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Plant Physiol, March 2000, Vol. 122, pp. 645-656
Purification of a Jojoba Embryo Wax Synthase, Cloning of its
cDNA, and Production of High Levels of Wax in Seeds of Transgenic
Arabidopsis
Kathryn D.
Lardizabal,*
James G.
Metz,
Tetsuo
Sakamoto,1
William C.
Hutton,
Michael R.
Pollard,2 and
Michael W.
Lassner3
Calgene Campus, Monsanto, 1920 Fifth Street, Davis, California
95616 (K.D.L., J.G.M., M.R.P., M.W.L.); Nippon Steel Corporation,
Tokyo, Japan (T.S.); and Monsanto, 800 N. Lindbergh Boulevard, St.
Louis, Missouri 63167 (W.C.H.)
Wax
synthase (WS, fatty acyl-coenzyme A [coA]: fatty alcohol
acyltransferase) catalyzes the final step in the synthesis of linear
esters (waxes) that accumulate in seeds of jojoba (Simmondsia chinensis). We have characterized and partially purified this enzyme from developing jojoba embryos. A protein whose presence correlated with WS activity during chromatographic fractionation was
identified and a cDNA encoding that protein was cloned. Seed-specific expression of the cDNA in transgenic Arabidopsis conferred high levels
of WS activity on developing embryos from those plants. The WS sequence
has significant homology with several Arabidopsis open reading frames
of unknown function. Wax production in jojoba requires, in addition to
WS, a fatty acyl-CoA reductase (FAR) and an efficient fatty acid
elongase system that forms the substrates preferred by the FAR. We have
expressed the jojoba WS cDNA in Arabidopsis in combination with cDNAs
encoding the jojoba FAR and a  -ketoacyl-CoA synthase (a component of
fatty acid elongase) from Lunaria annua.
13C-Nuclear magnetic resonance analysis of pooled whole
seeds from transgenic plants indicated that as many as 49% of the oil
molecules in the seeds were waxes. Gas chromatography analysis of
transmethylated oil from individual seeds suggested that wax levels may
represent up to 70% (by weight) of the oil present in those seeds.
1
Present address: Faculty of Engineering, Kyushu
University, Hakozaki 6-10-1, Fukuoka 812-8581, Japan.
2
Present address: Department of Botany and Plant
Pathology, Michigan State University, East Lansing, MI 48824-1312.
3
Present address: Maxygen Inc., 515 Galveston
Drive, Redwood City, CA 94063.
*
Corresponding author; e-mail kathy.lardizabal{at}monsanto.com; fax
530-792-2453.
© 2000 American Society of Plant Physiologists
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