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Plant Physiol, May 2000, Vol. 123, pp. 265-274
Disaccharide-Mediated Regulation of
Sucrose:Fructan-6-Fructosyltransferase, a Key Enzyme of Fructan
Synthesis in Barley Leaves1
Joachim
Müller,*
Roger A.
Aeschbacher,
Norbert
Sprenger,2
Thomas
Boller, and
Andres
Wiemken
Botanisches Institut der Universität Basel, Hebelstrasse 1, CH-4056 Basel, Switzerland
Previous work has indicated that sugar sensing may be important in
the regulation of fructan biosynthesis in grasses. We used primary
leaves of barley (Hordeum vulgare cv Baraka) to study the mechanisms involved. Excised leaf blades were supplied in the dark
with various carbohydrates. Fructan pool sizes and two key enzymes of
fructan biosynthesis, sucrose (Suc):Suc-1-fructosyltransferase (1-SST;
EC 2.4.1.99) and Suc:fructan-6-fructosyltransferase (6-SFT; EC
2.4.1.10) were analyzed. Upon supply of Suc, fructan pool sizes
increased markedly. Within 24 h, 1-SST activity was stimulated by
a factor of three and 6-SFT-activity by a factor of more than 20, compared with control leaves supplemented with mannitol (Mit). At the
same time, the level of mRNA encoding 6-SFT increased conspicuously.
These effects were increased in the presence of the invertase inhibitor
2,5-dideoxy-2,5-imino-D-mannitol. Compared with equimolar
solutions of Suc, glucose (Glu) and fructose stimulated 6-SFT activity
to a lesser extent. Remarkably, trehalose (Tre; Glc- -1 and
1--Glc) had stimulatory effects on 6-SFT activity and, to a somewhat
lesser extent, on 6-SFT mRNA, even in the presence of validoxylamine A,
a potent trehalase inhibitor. Tre by itself, however, in the presence
or absence of validoxylamine A, did not stimulate fructan accumulation.
Monosaccharides phosphorylated by hexokinase but not or weakly
metabolized, such as mannose (Man) or 2-deoxy-Glc, had no stimulatory
effects on fructan synthesis. When fructose or Man were supplied
together with Tre, fructan and starch biosynthesis were strongly
stimulated. Concomitantly, phospho-Man isomerase (EC 5.3.1.8) activity
was detected. These results indicate that the regulation of fructan
synthesis in barley leaves occurs independently of hexokinase and is
probably based on the sensing of Suc, and also that the structurally
related disaccharide Tre can replace Suc as a regulatory compound.
1
This work was supported by grants from the Swiss
National Science Foundation (to T.B. and A.W.) and by a fellowship from
the Roche Foundation (to J.M.).
2
Present adress: Carnegie Institute of
Washington, 260 Panama Street, Stanford, CA 94305.
*
Corresponding author; e-mail joachim.mueller{at}unibas.ch; fax
41-61-2672330.
© 2000 American Society of Plant Physiologists
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