Abstract
Large-scale sequencing of randomly selected cDNA clones was used to isolate numerous genes in rice (Oryza sativa L.). Total RNA used for cDNA synthesis was prepared from suspension-cultured cells of rice grown under stressed conditions, such as in saline or nitrogen-starvation conditions. A total of 780 cDNA clones were partially sequenced and about 15% could be identified as putative genes. In the library constructed under saline conditions, we identified several genes associated with signal transduction, such as protein kinase and small GTP-binding protein genes. Many stress-related genes were isolated from both the saline and nitrogen-starvation libraries. These results indicate that stress treatment of suspension-cultured cells makes it possible to efficiently isolate various types of plant genes. To examine the usefulness of such tagged cDNAs for the study of gene expression in a specific metabolic pathway, we analyzed mRNA levels of genes engaged in the ATP-generating pathways in cultured cells of rice under different stresses, such as 20% sucrose, salt stress, cold stress and nitrogen-starvation stress. The results suggest that the coordinated induction of several genes in key steps under stressed conditions may be essential for activation of the entire energy-producing pathway to maintain homeostasis in rice cells. Expressed sequence tags identified by random cDNA sequencing provide the opportunity to generate a transcript map of rice genes.
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Adams MD, Kelley JM, Gocayne JD, Dubnick M, Polymeropoulos MH, Xiao H, Merril CR, Wu A, Olde B, Moreno RF, Kerlavage AR, McCombie WR, Venter JC: Complementary DNA sequencing: expressed sequence tags and human genome project. Science 252: 1651–1656 (1991).
Adams MD, Dubnick M, Kerlavage AR, Moreno R, Kelley JM, Utterback TR, Nagle JW, Fields C, Venter JC: Sequence identification of 2375 human brain genes. Nature 355: 632–634 (1992).
Adams MD, Kerlavage AR, Fields C, Venter JC: 3400 new expressed sequence tags identify diversity of transcripts in human brain. Nature Genet 4: 256–267 (1993).
Höfte H, Desprez T, Amselem J, Chiapello H, Caboche M, Moisan A, Jourjon M-F, Charpenteau J-L, Berthomieu P, Guerrier Y, Giraudat J, Quigley F, Thomas F, Yu D-Y, Mache R, Raynal M, Cooke R, Grellet F, Delseny M, Parmentier Y, de Marcillac G, Gigot C, Fleck J, Philipps G, Axelos M, Bardet C, Tremousaygue D, Lescure B: An inventory of 1152 expressed sequence tags obtained by partial sequencing of cDNAs fromArabidopsis thaliana. Plant J 4: 1051–1061 (1993).
Höög C: Isolation of a large number of novel mammalian genes by a differential cDNA library screening strategy. Nucl Acids Res 19: 6123–6127 (1991).
Keith CS, Hoang DO, Barret BM, Feigelman B, Nelson MC, Thai H, Baysdorfer C: Partial sequence analysis of 130 randomly selected maize cDNA clones. Plant Physiol 101: 329–332 (1993).
Kidou S, Anai T, Umeda M, Aotsuka S, Tsuge T, Kato A, Uchimiya H: Molecular structure of ras-related small GTP-binding protein genes of rice plants and GTPase activities of gene products inEscherichia coli. FEBS Lett 332: 282–286 (1993).
McCombie WR, Adams MD, Kelley JM, FitzGerald MG, Uttetoack TR, Khan M, Dubnick M, Kerlavage AR, Venter JC, Fields C:Caenorhabditis elegans expressed sequence tags identify gene families and potential disease gene homologues. Nature Genet 1: 124–131 (1992).
McCouch SR, Kochert G, Yu ZH, Wang ZY, Khush GS, Coffman WR, Tanksley SD: Molecular mapping of rice chromosomes. Theor Appl Genet 76: 815–829 (1988).
Okubo K, Hori N, Matoba R, Niiyama T, Fukushima A, Kojima Y, Matsubara K: Large scale cDNA sequencing for analysis of quantitative and qualitative aspects of gene expression. Nature Genet 2: 173–179 (1992).
Palmiter RD: Magnesium precipitation of ribonucleoprotein complexes: expedient techniques for the isolation of undergraded polysomes and messenger ribonucleic acid. Biochemistry 13: 3606 (1974).
Park YS, Kwak JM, Kwon O-Y, Kim YS, Lee DS, Cho MJ, Lee HH, Nam HG: Generation of expressed sequence tags of random root cDNA clones ofBrassica napus by single-run partial sequencing. Plant Physiol 103: 359–370 (1993).
Saito A, Yano M, Kishimoto N, Nakagahra M, Yoshimura A, Saito K, Kuhara S, Ukai Y, Kawase M, Nagamine T, Yoshimura S, Ideta O, Ohsawa R, Hayano Y, Iwata N, Sugiura M: Linkage map of restriction fragment length polymorphism loci in rice. Jpn J Breed 41: 665–670 (1991).
Smith LM, Sanders JZ, Kaiser RJ, Hughes P, Dodd C, Connell CR, Heiner C, Kent SBH, Hood LE: Fluorescence detection in automated DNA sequence analysis. Nature 321: 674–679 (1986).
Toriyama K, Hinata K: Cell suspension and protoplate culture in rice. Plant Sci 41: 179–183 (1985).
Uchimiya H, Kidou S, Shimazaki T, Aotsuka S, Takamatsu S, Nishi R, Hashimoto H, Matsubayashi Y, Kidou N, Umeda M, Kato A: Random sequencing of cDNA libraries reveals a variety of expressed genes in cultured cells of rice (Oryza sativa L.). Plant J 2: 1005–1009 (1992).
Waterston R, Martin C, Craxton M, Huynh C, Coulson A, Hillier L, Durbin R, Green P, Shownkeen R, Halloran N, Metzstein M, Hawkins T, Wilson R, Berks M, Du Z, Thomas K, Thierry-Mieg J, Sulston J. A survey of expressed genes inCaenorhabditis elegans. Nature Gene 1: 114–123 (1992).
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Umeda, M., Hara, C., Matsubayashi, Y. et al. Expressed sequence tags from cultured cells of rice (Oryza sativa L.) under stressed conditions: analysis of transcripts of genes engaged in ATP-generating pathways. Plant Mol Biol 25, 469–478 (1994). https://doi.org/10.1007/BF00043875
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DOI: https://doi.org/10.1007/BF00043875