First published online September 6, 2002; 10.1104/pp.006189
Plant Physiol, October 2002, Vol. 130, pp. 561-576
Identification and Biochemical Characterization of Mutants in the
Proanthocyanidin Pathway in Arabidopsis1
Sharon
Abrahams,*
Gregory J.
Tanner,
Philip J.
Larkin, and
Anthony
R.
Ashton
Commonwealth Scientific and Industrial Research
Organization-Plant Industry, G.P.O. Box 1600, Canberra,
Australian Capital Territory 2601, Australia
Proanthocyanidin (PA), or condensed tannin, is a polymeric
flavanol that accumulates in a number of tissues in a wide variety of
plants. In Arabidopsis, we found that PA precursors (detected histochemically using OsO4) accumulate in the endothelial
cell layer of the seed coat from the two-terminal cell stage of embryo development onwards. To understand how PA is made, we screened mature
seed pools of T-DNA-tagged Arabidopsis lines to identify mutants
defective in the synthesis of PA and found six tds
(tannin-deficient seed) complementation groups defective
in PA synthesis. Mutations in these loci disrupt the amount
(tds1, tds2, tds3,
tds5, and tds6) or location and amount of
PA (tds4) in the endothelial cell layer. The PA
intermediate epicatechin has been identified in wild type and mutants
tds1, tds2, tds3, and
tds5 (which do not produce PA) and tds6
(6% of wild-type PA), whereas tds4 (2% of wild-type
PA) produces an unidentified dimethylaminocinnamaldehyde-reacting compound, indicating that the mutations may be acting on genes beyond
leucoanthocyanidin reductase, the first enzymatic reduction step
dedicated to PA synthesis. Two other mutants were identified, an allele
of tt7, which has a spotted pattern of PA deposition and
produces only 8% of the wild-type level of type PA as propelargonidin, and an allele of tt8 producing no PA. Spotted patterns
of PA deposition observed in seed of mutants tds4 and
tt7-3 result from altered PA composition and
distribution in the cell. Our mutant screen, which was not exhaustive,
suggests that the cooperation of many genes is required for successful
PA accumulation.
1
This work was supported by Pioneer Hi-Bred and
by Meat and Livestock Australia.
*
Corresponding author; e-mail sharon.abrahams{at}csiro.au; fax
6-02-62465000.
© 2002 American Society of Plant Physiologists
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