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Genetic
Linkage Mapping of Coneflower
Ahmad
Naseer Aziz
Institute of Agricultural and Environmental Research
Seminar Series
Tennessee State University, Nashville, TN
October 19, 2005
Coneflowers (Echinacea spp.) are important to both
the ornamental and medicinal herb industries, thus efficient methods for
their genetic analysis are needed. Production of molecular markers from
individual pollen grains can generate genetic data useful for many
analyses including linkage mapping without the need of performing
controlled pollinations. The limited amount of DNA found in a pollen
grain is not adequate for such analyses especially when genetic markers
are produced after restriction digest. Therefore, pollen DNA needs to be
increased by using primer extension pre-amplification (PEP) protocol. In
this research, we report on the use of a novel method for the
amplification of DNA from individual pollen grains and the subsequent
identification of AFLP (amplified fragment length polymorphism) markers.
Echinacea purpurea var. “Magnus” pollen genome was amplified with
modified PEP procedures to increase the sizes and numbers of AFLP
markers in subsequent AFLP analyses of these PEP products. The parental
genome was analyzed with AFLP procedure and 24 primer pairs were found
suitable. The PEP products from individual pollen grains were analyzed
by the selected 24 AFLP primer pairs. The parental AFLP markers are
being scored for their presence or absences in the pollen samples. This
is the first report on the production of AFLP markers from PEP products.
Currently, the linkage of parental AFLP markers is being analyzed to
construct a genetic map of Echinacea purpurea var. “Magnus”.
Currently, the reports on markers’ presence/absence have been imported
into JoinMap® 3.0 software (Plant Research International,
Wageningen, Netherlands) for subsequent genetic linkage analysis.
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