RFLP analysis in chrysanthemum. 1. Probe and primer development
1994
Wolff, K. | Peters-Van Rijn, J. | Hofstra, H. (TNO Nutrition and Food Research, Zeist (Netherlands). Dept. of Microbiology)
In order to study genetic variability at the DNA level in chrysanthemum (Dendranthema grandiflora Tzvelev) Pst1 and Hind3 genomic libraries were constructed. Probes from both libraries were tested for the presence of restriction fragment length polymorphisms (RFLPs). Of the probes from the Pst1 library 91% appeared to hybridize to low-copy genes, while only 35% of those from the Hind3 library appeared to do so. The Pst1 probes were used in further analyses as 79% of them showed RFLPs, whereas the Hind3 low-copy number probes gave only 14% polymorphic patterns. Because of the hexaploid character of chrysanthemum, complex patterns generally consisting of 6 to 12 fragments were visible on a Southern blot after hybridization. To simplify the genetic analysis, locus-specific polymerase chain reaction (PCR) primers were developed that gave simple polymorphic patterns in a number of cases. The RFLP probes and primers developed will be used in future marker-assisted selection in this polyploid crop.
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