Arabidopsis non-specific phospholipase C1: characterization and its involvement in response to heat stress.

Zuzana Krčková,Přemysl Pejchar,Daniela Kocourková,Olga Valentová,Jitka Brouzdová,Dominique Rainteau,Michal Daněk,Eric Ruelland,Jan Martinec

doi:10.3389/fpls.2015.00928

Abstract

The Arabidopsis non-specific phospholipase C1 (NPC) protein family is encoded by the genes NPC1 – NPC6. It has been shown that NPC4 and NPC5 possess phospholipase C activity; NPC3 has lysophosphatidic acid phosphatase activity. NPC3, 4 and 5 play roles in the responses to hormones and abiotic stresses. NPC1, 2 and 6 has not been studied functionally yet. We found that Arabidopsis NPC1 expressed in Escherichia coli possesses phospholipase C activity in vitro. This protein was able to hydrolyse phosphatidylcholine to diacylglycerol. NPC1-green fluorescent protein was localized to secretory pathway compartments in Arabidopsis roots. In the knock out T-DNA insertion line NPC1 (npc1) basal thermotolerance was impaired compared with wild-type (WT); npc1 exhibited significant decreases in survival rate and chlorophyll content at the seventh day after heat stress (HS). Conversely, plants overexpressing NPC1 (NPC1-OE) were more resistant to HS compared with WT. These findings suggest that NPC1 is involved in the plant response to heat.

Highlights

Phospholipases are recognized as key components of the phospholipid signaling network that regulates plant growth and development
The NPC1 produced in E. coli accumulated in inclusion bodies
We had to use a lower concentration of isopropyl β-thio-galactopyranoside (IPTG; 0.1 mM) and lower temperature (16◦C) for a longer induction time

Summary

Introduction

Phospholipases are recognized as key components of the phospholipid signaling network that regulates plant growth and development. This network includes, among other enzymes, PLC, PLD and phospholipases A1 and A2. These enzymes produce a set of second messenger molecules and lipid derivatives that are implicated in both plant metabolism and intracellular signaling. PC-PLCs, which are known in plants as NPC, are characterized by broader substrate ranges that include abundant PC (Nakamura et al, 2005; Pokotylo et al, 2013)

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