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Identification and analysis of safener-inducible expressed sequence tags in Populus using a cDNA microarray

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Abstract

Safeners are the chemicals used to protect plants from detrimental effects of herbicides, but their mode of action at the molecular level is not well understood. As an initial step towards understanding the molecular mechanism of safener action in trees, homologous genes in hybrid poplar (Populus nigra × Populus maximowiczii) that were induced by a safener were identified. We here describe the identification of differentially expressed genes in Populus that are induced by Concep-III, a herbicide safener. Expressed sequence tags (ESTs) enriched for transcriptionally induced genes were isolated by suppressive subtractive hybridization (SSH). The SSH library cDNA inserts were used to construct a cDNA microarray for high-throughput validation of the up-regulated expression of safener-induced genes. Single-pass and partial sequences of 1,344 safener-induced ESTs were assembled into 418 singletons and 328 clusters, but the putative functions of almost 53% of the ESTs are not known. Genes encoding proteins involved in all three different phases of safener action, viz., oxidation, conjugation, and sequestration, were found in the SSH library. Almost 75% of genes that showed greater than 2-fold expression upon safener treatment were redundant in the SSH library. The expression pattern for selected genes was validated by reverse transcription–polymerase chain reaction. A few safener-induced genes that were not previously reported to be induced by safeners, but which may have a role in herbicide metabolism, were identified. The newly identified genes could have potential for application in genetic engineering of plants for herbicide detoxification and tolerance.

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Abbreviations

EST :

Expressed sequence tag

SSH :

Suppressive subtractive hybridization

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Author notes

  1. A. S. Rishi

    Present address: Department of Plant Sciences, University of Arizona, 1140 E South Campus Drive, Forbes Building, Tucson, AZ 85721, USA

  2. Neil D. Nelson

    Present address: Forestry Sciences Laboratory, North Central Research Station, 5985 Hwy K, Rhinelander, WI 54501, USA

Authors and Affiliations

  1. Department of Biological Sciences, East Tennessee State University, Johnson City, TN 37614, USA

    Arun Goyal

  2. Biotechnology Initiative, Forestry and Forest Products Division, Center for Applied Research and Technology Development (CARTD), Natural Resources Research Institute, University of Minnesota-Duluth, Duluth, MN 55811, USA

    A. S. Rishi, Neil D. Nelson & Arun Goyal

  3. Department of Microbiology, University of Minnesota, Minneapolis, MN 55108, USA

    Shirin Munir & Vivek Kapur

Authors
  1. A. S. Rishi
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  2. Shirin Munir
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Correspondence to Arun Goyal.

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Rishi, A.S., Munir, S., Kapur, V. et al. Identification and analysis of safener-inducible expressed sequence tags in Populus using a cDNA microarray. Planta 220, 296–306 (2004). https://doi.org/10.1007/s00425-004-1356-9

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  • DOI: https://doi.org/10.1007/s00425-004-1356-9

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