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In vitro culture promotes partial autonomous endosperm development in unfertilized ovules of wild-type Arabidopsis thaliana var. Columbia

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Abstract

Partial endosperm development without paternal genome involvement was induced in unpollinated ovaries of wild-type Arabidopsis thaliana cultured in vitro. Unpollinated pistils were cultured on hormone-free Murashige and Skoog (MS) medium with addition of 6% sucrose and supplemented with: benzylaminopurine (BAP; 2 mg l−1) combined with naphthylacetic acid (NAA; 0.1 mg l−1), 2,4-dichlorophenoxyacetic acid (2,4-D; explants exposed to 1-h auxin shock 20 or 40 mg l−1, and transferred to hormone-free MS medium). Initiation of autonomous endosperm (AE) development was induced on all media used in 54 ovules from 39 cultured ovaries (26%), with an average frequency of 1.4 ovules/ovary. The highest frequency of partial endosperm formation occurred on media combining the two growth regulators BAP and NAA (59% of ovaries had ovules with AE), although endosperm development was also induced on hormone-free medium (in 20.5% of ovaries). The number of AE nuclei ranged from 2 to ~50, depending on the day of culture and medium; neither cellularization nor differentiation on specific regions typical for endosperm of wild-type Arabidopsis, were noted. Fertilization independent endosperm most probably originated from the secondary nucleus, but involvement of the polar nuclei could not be excluded, as indicated by nuclear size and structure. In vitro conditions did not influence egg cell proliferation. Gynogenic embryos were observed neither in the ovules with autonomous endosperm nuclei nor in ovules without endosperm induction.

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Acknowledgements

We thank the kind generosity of Professor Jolanta Małuszyńska from the Department of Plant Anatomy and Cytology of Silesian University in Katowice for providing experimental seeds and Prof. Halina Gabryś from the Department of Plant Biochemistry and Physiology of the Jagiellonian University in Cracow for control plants. Special thanks are due for Professor John Herr Jr. from the University of South Carolina in Columbia for his kind help in the ovule clearing technique and for his invaluable comments in preparing the manuscript. We also thank Professor Val Raghavan from the Ohio State University in Columbus for stimulating discussion regarding interpretation of the results.

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Authors and Affiliations

  1. Department of Genetics and Cytology, University of Gdańsk, Kładki 24, 80-822, Gdańsk, Poland

    Joanna Rojek & Jerzy Bohdanowicz

  2. Department of Plant Cytology and Embryology, Jagiellonian University, Grodzka 52, 31-044, Cracow, Poland

    Elżbieta Kuta

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  1. Joanna Rojek
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  2. Elżbieta Kuta
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  3. Jerzy Bohdanowicz
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Correspondence to Joanna Rojek.

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Rojek, J., Kuta, E. & Bohdanowicz, J. In vitro culture promotes partial autonomous endosperm development in unfertilized ovules of wild-type Arabidopsis thaliana var. Columbia. Sex Plant Reprod 18, 29–36 (2005). https://doi.org/10.1007/s00497-005-0246-z

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