Detection of somaclonal variation in cultured rice cells using digoxigenin-based random amplified polymorphic DNA

A procedure for the non-radioactive detection of random amplified polymorph ic DNA (RAPD) was developed and designated as digoxigenin (DIG)-based RAPD. Using this procedure, we analyzed somaclonal variation in cultured cells o f rice. Somaclonal variation was found to increase with culture age. More t han 50 polymorphic fragments were identified with the four primers tested. Random sequencing of 10 clones generated one intron, one 5'-noncoding, and eight non-redundant expressed sequences. A database search for homology sho wed that the eight exon sequences displayed a significant similarity to seq uences already stored in EMBL, GenBank and DDBJ. The sources of the known g enes ranged From microorganism to human, including three rice genes. The re sults showed that somaclonal variation might have occurred in transfer RNA, ribosomal protein, and other genes during cell culture.