Analysis of mRNAs that Accumulate in Response to Low Temperature Identifies a Thiol Protease Gene in Tomato

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Abstract
We have studied the induction of gene expression at low temperature by cloning mRNAs that accumulate when unripe tomato (Lycopersicon esculentum) fruit are incubated at 4.degree.C. Two cloned mRNAs, C14 and C17, accumulate relatively rapidly in response to cold treatment, while a third, C19, displays a delayed response. Significant levels of these mRNAs were not detected during fruit ripening at normal temperature. We have analyzed gene expression of different temperatures and detect half-maximal accumulation of the C14 and C17 mRNAs at 16.degree.C and 11.degree.C, respetively, and have observed that sustained gene expression requires continuous cold treatment. furthermore, the level of C14 and C17 gene expression in cold-tolerant (hybrid L. esculentum/Lycopersicon pimpinellifolium) fruit is different from that in cold-sensitive (L. esculentum) fruit. DNA sequence analysis indicates that the C14 mRNA encodes a polypeptide with a region that is homologous to the plant thiol proteases actinidin and papain and to the animal thiol protease cathespin H. We conclude from these experiments that low temperature selectively induces the expression of specific genes and that one such gene encodes a thiol protease.