Abstract
A cDNA library was constructed from leaves of a cold-acclimated Eucalyptus gunnii elite clone and 13,056 ESTs were sequenced. When the 5,457 unique sequences were compared with Arabidopsis, rice, and poplar datasets, more than 38% of no-hits were found and the best annotation was provided by the woody perennial poplar database. However, among the 62% of sequences that could be annotated, the vast majority (87.5%) was found to be common to the four compared plant species, which shows how highly conserved this gene pool is across the plant kingdom. When the distribution of the annotated sequences was studied according to the Gene Ontology classification, some typical features of the dehydrative stress transcriptome were observed. In particular, genes from sugar metabolism and, above all, those induced by stress or external stimuli were well represented (6% in total). In addition, the library was found to contain a substantial number of ESTs encoding putative transcription factors including CBF (CRT-Binding Factor). The enrichment of this library with stress-related genes was strongly suggested by the high redundancy (up to 300 ESTs) of several genes known to be involved in cell protection such as the PCP (Pollen Coat Protein), Lti6b (Low temperature induced) or metallothionein. It was further confirmed by demonstrating the cold induction of a set of representative genes through Real Time-PCR. All these characteristics make this Eucalyptus EST collection very suitable for investigating the molecular basis of cold tolerance in a woody perennial and for isolating important or new candidate frost tolerance genes.
Similar content being viewed by others
References
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215(3):403–410
Bausher M, Shatters R, Chaparro J, Dang P, Hunter W, Niedz R (2003) An expressed sequence tag (EST) set from Citrus sinensis L. Osbeck whole seedlings and the implications of further perennial source investigations. Plant Sci 165:415–422
Benedict C, Skinner JS, Meng R, Chang Y, Bhalerao R, Huner NPA, Finn CE, Chen THH, Hurry V (2006) The CBF1-dependent low temperature signalling pathway, regulon and increase in freeze tolerance are conserved in Populus spp. Plant, Cell and Environ 29:1259–1272
Brooker MIH (2000) A new classification of the genus Eucalyptus l’Her. (Myrtaceae). Aust Syst Bot 13:79–148
Cao S, Ye M, Jiang S (2005) Involvement of GIGANTEA gene in the regulation of the cold stress response in Arabidopsis. Plant Cell Reports 25:683–690
Chinnusamy V, Ohta M, Kanrar S, Lee BH, Hong X, Agarwal M, Zhu JK (2003) ICE1: a regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis. Genes Dev 17:1043–1054
Choi H, Hong J, Ha J, Kang J, Kim SY (2000) ABFs, a family of ABA-responsive element binding factors. J Biol Chem 275:1723–1730
Davletova S, Schlauch K, Coutu J, Mittler R (2005) The zinc-finger protein ZAT12 plays a central role in reactive oxygen and abiotic stress signaling in Arabidopsis. Plant Physiol 139:847–856
El Kayal W, Keller G, Debayles C, Kumar R, Weier D, Teulières C, Marque C (2006a) Regulation of tocopherol biosynthesis through transcriptional control of tocopherol cyclase during cold hardening in Eucalyptus gunnii. Physiol Plant 126:221–223
El Kayal W, Navarro M, Marque G, Keller G, Marque C, Teulières C (2006b) Expression profile of CBF-like transcriptional factor genes from Eucalyptus in response to cold. J Exp Bot 57:2455–2469
FAO (2000) Global forest resource assessment, main report. United Nations Food and Agriculture Organisation, Rome
Forment J, Gadea J, Huerta L, Abizanda L, Agusti J, Alamar S, Alos E, Andres F, Arribas R, Beltran JP, Berbel A, Blazquez MA, Brumos J, Canas LA, Cercos M, Colmenero-Flores JM, Conesa A, Estables B, Gandia M, Garcia-Martinez JL, Gimeno J, Gisbert A, Gomez G, Gonzalez-Candelas L, Granell A, Guerri J, Lafuente MT, Madueno F, Marcos JF, Marques MC, Martinez F, Martinez-Godoy MA, Miralles S, Moreno P, Navarro L, Pallas V, Perez-Amador MA, Perez-Valle J, Pons C, Rodrigo I, Rodriguez PL, Royo C, Serrano R, Soler G, Tadeo F, Talon M, Terol J, Trenor M, Vaello L, Vicente O, Vidal C, Zacarias L, Conejero V (2005) Development of a Citrus genome-wide EST collection and cDNA microarray as resources for genomic studies. Plant Mol Biol 57:375–391
Hjelm U, Ogren E (2003) Is photosynthetic acclimation to low temperature controlled by capacities for storage and growth at low temperature? Results from comparative studies of grasses and trees. Physiol Plant 119:113–120
Huang X, Madan A (1999) CAP3: a DNA sequence assembly program. Genome Res 9:868–877
Ji W, Li Y, Li J, Dai C-h, Wang X, Bai X, Cai H, Yang L, Zhu Y-m (2006) Generation and analysis of expressed sequence tags from NaCl-treated Glycine soja. BMC Plant Biol 6:4
Jones J, Dunsmuir P, Bedbrook J (1985) High level expression of introduced chimeric genes in regenerated transformed plants. EMBO J 4:2411–2418
Keller G (2006) Analyse du transcriptome de l’Eucalyptus pendant l’acclimatation au froid. Recherche de gènes candidats de la tolérance au gel. Doctorat de l’Université Paul Sabatier, Toulouse, France
Kim SY (2006) The role of ABF family bZIP class transcription factors in stress response. Physiol Plant 126:519–527
Kohler A, Blaudez D, Chalot M, Martin F (2004) Cloning and expression of multiple metallothioneins from hybrid poplar. New Phytol 164:83–93
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-DDCT method. Methods 25:402–408
Mehta PA, Sivaprakash K, Parani M, Venkataraman G, Parida AK (2005) Generation and analysis of expressed sequence tags from the salt-tolerant mangrove species Avicennia marina (Forsk) Vierh. Theor Appl Genet 110:416–424
Myburg A, Grattapaglia D, Tuskan G, Schmutz J, Rokhsar D, Barry K, Bristow J; The Eucalyptus Genome Network (EUCAGEN) (2008) Eucalyptus: sequencing a global tree genome for energy, fiber and wood, Genomic of energy and environnement, JGI Third Annual Meeting, march 26–28 2008, Walnut Creek, California, USA
Paux E, Tamasloukht M, Ladouce N, Sivadon P, Grima-Pettenati J (2004) Identification of genes preferentially expressed during wood formation in Eucalyptus. Plant Mol Biol 55:263–280
Pavy N, Paule C, Parsons L, Crow JA, Morency M-J, Cooke J, Johnson JE, Noumen E, Guillet-Claude C, Butterfield Y, Barber S, Yang G, Liu J, Stott J, Kirkpatrick R, Siddiqui A, Holt R, Marra M, Seguin A, Retzel E, Bousquet J, MacKay J (2005) Generation, annotation, analysis and database integration of 16,500 white spruce EST clusters. BMC Genomics 6:144
Poke FS, Vaillancourt RE, Potts BM, Reid JB (2005) Genomic research in Eucalyptus. Genetica 125:79–101
Porat R, Pasentsis K, Rozentzvieg D, Gerasopoulos D, Falara V, Samach A, Lurie S, Kanellis AK (2004) Isolation of a dehydrin cDNA from orange and grapefruit citrus fruit that is specifically induced by the combination of heat followed by chilling temperatures. Physiol Plant 120:256–264
Ruiter RK, van Eldik GJ, van Herpen MMA, Schrauwen JAM, Wullems GJ (1999) Hydration-dependent gene expression in Brassica oleracea anthers. Sex Plant Reprod 12:135–143
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, New York
Savitch LV, Leonardos ED, Krol M, Jansson S, Grodzinski B, Huner NPA, Öquist G (2002) Two different strategies for light utilization in photosynthesis in relation to growth and cold acclimation. Plant Cell Env 25:761–771
Shinwari ZK, Nakashima K, Miura S, Kasuga M, Seki M, Yamaguchi-Shinozaki K, Shinozaki K (1998) An Arabidopsis gene family encoding DRE/CRT binding proteins involved in low-temperature-responsive gene expression. Biochem Biophys Res Commun 250:161–170
Teulières C, Marque C (2007) Eucalyptus. In: Pua EC, Davey MR (eds) Transgenic crops, vol 60. Springer, Berlin, pp 387–406
Teulières C, Bossinger G, Moran G, Marque C (2007) Stress studies in Eucalyptus. In: Plant Stress, Global Science Books (eds),vol I(2), 197–215
Thomashow MF (1999) Plant cold acclimation: freezing tolerance genes and regulatory mechanisms. Annu Rev Plant Physiol Plant Mol Biol 50:571–599
Vannini C, Locatelli F, Bracale M, Magnani E, Marsoni M, Osnato M, Mattana M, Baldoni E, Coraggio I (2004) Overexpression of the rice Osmyb4 gene increases chilling and freezing tolerance of Arabidopsis thaliana plants. Plant J 37:115–127
Vogel JT, Zarka DG, Van Buskirk HA, Fowler SG, Thomashow MF (2005) Roles of the CBF2 and ZAT12 transcription factors in configuring the low temperature transcriptome of Arabidopsis. Plant J 41:195–211
Wei H, Dhanaraj AL, Rowland LJ, Fu Y, Krebs SL, Arora R (2005) Comparative analysis of expressed sequence tags from cold-acclimated and non-acclimated leaves of Rhododendron catawbiense Michx. Planta 221:406–416
Welling A, Palva ET (2006) Molecular control of cold acclimation in trees. Physiol Plant 127:167–181
Wissel K, Pettersson F, Berglund A, Jansson S (2003) What affects mRNA levels in leaves of field-grown aspen? A study of developmental and environmental influences. Plant Physiol 133:1190–1197
Wong CE, Li Y, Whitty BR, Diaz-Camino C, Akhter SR, Brandle JE, Golding GB, Weretilnyk EA, Moffatt BA, Griffith M (2005) Expressed sequence tags from the Yukon ecotype of Thellungiella reveal that gene expression in response to cold, drought and salinity shows little overlap. Plant Mol Biol 58:561–574
Acknowledgements
We thank AFOCEL for supplying the Eucalyptus cuttings. Research supports, salaries, and grants were provided by French Ministry of Research and Technology (MNRT), the “Midi Pyrénées French Council” and TEMBEC SA R&D KRAFT (St GAUDENS, France). Thanks to Victoria McBride for accurate English editing of the manuscript.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by J. Dean
Rights and permissions
About this article
Cite this article
Keller, G., Marchal, T., SanClemente, H. et al. Development and functional annotation of an 11,303-EST collection from Eucalyptus for studies of cold tolerance. Tree Genetics & Genomes 5, 317–327 (2009). https://doi.org/10.1007/s11295-008-0184-7
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11295-008-0184-7