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Phospholipase C (PLC) has been implicated in several stress responses, including drought. Overexpression (OE) of PLC has been shown to improve drought tolerance in various plant species. Arabidopsis contains nine PLC genes, which are subdivided into four clades. Earlier, OE of PLC3, PLC5 or PLC7 was found to increase Arabidopsis ' drought tolerance. Here, we confirm this for three other PLCs: PLC2 , the only constitutively expressed AtPLC; PLC4 , reported to have reduced salt tolerance and PLC9 , of which the encoded enzyme was presumed to be catalytically inactive. To compare each PLC and to discover any other potential phenotype, two independent OE lines of six At PLC genes, representing all four clades, were simultaneously monitored with the GROWSCREEN-FLUORO phenotyping platform, under both control- and mild-drought conditions. To investigate which tissues were most relevant to achieving drought survival, we additionally expressed At PLC5 using 13 different cell- or tissue-specific promoters. While no significant differences in plant size, biomass or photosynthesis were found between PLC lines and wild-type (WT) plants, all PLC -OE lines, as well as those tissue-specific lines that promoted drought survival, exhibited a stronger decrease in 'convex hull perimeter' (= increase in 'compactness') under water deprivation compared to WT. Increased compactness has not been associated with drought or decreased water loss before although a hyponastic decrease in compactness in response to increased temperatures has been associated with water loss. We propose that the increased compactness could lead to decreased water loss and potentially provide a new breeding trait to select for drought tolerance. [ABSTRACT FROM AUTHOR]

Phospholipase C (PLC) is a well-known signaling enzyme in metazoans that hydrolyzes phosphatidylinositol 4,5-bisphosphate (PIP2) to produce inositol 1,4,5-trisphosphate and diacylglycerol as second messengers involved in mutiple processes. Plants contain PLC too, but relatively little is known about its function there. The model system Arabidopsis thaliana contains nine PLC genes. Reversed genetics have implicated several roles for PLCs in plant development and stress signaling. Here, PLC5 is functionally addressed. Promoter–β-glucuronidase (GUS) analyses revealed expression in roots, leaves and flowers, predominantly in vascular tissue, most probably phloem companion cells, but also in guard cells, trichomes and root apical meristem. Only one plc5-1 knock-down mutant was obtained, which developed normally but grew more slowly and exhibited reduced primary root growth and decreased lateral root numbers. These phenotypes could be complemented by expressing the wild-type gene behind its own promoter. Overexpression of PLC5 (PLC5-OE) using the UBQ10 promoter resulted in reduced primary and secondary root growth, stunted root hairs, decreased stomatal aperture and improved drought tolerance. PLC5-OE lines exhibited strongly reduced phosphatidylinositol 4-monophosphate (PIP) and PIP2 levels and increased amounts of phosphatidic acid, indicating enhanced PLC activity in vivo. Reduced PIP2 levels and stunted root hair growth of PLC5-OE seedlings could be recovered by inducible overexpression of a root hair-specific PIP 5-kinase, PIP5K3. Our results show that PLC5 is involved in primary and secondary root growth and that its overexpression improves drought tolerance. Independently, we provide new evidence that PIP2 is essential for the polar tip growth of root hairs. [ABSTRACT FROM AUTHOR]