peroxidase gene polymorphism in lens and vicia species

1
Poster presentations / Current Opinion in Biotechnology 24S (2013) S48–S143 S127 highest amount of total polyphenolic compounds was detected in cultivar Negra; single polyphenols as follow: gallic acid 33.14 mg/L, caftaric acid 26.71 mg/L in pulp, (+) catechin 65.71 mg/L and () epi- catechin 61.48 mg/L in seeds and myricetin 5.18 mg/L, quercetin 3.78 mg/L and kaempferol 2.73 in grape skin. http://dx.doi.org/10.1016/j.copbio.2013.05.402 Response of Vitis vinifera cell cultures on stress caused by methyl jasmonate Martin Sák 1,2 , Ivana Dokupilová 3,4 , Daniel Mihálik 3 , Ján ˇ Sajbidor 1 1 Institute of Biotechnology and Food Science, Faculty of Chemical and Food Technology, Slovak University of Technology, Bratislava, Slovakia 2 Department of Analytical Chemistry, Plant Production Research Cen- ter, Pieˇ st’any, Slovakia 3 Department of Molecular Biology, Plant Production Research Center, Pieˇ st’any, Slovakia 4 Institute of Biochemistry, Nutrition and Health Protection, Faculty of Chemical and Food Technology, Slovak University of Technology, Bratislava, Slovakia E-mail address: [email protected] (M. Sák). Plant cell culture represent potential source of secondary metabo- lites which cannot be produced by microorganism and chemical synthesis. However, only a few of plant cell cultures produce these compounds in commercially amount. In vitro cell cultures of Vitis vinifera cv. Váh were inoculated with commercial elicitor methyl jasmonate for detection of polyphenol production depend on cultivation time and elicitor concentration. Influence of different elicitor amounts was evaluated by high performance liquid chro- matography with diode array detection in 1, 4, 6, and 8 days after elicitation. Cellular responses to different concentration of methyl jasmonate (0.2 mM, 0.4 mM and 0.8 mM) included rapid alkalinisa- tion, polyphenol production and cell death. Variously distributed phenolic compounds including (+) catechin, () epicatechin, p- coumaric acid, syringaldehyde, rutin and p-coumaric acid detected in cells were statistically evaluated depended on time and elicitor concentration. Overall, total production of polyphenols increased in treated cells compared with untreated; the addition of methyl jasmonate had a generally increasing effect to production and the highest amount was measured in 8 day after elicitation reaching 7.83 mg/g(fw). On the other hand control cells displayed a higher amount of viable cells. When elicitors were supplied, cell growth was reduced although cell viability did not show a massive cell lysis. The alkalinization of the culture medium measured during the each period of polyphenol sampling was almost constant in the control (difference ±0.01), but decreased about average 0.15 in treated cultures after of each cultivation day. http://dx.doi.org/10.1016/j.copbio.2013.05.403 Development important forms of soft spring wheat in North Kazakhstan Assiya Turganbayeva, Oksana Hapilina, Galina Shek, Almagul Kakimzhanova National Center for Biotechnology, Astana, Kazakhstan E-mail address: [email protected] (A. Turganbayeva). Spring wheat is the main exporting crop in Kazakhstan. The major share of grain sold on the world market is the grain of spring wheat grown in North Kazakhstan, where the area under this crop is 85%. The aim of this study was to create new varieties of wheat through biotechnology and conventional breeding with directional selection for resistance to drought and disease. Objects were vari- eties of wheat in Kazakhstan. In 2011, for state testing were given new varieties of spring wheat — Ak Orda and Kazakhstan 20. A new variety of Kazakhstan 20 refers to a kind of Lutescens. It was obtained by in vitro from hybrid of Akmola 2/Akmola 40. The line is drought resistant and responsive to moisture conditions. Aver- age yield for 2009–2011 on the steam background was 20.6 t/ha. In 2011, through the manufacturing variety trials exceeded the yield standard on 5.6 t/ha. A new variety of Ak Orda refers to Albidum variety. It was obtained by the cell selection method with culture filtrate of the Alternaria alternata of constant line of hybrid origin. In the manufacturing variety trials, exceeded the yield standard on 2.9 t/ha. The line is resistant to ‘black germ’. Studies from 2000 to 2011 on the selection in vitro with adding selective agents and test progeny of regenerated wheat revealed competitive lines to create new varieties of spring wheat. http://dx.doi.org/10.1016/j.copbio.2013.05.404 Peroxidase gene polymorphism in lens and Vicia species Duygu Sari 1 , Cengiz Toker 1 , Nedim Mutlu 2 1 Department of Field Crops, Faculty of Agriculture, Akdeniz University, TR-07070 Antalya, Turkey 2 Department of Agricultural Biotechnology, Faculty of Agriculture, Akdeniz University, TR-07070 Antalya, Turkey E-mail address: [email protected] (C. Toker). Peroxidases are organized into three classes as intracellular perox- idases, secretory fungal peroxidases and plant peroxidases. Plant peroxidases belong to a multigene family and play important roles in many stress-related interactions, such as pathogen infection, insect tolerance, salt tolerance, auxin degradation, cell wall ligni- fications, tissue suberization, and plant senescence. They posses highly conserved domains allowing oligonucleotide primers to be designed to amplify DNA sequences coding for peroxidases from plants with unsequenced genomes. Thanks to conserved structure, plant peroxidases are used for classification of plant species. In this study, peroxidase gene-based polymorphism among Lens species including L. culinaris Medik., L. orientalis (Boiss.) Ponert, L. tomento- sus Ladiz., L. odemensis Ladiz., L. lamottei Czefr., L. ervoides (Brign.) Grande and L. nigricans (M.Bieb.) Godr. and Vicia L. comprising Vicia montbretii (Fisch. and Mey.) Davis and Plitmann, and correlation between peroxidase markers were conducted. Thirty Lens and 2 Vicia species were evaluated using 22 peroxidase specific primers. PCR amplification indicated high polymorphism between species based on peroxidases. Especially for 7 primer combinations yielded higher polymorphisms than the others. These results indicated that peroxidase gene family can be used to study genotypic diversity and evolutionary relationships. http://dx.doi.org/10.1016/j.copbio.2013.05.405 Resistance gene analog polymorphism in Lens species Duygu Sari 1 , Nedim Mutlu 2 , Cengiz Toker 1 1 Department of Field Crops, Faculty of Agriculture, Akdeniz University, TR-07070 Antalya, Turkey 2 Department of Agricultural Biotechnology, Faculty of Agriculture, Akdeniz University, TR-07070 Antalya, Turkey E-mail address: [email protected] (C. Toker). Plant resistance genes (R-gene) play an important role in plant–pathogen recognition and these genes present several highly

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Page 1: Peroxidase gene polymorphism in lens and Vicia species

Poster presentations / Current Opinion in Biotechnology 24S (2013) S48–S143 S127

highest amount of total polyphenolic compounds was detected incultivar Negra; single polyphenols as follow: gallic acid 33.14 mg/L,caftaric acid 26.71 mg/L in pulp, (+) catechin 65.71 mg/L and (−) epi-catechin 61.48 mg/L in seeds and myricetin 5.18 mg/L, quercetin3.78 mg/L and kaempferol 2.73 in grape skin.

http://dx.doi.org/10.1016/j.copbio.2013.05.402

Response of Vitis vinifera cell cultures on stress caused bymethyl jasmonate

Martin Sák 1,2, Ivana Dokupilová 3,4, Daniel Mihálik 3, JánSajbidor 1

1 Institute of Biotechnology and Food Science, Faculty of Chemical andFood Technology, Slovak University of Technology, Bratislava, Slovakia2 Department of Analytical Chemistry, Plant Production Research Cen-ter, Piest’any, Slovakia3 Department of Molecular Biology, Plant Production Research Center,Piest’any, Slovakia4 Institute of Biochemistry, Nutrition and Health Protection, Facultyof Chemical and Food Technology, Slovak University of Technology,Bratislava, SlovakiaE-mail address: [email protected] (M. Sák).

Plant cell culture represent potential source of secondary metabo-lites which cannot be produced by microorganism and chemicalsynthesis. However, only a few of plant cell cultures producethese compounds in commercially amount. In vitro cell culturesof Vitis vinifera cv. Váh were inoculated with commercial elicitormethyl jasmonate for detection of polyphenol production dependon cultivation time and elicitor concentration. Influence of differentelicitor amounts was evaluated by high performance liquid chro-matography with diode array detection in 1, 4, 6, and 8 days afterelicitation. Cellular responses to different concentration of methyljasmonate (0.2 mM, 0.4 mM and 0.8 mM) included rapid alkalinisa-tion, polyphenol production and cell death. Variously distributedphenolic compounds including (+) catechin, (−) epicatechin, p-coumaric acid, syringaldehyde, rutin and p-coumaric acid detectedin cells were statistically evaluated depended on time and elicitorconcentration. Overall, total production of polyphenols increasedin treated cells compared with untreated; the addition of methyljasmonate had a generally increasing effect to production and thehighest amount was measured in 8 day after elicitation reaching7.83 mg/g(fw). On the other hand control cells displayed a higheramount of viable cells. When elicitors were supplied, cell growthwas reduced although cell viability did not show a massive celllysis. The alkalinization of the culture medium measured duringthe each period of polyphenol sampling was almost constant inthe control (difference ±0.01), but decreased about average 0.15 intreated cultures after of each cultivation day.

http://dx.doi.org/10.1016/j.copbio.2013.05.403

Development important forms of soft spring wheat in NorthKazakhstan

Assiya Turganbayeva, Oksana Hapilina, Galina Shek, AlmagulKakimzhanova

National Center for Biotechnology, Astana, KazakhstanE-mail address: [email protected] (A. Turganbayeva).

Spring wheat is the main exporting crop in Kazakhstan. The majorshare of grain sold on the world market is the grain of spring wheatgrown in North Kazakhstan, where the area under this crop is85%. The aim of this study was to create new varieties of wheat

through biotechnology and conventional breeding with directionalselection for resistance to drought and disease. Objects were vari-eties of wheat in Kazakhstan. In 2011, for state testing were givennew varieties of spring wheat — Ak Orda and Kazakhstan 20. Anew variety of Kazakhstan 20 refers to a kind of Lutescens. It wasobtained by in vitro from hybrid of Akmola 2/Akmola 40. The lineis drought resistant and responsive to moisture conditions. Aver-age yield for 2009–2011 on the steam background was 20.6 t/ha. In2011, through the manufacturing variety trials exceeded the yieldstandard on 5.6 t/ha. A new variety of Ak Orda refers to Albidumvariety. It was obtained by the cell selection method with culturefiltrate of the Alternaria alternata of constant line of hybrid origin.In the manufacturing variety trials, exceeded the yield standard on2.9 t/ha. The line is resistant to ‘black germ’. Studies from 2000 to2011 on the selection in vitro with adding selective agents and testprogeny of regenerated wheat revealed competitive lines to createnew varieties of spring wheat.

http://dx.doi.org/10.1016/j.copbio.2013.05.404

Peroxidase gene polymorphism in lens and Vicia species

Duygu Sari 1, Cengiz Toker 1, Nedim Mutlu 2

1 Department of Field Crops, Faculty of Agriculture, Akdeniz University,TR-07070 Antalya, Turkey2 Department of Agricultural Biotechnology, Faculty of Agriculture,Akdeniz University, TR-07070 Antalya, TurkeyE-mail address: [email protected] (C. Toker).

Peroxidases are organized into three classes as intracellular perox-idases, secretory fungal peroxidases and plant peroxidases. Plantperoxidases belong to a multigene family and play important rolesin many stress-related interactions, such as pathogen infection,insect tolerance, salt tolerance, auxin degradation, cell wall ligni-fications, tissue suberization, and plant senescence. They posseshighly conserved domains allowing oligonucleotide primers to bedesigned to amplify DNA sequences coding for peroxidases fromplants with unsequenced genomes. Thanks to conserved structure,plant peroxidases are used for classification of plant species. In thisstudy, peroxidase gene-based polymorphism among Lens speciesincluding L. culinaris Medik., L. orientalis (Boiss.) Ponert, L. tomento-sus Ladiz., L. odemensis Ladiz., L. lamottei Czefr., L. ervoides (Brign.)Grande and L. nigricans (M.Bieb.) Godr. and Vicia L. comprising Viciamontbretii (Fisch. and Mey.) Davis and Plitmann, and correlationbetween peroxidase markers were conducted. Thirty Lens and 2Vicia species were evaluated using 22 peroxidase specific primers.PCR amplification indicated high polymorphism between speciesbased on peroxidases. Especially for 7 primer combinations yieldedhigher polymorphisms than the others. These results indicated thatperoxidase gene family can be used to study genotypic diversityand evolutionary relationships.

http://dx.doi.org/10.1016/j.copbio.2013.05.405

Resistance gene analog polymorphism in Lens species

Duygu Sari 1, Nedim Mutlu 2, Cengiz Toker 1

1 Department of Field Crops, Faculty of Agriculture, Akdeniz University,TR-07070 Antalya, Turkey2 Department of Agricultural Biotechnology, Faculty of Agriculture,Akdeniz University, TR-07070 Antalya, TurkeyE-mail address: [email protected] (C. Toker).

Plant resistance genes (R-gene) play an important role inplant–pathogen recognition and these genes present several highly