Molecular analysis of ethylene signal transduction in tomato
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The plant hormone ethylene plays an important role in plant growth, development, and physiology. One of the critical components of the ethylene signal transduction pathway, ctr1 (constitutive triple response), was identified using a particularly useful seedling screen that takes advantage of the profound effects ethylene has on etiolated seedlings, known as triple response. CTR1 is one of six Arabidopsis MAPKKKs that are related to the Raf kinases, and acts as a negative regulator of ethylene response. In this study, isolation and characterization of a family of CTR1-like genes in tomato is reported. Based on amino acid alignments and phylogenetic analysis, the tomato CTR1-like (LeCTR) genes are more similar to Arabidopsis CTR1 (AtCTR1) than any other MAPKKK sequences in the Arabidopsis genome. The capacity of the LeCTR genes to function as negative regulators in ethylene signal transduction was tested through complementation of the Arabidopsis ctr1-8 mutant. Quantitative real-time PCR was carried out to generate an expression profile for the CTR1-like gene family during different stages of development marked by increased ethylene biosynthesis, including fruit ripening. The possibility of a multi-gene family of CTR1-like genes in other species besides tomato was examined through mining of EST and genomic sequence databases. Based on nucleotide and amino acid identity, At4g24480 is most similar to AtCTR1 and could potentially represent a CTR1-like gene in Arabidopsis. Arabidopsis plants carrying a T-DNA insert in the At4g24480 locus were examined for abnormal ethylene response phenotypes including sensitivity to other hormones, signal molecules and abiotic stresses. Two mutant alleles, ctr1-1 and ctr1-8, containing mutations that disrupt kinase activity and receptor association, respectively, were examined for sensitivity to these same treatments in an effort to better characterize ethylene hormone and non-hormone interactions. They also served as controls to determine if At4g24480 indeed possessed CTR1-like function. Arabidopsis and tomato represent species with very distinct fruit ripening/maturation programs. The critical dependence on ethylene for fruit ripening in tomato might have resulted in alteration or modification of the ethylene signal transduction pathway relative to Arabidopsis. Plans to characterize individual functions of the LeCTR genes through over-expression and reduced expression in tomato are outlined.
Adams-Phillips, Lori C. (2003). Molecular analysis of ethylene signal transduction in tomato. Doctoral dissertation, Texas A&M University. Texas A&M University. Available electronically from