Molecular cloning of a ripening-specific lipoxygenase and its expression during wild-type and mutant tomato fruit development.
A 94-kD protein that accumulates predominately in tomato (Ly-copersicon esculentum) fruit during ripening was purified, and antibodies specific for the purified protein were used to isolate cDNA clones from a red-ripe fruit cDNA library. A sequence analysis of these cDNAs and cross-reactivity of the 94-kD-specific antibodies to the soybean lipoxygenase (LOX) L-1, L-2, and L-3 proteins and soybean LOX L-1-specific antibodies to the 94-kD protein identified it as a member of the LOX gene family. Maximum levels of the 94-kD LOX mRNA and protein are present in breaker to ripe and red-ripe stages, respectively. Expression of 94-kD LOX in different tissues from mature green and red-ripe tomato fruits was found to be greatest in the radial walls of ripe fruit, but immunocytolocalization using tissue printing suggests that the highest accumulation of its protein occurs in locular jelly. None of 94-kD LOX is expressed in nonripening mutant fruits of any age. Never-ripe mutant fruit accumulate the 94-kD LOX mRNA to levels similar to those obtained in wild-type fruit, but fail to accumulate the 94-kD LOX protein. Collectively, the results show that expression of 94-kD LOX is regulated by the ripening process, and ethylene may play a role in its protein accumulation.
Kausch
K D
KD
Department of Horticulture, Purdue University, West Lafayette, Indiana 47907-1165, USA.
Handa
A K
AK
D003001:Cloning, Molecular; D018076:DNA, Complementary; D015723:Gene Library; D005838:Genotype; D008084:Lipoxygenase / Q000096:biosynthesis* / Q000302:isolation & purification; D018551:Solanum lycopersicum / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D005810:Multigene Family; D009154:Mutation; D010802:Phylogeny; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification
D018076:DNA, Complementary; D011994:Recombinant Proteins; D008084:Lipoxygenase