Overexpressing the Multiple-Stress Responsive Gene At1g74450 Reduces Plant Height and Male Fertility in Arabidopsis thaliana

Abstract
A subset of genes in Arabidopsis thaliana is known to be up-regulated in response to a wide range of different environmental stress factors. However, not all of these genes are characterized as yet with respect to their functions. In this study, we used transgenic knockout, overexpression and reporter gene approaches to try to elucidate the biological roles of five unknown multiple-stress responsive genes in Arabidopsis. The selected genes have the following locus identifiers: At1g18740, At1g74450, At4g27652, At4g29780 and At5g12010. Firstly, T-DNA insertion knockout lines were identified for each locus and screened for altered phenotypes. None of the lines were found to be visually different from wildtype Col-0. Secondly, 35S-driven overexpression lines were generated for each open reading frame. Analysis of these transgenic lines showed altered phenotypes for lines overexpressing the At1g74450 ORF. Plants overexpressing the multiple-stress responsive gene At1g74450 are stunted in height and have reduced male fertility. Alexander staining of anthers from flowers at developmental stage 1213 showed either an absence or a reduction in viable pollen compared to wildtype Col-0 and At1g74450 knockout lines. Interestingly, the effects of stress on crop productivity are most severe at developmental stages such as male gametophyte development. However, the molecular factors and regulatory networks underlying environmental stress-induced male gametophytic alterations are still largely unknown. Our results indicate that the At1g74450 gene provides a potential link between multiple environmental stresses, plant height and pollen development. In addition, ruthenium red staining analysis showed that At1g74450 may affect the composition of the inner seed coat mucilage layer. Finally, C-terminal GFP fusion proteins for At1g74450 were shown to localise to the cytosol.

Citation
Visscher, A. M., Belfield, E. J., Vlad, D., Irani, N., Moore, I., & Harberd, N. P. (2015). Overexpressing the Multiple-Stress Responsive Gene At1g74450 Reduces Plant Height and Male Fertility in Arabidopsis thaliana. PLOS ONE, 10(10), e0140368. doi:10.1371/journal.pone.0140368

Acknowledgements
This work was supported by King Abdullah University of Science and Technology [Award No. KUK-I1-002-03] and a Biotechnology and Biological Sciences Research Council grant [BB/F020759/1]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Publisher
PUBLIC LIBRARY SCIENCE

Journal
PLOS ONE

DOI
10.1371/journal.pone.0140368

PubMed ID
26485022

PubMed Central ID
PMC4619001

Additional Links
https://dx.plos.org/10.1371/journal.pone.0140368

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