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PLANT PHYSIOLOGY , Vol 106, Issue 1 109-118, Copyright © 1994 by American Society of Plant Biologists

MOLECULAR BIOLOGY AND GENE REGULATION

The Cloning of Two Tomato Lipoxygenase Genes and Their Differential Expression during Fruit Ripening

B. J. Ferrie, N. Beaudoin, W. Burkhart, C. G. Bowsher and S. J. Rothstein
Department of Molecular Biology and Genetics, University of Guelph, Guelph, Ontario, Canada N1G 2W1 (B.J.F., N.B., C.G.B., S.J.R.)

A membrane-associated lipoxygenase from breaker-stage fruit of tomato (Lycopersicon esculentum Mill.) was purified and partially sequenced. Using degenerate oligonucleotides corresponding to portions of this sequence, a cDNA was amplified by PCR and used to screen a breaker fruit cDNA library. Two clones, tomloxA and tomloxB, were isolated and one of these (tomloxA) corresponded to the isolated protein. Genomic clones were isolated and sequence data from these were used to obtain the 5[prime] ends of the cDNAs. The 2.8-kb cDNAs encode proteins that are similar in size and sequence to each other and to other plant lipoxygenases. DNA blot analysis indicated that tomato contains three or more genes that encode lipoxygenase. RNA blot analysis showed that tomloxA is expressed in germinating seeds as well as in ripening fruit, where it reached its peak during breaker stage. tomloxB appears to be fruit specific and is at its highest level in ripe fruit.


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