Abstract

Gametophytes of the fern Ceratopteris richardii develop into either hermaphrodites or males. As hermaphrodites develop, they secrete antheridiogen, or ACE, into the environment, inducing male development in undifferentiated gametophytes. Hermaphrodites are composed of archegonia, antheridia, rhizoids and a notch meristem, while males consist of antheridia and rhizoids. Much of the research on sexual and morphological development concerns gametophytes grown in sterile environments. Using biochemical and molecular techniques we identify a soil bacterium and explore its effects on sexual and rhizoid development. Hermaphrodite and male gametophytes were exposed to this bacterium and the effects on sexual development, rhizoid length and rhizoid number were explored. The bacterium was identified as a pseudomonad, Pseudomonas nitroreducens. Gametophytes grown in the presence of the pseudomonad were more likely to develop into hermaphrodites across all gametophyte densities. Across all gametophyte sizes, hermaphrodites had rhizoids that were 2.95× longer in the presence of the pseudomonad while males had rhizoids that were 2.72× longer in the presence of the pseudomonad. Both hermaphrodite and male gametophytes developed fewer rhizoids in the presence of the pseudomonad. Control hermaphrodites produced 1.23× more rhizoids across all gametophyte sizes. For male gametophytes grown in the absence of the pseudomonad, the rate of increase in the number of rhizoids was greater with increasing size in the control than the rate of increase in males grown in the presence of the pseudomonad. The pseudomonad may be acting on gametophyte sexual development via several potential mechanisms: degradation of ACE, changes in nutrient availability or phytohormone production. The pseudomonad may also increase rhizoid number through production of phytohormones or changes in nutrient availability.

Highlights

  • The soil environment that natural populations of plants are exposed to is quite complex (Torsvik et al 1996; Torsvik and Øvreås 2002)

  • Comparison of the 16S rRNA sequence from the soil bacteria isolate to the EZTaxon-e Database resulted in a sequence alignment of 29 out of 30 results to bacteria within the genus Pseudomonas, where the top sequence alignment was to Pseudomonas nitroreducens with a similarity score of 99.86 %

  • Effects of bacteria on C. richardii sex determination In order to determine whether the pseudomonad affected sexual development in C. richardii, an analysis of covariance (ANCOVA) was performed

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Summary

Introduction

The soil environment that natural populations of plants are exposed to is quite complex (Torsvik et al 1996; Torsvik and Øvreås 2002). Bacteria represent major players in the soil with cell numbers thought to approach 1 × 109–1.5 × 1010 per g of soil (Torsvik et al 1990; Ranjard et al 2000) constituting over 4000 unique genomes per g of soil (Torsvik et al 1990) These bacteria are known to participate heavily in nutrient cycling, as well as intimate crosstalk with sporophyte roots (Brencic and Winans 2005). Ferns offer a unique opportunity to explore such associations given that they offer a free-living gametophyte in common with bryophytes and a free-living sporophyte generation in common with seed plants In ferns these two generations may briefly share the same microenvironment though there are inherent differences between gametophyte rhizoids and sporophyte roots: rhizoids are multicellular, while the morphologically and functionally similar root hairs of the sporophyte are single celled (Jones and Dolan 2012)

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