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First published online October 27, 2006; 10.1104/pp.106.086363

Plant Physiology 142:1480-1492 (2006)
© 2006 American Society of Plant Biologists

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CELL BIOLOGY AND SIGNAL TRANSDUCTION

Arabidopsis Reversibly Glycosylated Polypeptides 1 and 2 Are Essential for Pollen Development1,[W]

Georgia Drakakaki, Olga Zabotina, Ivan Delgado2, 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 (G.D., O.Z., I.D., S.R., N.R.); and Department of Energy Plant Research Laboratory, Michigan State University, East Lansing, Michigan 48824 (K.K.)

Reversibly glycosylated polypeptides (RGPs) have been implicated in polysaccharide biosynthesis. To date, to our knowledge, no direct evidence exists for the involvement of RGPs in a particular biochemical process. The Arabidopsis (Arabidopsis thaliana) genome contains five RGP genes out of which RGP1 and RGP2 share the highest sequence identity. We characterized the native expression pattern of Arabidopsis RGP1 and RGP2 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 colocalized in the same compartments and coimmunoprecipitated 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 a 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.


1 This work was supported by the National Science Foundation (plant genome grant no. DBI–0211797).

2 Present address: Identigene, Inc., 5615 Kirby Dr., Ste. 800, Houston, TX 77005–2451.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Natasha Raikhel (nraikhel{at}ucr.edu).

[W] The online version of this article contains Web-only data.

www.plantphysiol.org/cgi/doi/10.1104/pp.106.086363

* Corresponding author; e-mail nraikhel{at}ucr.edu; fax 951–827–2155.

Received July 5, 2006; accepted October 17, 2006; published October 27, 2006.


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