeks they were planted in to plastic pots containing potting soil and maintained in the plant growth room. Approximately 3 months after the transfer to room conditions, the flowering of the regenerated plants could be observed. With this efficient protocol, disease and herbi-
New Biotechnology · Volume 29S · September 2012
cide free bulk plant material can be supplied throughout the year for pharmaceutical purposes. Keywords: Bellis perennis; In vitro culture; Micropropagation
Poster 5.0.61
http://dx.doi.org/10.1016/j.nbt.2012.08.499
G. Brice˜ no1,2,∗ , M.S. Fuentes4 , G. Palma2,3 , M.J. Amoroso4 , M.C. 1,2 Diez
Poster 5.0.60
Chlorpyrifos degradation by consortium of actinobacteria isolated from contaminated environment
1
Clustering of SSR markers located on 2AL for yellow rust resistance in hexaploid wheat (Triticum aestivum L.) Funda Senturk Akfirat1,∗ , Fahriye Ertugrul2 , Mehmet Cakir3 , Semra Hasancebi4 , Yildiz Aydin5 , Kadir Akan6 , Zafer Mert6 , Ahu Altinkut Uncuoglu7 1
Gebze Institute of Technology, Faculty of Science, Department of Molecular Biology and Genetics, Cayirova Campus, 41700 Gebze, Kocaeli, Turkey 2 TUBITAK, Agriculture, Forestry and Veterinary Research Group, Tunus Street, No: 80, 06100 Kavaklıdere, Ankara, Turkey 3 Murdoch University, State Agricultural Biotechnology Centre, Perth, Western Australia, Australia 4 TUBITAK, MRC, Genetic Engineering and Biotechnology Institute, P.O. Box 21, 41470 Gebze, Kocaeli, Turkey 5 Marmara University, Faculty of Science and Letters, Department of Biology, 34722 Kadikoy, Istanbul, Turkey 6 Central Research Institute of Field Crops, P.O. Box 226, Lodumlu, 06042 Ankara, Turkey 7 Marmara University, Faculty of Engineering, Department of Bioengineering, 34722 Kadikoy, Istanbul, Turkey Yellow rust, caused by Puccinia striiformis f. sp. tritici, is one of the most devastating diseases of wheat throughout the world. Molecular markers are powerful tools in marker-assisted selection, gene pyramiding and gene cloning of important crop traits especially for disease resistance. Previously, we identified Xgwm382 and Xgwm311 as diagnostic markers located on chromosome group 2 for disease resistance against yellow rust in Izgi2001 × ES14 F2 population by using 230 SSR primer pairs. In order to understand the chromosomal group location of these diagnostic markers in the same population, we selected 17 SSR markers mapped only in one genome of chromosome group 2 (A, B, D) around 1–21 cM distance to Xgwm382 based on the SSR consensus map of wheat. One out of 17 SSR markers, Xwmc658, that is associated with yellow rust resistance located only in 2AL and can potentially be used to select yellow rust resistant wheat germplasm. This work has highlighted the importance of the distal region of the long arm of wheat chromosome 2AL implicated in yellow rust resistance flanked by markers Xgwm382, Xgwm311 and Xwmc658. Keywords: 2AL; SSR markers; Yellow rust; Triticum aestivum L
Department of Chemical Engineering, Universidad de La Frontera, P.O. Box 54-D, Temuco, Chile 2 Department of Chemical Science, Universidad de La Frontera, Temuco, Chile 3 Scientific and Technological Bioresource Nucleus, Universidad de La Frontera, Chile 4 Planta Piloto de Procesos Industriales Microbiológicos (ProimiConicet), Avenida Belgrano y Pasaje Caseros, 4000 Tucumán, Argentina
The aim of this work was to demonstrate the efficiency of pure and mixed actinobacteria cultures isolated from Chilean (Streptomyces sp. Strain AC5 y AC7) and Argentinean (Streptomyces sp. A2, A5, A11, M7) contaminated sites for removing the insecticide chlorpyrifos (CP) from liquid medium. Preliminary actinobacteria tolerance assays were carried out in Petri plates containing solid minimal medium (MM) in the presence of CP at concentration of 1.66 mg L−1 . In most of cases, actinobacteria strains showed high tolerance levels to the pesticides. To determine the potential antagonistic effect of the actinobacteria strains, Petri dishes with solid starch casein medium were sown confronting individual strains with each other actinobacteria. No growth inhibition was observed, so, there were no responses to antagonism between the assayed strains. The ability of pure and mixed actinobacteria cultures to growth and remove CP in liquid MM was evaluated determining growth (dry weight), pesticide removal (gas chromatography) and pH variation. After 72 h of incubation, the CP removal was 67–89% when individual strains were evaluated, 83% and 96% CP removal was observed in the mixed culture formed by Streptomyces sp. A2-A5-A11-M7 and Streptomyces sp. AC5-AC7, respectively. Finally, with the consortium constituted by six strains, 92% of CP was removed, the biomass was duplicated and the pH increase from 7.0 to 8.3. These microorganisms showed high degradation capacity to remove organophosphorus pesticides such as chlorpyrifos. Therefore actinobacterias isolated from contaminated sites could be recommended for bioremediation pesticides. This study was supported by FONDECYT postdoctoral project N◦ 3100118 and the “Program of Scientific International Cooperation CONYCYT/MINCYT” 2009-111. http://dx.doi.org/10.1016/j.nbt.2012.08.501 Poster 5.0.62
http://dx.doi.org/10.1016/j.nbt.2012.08.500
Molecular characterization of environmental isolates resistant to aluminum Serim Gamze, Yılmaz Fadime∗ , Icgen Bulent, Ergene Aysun Kırıkkale University, Department of Biology, 71450 Kırıkkale, Turkey Pollution in industrial areas is a serious environmental concern, and interest in bacterial resistance to heavy metals is of practical significance. The present work was aimed to isolate and idenwww.elsevier.com/locate/nbt S179
tsuruhatensis with a GenBank accession number of AB075017. The cadmium-resistant isolate was shown to be resistant to other heavy metals like Al3+ , Li2+ , Ba2+ , Mn2+ , Pb2+ , Ag2+ , Sn2+ , Ni2+ , Zn2+ and Sr2+ . Resistance to the antibiotics ampicillin, netilmicin, oxacillin, penicillin and rifampin was also noted. Transformation and curing results revealed that the cadmium resistance ability of Delftia tsuruhatensis was chromosome-encoded. Keywords: Cadmium-resistant; 16S rDNA sequencing; Delftia tsuruhatensis; Heavy metal resistance http://dx.doi.org/10.1016/j.nbt.2012.08.496 Poster 5.0.57 Evaluation some seed quality characters of (Cupressus sempervirens var. horizantalis) species using accelerated aging test Fatemeh Ahmadloo1,∗ , Masoud Tabari2 1 2
Faculty of Natural Resources, Tarbiat Modares University, Nour, Iran College of Natural Resources, Tarbiat Modares University, Nour, Iran
The accelerated aging test is used for estimating a vigor index of seed lots and also for estimating their storage potential. Current research carried out for considering the effect accelerated ageing on some of the physiological characteristics of Cupressus sempervirens var. horizantalis seeds including germination percent, mean germination time, germination speed, germination energy and vigor index. The design was set up as a Completely Randomized Design (CRD) with five treatment and four replications 100 per petridish. The seed samples were exposed to 40◦ C and relative humidity of 100 percent for zero, 24, 48, 72, 96 and 120 h. Then seeds were kept for 37 days in germinator (16 h photoperiod at 1000 lux and 20◦ C). For measuring Electrical Conductivity (EC) test and Tetrazolium test (TZ), seeds treated exposed to 25◦ C in incubator for 24 h. The results indicated that increasing aging period significantly reduced most of seed physiological characteristics. The highest germination percent 81.1% and speed of germination 8.4 n/d was observed in control treatment and the lowest for 120 h (11.55% and 1.1 n/d respectively). Seed vigor strongly decreased and it was more affecting accelerated ageing. The EC values were the least and highest in control sample and all of accelerated ageing treatments, respectively. The viability chart illustrates the relationship between initial germination, seed deterioration rate and storage condition over time. Therefore, proper storage conditions of C. sempervirens seeds are important before sowing of the seeds next year. Keywords: Accelerated aging test; Electrical conductivity; Germination; Seed vigor http://dx.doi.org/10.1016/j.nbt.2012.08.497
S178
www.elsevier.com/locate/nbt
Poster 5.0.58 Inhibition of Escherichia coli by essential oil extracted from shiitake mushroom Fatma Ozen1,∗ , F. Yesim Ekinci1,2 , May Korachi1 1 2
Yeditepe University, Institute of Biotechnology, Istanbul, Turkey Yeditepe University, Department of Food Engineering, Istanbul, Turkey
Shiitake (Lentinula edodes) is an edible mushroom which is a popular food source in Asia, USA and Europe. The aim of this study was to investigate the antimicrobial activity of Shiitake essential oil extracts against Enterococcus faecalis ATCC 19433, Escherichia coli ATCC 25922, Listeria monocytogenes ATCC 15313, Shigella flexneri ATCC 12022, Staphylococcus aureus ATCC 25923, Yersinia enterocolitica ATCC 23715. Essential oil extracts were analysed by gas chromatography–mass spectrometry (GC–MS) for its qualitative and quantitative composition. Antimicrobial activity and minimum inhibitory concentrations (MIC) were performed (concentrations 0.24 mg/l to 30.7 mg/l). Shiitake essential oil demonstrated antimicrobial activity against E. coli (>15 ± 5.6 mm), L. monocytogenes (>9.5 ± 2.1 mm) and Y. enterocolitica (>9.5 ± 2.1 mm). The MIC value of essential oil for E. coli, Y. enterocolitica and L. monocytogenes were found at concentration of 1.9 mg/l. Keywords: Shiitake; Essential oil; Disc diffusion assay; MIC; Food-borne pathogens http://dx.doi.org/10.1016/j.nbt.2012.08.498 Poster 5.0.59 Efficient in vitro regeneration of common daisy, a medicinal plant F. Pehlivan Karakas∗ , A. Ucar Turker Department of Biology, Faculty of Arts and Sciences, Abant Izzet Baysal University, Bolu 14280, Turkey Bellis perennis L. (common daisy) is a medicinal plant that has been used to treat common cold, wounds, stomachache, eye diseases, eczema, gastritis, cancer, diarrhea, rheumatism, inflammation and infections of the upper respiratory tract. A highly efficient and rapid regeneration system via multiple shoot formation was developed for common daisy. Explants (leaf, pedicel, petiole and root segments) excised from sterile field-grown plants were cultured on medium supplemented with different concentrations and combinations of various plant growth regulators. The pedicel explants formed more shoots than other explants. The root explants did not form any shoot regeneration on any media. The best shoot proliferation was obtained from pedicel explants cultured on media with 0.5 mg/l TDZ and 0.5 mg/l IAA. Regenerated shoots were transferred to rooting media containing different concentrations of IAA, IBA, NAA or 2,4-D. Most shoots developed roots on medium with 1.0 mg/l IAA. Rooted explants were transferred to vermiculate in Magenta containers for acclimatization and after 3 weeks they were planted in to plastic pots containing potting soil and maintained in the plant growth room. Approximately 3 months after the transfer to room conditions, the flowering of the regenerated plants could be observed. With this efficient protocol, disease and herbi-
New Biotechnology · Volume 29S · September 2012
cide free bulk plant material can be supplied throughout the year for pharmaceutical purposes. Keywords: Bellis perennis; In vitro culture; Micropropagation
Poster 5.0.61
http://dx.doi.org/10.1016/j.nbt.2012.08.499
G. Brice˜ no1,2,∗ , M.S. Fuentes4 , G. Palma2,3 , M.J. Amoroso4 , M.C. 1,2 Diez
Poster 5.0.60
Chlorpyrifos degradation by consortium of actinobacteria isolated from contaminated environment
1
Clustering of SSR markers located on 2AL for yellow rust resistance in hexaploid wheat (Triticum aestivum L.) Funda Senturk Akfirat1,∗ , Fahriye Ertugrul2 , Mehmet Cakir3 , Semra Hasancebi4 , Yildiz Aydin5 , Kadir Akan6 , Zafer Mert6 , Ahu Altinkut Uncuoglu7 1
Gebze Institute of Technology, Faculty of Science, Department of Molecular Biology and Genetics, Cayirova Campus, 41700 Gebze, Kocaeli, Turkey 2 TUBITAK, Agriculture, Forestry and Veterinary Research Group, Tunus Street, No: 80, 06100 Kavaklıdere, Ankara, Turkey 3 Murdoch University, State Agricultural Biotechnology Centre, Perth, Western Australia, Australia 4 TUBITAK, MRC, Genetic Engineering and Biotechnology Institute, P.O. Box 21, 41470 Gebze, Kocaeli, Turkey 5 Marmara University, Faculty of Science and Letters, Department of Biology, 34722 Kadikoy, Istanbul, Turkey 6 Central Research Institute of Field Crops, P.O. Box 226, Lodumlu, 06042 Ankara, Turkey 7 Marmara University, Faculty of Engineering, Department of Bioengineering, 34722 Kadikoy, Istanbul, Turkey Yellow rust, caused by Puccinia striiformis f. sp. tritici, is one of the most devastating diseases of wheat throughout the world. Molecular markers are powerful tools in marker-assisted selection, gene pyramiding and gene cloning of important crop traits especially for disease resistance. Previously, we identified Xgwm382 and Xgwm311 as diagnostic markers located on chromosome group 2 for disease resistance against yellow rust in Izgi2001 × ES14 F2 population by using 230 SSR primer pairs. In order to understand the chromosomal group location of these diagnostic markers in the same population, we selected 17 SSR markers mapped only in one genome of chromosome group 2 (A, B, D) around 1–21 cM distance to Xgwm382 based on the SSR consensus map of wheat. One out of 17 SSR markers, Xwmc658, that is associated with yellow rust resistance located only in 2AL and can potentially be used to select yellow rust resistant wheat germplasm. This work has highlighted the importance of the distal region of the long arm of wheat chromosome 2AL implicated in yellow rust resistance flanked by markers Xgwm382, Xgwm311 and Xwmc658. Keywords: 2AL; SSR markers; Yellow rust; Triticum aestivum L
Department of Chemical Engineering, Universidad de La Frontera, P.O. Box 54-D, Temuco, Chile 2 Department of Chemical Science, Universidad de La Frontera, Temuco, Chile 3 Scientific and Technological Bioresource Nucleus, Universidad de La Frontera, Chile 4 Planta Piloto de Procesos Industriales Microbiológicos (ProimiConicet), Avenida Belgrano y Pasaje Caseros, 4000 Tucumán, Argentina
The aim of this work was to demonstrate the efficiency of pure and mixed actinobacteria cultures isolated from Chilean (Streptomyces sp. Strain AC5 y AC7) and Argentinean (Streptomyces sp. A2, A5, A11, M7) contaminated sites for removing the insecticide chlorpyrifos (CP) from liquid medium. Preliminary actinobacteria tolerance assays were carried out in Petri plates containing solid minimal medium (MM) in the presence of CP at concentration of 1.66 mg L−1 . In most of cases, actinobacteria strains showed high tolerance levels to the pesticides. To determine the potential antagonistic effect of the actinobacteria strains, Petri dishes with solid starch casein medium were sown confronting individual strains with each other actinobacteria. No growth inhibition was observed, so, there were no responses to antagonism between the assayed strains. The ability of pure and mixed actinobacteria cultures to growth and remove CP in liquid MM was evaluated determining growth (dry weight), pesticide removal (gas chromatography) and pH variation. After 72 h of incubation, the CP removal was 67–89% when individual strains were evaluated, 83% and 96% CP removal was observed in the mixed culture formed by Streptomyces sp. A2-A5-A11-M7 and Streptomyces sp. AC5-AC7, respectively. Finally, with the consortium constituted by six strains, 92% of CP was removed, the biomass was duplicated and the pH increase from 7.0 to 8.3. These microorganisms showed high degradation capacity to remove organophosphorus pesticides such as chlorpyrifos. Therefore actinobacterias isolated from contaminated sites could be recommended for bioremediation pesticides. This study was supported by FONDECYT postdoctoral project N◦ 3100118 and the “Program of Scientific International Cooperation CONYCYT/MINCYT” 2009-111. http://dx.doi.org/10.1016/j.nbt.2012.08.501 Poster 5.0.62
http://dx.doi.org/10.1016/j.nbt.2012.08.500
Molecular characterization of environmental isolates resistant to aluminum Serim Gamze, Yılmaz Fadime∗ , Icgen Bulent, Ergene Aysun Kırıkkale University, Department of Biology, 71450 Kırıkkale, Turkey Pollution in industrial areas is a serious environmental concern, and interest in bacterial resistance to heavy metals is of practical significance. The present work was aimed to isolate and idenwww.elsevier.com/locate/nbt S179