Recovery of Citrus Somatic Hybrids Tolerant to Phoma Tracheiphila Toxin, Combining Selection and Identification by RAPD Markers

Autor: Alessandra Gentile, Z. N. Deng, Aliza Vardi, Elisabetta Nicolosi, F. Domina, E. Tribulato
Rok vydání: 1995
Předmět:
Zdroj: Current Issues in Plant Molecular and Cellular Biology ISBN: 9789401041355
DOI: 10.1007/978-94-011-0307-7_23
Popis: Phoma tracheiphila is a pathogen that causes a serious tracheomycotic disease in commercial lemon cultivars. Somatic hybridization offers the possibility to transfer the trait for tolerance into susceptible cultivars. One of the limitations of this approach is selection and identification of nuclear hybrids at an early stage. Protoplasts isolated from calli of ‘Murcott’ tangor (tolerant) and ‘Messina’ lemon (susceptible) were fused (PEG treatment). Prior to fusion, ‘Murcott’ protoplasts were treated with 0.25 mM iodoacetate. Fusion treated protoplasts were plated in liquid medium for 40 days and then exposed to 0.6 μM of P. tracheiphila toxin. Total DNA was extracted from the selected calli and corresponding embryos and analyzed by RAPDs. With the selected primers, hybrids were successfully identified. In the survey of coculture with 1 μM toxin, the hybrid callus lines showed tolerance to the toxin. Immunoblot analysis of the recovered hybrid calli indicated the presence of chitinase among the extracellular proteins. The combined methods, selection and identification by RAPD markers, resulted in 90% recovery of nuclear hybrids tolerant to the P. tracheiphila toxin.
Databáze: OpenAIRE