Sequence-specific PCR markers were derived from one new and eight previously
identified random amplified polymorphic DNA (RAPD) markers for the purpose of
identifying grape (Vitis) rootstocks. The markers were developed because the RAPD
assay was found to be inconsistent and the original RAPD markers unreliable. Southern
hybridization analysis of the RAPD gels with cloned RAPD bands as probes revealed
deficiencies of scoring RAPD bands based solely on ethidium bromide staining. In one
case, bands of the same size generated by the same primer in different rootstocks -- normally
scored as the same marker -- failed to cross-hybridize, implying a lack of
homology between the bands. In another case, a band scored as absent based on
ethidium bromide staining was detected by hybridization. One of nine RAPD bands was
cloned in the present study. All nine were partially sequenced and sequence-specific
pairs of primers were synthesized for each for use under stringent PCR conditions. Three
of the primer pairs generated products only from the rootstocks from which the original
RAPD bands had been cloned; three others produced products from additional rootstocks
while still generating polymorphisms; two others generated apparent length variants from
some accessions; and one primer pair resulted in a loss of polymorphism. Based on the
eight polymorphic markers, five of nine rootstocks could be unambiguously
distinguished. / Graduation date: 1996
| Identifer | oai:union.ndltd.org:ORGSU/oai:ir.library.oregonstate.edu:1957/34994 |
| Date | 30 June 1995 |
| Creators | Xu, Hong, 1968- |
| Contributors | Bakalinsky, Alan Tagore |
| Source Sets | Oregon State University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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