Identification and expression analysis of a novel phytocystatin in developing and germinating seeds of triticale (×Triticosecale Wittm.)

Joanna Simińska, Wiesław Bielawski

Abstract


In this paper the complete cDNA sequence of a newly identified triticale phytocystatin, TrcC-7, was analyzed. Because TrcC-7 transcripts were present in seeds, we hypothesized that it may regulate storage protein accumulation and degradation. Therefore, changes in mRNA and protein levels during the entire period of seed development and germination were examined. Expression of TrcC-7 increased during development and decreased at the end of maturation and subsequently increased during seed germination. Based on these results, TrcC-7 likely regulates cysteine proteinase activity during the accumulation and mobilization of storage proteins.

Keywords


phytocystatin; cysteine proteinase inhibitor; seed development; germination

Full Text:

PDF

References


Grudkowska M, Zagdańska B. Multifunctional role of plant cysteine proteinases. Acta Biochim Polon. 2004;51:609–624.

Szewińska J, Zdunek-Zastocka E, Pojmaj M, Bielawski W. Molecular cloning and expression analysis of triticale phytocystatins during development and germination of seeds. Plant Mol Biol Rep. 2012;30:867–877. http://dx.doi.org/10.1007/s11105-011-0384-4

Prabucka B, Drzymała A, Grabowska A. Molecular cloning and expression analysis of the main gliadin-degrading cysteine endopeptidase EP8 from triticale. J Cereal Sci. 2013;58:284–289. http://dx.doi.org/10.1016/j.jcs.2013.06.004

Szewińska J, Prabucka B, Krawczyk M, Mielecki M, Bielawski W. The participation of phytocystatin TrcC-4 in the activity regulation of EP8, the main prolamin degrading cysteine endopeptidase in triticale seeds. Plant Growth Regul. 2013;69:131–137. http://dx.doi.org/10.1007/s10725-012-9756-5

Chomczynski P, Sacchi N. Single-step method of total RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987;162:156–159. http://dx.doi.org/10.1016/0003-2697(87)90021-2

Untergrasser A, Cutcutache I, Koressaar T, Ye J, Faircloth BC, Remm M, Rozen SG. Primer3 – new capabilities and interfaces. Nucleic Acids Res. 2012;40(15):e115. http://dx.doi.org/10.1093/nar/gks596

Katoh K, Standley DM. MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Mol Biol Evol. 2013;30:772–780. http://dx.doi.org/10.1093/molbev/mst010

Goujon M, McWilliam H, Li W, Valentin F, Squizzato S, Paern J, Lopez R. A new bioinformatics analysis tools framework at EMBL-EBI. Nucleic Acids Res. 2010;38(2 suppl):W695–W699. http://dx.doi.org/10.1093/nar/gkq313

Gasteiger E, Hoogland C, Gattiker A, Duvaud S, Wilkins MR, Appel RD, Bairoch A. Protein identification and analysis tools on the ExPASy server. In: Walker JM, editor. The proteomics protocols handbook. New York, NY: Humana Press; 2005. p. 571–607. http://dx.doi.org/10.1385/1-59259-890-0:571

Petersen TN, Brunak S, von Heijne G, Nielsen H. SignalP 4.0: discriminating signal peptides from transmembrane regions. Nat Methods. 2011;8:785–786. http://dx.doi.org/10.1038/nmeth.1701

Martinez M, Cambra I, Carrillo L, Diaz-Mendoza M, Diaz I. Characterization of the entire cystatin gene family in barley and their target cathepsin L-like cysteine-proteases, partners in the hordein mobilization during seed germination. Plant Physiol. 2009;151:1531–1545. http://dx.doi.org/10.1104/pp.109.146019

Abe K, Emori Y, Kondo H, Suzuki K, Arai S. Molecular cloning of a cysteine proteinase inhibitor of rice (oryzacystatin). Homology with animal cystatins and transient expression in the ripening process of rice seeds. J Biol Chem. 1987;262:16793–16797.

Kondo H, Abe K, Nishimura I, Watanabe H, Emori Y, Arai S. Two distinct cystatin species in rice seeds with different specificities against cysteine proteinases. Molecular cloning, expression, and biochemical studies on oryzacystatin-II. J Biol Chem. 1990;265:15832–15837.

Massonneau A, Condamine P, Wisniewski JP, Zivy M, Rogowsky PM. Maize cystatins respond to developmental cues, cold stress and drought. Biochim Biophys Acta. 2005;1729:186–199. http://dx.doi.org/10.1016/j.bbaexp.2005.05.004

Corre-Menguy F, Cejudo FJ, Mazubert C, Vidal J, Lelandais-Brière C, Torres G, Rode A, Hartmann C. Characterization of the expression of a wheat cystatin gene during caryopsis development. Plant Mol Biol. 2002;50:687–698. http://dx.doi.org/10.1023/A:1019906031305

Kuroda M, Kiyosaki T, Matsumoto I, Misaka T, Arai S, Abe K. Molecular cloning, characterization, and expression of wheat cystatins. Biosci Biotechnol Biochem. 2001;65:22–28. http://dx.doi.org/10.1271/bbb.65.22

Abraham Z, Martinez M, Carbonero P, Diaz I. Structural and functional diversity within the cystatin gene family of Hordeum vulgare. J Exp Bot. 2006;57:4245–4255. http://dx.doi.org/10.1093/jxb/erl200

Dutt S, Singh VK, Marla SS, Kumar A. In silico analysis of sequential, structural and functional diversity of wheat cystatins and its implication in plant defense. Genomics Proteomics Bioinformatics. 2010;8:42–56. http://dx.doi.org/10.1016/S1672-0229(10)60005-8

Womack JS, Randall J, Kemp JD. Identifcation of a signal peptide for oryzacystatin-I. Planta. 2000;210:844–847. http://dx.doi.org/10.1007/s004250050688




DOI: https://doi.org/10.5586/asbp.2015.011

Journal ISSN:
  • 2083-9480 (online)
  • 0001-6977 (print; ceased since 2016)
This is an Open Access journal, which distributes its content under the terms of the Creative Commons Attribution License, which permits redistribution, commercial and non-commercial, provided that the content is properly cited.
The journal is a member of the Committee on Publication Ethics (COPE) and aims to follow the COPE’s principles.
The journal publisher is a member of the Open Access Scholarly Publishers Association.
The journal content is indexed in Similarity Check, the Crossref initiative to prevent scholarly and professional plagiarism.
Publisher
Polish Botanical Society