Abstract
Deschampsia antarctica (Poaceae) is one of the two vascular plants known to have colonized the Antarctic region. Studies examining the biosynthesis of flavonoids, compounds which plants use, for example, for protection against overexposure to UV light or as antioxidants that scavenge free radicals and other oxidative species, in D. antarctica may provide clues to its success in that extreme environment. We characterized the family of genes encoding chalcone isomerase (CHI EC 5.5.1.6), an important enzyme involved in flavonoid biosynthesis, in D. antarctica. Sequence analysis of the three family members revealed differences in numbers of introns and lengths of coding regions among the three and suggest that DaCHI3 is likely a pseudogene (ψDaChi2). Salinity stress resulted in differential mRNA expression of the DaCHI genes with ψDaCHI2 exhibiting the earliest response (3-h post-treatment), induced by as much as sevenfold, while DaCHI1 and DaCHI2 mRNAs accumulated later (3d and 5d post-treatment, respectively) and, in the case of DaCHI2, with a response of nearly sixfold. We discuss how differences in the proposed gene structures, deduced protein characteristics, and mRNA expression patterns suggest that the members of this gene family may have unique functions in the phenylpropanoid pathway in D. antarctica.
Similar content being viewed by others
References
Abagyan R, Totrov M, Kuznetsov D (1994) Icm—a new method for protein modeling and design: applications to docking and structure prediction from the distorted native conformation. J Comput Chem 15:488–506
Ahmed N, Maekawa M, Utsugi S, Rikiishia K, Ahmad A, Noda K (2006) The wheat Rc gene for red coleoptile colour codes for a transcriptional activator of late anthocyanin biosynthesis genes. J Cereal Sci 44:54–58
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215(3):403–410
Altschul S, Madden T, Schäffer A, Zhang J, Zhang Z et al (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402
Baker NA, Sept D, Joseph S, Holst MJ, McCammon JA (2001) Electrostatics of nanosystems: application to microtubules and the ribosome. Proc Natl Acad Sci USA 98(18):10037–10041
Bednar RA, Hadcock JR (1988) Purification and characterization of chalcone isomerase from soybeans. J Biol Chem 263:9582–9588
Bennetzen JL (2000) Transposable element contributions to plant gene and genome evolution. Plant Mol Biol 42:251–269
Benson D, Boguski M, Lipman DJ, Ostell J, Ouellette B (1998) GenBank. Nucleic Acids Res 26(1):1–7
Brendel V, Zhu W (2002) Computational modeling of gene structure in Arabidopsis thaliana. Plant Mol Biol 48:49–58
Burbulis IE, Winkel-Shirley B (1999) Interactions among enzymes of the Arabidopsis flavonoid biosynthetic pathway. Proc Natl Acad Sci USA 96(22):12929–12934
Chutipaijit S, Cha-um S, Sompornpailin K (2009) Differential accumulations of proline and flavonoids in Indica Rice varieties against salinity. Pak J Bot 41(5):2497–2506
Convey P, Smith RIL (2006) Responses of terrestrial Antarctic ecosystems to climate change. Plant Ecol 182:1–10
Dixon R, Steele C (1999) Flavonoids and isoflavonoids—a gold mine for metabolic engineering. Trends Plant Sci 4(10):394–400
Döring HP, Starlinger P (1986) Molecular genetics of transposable elements in plants. Ann Rev Genet 20:175–200
Druka A, Kudrna D, Rostoks N, Brueggeman R, von Wettstein D et al (2003) Chalcone isomerase gene from rice (Oryza sativa) and barley (Hordeum vulgare): physical, genetic and mutation mapping. Gene 302(1–2):171–178
Fowbert JA, Smith RIL (1994) Rapid population increase in native vascular plants in the Argentine Islands, Antarctic Peninsula. Arct Antarct Alp Res 26:290–296
Grace SC, Logan BA (2000) Energy dissipation and radical scavenging by the plant phenylpropanoid pathway. Philos Trans R Soc Lond B Biol Sci 355(1402):1499–1510
Guern J, Mathieu Y, Kurkdjian A, Manigault P, Manigault J, Gillet B, Beloeil C, Lallemand JY (1989) Regulation of vacuolar pH of plant cells. II. A 31 P NMR study of the modifications of vacuolar pH in isolated vacuoles induced by proton pumping and cation/H+exchanges. Plant Physiol 89:27–36
Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98
Humphrey W, Dalke A, Schulten K (1996) VMD: visual molecular dynamics. J Mol Graph 14(1):33–38
Irani NG, Hernandez JM, Grotewold E (2003) Regulation of anthocyanin pigmentation. Recent Adv Phytochem 38:59–78
Jaiswal P, Ni J, Yap I, Ware D, Spooner W et al (2006) Gramene: a bird’s eye view of cereal genomes. Nucleic Acids Res 34:717–723
Jez JM, Noel JP (2002) Reaction mechanism of chalcone isomerase. pH dependence, diffusion control, and product binding differences. J Biol Chem 277:1361–1369
Jez JM, Bowman ME, Noel JP (2002) Role of hydrogen bonds in the reaction mechanism of chalcone isomerase. Biochemistry 41(16):5168–5176
Jez JM, Bowman ME, Dixon RA, Noel JP (2000) Structure and mechanism of the evolutionarily unique plant enzyme chalcone isomerase. Nat Struct Biol 7:786–791
Kim S, Jones R, Yoo KS, Pike LM (2004) Gold color in onions (Allium cepa): a natural mutation of the chalcone isomerase gene resulting in a premature stop codon. Mol Genet Genomics 272(4):411–419
Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA et al (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948
Laskowski RA, Macarthur MW, Moss DS, Thornton JM (1993) Procheck: a program to check the stereochemical quality of protein structures. J Appl Crystallogr 26:283–291
Lawton-Rauh A (2003) Evolutionary dynamics of duplicated genes in plants. Mol Phylogenet Evol 29(3):396–409
Li F, Jin Z, Qu W, Zhao D, Ma F (2006) Cloning of a cDNA encoding the Saussurea medusa chalcone isomerase and its expression in transgenic tobacco. Plant Physiol Biochem 44(7–9):455–461
Long M, Betrán E, Thornton K, Wang W (2003) The origin of new genes: glimpses from the young and old. Nat Rev Genet 4(11):865–875
Melo F, Devos D, Depiereux E, Feytmans E (1997) ANOLEA: a www server to assess protein structures. Proc Int Conf Intell Syst Mol Biol 5:187–190
Morris G, Goodsell D, Halliday R, Huey R, Hart W et al (1999) Automated docking using a Lamarckian genetic algorithm and an empirical binding free energy function. J Comput Chem 19:1639–1662
Muir SR, Collins GJ, Robinson S, Hughes S, Bovy A et al (2001) Overexpression of petunia chalcone isomerase in tomato results in fruits containing increased levels of flavonols. Nat Biotechnol 19:470–474
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–479
Pardo JM, Cubero B, Leidi EO, Quintero FJ (2006) Alkali cation exchangers: roles in cellular homeostasis and stress tolerance. J Exp Bot 57(5):1181–1199
Patthy L (1999) Genome evolution and the evolution of exon-shuffling-a review. Gene 238(1):103–114
Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Res 29:2002–2007
Phillips JC, Braun R, Wang W, Gumbart J, Tajkhorshid E et al (2005) Scalable molecular dynamics with NAMD. J Comput Chem 26:1781–1802
Porebski SL, Bailey G, Baum RB (1997) Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components. Plant Mol Biol Rep 15(1):8–15
Prieto H, Utz D, Castro A, Aguirre C, González-Agüero M et al (2007) Browning in Annona cherimola fruit: role of polyphenol oxidase (PPO) and characterization of a coding sequence of the enzyme. J Agric Food Chem 55(22):9208–9218
Prince VE, Pickett FB (2002) Splitting pairs: diverging fates of duplicated genes. Nat Rev Genet 3:827–837
Quattrocchio F, Baudry A, Lepiniec L, Grotewold E (2006) The regulation of flavonoid biosynthesis. In: Grotewold E (ed) The science of flavonoids. Springer, New York, pp 97–122
Ralston L, Subramanian S, Matsuno M, Yu O (2005) Partial reconstruction of flavonoid and isoflavonoid biosynthesis in yeast using soybean type I and type II chalcone isomerases. Plant Physiol 137:1375–1388
Reuber S, Jende-Strid B, Wray V, Weissenböck G (1997) Accumulation of the chalcone isosalipurposide in primary leaves of barley flavonoid mutants indicates a defective chalcone isomerase. Physiol Plant 101:827–832
Robberecht R, Caldwell MM (1983) Protective mechanisms and acclimation to solar ultraviolet-B radiation in Oenothera stricta. Plant Cell Environ 6:477–485
Rozema J, Bjorn LO, Bornman JF, Gaberscik A, Haeder DP et al (2002) The role of UV-B radiation in aquatic and terrestrial ecosystems: an experimental and functional analysis of the evolution of UV-absorbing compounds. J Photochem Photobiol B 66:2–12
Sali A, Blundell TL (1993) Comparative protein modelling by satisfaction of spatial restraints. J Mol Biol 234:779–815
Shih CH, Chu H, Tang LK, Sakamoto W, Maekawa M et al (2008) Functional characterization of key structural genes in rice flavonoid biosynthesis. Planta 228(6):1043–1054
Shimada N, Aoki T, Sato S, Nakamura Y, Tabata S et al (2003) A cluster of genes encodes the two types of chalcone isomerase involved in the biosynthesis of general flavonoids and legume-specific 5-deoxy(iso)flavonoids in Lotus japonicus. Plant Physiol 131:941–951
Solovchenko A, Schmitz-Eiberger M (2003) Significance of skin flavonoids for UV-B-protection in apple fruits. J Exp Bot 54(389):1977–1984
Sparvoli F, Martin C, Scienza A, Gavazzi G, Tonelli C (1994) Cloning and molecular analysis of structural genes involved in flavonoid and stilbene biosynthesis in grape (Vitis vinifera L.). Plant Mol Biol 24(5):743–755
Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25(24):4876–4882
Usuka J, Brendel V (2000) Gene structure prediction by spliced alignment of genomic DNA with protein sequences: increased accuracy by differential splice site scoring. J Mol Biol 297:1075–1085
van Tunen AJ, Koes RE, Spelt CE, van der Krol AR, Stuitje AR et al (1988) Cloning of the two chalcone flavanone isomerase genes from Petunia hybrida: coordinate, light-regulated and differential expression of flavonoid genes. EMBO J 7:1257–1263
Walsh JB (1995) How often do duplicated genes evolve new functions? Genetics 139(1):421–428
Wiederstein M, Sippl MJ (2007) ProSA-web: interactive web service for the recognition of errors in three-dimensional structures of proteins. Nucleic Acids Res 35:W407–W410
Zamora P, Rasmussen S, Pardo A, Prieto H, Zuñiga GE (2010) Antioxidant responses of in vitro shoots of Deschampsia antarctica to Polyethylene glycol treatment. Antarct Sci 22(2):163–169
Acknowledgments
This research was funded through a Chilean Antarctic Institute Fellowship in support of doctoral theses focused on Antarctic studies to PZ, Fondecyt Grant 11085002, Programa Bicentenario de Ciencia y Tecnología grant PDA-23, the ICM program overseen by Dr. G. Zapata-Torres and DICYT (Universidad de Santiago de Chile).
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Zamora, P., Pardo, A., Fierro, A. et al. Molecular characterization of the chalcone isomerase gene family in Deschampsia antarctica . Polar Biol 36, 1269–1280 (2013). https://doi.org/10.1007/s00300-013-1346-0
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00300-013-1346-0