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Activation tagging is an effective gene tagging system in Populus

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Abstract

Knowledge of the functional relationship between genes and organismal phenotypes in perennial plants is extremely limited. Using a population of 627 independent events, we assessed the feasibility of activation tagging as a forward genetics tool for Populus. Mutant identification after 2 years of field testing was nearly sevenfold (6.5%) higher than in greenhouse studies that employed Arabidopsis and identical transformation vectors. Approximately two thirds of all mutant phenotypes were not seen in vitro and in the greenhouse; they were discovered only after the second year of field assessment. The trees’ large size (5-10 m in height), perennial growth, and interactions with the natural environment are factors that are thought to have contributed to the high rate of observable phenotypes in the field. The mutant phenotypes affected a variety of morphological and physiological traits, including leaf size and morphology, crown architecture, stature, vegetative dormancy, and tropic responses. Characterization of the insertion in more than 100 events with and without mutant phenotypes showed that tags predominantly (70%) inserted in a 13-Kbp region up- and downstream of the genes’ coding regions with approximately even distribution among the 19 chromosomes. Transcriptional activation was observed in many proximal genes studied. Successful phenotype recapitulation was observed in 10 of 12 retransformed genes tested, indicating true tagging and a functional relationship between the genes and observed phenotypes for most activation lines. Our studies indicate that in addition to associating mapping and QTL approaches, activation tagging can be used successfully as an effective forward gene discovery tool in Populus.

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Acknowledgments

This work was supported in part by grants from the US Department of Energy Poplar Genome Based Research for Carbon Sequestration in Terrestrial Ecosystems Program (DE-FG02-06ER64185, DE-FG02-05ER64113); the Consortium for Plant Biotechnology Research, Inc. (GO12026-203A); USDA-CSREES, Biotechnology Risk Assessment Research Grants Program (2004-35300-14687); and Genome Canada. We thank Chamini Illangasinghe and Rama Joshi for production of binary constructs and RT-PCR analyses.

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Authors and Affiliations

  1. School of Forest Resources and Environmental Science, Michigan Technological University, Houghton, MI, 49931-1295, USA

    Victor Busov, Yordan Yordanov & Jiqing Gou

  2. Department of Forestry and Natural Resources, Purdue University, West Lafayette, IN, 47907-2061, USA

    Richard Meilan

  3. Department of Forest Ecosystems and Society, Oregon State University, Corvallis, OR, 97331-5752, USA

    Cathleen Ma & Steven Strauss

  4. Department of Biology, Queen’s University, Kingston, ON, K7L 3N6, Canada

    Sharon Regan

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  1. Victor Busov
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  2. Yordan Yordanov
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  4. Richard Meilan
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Corresponding author

Correspondence to Victor Busov.

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Communicated by W. Boerjan

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Table S1

Insertion sites of activation tagging vector in the Populus genome (XLS 84 kb)

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Busov, V., Yordanov, Y., Gou, J. et al. Activation tagging is an effective gene tagging system in Populus . Tree Genetics & Genomes 7, 91–101 (2011). https://doi.org/10.1007/s11295-010-0317-7

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  • DOI: https://doi.org/10.1007/s11295-010-0317-7

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