Volume-based pollen size analysis: an advanced method to assess somatic and gametophytic ploidy in flowering plants
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Volume-based pollen size analysis: an advanced method to assess somatic and gametophytic ploidy in flowering plants Nico De Storme • Linda Zamariola • Martin Mau • Timothy F. Sharbel • Danny Geelen
Received: 19 October 2012 / Accepted: 31 December 2012 Ó Springer-Verlag Berlin Heidelberg 2013
Abstract Pollen size is often used as a biological parameter to estimate the ploidy and viability of mature pollen grains. In general, pollen size quantification is performed one- or two-dimensionally using image-based diameter measurements. As these approaches are elaborate and time consuming, alternative approaches that enable a quick, reliable analysis of pollen size are highly relevant for plant research. In this study, we present the volumebased particle size analysis technique as an alternative method to characterize mature pollen. Based on a comparative assay using different plant species (including tomato, oilseed rape, kiwifruit, clover, among others), we found that volume-based pollen size measurements are not biased by the pollen shape or position and substantially reduce non-biological variation, allowing a more accurate determination of the actual pollen size. As such, volumebased particle size techniques have a strong discriminative power in detecting pollen size differences caused by alterations in the gametophytic ploidy level and therefore allow for a quick and reliable estimation of the somatic
Communicated by Scott Russell.
Electronic supplementary material The online version of this article (doi:10.1007/s00497-012-0209-0) contains supplementary material, which is available to authorized users. N. De Storme L. Zamariola D. Geelen (&) Department of Plant Production, Faculty of Bioscience Engineering, University of Ghent, Coupure Links 653, 9000 Ghent, Belgium e-mail: [email protected] M. Mau T. F. Sharbel Leibniz Institut fu¨r Pflanzengenetik und Kulturpflanzenforschung, Corrensstraße 3, 06466 Stadt Seeland, OT Gatersleben, Germany
ploidy level. Based on observations in Arabidopsis thaliana gametophytic mutants and differentially reproducing Boechera polyantha lines, we additionally found that volume-based pollen size analysis provides quantitative and qualitative data about alterations in male sporogenesis, including aneuploid and diploid gamete formation. Volume-based pollen size analysis therefore not only provides a quick and easy methodology to determine the somatic ploidy level of flowering plants, but can also be used to determine the mode of reproduction and to quantify the level of diplogamete formation. Keywords Pollen size Volume-based particle size analysis Ploidy determination 2n Gametes Microsporogenesis Arabidopsis thaliana Boechera polyantha
Introduction Due to high prevalence of polyploidy, the development of reliable and simple techniques that allow for a rapid determination of the somatic or gametophytic ploidy level is of great importance for several aspects of plant research. Indeed, in plants, variations in somatic ploidy level are not only
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