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Published on October 27, 2006; 10.1104/pp.106.086363

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Received July 5, 2006
Accepted October 17, 2006

Arabidopsis RGP1 and RGP2 are Essential for Pollen Development

Georgia Drakakaki , Olga Zabotina , Ivan Delgado , Stéphanie Robert , Kenneth Keegstra , and Natasha Raikhel *

The Center for Plant Cell Biology and Department of Botany and Plant Sciences, University of California, Riverside, California 92521
Department of Energy Plant Research Laboratory, Michigan State University, East Lansing, Michigan, 48824

* Corresponding author; email: nraikhel{at}ucr.edu.

Reversibly Glycosylated Polypeptides (RGPs) have been implicated in polysaccharide biosynthesis. To date, no direct evidence exists for the involvement of RGPs in a particular biochemical process. The Arabidopsis genome contains five RGP genes out of which RGP1 and RGP2 share the highest sequence identity. We characterized the native expression pattern of Arabidopsis RGPs 1 and 2 and used reverse genetics to investigate their respective functions. Although both genes are ubiquitously expressed, the highest levels are observed in actively growing tissues, and in mature pollen in particular. RGPs showed cytoplasmic and transient association with Golgi. In addition both proteins co-localized in the same compartments and co-immunoprecipitated from plant cell extracts. Single gene disruptions did not show any obvious morphological defects under greenhouse conditions, whereas the double-insertion mutant could not be recovered. We present evidence that the double mutant is lethal, and demonstrate the critical role of RGPs, particularly in pollen development. Detailed analysis demonstrated, that mutant pollen development is associated with abnormally enlarged vacuoles and poorly defined inner cell wall layer, which consequently results in disintegration of the pollen structure during pollen mitosis I. Taken together, our results indicate that RGP1 and RGP2 are required during microspore development and pollen mitosis, either affecting cell division and/or vacuolar integrity.




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