Asunto(s)
Perfilación de la Expresión Génica/métodos , Genes de Plantas , Plantas/genética , ARN de Planta/análisis , Clonación Molecular/métodos , Electroforesis en Gel de Poliacrilamida/métodos , Etilenos/biosíntesis , Frutas/genética , Hidrolasas/genética , Hibridación de Ácido Nucleico/métodos , Desarrollo de la Planta , Sondas ARN/síntesis químicaRESUMEN
Structural and enzymatic aspects of leaf and fruit abscission in explants treated with exogenous ethylene were studied and compared with those previously observed under field conditions. Light and electron microscopy observations reconfirm that differences in abscission occur in leaf and fruit, and in explants cell separation is accelerated, with a more dramatic degradation of the cell walls. In fruit, digestion starts from the middle lamella, and subsequently extends to the entire parietal mass. In the leaf the process involves the primary cell wall, leaving, at the end, undigested materials. In fruit the lytic activity is sustained by endocellulase, and by exo- as well as endopolygalacturonase. In the leaf only endocellulase activity has been found. Cell enlargement phenomena occur subsequent to abscission zone activation in both leaf and fruit explants, involving the cells of either the separation layer or the adjacent region.
RESUMEN
Ethylene-induced abscission in leaf and fruit explants of peach involves different enzymes. In leaves abscission is accompanied by increased occurrence of cellulase forms differing in isoelectric point (pI 6.5 and 9.5). A polypeptide with a molecular mass of 51 kDa gives in a western blot a strong cross-reaction with an antibody raised against a maturation cellulase from avocado fruit. Cellulase activity is also found in abscising fruit explants but the amount is very low compared to that of the leaf explants. A northern analysis with a cellulase clone from avocado reveals the presence of two hybridizing mRNAs with a size of 2.2 kb and 1.8 kb, respectively. The steady-state level of the 2.2 kb mRNA is significantly increased by treatment with ethylene. Polygalacturonases are not detected in abscising leaves, but are strongly induced by ethylene in fruit explants. Of the three forms found, two are exopolygalacturonases while the third is an endoenzyme. Ethylene activates preferentially the endoenzyme and the basic exoenzyme but depresses the acid exopolygalacturonases. A northern analysis carried out with a cDNA coding for tomato endopolygalacturonase shows hybridization only with one endopolygalacturonase mRNA form in the fruit abscission zone. Treatment with ethylene causes an increase in the steady-state level of this mRNA. The differences in the enzyme patterns observed in fruit and leaf abscission zones and a differential enzyme induction suggest the feasibility to regulate fruit abscission in peach with the aid of antisense RNA genes.