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First published online February 27, 2003; 10.1104/pp.102.016840

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Plant Physiol, March 2003, Vol. 131, pp. 1487-1495

Tobacco Transgenic Lines That Express Fenugreek Galactomannan Galactosyltransferase Constitutively Have Structurally Altered Galactomannans in Their Seed Endosperm Cell Walls1

J.S. Grant Reid,* Mary E. Edwards, Cathryn A. Dickson, Catherine Scott, and Michael J. Gidley

Department of Biological Sciences, University of Stirling, Stirling FK9 4LA, United Kingdom (J.S.G.R., M.E.E., C.A.D., C.S.); and Unilever Research Laboratory, Colworth House, Sharnbrook, Bedford MK44 1LQ, United Kingdom (M.J.G.)

Galactomannans [(1right-arrow6)-alpha -D-galactose (Gal)-substituted (1right-arrow4)-beta -D-mannans] are major cell wall storage polysaccharides in the endosperms of some seeds, notably the legumes. Their biosynthesis in developing legume seeds involves the functional interaction of two membrane-bound glycosyltransferases, mannan synthase (MS) and galactomannan galactosyltransferase (GMGT). MS catalyzes the elongation of the mannan backbone, whereas GMGT action determines the distribution and amount of Gal substitution. Fenugreek (Trigonella foenum-graecum) forms a galactomannan with a very high degree of Gal substitution (Man/Gal = 1.1), and its GMGT has been characterized. We now report that the endosperm cell walls of the tobacco (Nicotiana tabacum) seed are rich in a galactomannan with a very low degree of Gal substitution (Man/Gal about 20) and that its depositional time course is closely correlated with membrane-bound MS and GMGT activities. Furthermore, we demonstrate that seeds from transgenic tobacco lines that express fenugreek GMGT constitutively in membrane-bound form have endosperm galactomannans with increased average degrees of Gal substitution (Man/Gal about 10 in T1 generation seeds and about 7.5 in T2 generation seeds). Membrane-bound enzyme systems from transgenic seed endosperms form galactomannans in vitro that are more highly Gal substituted than those formed by controls under identical conditions. To our knowledge, this is the first report of structural manipulation of a plant cell wall polysaccharide in transgenic plants via a biosynthetic membrane-bound glycosyltransferase.

1 This work was supported by the Biotechnology and Biological Sciences Research Council (UK; research grant).

* Corresponding author; e-mail j.s.g.reid{at}stir.ac.uk; fax 44-1786-464994.

© 2003 American Society of Plant Biologists


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