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JXB Advance Access originally published online on April 8, 2004
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Journal of Experimental Botany, Vol. 55, No. 399, pp. 961-973, May 1, 2004
© 2004 Oxford University Press

Cell and Molecular Biology, Biochemistry and Molecular Physiology

A physical, enzymatic, and genetic characterization of perturbations in the seeds of the brownseed tomato mutants

Received 28 August 2003; Accepted 20 January 2004

A. Bruce Downie1,*, Lynnette M. A. Dirk1, Qilong Xu1, Jennifer Drake2, Deqing Zhang1, Manjul Dutt1, Alan Butterfield2, Robert R. Geneve1, J. Willis Corum, III1, Karl G. Lindstrom3 and John C. Snyder1

1 Department of Horticulture, Plant Science Building, University of Kentucky, 1405 Veterans Drive, Lexington, KY 40546-0312, USA
2 Center of Membrane Sciences, Department of Chemistry, University of Kentucky, Lexington, KY 40546, USA
3 University of Kentucky, Advanced Genetics Technology Center, University of Kentucky, Lexington, KY 40546, USA

* To whom correspondence should be addressed. Fax: +1 859 257 7874. E-mail: adownie{at}uky.edu

The brownseed mutants (bs1, bs2, and bs4) of tomato all possess dark testae and deleteriously affect seed germination speed and/or final percentage. Poor germination performance of the bs1 but not the bs4 mutant, was due to greater impediment to radicle egress. Testa toughening (bs1) was prevented by drying in N2. However, poor germination speed was hardly affected by drying. GA4+7 did not ameliorate germination percentage or speed (bs1, bs2), whereas bs4 seeds commenced radicle protrusion sooner and had a greater germination percentage. bs1 mutant seeds have two times more catalase activity while those of bs4 contained six times more peroxidase and almost two times more catalase activity than WTs. bs4 release only half of the reactive oxygen species into the media than WT during imbibition. EPR detected the presence of free radicals in bs1 and its WT. bs mutants were epistatic to 12 anthocyaninless mutations, at least some of which produce seeds of lighter than usual testa colour. Macro-arrays of subtractive, suppressive PCR products identified differentially regulated transcripts between seeds of bs4 and WT. EST identity suggests bs4 does not exit the developmental programme upon attaining maturity.

Key words: Catalase, electron paramagnetic resonance (EPR), free radicals, germination, Lycopersicon esculentum, macro-array, peroxidase, seed, subtractive-suppressive PCR, testa.


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