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Plant Physiol, December 2001, Vol. 127, pp. 1764-1772
Analysis of the Raffinose Family Oligosaccharide Pathway in Pea
Seeds with Contrasting Carbohydrate Composition1
Thomas
Peterbauer,
Leslaw B.
Lahuta,
Andreas
Blöchl,
Jan
Mucha,
David A.
Jones,
Cliff L.
Hedley,
Richard J.
Gòrecki, and
Andreas
Richter*
Chemical Physiology of Plants, Institute of Ecology, University of
Vienna, A-1090 Vienna, Austria (T.P., A.B., A.R.); Department of Plant
Physiology and Biotechnology, University of Warmia and Mazury, 10-727
Olsztyn, Poland (L.B.L., R.J.G.); Department of Applied Genetics, John
Innes Centre, Norwich NR4 7UH, United Kingdom (D.A.J., C.L.H.); and
Centre for Applied Genetics, University of Agricultural Sciences
Vienna, A-1190 Vienna, Austria (J.M.)
Raffinose family oligosaccharides (RFOs) are synthesized by
a set of galactosyltransferases, which sequentially add galactose units
from galactinol to sucrose. The accumulation of RFOs was studied
in maturing seeds of two pea (Pisum sativum) lines with contrasting RFO composition. Seeds of the line SD1
accumulated stachyose as the predominant RFO, whereas verbascose, the
next higher homolog of stachyose, was almost absent. In seeds of the line RRRbRb, a high level of verbascose was accumulated alongside with
stachyose. The increase in verbascose in developing RRRbRb seeds was
associated with galactinol-dependent verbascose synthase activity. In
addition, a galactinol-independent enzyme activity was detected, which
catalyzed transfer of a galactose residue from one stachyose molecule
to another. The two enzyme activities synthesizing verbascose showed an
optimum at pH 7.0. Both activities were almost undetectable in
SD1. Maximum activity of stachyose synthase was about
4-fold higher in RRRbRb compared with SD1, whereas the
activities of galactinol synthase and raffinose synthase were only
about 1.5-fold higher in RRRbRb. The levels of galactinol synthase and
stachyose synthase activity were reflected by steady-state levels of
corresponding mRNAs. We suggest that the accumulation of verbascose in
RRRbRb was controlled by a coordinated up-regulation of the last steps
of verbascose biosynthesis.
1
This work was supported by the Austrian Science
Foundation (grant no. P13955-BIO).
*
Corresponding author; e-mail Andreas.Richter{at}univie.ac.at;
fax 43-1-4277-9542.
© 2001 American Society of Plant Physiologists
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