New Zealand Journal of Crop and Horticultural Science abstracts
Detection of variation among and within asparagus hybrids
using random amplified DNA (RAPD) markers
W. O. HOLLINGSWORTH
C. B. CHRISTIE
M. A. NICHOLS
H. F. NEILSON
Department of Plant Science
Massey University
Private Bag 11 222
Palmerston North, New Zealand
Abstract The reliability of random amplified polymorphic DNA
(RAPD) techniques to amplify polymorphisms in the asparagus (Asparagus
officinallis L.) genome was investigated. DNA fragments generated by
10-base primers were separated on 1.5% agarose or 8% polyacrylamide gels, and
the sensitivity of ethidium bromide and silver staining of amplified DNA
products analysed on these gels was compared. Resolution of DNA bands on
polyacrylamide gels was superior to that on agarose gels. Silver staining was
more sensitive than ethidium bromide staining. The gel type used to separate
DNA bands, and the staining technique used influenced the number of bands
visualised for each DNA profile generated. The six asparagus cultivars used in
this study were distinguished by unique banding patterns generated by each
primer. OPC-12 for example generated polymorphic markers unique to three of the
cultivars investigated. These markers were: ASP (500, 400, and 300 bp), TU (700
bp), and (PC 550 bp). Our investigation indicates that RAPD markers can be used
to characterise asparagus cultivars, and that the technique is sensitive enough
to reveal differences within seed-raised commercial cultivars. RAPD technology
has the potential to detect somaclonal variation occurring during
micropropagation.
Keywords Asparagus officinalis; primers;
micropropagation; RAPDs; ethidium bromide; silver stain; agarose;
polyacrylamide
H97-17
Received 9 May 1997; accepted 24 November 1997
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