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Senescence is a complex process of aging in which most organs in plants undergo a series of cellular and biochemical changes, ultimately leading to loss of function. Ethylene is the major plant hormone implicated in the regulation of senescence. However, little is known about the role of the ethylene signal transduction machinery in initiating leaf senescence. To better understand the molecular basis of ethylene signaling in senescence, we attempted to isolate new components of the signaling pathway by screening mutants deficient in ethylene biosynthesis and signaling. Here, we describe the isolation of two mutants in which the ethylene synthesis rate was much lower than wild type and the three mutants were found to be responsive to induction of leaf senescence by darkness. Genetic analysis revealed that the ethylene biosynthetic mutants were located on a single genetic locus, the ethylene insensitive 1 (ein1) locus. Both ein1 and ein2 exhibited a mutant phenotype in response to darkness and 1-aminocyclopropane-1-carboxylic acid (ACC) treatment. The ein2 mutant also exhibited a mutant phenotype in response to the RNA interference (RNAi) of ein2. The double mutant exhibited a far more severe phenotype. Our results indicate that the two ethylene biosynthetic genes, EIN1 and EIN2, and
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