Plant peroxisomes are degraded by starvation-induced and constitutive autophagy in tobacco BY-2 suspension-cultured cells.

Olga V Voitsekhovskaja,Andreas Schiermeyer,Sigrun Reumann

doi:10.3389/fpls.2014.00629

Abstract

Very recently, autophagy has been recognized as an important degradation pathway for quality control of peroxisomes in Arabidopsis plants. To further characterize the role of autophagy in plant peroxisome degradation, we generated stable transgenic suspension-cultured cell lines of heterotrophic Nicotiana tabacum L. cv. Bright Yellow 2 expressing a peroxisome-targeted version of enhanced yellow fluorescent protein. Indeed, this cell line model system proved advantageous for detailed cytological analyses of autophagy stages and for quantification of cellular peroxisome pools under different culturing conditions and upon inhibitor applications. Complementary biochemical, cytological, and pharmacological analyses provided convincing evidence for peroxisome degradation by bulk autophagy during carbohydrate starvation. This degradation was slowed down by the inhibitor of autophagy, 3-methyladenine (3-MA), but the 3-MA effect ceased at advanced stages of starvation, indicating that another degradation mechanism for peroxisomes might have taken over. 3-MA also caused an increase particularly in peroxisomal proteins and cellular peroxisome numbers when applied under nutrient-rich conditions in the logarithmic growth phase, suggesting a high turnover rate for peroxisomes by basal autophagy under non-stress conditions. Together, our data demonstrate that a great fraction of the peroxisome pool is subject to extensive autophagy-mediated turnover under both nutrient starvation and optimal growth conditions. Our analyses of the cellular pool size of peroxisomes provide a new tool for quantitative investigations of the role of plant peroxisomes in reactive oxygen species metabolism.

Highlights

Plant peroxisomes perform many important functions including metabolism of reactive oxygen species (ROS), photorespiration, lipid metabolism, synthesis of plant hormones, polyamine metabolism, and ureate catabolism (Hayashi and Nishimura, 2003; del Rio et al, 2006; Reumann and Weber, 2006; Graham, 2008; Moschou et al, 2008; Kaur et al, 2009; Palma et al, 2009)
By comprehensive biochemical and cytological analyses we demonstrate in this study that peroxisomes in BY-2 cells are degraded by autophagy under nutrient starvation
Recently, several research groups independently published conclusive evidence that Arabidopsis peroxisomes are degraded by autophagy and that this process is important for quality control of peroxisomes (Farmer et al, 2013; Kim et al, 2013; Shibata et al, 2013; Yoshimoto et al, 2014)

Summary

Introduction

Plant peroxisomes perform many important functions including metabolism of reactive oxygen species (ROS), photorespiration, lipid metabolism, synthesis of plant hormones, polyamine metabolism, and ureate catabolism (Hayashi and Nishimura, 2003; del Rio et al, 2006; Reumann and Weber, 2006; Graham, 2008; Moschou et al, 2008; Kaur et al, 2009; Palma et al, 2009). Plant peroxisomes play essential roles in photomorphogenesis, embryogenesis, and seed germination and in plant pathogen defense mechanisms. Peroxisome biogenesis has been studied in considerable detail both mechanistically and at the molecular level. Contrary to plant peroxisome biogenesis, our knowledge about mechanisms of peroxisome degradation in plant cells has remained scarce

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