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Comparative analysis of expressed sequence tags from cold-acclimated and non-acclimated leaves of Rhododendron catawbiense Michx

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Abstract

An expressed sequence tag (EST) analysis approach was undertaken to identify major genes involved in cold acclimation of Rhododendron, a broad-leaf, woody evergreen species. Two cDNA libraries were constructed, one from winter-collected (cold-acclimated, CA; leaf freezing tolerance −53°C) leaves, and the other from summer-collected (non-acclimated, NA; leaf freezing tolerance −7°C) leaves of field-grown Rhododendron catawbiense plants. A total of 862 5′-end high-quality ESTs were generated by sequencing cDNA clones from the two libraries (423 from CA and 439 from NA library). Only about 6.3% of assembled unique transcripts were shared between the libraries, suggesting remarkable differences in gene expression between CA and NA leaves. Analysis of the relative frequency at which specific cDNAs were picked from each library indicated that four genes or gene families were highly abundant in the CA library including early light-induced proteins (ELIP), dehydrins/late embryogenesis abundant proteins (LEA), cytochrome P450, and beta-amylase. Similarly, seven genes or gene families were highly abundant in the NA library and included chlorophyll a/b-binding protein, NADH dehydrogenase subunit I, plastidic aldolase, and serine:glyoxylate aminotransferase, among others. Northern blot analyses for seven selected abundant genes confirmed their preferential expression in either CA or NA leaf tissues. Our results suggest that osmotic regulation, desiccation tolerance, photoinhibition tolerance, and photosynthesis adjustment are some of the key components of cold adaptation in Rhododendron.

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Abbreviations

CA:

Cold acclimated

ELIP:

Early light-induced protein

EST:

Expressed sequence tag

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

LEA:

Late embryogenesis abundant protein

NA:

Non-acclimated

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Acknowledgements

We greatly appreciate the help of Drs. Robert Bogden, Yujia Wu and Jerry Higbee of Amplicon Express, Pullman, WA, USA for construction of cDNA library and EST sequencing. We are thankful to Ms. Li Sun for her assistance in data analysis and the preparation of the manuscript. This journal paper of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa, Project No. 3601 was supported by Hatch Act and State of Iowa funds.

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Authors and Affiliations

  1. Department of Horticulture, Iowa State University, Ames, IA, 50011, USA

    Hui Wei, Anik L. Dhanaraj & Rajeev Arora

  2. USDA-ARS, Fruit Lab, Beltsville, MD, 20705, USA

    Anik L. Dhanaraj & Lisa J. Rowland

  3. Interdepartmental Genetics Graduate Program, Department of Genetics, and Developmental and Cellular Biology, Iowa State University, Ames, IA, 50011, USA

    Yan Fu

  4. David G. Leach Research Station of the Holden Arboretum, Mentor, OH, 44094, USA

    Stephen L. Krebs

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  1. Hui Wei
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Correspondence to Rajeev Arora.

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Wei, H., Dhanaraj, A.L., Rowland, L.J. et al. Comparative analysis of expressed sequence tags from cold-acclimated and non-acclimated leaves of Rhododendron catawbiense Michx. Planta 221, 406–416 (2005). https://doi.org/10.1007/s00425-004-1440-1

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