Methods of monitoring genomic integrity and detecting genomic destabilization of plant cells in tissue culture

摘要

The invention provides a method for obtaining molecular markers for use as a diagnostic and quality control tool to identify genomic polymorphisms that arise during the process of tissue culture of in vitro propagated plants. By using a representational difference analysis (RDA) adapted for plant genomes, a set of nucleic acid difference sequences between normal and off-type plant genomes are obtained. The invention further provides a method for isolating sets of variant sequences which are common to many naturally occurring or tissue culture-generated off-types of the same cultivar or species, in addition to variant sequences present in all off-types, regardless of the phenotypic mutation, and/or in all off-types that exhibit the same mutation. Detection of somaclonal variation by the method of the invention may present an opportunity to optimize tissue culture conditions and to optimize plant multiplication rates without producing a significant number of off-types.