ISHS Acta Horticulturae 663: XI Eucarpia Symposium on Fruit Breeding and Genetics
THE TECHNOLOGY OF CDNA ARRAY FILTER HYBRIDISATION TO IDENTIFY DIFFERENTIALLY WOOD EXPRESSED-CDNAS INVOLVED IN COMPATIBLE AND INCOMPATIBLE GRAFTING IN PEACH |
| Authors: | M.A. Palombi, H.A Becker, C. Damiano |
| Keywords: | Prunus, grafting, filter hybridisation, array, gene expression |
| Abstract:
Cultivar stability in fruit trees is ensured by grafting onto particular rootstocks. The major limiting factor in a graft union is the incompatibility reaction (complete or incomplete) between scions and rootstocks. In this work, we analyse changes in mRNA patterns, during the first step of grafting (until callus formation) to isolate specific cDNAs involved in graft (in)compatibility in peach. In order to study many genes simultaneously, we used the technology of cDNA array filter hybridisation. Two cDNA libraries were constructed, one from wood of the peach rootstock GF677 and one from wood of the cultivar ‘Calypso’. No less than 2500 cDNA clones for each library were obtained and, after amplification, were spotted on nylon filters which also included serial dilutions of two non peach-sequences, represented by cDNA clones for the human nebulin and desmin genes. 33P labelled cDNA probes were prepared starting from poly(A)+ RNA, purified at different times after callus formation, from both compatible and incompatible peach grafting combinations. Array data analysis for expression patterns allowed the identification of different cDNA clones putatively involved in the first step of compatible and incompatible peach grafting. Sequencing of the cDNA clones must be completed but preliminary results show that cDNA filter array can be a powerful tool for gene discovery and gene expression in peach grafting. | |
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