EXO70C2 is a key regulatory factor for optimal tip growth of pollen

Abstract
The exocyst, an eukaryotic tethering complex, co-regulates targeted exocytosis as an effector of small GTPases in polarized cell growth. In land plants, several exocyst subunits are encoded by double or triple paralogs, culminating in tens of EXO70 paralogs. Out of 23 Arabidopsis EXO70 isoforms, we analyzed seven isoforms expressed in pollen. Genetic and microscopic analyses of single mutants in EXO70A2, C1, C2, F1, H3, H5, and H6 genes revealed that only a loss-of-function EXO70C2 allele resulted in a significant male-specific transmission defect (segregation 40%:51%:9%) due to aberrant pollen tube growth. Mutant pollen tubes grown in vitro exhibited enhanced growth rate and a decreased thickness of the tip cell wall, causing tip bursts. However, exo70C2 pollen tubes could frequently recover and restart their speedy elongation, resulting in a repetitive stop-and-go growth dynamics. A pollen-specific depletion of the closest paralog, EXO70C1, using ami-RNA in the exo70C2 mutant background resulted in a complete pollen-specific transmission defect, suggesting redundant functions of EXO70C1 and EXO70C2. Both EXO70C1 and EXO70C2, GFP-tagged and expressed under their native promoters, localized in the cytoplasm of pollen grains, pollen tubes, and also root trichoblast cells. Expression of EXO70C2-GFP complemented aberrant growth of exo70C2 pollen tubes. The absent EXO70C2 interactions with core exocyst subunits in the yeast two-hybrid assay, cytoplasmic localization, and genetic effect suggest an unconventional EXO70 function possibly as a regulator of exocytosis outside the exocyst complex. In conclusion, EXO70C2 is a novel factor contributing to the regulation of optimal tip growth of Arabidopsis pollen tubes.

Citation
Synek L, Vukasinovic N, Kulich I, Hala M, Aldorfova K, et al. (2017) EXO70C2 is a key regulatory factor for optimal tip growth of pollen. Plant Physiology: pp.01282.2016. Available: http://dx.doi.org/10.1104/pp.16.01282.

Acknowledgements
This work was supported by the Czech Science Foundation (grant no. 15-24711S). Part of the V.Z. and I.K. income is covered by the Ministry of Education of the Czech Republic (project no. NPUI LO1417).

Publisher
American Society of Plant Biologists (ASPB)

Journal
Plant Physiology

DOI
10.1104/pp.16.01282

Additional Links
http://www.plantphysiol.org/content/early/2017/03/30/pp.16.01282

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