Oleosin expression and trafficking during oil body biogenesis in tobacco leaf cells.
We have established a versatile method for studying the interaction of the oleosin gene product with oil bodies during oil body biogenesis in plants. Our approach has been to transiently express a green fluorescent protein (GFP)-tagged Arabidopsis oleosin gene fusion in tobacco leaf cells containing bona fide oil bodies and then to monitor oleosin-GFP expression using real-time confocal laser scanning microscopy. We show that normally non-oil-storing tobacco leaf cells are able to synthesize and then transport oleosin-GFP fusion protein to leaf oil bodies. Synthesis and transport of oleosin-GFP fusion protein to oil bodies occurred within the first 6 h posttransformation. Oleosin-GFP fusion protein exclusively associated with the endoplasmic reticulum and was trafficked in a Golgi-independent manner at speeds approaching 0.5 microm sec(-1) along highly dynamic endoplasmic reticulum positioned over essentially static polygonal cortical endoplasmic reticulum. Our data indicate that oil body biogenesis can occur outside of the embryo and that oleosin-GFP can be used to monitor early events in oil body biogenesis in real-time.
Wahlroos
Tony
T
University of Turku, Joint Biotechnology Laboratory, Turku, Finland. tomili@utu.fi
Soukka
Jori
J
Denesyuk
Alexander
A
Wahlroos
Rina
R
Korpela
Timo
T
Kilby
Nigel J
NJ
D017360:Arabidopsis / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000096:biosynthesis* / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D018515:Plant Leaves / Q000378:metabolism*; D010938:Plant Oils / Q000378:metabolism; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013997:Time Factors; D014026:Nicotiana / Q000378:metabolism*
D029681:Arabidopsis Proteins; D008164:Luminescent Proteins; D010938:Plant Oils; D011993:Recombinant Fusion Proteins; C079353:oleosin protein, Arabidopsis; D049452:Green Fluorescent Proteins