A Diagnostic Test to Confirm Interspecific Allium Hybrids Using Random Amplified Polymorphic DNA from Crude Leaf DNA Extracts

Abstract

A crude extraction procedure for DNA was modified and used for small leaf samples (2-4 mg) in Allium. Six random decamers (OPD 5, 7, 14, 15, 18 and 19) generated characteristic random amplified polymorphic DNA (RAPD) profiles from leaf DNA of Allium tuberosum Rottl. ex Spreng. 'Tender pole' obtained using a crude micro extraction process These RAPD profiles were roughly similar to those of DNA purified using a sodium dodecyl sulfate (SDS) extraction procedure.
Crude extracts of leaf DNA were prepared from three accessions of A. tuberosum 'Tender pole', 'five 'Yoshitake hybrids' (A. chinense G. Don × A. thunbergii G. Don) and seven putative trispecific hybrids between these two groups. DNA amplification with four random decamers (OPD 14, 15, 18 and 19) by polymerase chain reaction produced a total of 186 bands : 49 bands were found only in the RAPD profiles of the putative hybrid and the corresponding male parent (M bands), whereas 58 bands were found in both female parent and trispecific hybrid plants (F bands). One of the seven putative hybrids had no M band, indicating that it had no relationship to its putative male parent and that it may have been the product of an unwanted pollination.