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Wuschel overexpression promotes somatic embryogenesis and induces organogenesis in cotton (Gossypium hirsutum L.) tissues cultured in vitro

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This work shows that overexpression of the WUS gene from Arabidopsis enhanced the expression of embryogenic competence and triggered organogenesis from some cells of the regenerated embryo-like structures.

Abstract

Agrobacterium-mediated genetic transformation of cotton was described in the late 1980s, but is still time consuming and largely genotype dependant due to poor regeneration. To help solve this bottleneck, we over-expressed the WUSCHEL (WUS) gene, a homeobox transcription factor cloned in Arabidopsis thaliana, known to stimulate organogenesis and/or somatic embryogenesis in Arabidopsis tissues cultured in vitro. The AtWUS gene alone, and AtWUS gene fused to the GFP marker were compared to the GFP gene alone and to an empty construct used as a control. Somatic embryogenesis was improved in WUS expressed calli, as the percentage of explants giving rise to embryogenic tissues was significantly higher (×3) when WUS gene was over-expressed than in the control. An interesting result was that WUS embryogenic lines evolved in green embryo-like structures giving rise to ectopic organogenesis never observed in any of our previous transformation experiments. Using our standard in vitro culture protocol, the overexpression of AtWUS in tissues of a recalcitrant variety did not result in the production of regenerated plants. This achievement will still require the optimization of other non-genetic factors, such as the balance of exogenous phytohormones. However, our results suggest that targeted expression of the WUS gene is a promising strategy to improve gene transfer in recalcitrant cotton cultivars.

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Acknowledgments

We thank P. Hilson, P. Laufs for their critical reading of the manuscript and J. Scarlett for checking English language. We are grateful to JC. Palauqui and F. Bonnot for their generous help in, respectively, confocal Imaging and statistical analysis. Seeds used to initiate our own seed stocks have been provided by CIRAD. This research was supported in part by Agence Nationale de la Recherche (ANR) agreement 05W 37, EUREKA project 3395 F 1203.

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Authors and Affiliations

  1. INRA, UMR1318, Institut Jean-Pierre Bourgin, RD10, 78000, Versailles, France

    O. Bouchabké-Coussa, M. Obellianne, D. Linderme, E. Montes, A. Maia-Grondard, F. Vilaine & C. Pannetier

  2. CIRAD, UR-SCA, 78026, Versailles, France

    O. Bouchabké-Coussa, M. Obellianne, D. Linderme & C. Pannetier

  3. CIRAD UMR AGAP, 78026, Versailles, France

    C. Pannetier

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  1. O. Bouchabké-Coussa
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Correspondence to C. Pannetier.

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Communicated by L. Jouanin.

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Bouchabké-Coussa, O., Obellianne, M., Linderme, D. et al. Wuschel overexpression promotes somatic embryogenesis and induces organogenesis in cotton (Gossypium hirsutum L.) tissues cultured in vitro. Plant Cell Rep 32, 675–686 (2013). https://doi.org/10.1007/s00299-013-1402-9

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