REGULATION OF A PETAL-SPECIFIC ETHYLENE-INDUCED 70-KDA PROTEIN FROM DIANTHUS-CARYOPHYLLUS

Ethylene enhances senescence of carnation (Dianthus caryophyllus L.) flowers. Sephacryl S-200 gel filtration followed by SDS-PAGE of extracts of petals exposed to ethylene revealed the increase or appearance of proteins with molecular masses of 70, 60, 35 and 33 kDa, and a concomitant decrease in proteins with molecular masses of 62, 45, 34, 30 and 26 kDa. The 70-kDa protein, which was not detected in untreated flowers, was isolated and polyclonal antibodies were prepared against it. Its N-terminal sequence (20 amino acids) was identical to that of a portion of the deduced protein encoded by cDNA clone pSR12 recently isolated by others from carnation petals. Western blot analyses of petal extracts after various exposure times to ethylene alone or in the presence of translation or transcription inhibitors suggested that accumulation of the 70-kDa protein is due primarily to ethylene-induced synthesis. The apparent induction by ethylene was further supported by data indicating that the protein is not formed in flowers pre-treated with silver thiosulfate, an inhibitor of ethylene action, prior to exposure to ethylene. Ethylene induction of the protein was found only in petal tissue, and the protein was not detected in other carnation plant organs. Immunological studies revealed that the protein is specifically expressed in carnation petals and not in petals of several other flowers. Southern and northern analyses using an SR12 cDNA probe indicated that the genomic DNA of petunia, carnation, and gerbera contain fragments homologous to SR12. Gene expression products were however, only detected in petals of ethylene-treated carnation flowers.