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Functional characterization of an acidic SK3 dehydrin isolated from an Opuntia streptacantha cDNA library

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Abstract

Cactus pears are succulent plants of the Cactaceae family adapted to extremely arid, hot and cold environments, making them excellent models for the study of molecular mechanisms underlying abiotic stress tolerance. Herein, we report a directional cDNA library from 12-month-old cladodes of Opuntia streptacantha plants subjected to abiotic stresses. A total of 442 clones were sequenced, representing 329 cactus pear unigenes, classified into eleven functional categories. The most abundant EST (unigen 33) was characterized under abiotic stress. This cDNA of 905 bp encodes a SK3-type acidic dehydrin of 248 amino acids. The OpsDHN1 gene contains an intron inserted within the sequence encoding the S-motif. qRT-PCR analysis shows that the OpsDHN1 transcript is specifically accumulated in response to cold stress, and induced by abscisic acid. Over-expression of the OpsDHN1 gene in Arabidopsis thaliana leads to enhanced tolerance to freezing treatment, suggesting that OpsDHN1 participates in freezing stress responsiveness. Generation of the first EST collection for the characterization of cactus pear genes constitutes a useful platform for the understanding of molecular mechanisms of stress tolerance in Opuntia and other CAM plants.

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Abbreviations

ABA:

Abscisic acid

CAM:

Crassulacean acid metabolism

ORF:

Open reading frame

OpsDHN1 :

Opuntia streptacantha dehydrin 1

qRT-PCR:

Quantitative reverse transcriptase-polymerase chain reaction

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Acknowledgments

This work was supported by SAGARPA (2004-C01-216) and the CONACYT (Investigación Ciencia Básica 2008-103106) fundings. We are grateful to Dr. Paul Riley for a grammatical review.

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Authors and Affiliations

  1. Division de Biologia Molecular, Instituto Potosino de Investigacion Cientifica y Tecnologica, Camino a la Presa de San Jose 2055, Apartado Postal 3-74 Tangamanga, C.P. 78216, San Luis Potosi, SLP, Mexico

    A. E. Ochoa-Alfaro, M. B. Pérez-Morales, P. Delgado-Sánchez, C. L. Cuevas-Velazquez & J. F. Jiménez-Bremont

  2. Facultad de Ciencias, Universidad Autonoma de San Luis Potosi, Av. Salvador Nava s/n, Zona Universitaria, C.P. 78290, San Luis Potosi, SLP, Mexico

    M. Rodríguez-Kessler

  3. Agricultura en Zonas Aridas, Centro de Investigaciones Biologicas del Noreste Mar Bermejo No. 195, Col. Playa Palo de Santa Rita, Apartado Postal 128, C.P. 23090, La Paz, BCS, Mexico

    G. Gómez-Anduro

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  1. A. E. Ochoa-Alfaro
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  2. M. Rodríguez-Kessler
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  3. M. B. Pérez-Morales
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  4. P. Delgado-Sánchez
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  6. G. Gómez-Anduro
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  7. J. F. Jiménez-Bremont
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Correspondence to J. F. Jiménez-Bremont.

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Suppl. Fig. S1 Prediction of secondary structure of the OpsDHN1 protein. a Prediction of disordered residues along OpsDHN1 sequence, b Disordered profile plot of OpsDHN1 protein. The plot shows position in the sequence against probability of being disordered. In total, 224 of the 248 amino acid residues are predicted to be disordered at the default threshold. The horizontal line is the order/disorder threshold for the default false positive rate of 5%. Secondary structure prediction was predicted using the Disopred2 tool from the Bloomsbury Center for Bioinformatics (http://bioinf.cs.ucl.ac.uk/disopred/)

Suppl. Fig. S2 Pairwise alignment of a genomic and the cDNA sequences containing the OpsDHN1 gene. The start and stop codons are shadowed. The intron borders GT/AG are bold indicated. Prediction of the branch point region (CURAY, underlined) located close to the intron acceptor site AG, was performed using the NetPlantGene Server (http://www.cbs.dtu.dk/services/NetPGene/)

Suppl. Fig. S3 Twelve-month-old O. streptacantha plants were grown either in semi-hydroponic conditions or in a commercial soil mixture, and subjected to different abiotic stresses during 17 d: control, salinity (N), heat-salinity (HN) (semi-hydroponic); control, cold (C), heat-cold (HC), heat-drought (HD) and drought-heat-cold (DHC) (soil mixture)

Suppl. Fig. S4 a Phenotype of the 35S::OpsDHN1-1 transgenic line and the wild type (Col-0) after 21 d of recovery from salt stress treatment (150 mM NaCl). b Survival rate of the 35S::OpsDHN1-1 transgenic line and Col-0 plants after 21 d of NaCl stress recovery. Data are mean ± SE (n = 10) from 5 replicates. The experiments were conducted twice obtaining similar results

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Ochoa-Alfaro, A.E., Rodríguez-Kessler, M., Pérez-Morales, M.B. et al. Functional characterization of an acidic SK3 dehydrin isolated from an Opuntia streptacantha cDNA library. Planta 235, 565–578 (2012). https://doi.org/10.1007/s00425-011-1531-8

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