Patent ID: 6500616
Filing Date: 2002-12-31
Classification: A01H,C12N,C12Q

Abstract:
A method for detecting genomic destabilization arising during tissue culture of plant cells, comprising the steps of:(a) isolating DNA from (i) a normal plant, and (ii) a plurality of off-type plants of the sane species as the normal plant, wherein each of the off-type plants has a different phenotypic mutation; (b) performing representational difference analysis to obtain a plurality of DNA subtraction products corresponding to a plurality of genetic sequence differences between the DNAs of the off-type plants and the DNA of the normal plant, wherein the plurality of genetic sequence differences include silent mutations that are not phenotypically expressed in the off-type plants; (c) isolating DNA from a sample of plant cells prior to tissue culture, wherein the plant cells are of the same species as the plants in step (a); (d) using the plurality of DNA subtraction products as genetic probes to detect existing genetic polymorphisms in the DNA from the sample of plant cells; (e) growing the plant cells in tissue culture; (f) isolating DNA from a sample of the plant cell in tissue culture; and (g) using the plurality of DNA subtraction products as genetic probes to detect a new genetic polymorphism arising in the plant cell DNA during tissue culture, wherein the new genetic polymorphism is not present in the DNA of steps (c) and (d) and wherein a plant regenerated from a plant cell comprising the new genetic polymorphism is phenotypically normal, wherein the presence of the new genetic polymorphism is indicative of genomic destabilization arising during tissue culture of plant cells.