Abstract

Summary The establishment of pollen–pistil compatibility is strictly regulated by factors derived from both male and female reproductive structures. Highly diverse small cysteine‐rich proteins (CRPs) have been found to play multiple roles in plant reproduction, including the earliest stages of the pollen–stigma interaction. Secreted CRPs found in the pollen coat of members of the Brassicaceae, the pollen coat proteins (PCPs), are emerging as important signalling molecules that regulate the pollen–stigma interaction.Using a combination of protein characterization, expression and phylogenetic analyses we identified a novel class of Arabidopsis thaliana pollen‐borne CRPs, the PCP‐Bs (for pollen coat protein B‐class) that are related to embryo surrounding factor (ESF1) developmental regulators. Single and multiple PCP‐B mutant lines were utilized in bioassays to assess effects on pollen hydration, adhesion and pollen tube growth.Our results revealed that pollen hydration is severely impaired when multiple PCP‐Bs are lost from the pollen coat. The hydration defect also resulted in reduced pollen adhesion and delayed pollen tube growth in all mutants studied.These results demonstrate that At PCP‐Bs are key regulators of the hydration ‘checkpoint’ in establishment of pollen–stigma compatibility. In addition, we propose that interspecies diversity of PCP‐Bs may contribute to reproductive barriers in the Brassicaceae.

Highlights

  • Pollination in angiosperms involves multiple phases of interaction between female reproductive tissues of the gynoecium and the male reproductive unit, pollen, and subsequently the pollen tube it produces on germination (Hiscock & Allen, 2008)

  • By utilizing T-DNA insertion lines carrying single, double and triple AtPCP-B gene knockouts, we examined the impact of these mutations on pollen morphology and the pollen–stigma interaction

  • In a previous study that characterized the S-locus glycoprotein (SLG)-binding pollen coat protein pollen coat proteins (PCPs)-A1 from Brassica oleracea (Doughty et al, 1998) two polypeptides were found to copurify with PCP-A1

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Summary

Introduction

Pollination in angiosperms involves multiple phases of interaction between female reproductive tissues of the gynoecium and the male reproductive unit, pollen, and subsequently the pollen tube it produces on germination (Hiscock & Allen, 2008). The process is highly selective such that the majority of heterospecific pollen fails to effect syngamy and intraspecific pollination can be blocked in those species which possess selfincompatibility (SI) systems. Such prezygotic reproductive barriers are evolutionarily advantageous as they limit wasted mating opportunities, contribute to reproductive isolation and facilitate outbreeding when SI is present (Yost & Kay, 2009; Smith et al, 2013). Such diversity is considered to be important in maintaining species barriers (Swanson & Vacquier, 2002; Takeuchi & Higashiyama, 2012)

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