PAG-I Plant Genome I Conference

Town & Country Conference Center, San Diego, CA, November, 1992.


PG-I: 52pg1

MOLECULAR MARKER AND QUANTITATIVE TRAIT MAPPING IN DOUGLAS-FIR

K.D. Jermstad 1, A.M. Reem 1, N.C. Wheeler 2 and D.B. Neale 1 1 Institute of Forest Genetics, Pacific Southwest Research Station, US Forest Service. 2 Wayerhaeuser Company


We are constructing a genetic linkage -map for Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco) based on restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) markers. The mapping population is a 3-generation outbred pedigree constructed by Weyerhaeuser Company. This cross was selected because we expect it is segregating for loci which determine the time of bud flush and bud set. We will attempt to map QTLs for these important traits once the genetic map is completed. Segregating RFLP loci are identified by hybridizing southern blots containing HindIII-restricted DNAs from the 4 grandparents and 2 Fl parents with three types of DNA probes: 1) Douglas-fir cDNA probes, 2) Douglas-fir genomic-DNA probes and 3) mapped RFLP probes from loblolly pine (Pinus taeda L.) (Devey et al, 1991. TAG vol. 83:238-242). Five of six Douglas-fir cDNAs hybridized and showed polymorphism in the parents. Probe pCabDF1 revealed a single locus with two alleles. The four remaining probes showed polymorphisms at each of two loci. A small proportion (32%) of Douglas-fir genomic-DNA probes were polymorphic. All of these are segregating in the cross and thus can be mapped. A large proportion (75%) of loblolly pine cDNA probes hybridized to Douglas-fir genomic DNA; 50.5% of the probes detected polymorphism and segregate in the F2. In summary, an estimate of 121 segregating loci have been identified thus far in parental screening. We are also mapping RAPD markers in the three-generation pedigree. Segregating RAPDs are identified based on six megagametophytes (lN) from each of the two F1s. Preliminary screening of six megagametophytes from each mother tree with 119 primers showed 66 segregating loci with an average of 0.55 loci per primer. Primers that show segregation are used on a sample of 23 megagametophytes from each parent. Segregating RAPDs will then be mapped in the F2 population. The F2s will be clonally propagated for future use in quantitative trait mapping studies and also to provide a lasting source of material for RFLP-RAPD mapping.


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