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Plant Physiol, August 2000, Vol. 123, pp. 1623-1634
Identification of a CYP84 Family of Cytochrome P450-Dependent
Mono-Oxygenase Genes in Brassica napus and Perturbation of
Their Expression for Engineering Sinapine Reduction in the
Seeds1
Ramesh B.
Nair,
Richard W.
Joy IV,2
Eugen
Kurylo,
Xiaohong
Shi,
Joan
Schnaider,3
Raju S.S.
Datla,
Wilf
A.
Keller, and
Gopalan
Selvaraj*
Plant Biotechnology Institute, National Research Council of Canada,
Saskatoon, Saskatchewan, Canada, S7N 0W9
CYP84 is a recently identified family of cytochrome P450-dependent
mono-oxygenases defined by a putative ferulate-5-hydroxylase (F5H) from
Arabidopsis. Until recently F5H has been thought to catalyze the
hydroxylation of ferulate to 5-OH ferulate en route to sinapic acid.
Sinapine, a sinapate-derived ester in the seeds, is antinutritional and
a target for elimination in canola meal. We have isolated three
F5H-like genes (BNF5H1-3) from a
cultivated Brassica napus, whose amphidiploid progenitor is
considered to have arisen from a fusion of the diploids
Brassica rapa and Brassica oleracea. Two cultivated varieties of the
diploids were also found to contain BNF5H3 and
additionally either BNF5H1 or BNF5H2,
respectively. Whereas all three are >90% identical in their coding
sequence, BNF5H1 and BNF5H2 are closer to
each other than to BNF5H3. This and additional data
suggest that the two groups of genes have diverged in an ancestor of
the diploids. B. napus showed maximal F5H
expression in the stems, least in the seeds, and subtle differences among the expression profiles of the three genes elsewhere. Transgenic B. napus with cauliflower mosaic virus 35S-antisense
BNF5H contained up to 40% less sinapine, from 9.0 ± 0.3 mg in the controls to 5.3 ± 0.3 mg g 1 seed.
F5H from Arabidopsis and a similar enzyme from sweetgum (Liquidamber styraciflua) has recently been shown to have
coniferaldehyde hydroxylase activity instead of F5H activity. Thus the
supply of 5-OH coniferaldehyde or 5-OH ferulate has a bearing on
sinapine accumulation in canola seeds.
1
This is National Research Council of Canada
publication no. 43,773.
2
Present address: WestCan Malting, P.O. Box 113, Alix,
AB, Canada, T0C 0B0.
3
Present address: Aventis Crop Science, 203-407 Downey
Road, Saskatoon, SK, Canada, S7N 4L8.
*
Corresponding author; e-mail gopalan.selvaraj{at}nrc.ca; fax
306-975-4839.
© 2000 American Society of Plant Physiologists
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