Characterization of Novel Aperture Proteins in Arabidopsis thaliana
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Date
2018-05
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Publisher
The Ohio State University
Abstract
In many species, exine, the outer wall of pollen grains, is deposited non-uniformly on
the pollen surface. The gaps left on the pollen surface by the absence of exine deposition are
known as apertures. These apertures facilitate emergence of the pollen tubes, making them
important for male fertility in many plants. The positioning of the apertures is highly conserved
within species, yet it varies widely between species, making apertures an ideal model for
studying how cells control formation of distinct extracellular domains. Very little is known
about how the locations of apertures are specified and how apertures are formed. Dobritsa
lab's current work involves identification and characterization of novel proteins in Arabidopsis
required for aperture formation. My project focused on two of the mutants recently found
during a mutagenesis screen, both of which completely lack apertures. The candidate genes
responsible for these mutations, called STRUBBELIG-RECEPTOR FAMILY 2 (SRF2) and
INAPERTURATE POLLEN 2 (INP2), were isolated, fused with YFP in some cases, and introduced
into their respective mutants in Arabidopsis using Agrobacterium-mediated transformation. I
created the constructs and confirmed the identity of these genes by phenotypic rescue of their
mutants. The expression and localization of the SRF2 protein was characterized using
fluorescence of the YFP tag. In addition, I also tested the effects of inactivation of ELMOD-A, a
close paralog of a third gene involved in aperture formation, called MACARON, by mutating
ELMOD-A with CRISPR and demonstrated that ELMOD-A is also involved in pollen aperture
formation.