Immunodetection and Characterization of Tomato Endo-β-1,4-Glucanase Cel1 Protein in Flower Abscission Zones

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Abstract
Tomato (Lycopersicon esculentum Mill.) endo-β-1,4-glucanase Cel1 mRNA accumulation was previously correlated with abscission of flower explants. Cel1 antibodies were raised against a fusion protein encoding a portion of the Cel1 polypeptide and was shown to react specifically with three polypeptides with molecular masses ranging between 51 and 53 kD in flower abscission zones induced to abscise. All three polypeptides were clearly suppressed in two transgenic lines expressing an antisense Cel1 gene that specifically suppressed the accumulation of Cel1 mRNA, indicating that all three polypeptides are products of the Cel1 gene. Cel1 protein accumulation was correlated with flower abscission. Breakstrength and Cel1 protein content were also analyzed in flower explants, indicating that Cel1 protein accumulation is correlated with the final stages of flower shedding, which suggests that Cel1 is involved in the late stage of abscission. These results support the involvement of Cel1 in the abscission of flower explants and suggest that other hydrolase activities also participate in that process.