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Generation of Transgenic Spores of the Fern Ceratopteris richardii to Analyze Ca2+ Transport Dynamics During Gravity-Directed Polarization

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Abstract

Spores from the fern, Ceratopteris richardii, have been used to study gravity-directed cell polarization for over three decades. This system is ideal for these studies because it has a highly predictable growth and developmental pattern and primarily responds to the mechanical force of gravity during polarization. Early studies on the development in this system showed that during the first 24 h of germination, Ceratopteris spores establish a Ca2+ concentration differential along the outer periphery of the cell that is defined by the uptake of Ca2+ through channels at the bottom of the spore and an efflux of Ca2+ through pumps at the top. This 100-fold [Ca2+] differential is sensitive to the direction and magnitude of the gravitational force. In a low-gravity environment or when the uptake of Ca2+ is blocked, spore polarization becomes random. These results support the hypothesis that the uptake of Ca2+ is necessary for gravity-directed polarization in Ceratopteris spores. For many years, studies of Ceratopteris were limited by the inability to produce stable transformants. However, recent protocols have led to the first stably transformed lines in Ceratopteris richardii. This work reviews and discusses the latest studies of gravity-induced changes in Ca2+ transport dynamics in Ceratopteris spores, the use of transformation protocols to overexpress or knock out genes relevant to the transport of Ca2+ in Ceratopteris, and the potential value of mutants to more thoroughly understand the role of Ca2+ transport dynamics in gravity-directed polarization of spores.

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Acknowledgments

We acknowledge the critically valuable assistance of Dr. Benjamin Smith at the University of Oklahoma who did the FLIM imaging and video assembly and Dr. Greg Clark at the University of Texas at Austin in facilitating all the research described here, which was supported by grants NNX13AM54G and NNX15AB85A from NASA awarded to S. Roux and G. Clark.

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Correspondence to Stanley J. Roux .

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Cannon, A.E., Salmi, M.L., Cantero, A., Roux, S.J. (2018). Generation of Transgenic Spores of the Fern Ceratopteris richardii to Analyze Ca2+ Transport Dynamics During Gravity-Directed Polarization. In: Fernández, H. (eds) Current Advances in Fern Research. Springer, Cham. https://doi.org/10.1007/978-3-319-75103-0_14

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