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Contrasting cDNA-AFLP profiles between crown and leaf tissues of cold-acclimated wheat plants indicate differing regulatory circuitries for low temperature tolerance

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Abstract

Low-temperature (LT) tolerance in winter wheat (Triticum aestivum L.) is an economically important but complex trait. Four selected wheat genotypes, a winter hardy cultivar, Norstar, a tender spring cultivar, Manitou and two near-isogenic lines with Vrn-A1 (spring Norstar) and vrn-A1 (winter Manitou) alleles of Manitou and Norstar were cold-acclimated at 6°C and crown and leaf tissues were collected at 0, 2, 14, 21, 35, 42, 56 and 70 days of cold acclimation. cDNA-AFLP profiling was used to determine temporal expression profiles of transcripts during cold-acclimation in crown and leaf tissues, separately to determine if LT regulatory circuitries in crown and leaf tissues could be delineated using this approach. Screening 64 primer combinations identified 4,074 and 2,757 differentially expressed transcript-derived fragments (TDFs) out of which 38 and 16% were up-regulated as compared to 3 and 6% that were down-regulated in crown and leaf tissues, respectively. DNA sequencing of TDFs revealed sequences common to both tissues including genes coding for DEAD-box RNA helicase, choline-phosphate cytidylyltransferase and delta-1-pyrroline carboxylate synthetase. TDF specific to crown tissues included genes coding for phospahtidylinositol kinase, auxin response factor protein and brassinosteroid insensitive 1-associated receptor kinase. In leaf, genes such as methylene tetrahydrofolate reductase, NADH-cytochrome b5 reductase and malate dehydrogenase were identified. However, 30 and 14% of the DNA sequences from the crown and leaf tissues, respectively, were hypothetical or unknown proteins. Cluster analysis of up-, down-regulated and unique TDFs, DNA sequence and real-time PCR validation, infer that mechanisms operating in crown and leaf tissue in response to LT are differently regulated and warrant further studies.

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Acknowledgments

Financial support from Genome Canada/Genome Prairie (DBF and RNC), Ducks Unlimited (DBF), Canada Research Chairs (RNC) and Canada Foundation for Innovation (RNC) is gratefully acknowledged. The excellent technical assistance of Upekha Basnayaka, Jenna Drinkwater, Leah Madsen, Dr. Ximing Luo, Garcia Schellhorn and Twyla Chastain is greatly appreciated. The excellent help of summer students Pubudu Basnayaka and Daniel Hiebert is also greatly appreciated.

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  1. Department of Plant Sciences, College of Agriculture and Bioresources, University of Saskatchewan, 51 Campus Drive, Saskatoon, SK, S7N 5A8, Canada

    Seedhabadee Ganeshan, Pallavi Sharma, D. Brian Fowler & Ravindra N. Chibbar

  2. Food and Bioproduct Sciences, College of Agriculture and Bioresources, University of Saskatchewan, 51 Campus Drive, Saskatoon, SK, S7N 5A8, Canada

    Lester Young

  3. Centre for Rural Development and Technology, Indian Institute of Technology, Delhi, Hauz Khas, New Delhi, India

    Ashwani Kumar

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  1. Seedhabadee Ganeshan
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Correspondence to Ravindra N. Chibbar.

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Seedhabadee Ganeshan and Pallavi Sharma contributed equally to this manuscript.

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Ganeshan, S., Sharma, P., Young, L. et al. Contrasting cDNA-AFLP profiles between crown and leaf tissues of cold-acclimated wheat plants indicate differing regulatory circuitries for low temperature tolerance. Plant Mol Biol 75, 379–398 (2011). https://doi.org/10.1007/s11103-011-9734-8

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