Weeping piñon (Pinus pinceana) has a restricted and fragmented range, trees are widely scattered within populations, and reproduction is limited. Nevertheless, genetic diversity was high; based on 27 isozyme loci in 18 enzyme systems, unbiased expected heterozygosity averaged 0.174. Differentiation also was high (F(ST) = 0.152), reflecting isolation between southern, central, and northern fragments of the range. Among populations in the northern fragment, F(ST) was only 0.056, and the number of migrants per generation (Nm) was 4.21, which should preclude fixation. Nm between central and southern populations or between them and populations in the northern fragment was lower, 0.99-1.66, indicating a degree of genetic isolation. Multilocus outcrossing rates (t(m)) ranged from 0.836 in the south to 0.897 in the north. Therefore, selfing is low but statistically significant. The equilibrium inbreeding coefficient (F(e)) calculated from t(m) was in good agreement with observed inbreeding coefficients, suggesting that weeping piñon may be near equilibrium with respect to inbreeding and selection against selfed trees. Weeping piñon was variable at all loci polymorphic in maxipiñon (Pinus maximartinezii) and, therefore, qualifies as a possible progenitor of maxipiñon. Because of the high level of diversity, reasonable levels of gene flow within the northern fragment of weeping piñon's range, high rates of outcrossing, and, perhaps, only weak selection against inbred trees, protection in reserves would be a viable option for conservation.
Martínez spruce (Picea martínezii T.F. Patterson) is a conifer currently passing through a bottleneck, reduced to a few relict populations totaling less than 800 trees. We used isozyme markers to analyze the mating system and survey the level of genic diversity in two populations. The mating system was characterized by a high frequency of selfing. The multilocus outcrossing rates (tm) and 95% confidence intervals were only 0.399 (0.197 < tm < 0.601) for the smallest population and 0.589 (0.475 < tm < 0.703) or 0.685 (0.465 < tm < 0.905), depending on year, for the largest. These are among the lowest rates of outcrossing observed in conifers. The fixation indices for the two populations were -0.058 and 0.121, less than expected for such high levels of selfing. Expected heterozygosity, unbiased He, based on 22 loci in 13 enzyme systems, was 0.121 and 0.101 in the two populations. The proportion of the total genic diversity between populations, FST, was 2.4%. Nm, the number of migrants per generation, was about 1.00 or 10.17, depending on the method of estimation. The time since the two populations were isolated was estimated from Nei's genetic distance as only 150 to 15 000 years, which is consistent with a hypothesis of population collapse during late Pleistocene or Holocene warming. We discuss the implications for conservation.
Maxipiñon (Pinus maximartinezii Rzedowski), which is confined to a single population of approximately 2000 to 2500 mature trees, covers about 400 ha in southern Zacatecas, Mexico. Genetic diversity measured by expected heterozygosity was 0.122, which is moderate for pines. However, percentage polymorphic loci was low, 30.3%. The fixation index (F) of 0.081 indicated only slight heterozygote deficiency. Mating system analysis indicated a significant but low level of selling; the multilocus outcrossing rate, t , was 0.816. The mean of single locus estimates, t , was smaller (0.761), perhaps suggesting mating among relatives, although the difference between t and t was not statistically significant. The most striking features of maxipiñon's genetic structure were that no polymorphic locus had more than two alleles and most alleles at polymorphic loci were at intermediate frequencies. This is in contrast to other pines, which often have three to five or more alleles at some loci and in which the distribution of allele frequencies is U-shaped, most alleles being present at frequencies less than 10% or greater than 90%. A population with only two alleles per locus and at intermediate frequencies could occur if the population had been reduced to an extreme bottleneck and then expanded rapidly before random drift modified allele frequencies. A novel origin from a hybridization event would also explain the results. Significant gametic disequilibrium was detected at several pairs of loci in both maternal and paternal gametes. The presence of disequilibrium is in agreement with an origin from an extreme bottleneck, perhaps even a single seed. Furthermore, it demands that the event be relatively recent. The number of generations, as calculated from the observed mean disequilibrium, suggested that maxipiñon derived from an extreme bottleneck four to five generations ago, which is less than 1000 years in this species.
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