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Isolation and characterization of three cold acclimation-responsive dehydrin genes from Eucalyptus globulus

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Abstract

The molecular and physiological work related to cold hardiness in Eucalyptus globulus and the cold-responsive dehydrins is reported. The identification and full-length gene sequence of three dehydrins of 10, 20, and 30 kDa and the comparison of their promoters regarding to potential stress and hormone response elements in E. globulus are shown. The categorization of cold-responsive proteins as dehydrin was based on the similarity in amino acid composition with selected sequenced peptides from chilling-responsive dehydrin reported for other woody plants and the increasing of gene expression level during cold acclimation. The transcript accumulation for these three dehydrin genes increased with cold acclimation and decreased with deacclimation in leaf and stem tissues, being higher in a freezing-resistant genotype of E. globulus compared to a sensitive genotype. By western blot, five dehydrin peptides were identified which increased their expression, under cold stress in leaf and stem tissues. These results provide valuable information about cold acclimation and gene regulation in eucalypt genotypes that differ in their ability to tolerate frost temperature.

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Acknowledgments

The authors would like to thank Dr. Leon Bravo, Valeria Neira, Claudia Flores, and Daniela Salgado for their assistance in the establishment of growth chambers assays. The authors would also like to express their appreciation to Forestal Mininco S.A for providing the plant material, a grant from MECESUP, and a scholarship from Conicyt for the financial support to MF, including a research visit to Dr. Rajeev Arora's lab at Iowa State University. This research was supported by Genomica Forestal, grant from CORFO 05CTE04-03 and partially by Conicyt (grant no. 24100220).

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Authors and Affiliations

  1. Ciencias Forestales, Universidad de Concepcion, Concepcion, Chile

    Marta Fernandez & Sofia Valenzuela Águila

  2. Biotechnology Center, University of Concepcion, Concepcion, Chile

    Sofia Valenzuela Águila

  3. Department of Horticulture, Iowa State University, Ames, IA, USA

    Rajeev Arora & Keting Chen

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  1. Marta Fernandez
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  2. Sofia Valenzuela Águila
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  3. Rajeev Arora
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  4. Keting Chen
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Correspondence to Sofia Valenzuela Águila.

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Communicated by R. Sederoff

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Table S1

Description of three DHN sequences found in the cDNA library prepared from plants of E. globulus subjected to low temperature (nonfreezing) condition. Accession numbers of EuglDHNs nucleotide sequences are listed. The number of deduced amino acids residue, the protein pI, and the molecular weight of each EuglDHN protein are shown (DOC 31 kb)

Table S2

The collection of cis-regulatory elements in the promoter of three dehydrin genes of E. globulus (DOC 33.0 kb)

Supplementary Fig. S1

SDS-PAGE (12.5%) of protein extract from freezing-resistant and -sensitive genotypes during NA nonacclimated, CABF cold acclimated before night frost, CAAF cold acclimated after night frost, and DA deacclimated. Arrows indicate the relative migration of five peptides identified by western blot analysis. a Protein extract from leaf tissue in the upper panel and stem. b Protein extract from stem tissue in the lower panel (DOC 226 kb)

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Fernandez, M., Valenzuela Águila, S., Arora, R. et al. Isolation and characterization of three cold acclimation-responsive dehydrin genes from Eucalyptus globulus . Tree Genetics & Genomes 8, 149–162 (2012). https://doi.org/10.1007/s11295-011-0429-8

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