Plant Genome I Conference
Town & Country Conference Center, San Diego, CA, November, 1992.
PG-I: LINKAGE MAP OF SLASH PINE BASED ON MEGAGAMETIC DNA
LINKAGE MAP OF SLASH PINE BASED ON MEGAGAMETIC DNA.
J.P. van Buijtenen*, X. Kong*, E. Funkhouser*, W.L. Nance**,
C.D. Nelson**, L.S. Nelson**, G.N. Johnson**. * Texas
Agricultural Experiment Station, Texas A&M University, College
Station, TX 77843 ** Forestry Sciences Laboratory, USDA
Forest Service, Gulfport, MS 39505.
DNA from sixty-four megagametophytes of slash pine D4PC40
was extracted using a CTAB extraction procedure. The DNA was
used to develop a genetic map using the RAPD marker technique.
Since each megagametophyte is equivalent to a single haploid
gamete, this technique works extremely well, since the absence or
presence of each marker can be observed directly. The RAPD
reactions were performed using the automated facilities of the
USFS laboratory at Gulfport, MS, which made it possible to run
1536 PCR reactions per day. A total of 384 primers were screened
of which 94 were used for mapping purpose. Two primers were run
in duplicate for a total of 96. The PCR reactions resulted in
209 scoreable polymorphic bands of which 24 were eliminated,
because the segregation ratio deviated significantly from 1:1 (a
= .05). The remaining 185 loci formed 19 linkage groups of three
or more loci and four pairs. One locus was not linked to any
others. The resulting map covered 1776 cM or about 59 percent of
the estimated 3000 cM genome. This is less than expected and
probably caused by the tendency for many of the loci to cluster
into fairly tightly linked groups.
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