- Email: [email protected]
Isolation, identification and expression analysis of salt-responsive genes in Oryza sativa var. MR219 using PCR-based suppression subtractive hybridization
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Abstracts / Current Opinion in Biotechnology 22S (2011) S15–S152
stress. In the present study, six different wheat (Triticum aestivum L.) genotypes including two tolerant sister lines (Ducula-1 and Ducula-4), two standard cultivars widely grown in the Eastern Mediterranean (‘Basribey’ and ‘Golia’) and two susceptible cultivars (‘Seri-82’ and ‘Dogankent’) were subjected to different waterlogging treatments and their proline and chlorophyll contents were measured by spectrophotometer. About 25 anaerobically induced genes were analyzed using real time polymerase chain reaction (RT-PCR) method. Wheat seedlings were subjected to waterlogging treatments of 0, 0.5, 1, 2, 4, 6, 8, 12, 24, 48, 72 h and after each treatment, leaf samples were collected for chlorophyll, carotenoid, proline contents and gene expression analyses. A total of 32 primer pairs obtained from 25 anaerobically induced genes were used for RT-PCR. Total chlorophyll content decreased while proline content increased as waterlogging duration was prolonged. Gene expression differences among varieties under waterlogging and control conditions will be also presented. doi:10.1016/j.copbio.2011.05.476
P53 Isolation, identification and expression analysis of saltresponsive genes in Oryza sativa var. MR219 using PCRbased suppression subtractive hybridization Zamri Zainal 1 , Poya Hedayati 2 , Ismanizan Ismail 1 , Che Radziah Mohd Zain 2 , Zuraida A Rahman 3 1
Institue of Systems Biology, Universiti Kebangsaan, Malaysia School of Biosciences and Biotechnology, Universiti Kebangsaan, Malaysia 3 Biotechnology Research, MARDI, Malaysia 2
P54 Isolation of the homologoues of genes encoding CBF1/DREB1 proteins from Olive (Olea europaea) Beste Tacal, Gizem Hacimuto, Senay Vural Korkut Department of Biology, Faculty of Science and Arts, Yildiz Technical University, Istanbul, Turkey E-mail address: [email protected] (B. Tacal) Olive is an economically important plant cultivated in extensive areas in the Mediterranean countries. The plant may be subjected to heat and drought stress during the summer and chilling stress during the winter. Although it can stand long periods of water deficiency, it is moderately tolerant to cold environmental stress conditions including drought and cold have been found to induce expression of many genes in plants. DRE/CRT elements (dehydration responsive elements/C repeats) with the core sequence G/ACCGAC have been identified in the promoters of most stress inducible genes Arabidopsis transcriptional activators CBF/DREB1 bind to these cis-acting elements and control stress dependent gene expression. The CBF/DREB1 proteins belong to the plant specific AP2/ERF (APETALA2/ethylene responsive element binding factor) family of DNA-binding proteins. Although homologues of genes encoding CBF1/DREB1 proteins have been isolated and characterised in many plants, these genes have not been determined in olive. Reverse Transcription PCR and Rapid Amplification of cDNA Ends (RACE) was used to obtain full length cDNA. In this study a 300 bp fragment that shows homology to AP2 domain of CBF1/DREB1 genes was cloned. doi:10.1016/j.copbio.2011.05.478
E-mail address: [email protected] (Z. Zainal)
P55
Salinity stress is one of the most important environmental factors that impose huge reduction in crop growth and yield. Thus, there is a need of continued effort to understand the salt tolerance mechanism to enable development of salt tolerant plants. The present study was undertaken to generate information on salt stress responsive genes in Oryza sativa var. MR219, using suppression subtractive hybridization technique. cDNA library was constructed after exposing the two weeks old rice seedlings to 150 mM NaCl for 2, 4, 8, 12 and 24 h. A total of 576 high qualities of ESTs were obtained. BLASTX search revealed overexpression of 326 unigenes. Majority of these EST sequences are homologous to the previously known salt-responsive genes. Based on GO annotation, 326 EST sequences were divided in to three groups: 143 EST’s were located under biological process, 107 EST’s under cellular component and 76 were annotated under molecular functions, Using RT-PCR, we examined the expression of two candidate genes (MR219SAP8 and MR219salT) and confirmed that their expression levels were increased with different salt concentration. Further functional characterization of the genes that overexpress under salt stress will be of particular importance in understanding the complex mechanism associated with salt tolerance
Cloning of -carbonic anhydrase cDNA from Oleae europeae exposed to cold stress
doi:10.1016/j.copbio.2011.05.477
Munise Yurtsever, Senay Vural Korkut Department of Biology, Faculty of Science and Arts, Yildiz Technical University, Davutpasa Campus, Esenler/Istanbul, Turkey E-mail address: [email protected] (M. Yurtsever) Olive (Olea europeae) is an economically important plant grown extensively in most countries in Mediterranean Region. As well as consumption of fruits as food and for oil production different strategies may be developed to use olive based resources in biotechnology, one of which could be cloning and expression of enzyme encoding genes from various tissues of the plant. Carbonic anhydrase (CA) is a zinc-containing enzyme that catalyzes the reversible hydration of carbon dioxide (CO2 + H2 O ↔ HCO3 − + H+ ) and is widely distributed in animals, plants, archaebacteria, and eubacteria. CAs are involved in a variety of biological processes including pH regulation, CO2 transfer, ion exchange, respiration, biosynthesis, and photosynthetic CO2 fixation. Expression of CA transcript is differentially up regulated in response to various abiotic stresses. CA cDNA clone was obtained from leaves of Oleae europeae exposed to cold stress by reverse transcription PCR. 392 bp fragment of (-CA cDNA was cloned from leaves of O. europeae exposed to cold stress by homologous cloning. The sequence shared significant homology with a region of O. europeae (-CA contig sequence which is found in NCBI databases. Contig sequence information was used to obtained full length cDNA clone. doi:10.1016/j.copbio.2011.05.479
Abstracts / Current Opinion in Biotechnology 22S (2011) S15–S152
stress. In the present study, six different wheat (Triticum aestivum L.) genotypes including two tolerant sister lines (Ducula-1 and Ducula-4), two standard cultivars widely grown in the Eastern Mediterranean (‘Basribey’ and ‘Golia’) and two susceptible cultivars (‘Seri-82’ and ‘Dogankent’) were subjected to different waterlogging treatments and their proline and chlorophyll contents were measured by spectrophotometer. About 25 anaerobically induced genes were analyzed using real time polymerase chain reaction (RT-PCR) method. Wheat seedlings were subjected to waterlogging treatments of 0, 0.5, 1, 2, 4, 6, 8, 12, 24, 48, 72 h and after each treatment, leaf samples were collected for chlorophyll, carotenoid, proline contents and gene expression analyses. A total of 32 primer pairs obtained from 25 anaerobically induced genes were used for RT-PCR. Total chlorophyll content decreased while proline content increased as waterlogging duration was prolonged. Gene expression differences among varieties under waterlogging and control conditions will be also presented. doi:10.1016/j.copbio.2011.05.476
P53 Isolation, identification and expression analysis of saltresponsive genes in Oryza sativa var. MR219 using PCRbased suppression subtractive hybridization Zamri Zainal 1 , Poya Hedayati 2 , Ismanizan Ismail 1 , Che Radziah Mohd Zain 2 , Zuraida A Rahman 3 1
Institue of Systems Biology, Universiti Kebangsaan, Malaysia School of Biosciences and Biotechnology, Universiti Kebangsaan, Malaysia 3 Biotechnology Research, MARDI, Malaysia 2
P54 Isolation of the homologoues of genes encoding CBF1/DREB1 proteins from Olive (Olea europaea) Beste Tacal, Gizem Hacimuto, Senay Vural Korkut Department of Biology, Faculty of Science and Arts, Yildiz Technical University, Istanbul, Turkey E-mail address: [email protected] (B. Tacal) Olive is an economically important plant cultivated in extensive areas in the Mediterranean countries. The plant may be subjected to heat and drought stress during the summer and chilling stress during the winter. Although it can stand long periods of water deficiency, it is moderately tolerant to cold environmental stress conditions including drought and cold have been found to induce expression of many genes in plants. DRE/CRT elements (dehydration responsive elements/C repeats) with the core sequence G/ACCGAC have been identified in the promoters of most stress inducible genes Arabidopsis transcriptional activators CBF/DREB1 bind to these cis-acting elements and control stress dependent gene expression. The CBF/DREB1 proteins belong to the plant specific AP2/ERF (APETALA2/ethylene responsive element binding factor) family of DNA-binding proteins. Although homologues of genes encoding CBF1/DREB1 proteins have been isolated and characterised in many plants, these genes have not been determined in olive. Reverse Transcription PCR and Rapid Amplification of cDNA Ends (RACE) was used to obtain full length cDNA. In this study a 300 bp fragment that shows homology to AP2 domain of CBF1/DREB1 genes was cloned. doi:10.1016/j.copbio.2011.05.478
E-mail address: [email protected] (Z. Zainal)
P55
Salinity stress is one of the most important environmental factors that impose huge reduction in crop growth and yield. Thus, there is a need of continued effort to understand the salt tolerance mechanism to enable development of salt tolerant plants. The present study was undertaken to generate information on salt stress responsive genes in Oryza sativa var. MR219, using suppression subtractive hybridization technique. cDNA library was constructed after exposing the two weeks old rice seedlings to 150 mM NaCl for 2, 4, 8, 12 and 24 h. A total of 576 high qualities of ESTs were obtained. BLASTX search revealed overexpression of 326 unigenes. Majority of these EST sequences are homologous to the previously known salt-responsive genes. Based on GO annotation, 326 EST sequences were divided in to three groups: 143 EST’s were located under biological process, 107 EST’s under cellular component and 76 were annotated under molecular functions, Using RT-PCR, we examined the expression of two candidate genes (MR219SAP8 and MR219salT) and confirmed that their expression levels were increased with different salt concentration. Further functional characterization of the genes that overexpress under salt stress will be of particular importance in understanding the complex mechanism associated with salt tolerance
Cloning of -carbonic anhydrase cDNA from Oleae europeae exposed to cold stress
doi:10.1016/j.copbio.2011.05.477
Munise Yurtsever, Senay Vural Korkut Department of Biology, Faculty of Science and Arts, Yildiz Technical University, Davutpasa Campus, Esenler/Istanbul, Turkey E-mail address: [email protected] (M. Yurtsever) Olive (Olea europeae) is an economically important plant grown extensively in most countries in Mediterranean Region. As well as consumption of fruits as food and for oil production different strategies may be developed to use olive based resources in biotechnology, one of which could be cloning and expression of enzyme encoding genes from various tissues of the plant. Carbonic anhydrase (CA) is a zinc-containing enzyme that catalyzes the reversible hydration of carbon dioxide (CO2 + H2 O ↔ HCO3 − + H+ ) and is widely distributed in animals, plants, archaebacteria, and eubacteria. CAs are involved in a variety of biological processes including pH regulation, CO2 transfer, ion exchange, respiration, biosynthesis, and photosynthetic CO2 fixation. Expression of CA transcript is differentially up regulated in response to various abiotic stresses. CA cDNA clone was obtained from leaves of Oleae europeae exposed to cold stress by reverse transcription PCR. 392 bp fragment of (-CA cDNA was cloned from leaves of O. europeae exposed to cold stress by homologous cloning. The sequence shared significant homology with a region of O. europeae (-CA contig sequence which is found in NCBI databases. Contig sequence information was used to obtained full length cDNA clone. doi:10.1016/j.copbio.2011.05.479