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Eukaryotic Cell, August 2004, p. 966-975, Vol. 3, No. 4
1535-9778/04/$08.00+0     DOI: 10.1128/EC.3.4.966-975.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Dehydration-Specific Induction of Hydrophilic Protein Genes in the Anhydrobiotic Nematode Aphelenchus avenae

John A. Browne,1 Katharine M. Dolan,1 Trevor Tyson,1 Kshamata Goyal,2 Alan Tunnacliffe,2 and Ann M. Burnell1*

Institute of Bioengineering and Agroecology, Department of Biology, National University of Ireland Maynooth, Maynooth, County Kildare, Ireland,1 Institute of Biotechnology, University of Cambridge, Cambridge, United Kingdom2

Received 4 March 2004/ Accepted 1 June 2004

Some organisms can survive exposure to extreme desiccation by entering a state of suspended animation known as anhydrobiosis. The free-living nematode Aphelenchus avenae can be induced to enter the anhydrobiotic state by exposure to a moderate reduction in relative humidity. During this preconditioning period, the nematode accumulates large amounts of the disaccharide trehalose, which is thought to be necessary, but not sufficient, for successful anhydrobiosis. To identify other adaptations that are required for anhydrobiosis, we developed a novel SL1-based mRNA differential display technique to clone genes that are upregulated by dehydration in A. avenae. Three such genes, Aav-lea-1, Aav-ahn-1, and Aav-glx-1, encode, respectively, a late embryogenesis abundant (LEA) group 3 protein, a novel protein that we named anhydrin, and the antioxidant enzyme glutaredoxin. Strikingly, the predicted LEA and anhydrin proteins are highly hydrophilic and lack significant secondary structure in the hydrated state. The dehydration-induced upregulation of Aav-lea-1 and Aav-ahn-1 was confirmed by Northern hybridization and quantitative PCR experiments. Both genes were also upregulated by an osmotic upshift, but not by cold, heat, or oxidative stress. Experiments to investigate the relationship between mRNA levels and protein expression for these genes are in progress. LEA proteins occur commonly in plants, accumulating during seed maturation and desiccation stress; the presence of a gene encoding an LEA protein in an anhydrobiotic nematode suggests that some mechanisms of coping with water loss are conserved between plants and animals.

* Corresponding author. Mailing address: Institute of Bioengineering and Agroecology, Department of Biology, National University of Ireland Maynooth, Maynooth, Co. Kildare, Ireland. Phone: 353 1 7083840. Fax: 353 1 7083845. E-mail: ann.burnell{at}may.ie.

Eukaryotic Cell, August 2004, p. 966-975, Vol. 3, No. 4
1535-9778/04/$08.00+0     DOI: 10.1128/EC.3.4.966-975.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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