Differentiation and functional connection of vascular elements in compatible and incompatible pear/quince internode micrografts

Micrografts of internodes excised from in vitro grown pear plants(Pyrus communis L. cv. 'Bosc' (B) and cv. 'Butiffa Hardy'(BH)) and quince (Cydonia oblonga Mill. East Malling clone C (EMC)), were cultured aseptically to test the effectiveness of their functional vascular reconnection in relation to incompatibility-compatibility relationships that these genotypes exhibit in the field. The incompatible heterograft (B/EMC) showed a marked delay in internode cohesion compared with the autografts (both B/B and BH/BH) and the compatible heterograft (BH/EMC). Even when fused, the translocation of [C-14]-sorbitol from upper to lower internode was lower in B/EMC micrografts than in the other combinations. Epifluorescence Studies performed with carboxyfluorescin, a specific phloem probe, indicated that the limited translocation was caused by a delay in the establishment of functional phloem continuity between the two internodes. In the B/EMC combination, new differentiated tracheary elements (TE) in the parenchyma tissue at the graft interface between the two internodes were not detected until 30 days after grafting, whereas in the BH/EMC heterograft and both autografts, new xylem connections appeared to cross the interface 20 days after grafting. Immunohistochemical detection (terminal nick-end labeling assay) of the number of cells undergoing nuclear DNA fragmentation at the graft interface confirmed that the limited and delayed TE differentiation in B/EMC heterografts was associated with a decrease in the activity of programmed cell death processes involved in the differentiation of TE.