Differential induction of phenylalanine ammonia-lyase gene expression in response to in vitro callus unions of Prunus spp.

Abstract

Phenylalanine ammonia-lyase (PAL) is a key enzyme in the synthesis of phenolic compounds, which play a prominent role in graft union formation, including the marked effects of their accumulation in incompatibility response. The purpose of this study was to assess changes in the abundance of PAL mRNA during graft union development. Partial cDNA clones encoding the enzyme were isolated from in vitro callus tissue in the apricot (Prunus armeniaca L.) cultivar Moniqui and the plum rootstock Marianna 2624 (Prunus munsoniana x Prunus cerasifera). The deduced partial amino acid sequence showed high homology with PAL genes from other plant species. We monitored PAL expression 5, 10, 15 and 20 days after the establishment of in vitro callus unions. The levels of PAL mRNA increased 5 days after grafting in both compatible and incompatible unions. Nevertheless, significant differences were observed at the transcript level through both types of combinations from the second week. The results showed a higher level of PAL transcription in graft unions of incompatible partners, where a lack of adaptation between stock and scion takes place. The level of scion-stock compatibility was related to the PAL expression pattern. In addition, cell walls of the callus cells were not stained by phloroglucinol-HCl, indicating that the increased PAL expression did not result in the formation of lignin. However, staining with Naturstoff reagent A confirmed the highest accumulation of soluble and wall-bound phenolic compounds at the graft interface of incompatible unions.