Petunia phospholipase c1 is involved in pollen tube growth.
Although pollen tube growth is essential for plant fertilization and reproductive success, the regulators of the actin-related growth machinery and the cytosolic Ca2+ gradient thought to determine how these cells elongate remain poorly defined. Phospholipases, their substrates, and their phospholipid turnover products have been proposed as such regulators; however, the relevant phospholipase(s) have not been characterized. Therefore, we cloned cDNA for a pollen-expressed phosphatidylinositol 4,5-bisphosphate (PtdInsP2)-cleaving phospholipase C (PLC) from Petunia inflata, named Pet PLC1. Expressing a catalytically inactive form of Pet PLC1 in pollen tubes caused expansion of the apical Ca2+ gradient, disruption of the organization of the actin cytoskeleton, and delocalization of growth at the tube tip. These phenotypes were suppressed by depolymerizing actin with low concentrations of latrunculin B, suggesting that a critical site of action of Pet PLC1 is in regulating actin structure at the growing tip. A green fluorescent protein (GFP) fusion to Pet PLC1 caused enrichment in regions of the apical plasma membrane not undergoing rapid expansion, whereas a GFP fusion to the PtdInsP2 binding domain of mammalian PLC delta1 caused enrichment in apical regions depleted in PLC. Thus, Pet PLC1 appears to be involved in the machinery that restricts growth to the very apex of the elongating pollen tube, likely through its regulatory action on PtdInsP2 distribution within the cell.
Dowd
Peter E
PE
Department of Biochemistry and Molecular Biology, Pensylvania State University, University Park, Pensylvania 16802, USA.
Coursol
Sylvie
S
Skirpan
Andrea L
AL
Kao
Teh-hui
TH
Gilroy
Simon
S
D000199:Actins / Q000378:metabolism; D020013:Calcium Signaling; D002462:Cell Membrane / Q000378:metabolism; D015870:Gene Expression; D006056:Golgi Apparatus / Q000378:metabolism; D008969:Molecular Sequence Data; D032306:Petunia / Q000201:enzymology*; D043265:Phosphatidylinositol Diacylglycerol-Lyase / Q000378:metabolism; D011058:Pollen / Q000166:cytology / Q000201:enzymology* / Q000254:growth & development*; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D010738:Type C Phospholipases / Q000737:chemistry / Q000378:metabolism*
D000199:Actins; D011993:Recombinant Fusion Proteins; D010738:Type C Phospholipases; C432456:phospholipase C1; D043265:Phosphatidylinositol Diacylglycerol-Lyase