Cell wall polysaccharides are mislocalized to the Vacuole in echidna mutants.
During cell wall biosynthesis, the Golgi apparatus is the platform for cell wall matrix biosynthesis and the site of packaging, of both matrix polysaccharides and proteins, into secretory vesicles with the correct targeting information. The objective of this study was to dissect the post-Golgi trafficking of cell wall polysaccharides using echidna as a vesicle traffic mutant of Arabidopsis thaliana and the pectin-secreting cells of the seed coat as a model system. ECHIDNA encodes a trans-Golgi network (TGN)-localized protein, which was previously shown to be required for proper structure and function of the secretory pathway. In echidna mutants, some cell wall matrix polysaccharides accumulate inside cells, rather than being secreted to the apoplast. In this study, live cell imaging of fluorescent protein markers as well as transmission electron microscopy (TEM)/immunoTEM of cryofixed seed coat cells were used to examine the consequences of TGN disorganization in echidna mutants under conditions of high polysaccharide production and secretion. While in wild-type seed coat cells, pectin is secreted to the apical surface, in echidna, polysaccharides accumulate in post-Golgi vesicles, the central lytic vacuole and endoplasmic reticulum-derived bodies. In contrast, proteins were partially mistargeted to internal multilamellar membranes in echidna. These results suggest that while secretion of both cell wall polysaccharides and proteins at the TGN requires ECHIDNA, different vesicle trafficking components may mediate downstream events in their secretion from the TGN.
Department of Botany, University of British Columbia, Vancouver, BC, Canada V6T 1Z4.
Heather E
HE
McFarlane
Yoichiro
Y
Watanabe
Delphine
D
Gendre
Kimberley
K
Carruthers
Gabriel
G
Levesque-Tremblay
George W
GW
Haughn
Rishikesh P
RP
Bhalerao
Lacey
L
Samuels
D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D002473:Cell Wall / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D009154:Mutation; D010368:Pectins / Q000378:metabolism*; D010641:Phenotype; D062087:Plant Mucilage / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism*; D012639:Seeds / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D022161:Transport Vesicles / Q000378:metabolism*; D014617:Vacuoles / Q000378:metabolism*; D021601:trans-Golgi Network / Q000378:metabolism*
D062087:Plant Mucilage; D011134:Polysaccharides; D010368:Pectins
Arabidopsis; Polysaccharide; Secretion; Seed coat; Trans-Golgi network