- Email: [email protected]
Effect of salicylic acid and its derivatives on Alternaria brassicae induced pathogenesis and in vitro phosphorylation studies in Brassica juncea
S230
Abstracts / Journal of Biotechnology 136S (2008) S217–S231
To analyze the expression patterns of PNDREB1 in peanut, particularly under abiotic stresses treatments, RT-PCR analysis was performed. The results showed that the PNDREB1 were expressed constitutively. Furthermore, it could be induced by dehydration and 4 ◦ C low temperature, with slight induced by 250 mM NaCl treated for 2 h. In order to confirm the functions of AP2 domain in PNDREB1 biochemically and physiologically, the yeast one-hybrid system was carried out, meantime, the binary vectors of PNDREB1 were constructed for peanut and Arabidopsis transformation. The preliminary analysis indicating PNDREB1 participated in stress signal transduction pathway, and the further verification of PNDREB1’s function is still undergoing.
Liu, Y.G., Whittier, R.F., 1995. Thermal asymmetric interlaced PCR: automatable amplification and sequencing of insert end fragments from P1 and YAC clones for chromosome walking. Genomics 25, 674–681.
doi:10.1016/j.jbiotec.2008.07.488 IV1-YP-002 Effect of salicylic acid and its derivatives on Alternaria brassicae induced pathogenesis and in vitro phosphorylation studies in Brassica juncea Leena Singh 1,∗ , Gohar Taj 1 , Govind Krishan Garg 2 1
Reference Agarwal, P.K., Agarwal, P., Reddy, M.K., Sopory, S.K., 2006. Role of DREB transcription factors in abiotic and biotic stress tolerance in plants. Plant Cell Rep. 25, 1263–1274.
doi:10.1016/j.jbiotec.2008.07.487 IV1-P-032 Isolation of genomic sequences flanking the fibroin gene of Antheraea pernyi by using thermal asymmetric interlaced PCR Liji Jin 1,∗ , Weiping Xu 1,2 , Wenli Li 1 , Yongping Xu 1 , Lijia An 1 1
Department of Bioscience and Biotechnology, Dalian University of Technology, Dalian 116024, China 2 Alberta Agriculture and Rural Development, Agriculture Center, Lethbridge, Alberta T1J4V6, Canada E-mail address: [email protected] (L. Jin). In order to integrate foreign gene into the fibroin region of silkworm genome by homologous recombination, vectors were requested to be constructed with foreign gene bordering with DNA segments adjacent to the known fibroin gene (Li et al., 2003). In the present study, thermal asymmetric interlaced (TAIL) PCR (Liu and Whittier, 1995) was utilized to amplify the unknown flanking region of the fibroin gene of Antheraea pernyi. Nested sequencespecific primers were designed with high annealing temperature, and used together with the existing arbitrary degenerated primers of low annealing temperature. Each of the TAIL-PCR cycles consisted of two high stringency (annealing at ≥64 ◦ C) and one low stringency (annealing at ≤44 ◦ C) PCR cycles, which gradually prevailed the specific amplification over non-specific ones. Totally, three consecutive TAIL-PCR were carried out to amplify the 5 and 3 flanking region. A 994 bp fragment of 5 end and a 1438 bp fragment of 3 end adjacent to the known fibroin gene were isolated. Sequence analysis demonstrated that there was no highly conserved region of the isolated sequences with other silkworm genome DNA. The amplification of unknown flanking region of fibroin gene may facilitate the construction of homologous recombination vectors and the following genomic modification of A. pernyi fibroin gene. The rapid isolation of the flanking sequences of the fibroin gene demonstrated the TAIL-PCR followed was an efficient method for the isolation of unknown fragments adjacent to known sequences.
Department of Molecular Biology and Genetic Engineering, Gobind Ballab Pant University of Agriculture & Technology, Pantnagar, Udham Singh Nagar, Uttrakhand 263145, India 2 Krishidhan Seeds Ltd. Jalna, Maharastra, India E-mail address: sci [email protected] (L. Singh). Plants respond to different biotic and abiotic stresses by inducing various protein kinases and other signaling cascades with varying degrees of cross-talks. Salicylic acid and its derivatives have widely known to have induced Systemic Acquired Resistance (SAR) in different plants and have key roles in disease resistance (Shirasu et al., 1997). In our studies salicylic acid (SA) and its three derivatives methyl-salicylic acid, acetyl-salicylic acid and sulpho-salicylic acid have shown different degrees of SAR induction in different Brassica genotypes. In this study we have found that Alternaria brassicae challenge, along with derivatives of SA induces SAR and in vitro phosphorylation in Brassica juncea. The observation that disease progression and SAR induction both involves in vitro phosphorylation (Zhang and Klessig, 1997) suggests that phosphorylation is one of the most common and widely exploited of the mechanisms in plant defense signaling and protein kinases are the most probable candidate molecules involved in B. juncea and A. brassicae pathogenesis.
References Shirasu, K., Nakajima, H., Rajasekhar, V.K., Dixon, R.A., Lamb, c., 1997. Salicylic acid potentiates an agonist-dependent gain control that amplifies pathogen signals in the activation of defense mechanisms. Plant Cell 9, 261–270. Zhang, S., Klessig, D.F., 1997. Salicylic acid activates a 48 kD MAPKinase in tobacco. Plant J. 9, 809–824.
doi:10.1016/j.jbiotec.2008.07.489 IV1-YP-028 Compared pollen sterility of different rice cultivars in low temperature condition Jaehak Kim 1,∗ , Chenghai Jin 1 , Hua Jin 1,2 , Xiaoxia Gou 1 , Youngboum Shin 1 , Jeongil Lee 3 , Jongdoo Yea 3 , Soon-Kwan Hong 1,4 1
Department of Plant Biotechnology, Kangwon National University, Chuncheon, 200-701, Republic of Korea 2 Key Laboratory of Biotechnology and Bioresources Utilization, Dalian Nationalities University, Dalian, Liaoning, 116600, PR China 3 National Institute of Crop Science, RDA, 441-707, Republic of Korea 4 Institute of Bioscience and Biotechnology, Kangwon National University, Chuncheon, 200-701, Republic of Korea
References
E-mail address: [email protected] (S.-K. Hong).
Li, W.L., Jin, L.J., An, L.J., 2003. Construction of targeting vector and expression of green fluorescent protein in the silkworm, Antheraea pernyi. DNA Cell Biol. 22, 441–446.
Rice plants are susceptible to low temperature during the young microspore stage. Low temperature at this time increases spikelet sterility which can cause massive yield loss (Daniela et al., 2003). In
Abstracts / Journal of Biotechnology 136S (2008) S217–S231
To analyze the expression patterns of PNDREB1 in peanut, particularly under abiotic stresses treatments, RT-PCR analysis was performed. The results showed that the PNDREB1 were expressed constitutively. Furthermore, it could be induced by dehydration and 4 ◦ C low temperature, with slight induced by 250 mM NaCl treated for 2 h. In order to confirm the functions of AP2 domain in PNDREB1 biochemically and physiologically, the yeast one-hybrid system was carried out, meantime, the binary vectors of PNDREB1 were constructed for peanut and Arabidopsis transformation. The preliminary analysis indicating PNDREB1 participated in stress signal transduction pathway, and the further verification of PNDREB1’s function is still undergoing.
Liu, Y.G., Whittier, R.F., 1995. Thermal asymmetric interlaced PCR: automatable amplification and sequencing of insert end fragments from P1 and YAC clones for chromosome walking. Genomics 25, 674–681.
doi:10.1016/j.jbiotec.2008.07.488 IV1-YP-002 Effect of salicylic acid and its derivatives on Alternaria brassicae induced pathogenesis and in vitro phosphorylation studies in Brassica juncea Leena Singh 1,∗ , Gohar Taj 1 , Govind Krishan Garg 2 1
Reference Agarwal, P.K., Agarwal, P., Reddy, M.K., Sopory, S.K., 2006. Role of DREB transcription factors in abiotic and biotic stress tolerance in plants. Plant Cell Rep. 25, 1263–1274.
doi:10.1016/j.jbiotec.2008.07.487 IV1-P-032 Isolation of genomic sequences flanking the fibroin gene of Antheraea pernyi by using thermal asymmetric interlaced PCR Liji Jin 1,∗ , Weiping Xu 1,2 , Wenli Li 1 , Yongping Xu 1 , Lijia An 1 1
Department of Bioscience and Biotechnology, Dalian University of Technology, Dalian 116024, China 2 Alberta Agriculture and Rural Development, Agriculture Center, Lethbridge, Alberta T1J4V6, Canada E-mail address: [email protected] (L. Jin). In order to integrate foreign gene into the fibroin region of silkworm genome by homologous recombination, vectors were requested to be constructed with foreign gene bordering with DNA segments adjacent to the known fibroin gene (Li et al., 2003). In the present study, thermal asymmetric interlaced (TAIL) PCR (Liu and Whittier, 1995) was utilized to amplify the unknown flanking region of the fibroin gene of Antheraea pernyi. Nested sequencespecific primers were designed with high annealing temperature, and used together with the existing arbitrary degenerated primers of low annealing temperature. Each of the TAIL-PCR cycles consisted of two high stringency (annealing at ≥64 ◦ C) and one low stringency (annealing at ≤44 ◦ C) PCR cycles, which gradually prevailed the specific amplification over non-specific ones. Totally, three consecutive TAIL-PCR were carried out to amplify the 5 and 3 flanking region. A 994 bp fragment of 5 end and a 1438 bp fragment of 3 end adjacent to the known fibroin gene were isolated. Sequence analysis demonstrated that there was no highly conserved region of the isolated sequences with other silkworm genome DNA. The amplification of unknown flanking region of fibroin gene may facilitate the construction of homologous recombination vectors and the following genomic modification of A. pernyi fibroin gene. The rapid isolation of the flanking sequences of the fibroin gene demonstrated the TAIL-PCR followed was an efficient method for the isolation of unknown fragments adjacent to known sequences.
Department of Molecular Biology and Genetic Engineering, Gobind Ballab Pant University of Agriculture & Technology, Pantnagar, Udham Singh Nagar, Uttrakhand 263145, India 2 Krishidhan Seeds Ltd. Jalna, Maharastra, India E-mail address: sci [email protected] (L. Singh). Plants respond to different biotic and abiotic stresses by inducing various protein kinases and other signaling cascades with varying degrees of cross-talks. Salicylic acid and its derivatives have widely known to have induced Systemic Acquired Resistance (SAR) in different plants and have key roles in disease resistance (Shirasu et al., 1997). In our studies salicylic acid (SA) and its three derivatives methyl-salicylic acid, acetyl-salicylic acid and sulpho-salicylic acid have shown different degrees of SAR induction in different Brassica genotypes. In this study we have found that Alternaria brassicae challenge, along with derivatives of SA induces SAR and in vitro phosphorylation in Brassica juncea. The observation that disease progression and SAR induction both involves in vitro phosphorylation (Zhang and Klessig, 1997) suggests that phosphorylation is one of the most common and widely exploited of the mechanisms in plant defense signaling and protein kinases are the most probable candidate molecules involved in B. juncea and A. brassicae pathogenesis.
References Shirasu, K., Nakajima, H., Rajasekhar, V.K., Dixon, R.A., Lamb, c., 1997. Salicylic acid potentiates an agonist-dependent gain control that amplifies pathogen signals in the activation of defense mechanisms. Plant Cell 9, 261–270. Zhang, S., Klessig, D.F., 1997. Salicylic acid activates a 48 kD MAPKinase in tobacco. Plant J. 9, 809–824.
doi:10.1016/j.jbiotec.2008.07.489 IV1-YP-028 Compared pollen sterility of different rice cultivars in low temperature condition Jaehak Kim 1,∗ , Chenghai Jin 1 , Hua Jin 1,2 , Xiaoxia Gou 1 , Youngboum Shin 1 , Jeongil Lee 3 , Jongdoo Yea 3 , Soon-Kwan Hong 1,4 1
Department of Plant Biotechnology, Kangwon National University, Chuncheon, 200-701, Republic of Korea 2 Key Laboratory of Biotechnology and Bioresources Utilization, Dalian Nationalities University, Dalian, Liaoning, 116600, PR China 3 National Institute of Crop Science, RDA, 441-707, Republic of Korea 4 Institute of Bioscience and Biotechnology, Kangwon National University, Chuncheon, 200-701, Republic of Korea
References
E-mail address: [email protected] (S.-K. Hong).
Li, W.L., Jin, L.J., An, L.J., 2003. Construction of targeting vector and expression of green fluorescent protein in the silkworm, Antheraea pernyi. DNA Cell Biol. 22, 441–446.
Rice plants are susceptible to low temperature during the young microspore stage. Low temperature at this time increases spikelet sterility which can cause massive yield loss (Daniela et al., 2003). In