HETEROZYGOSITY AND DEVELOPMENTAL STABILITY UNDER INBREEDING AND CROSSBREEDING IN PINUS ATTENUATA

The relationship between stability of annual trunk growth and heterozygosity at 24 polymorphic isozyme loci was studied in 10-year-old trees of knobcone pine (Pinus attenuata Lemm.) that were the products of contrasting systems of mating, self-, and interpopulation cross-pollination. Heterozygosity and variability of trunk growth were strongly related only when inbreds and crossbreds were compared; the crossbreds showed greater residual variability on an absolute scale, and greater responsiveness to climate on both absolute and relative scales. Within the inbreds there was no evidence of a relationship between heterozygosity and variability. Within the crossbreds, only one trait, a measure of relative trunk growth rate, showed a relationship with heterozygosity, and indicated greater variability of the more heterozygous trees. These results, and others in the literature, suggest that the relationship of heterozygosity to homeostasis for fitness components is neither simple nor monotonic; it varies between scales of measurement, genetic backgrounds, and environments.     

Testing the effects of heterozygosity on growth rate plasticity in the seaweed Gracilaria chilensis (Rhodophyta)

Heterozygosity has been positively associated with fitness and population survival. However, the relationship between heterozygosity and adaptive phenotypic plasticity (i.e., plasticity which results in fitness homeostasis or improvement in changing environments) is unclear and has been poorly explored in seaweeds. In this study, we explored this relationship in the clonal red seaweed, Gracilaria chilensis by conducting three growth rate plasticity experiments under contrasting salinity conditions and by measuring heterozygosity with five microsatellite DNA markers. Firstly, we compared growth rate plasticity between the haploid and diploid phases. Secondly, we compared growth rate plasticity between diploids with different numbers of heterozygous loci. Finally, we compared growth rate plasticity between diploid plants from two populations that are expected to exhibit significant differences in heterozygosity. We found that, (i) diploids displayed a higher growth rate and lower growth rate plasticity than haploids, (ii) diploids with a higher number of heterozygous loci displayed lower growth rate plasticity than those exhibiting less heterozygosity, and (iii) diploid sporophytes from the population with higher heterozygosity displayed lower growth rate plasticity than those with lower heterozygosity. Accordingly, this study suggests that heterozygosity is inversely related to growth rate plasticity in G. chilensis. However, better genetic tools in seaweeds are required for a more definitive conclusion on the relationship between heterozygosity and phenotypic plasticity.     

OBSERVATIONS ON THE ECOLOGY AND EVOLUTION OF QUAKING ASPEN,POPULUS TREMULOIDES,IN THE COLORADO FRONT RANGE

Three protein polymorphisms and annual width increments measured from cores were used to study the ecology and evolution of quaking aspen, Populus tremuloides Michx., in the Front Range of Colorado. Correlations among elevation, age and diameter of the largest standing ramet, and sex and mean growth rate of 106 clones are reported and discussed. The variance of growth rate was apportioned within and among ramets of a clone; the majority of the variance is environmental, with broad heritability estimated as less than or equal to .32. Analyses of gene and genotypic frequencies of polymorphic proteins revealed little differentiation with elevation, but substantial differences between the sexes. For one of three pairwise tests, the genotypes are not distributed independently of one another; linkage disequilibrium between a peroxidase locus and phosphohexose isomerase is significantly different from zero. Multiple regression revealed a positive correlation between mean growth rate and degree of heterozygosity.     

Analysis of the relationship between allozyme heterozygosity and fitness in the rare Gentiana pneumonanthe L.

Especially for rare species occurring in small populations, which are prone to loss of genetic variation and inbreeding, detailed knowledge of the relationship between heterozygosity and fitness is generally lacking. After reporting on allozyme variation and fitness in relation to population size in the rare plant Gentiana pneumonanthe, we present a more detailed analysis of the association between heterozygosity and individual fitness. The aim of this study was to test whether increased fitness of more heterozygous individuals is explained best by the ‘inbreeding’ hypothesis or by the ‘overdominance’ hypothesis. Individual fitness was measured during 8 months of growth in the greenhouse as the performance for six life-history parameters. PCA reduced these parameters to four main Fitness Components. Individual heterozygosity was scored for seven polymorphic allozyme loci. For some of these loci (e.g. Aat3, Pgm1 and 6Pgdh2) heterozygotes showed a significantly higher relative fitness than homozygotes. To test the inbreeding model, regression analyses were performed between each Fitness Component and the number of heterozygous loci per individual. Multiple regressions with the adaptive distance of five loci as independent variables were used to test the overdominance model. Only the inbreeding model was a statistically significant explanation for the relationship between heterozygosity and fitness in G. pneumonanthe. The number of heterozygous loci was significantly negatively correlated with the coefficients of variation of three of the six initially measured fitness parameters. This suggests a lower developmental stability among more homozygous plants and may explain the higher phenotypic variation in small populations of the species observed earlier. The importance of the results for conservation biology is discussed.     

Allozyme variation in populations, full-sib families and selfed lines in Betula pendula Roth

Changes in genetic variability in populations (stand origins), full-sib (FS) families and three generations of selfed lines of Betula pendula were observed based on 15 allozyme loci. Growth vigour, measured as stem volume, and its relationship with heterozygosity was studied to determine the effect of inbreeding. Pooled FS families showed a higher percentage of polymorphic loci (P) and allelic numbers per locus (A) than those of natural populations, but no difference in heterozygosity. There was no difference in allozyme variability between fast-and slow-growing family groups, and heterozygosity was not correlated with stem volume among FS families. Allozyme variability was significantly decreased in advancing generations of selfing, and the further the selfing generation, the lower the heterozygosity and the slower the growth. Observed heterozygosity after advancing generations of inbreeding was increasingly higher than expected, indicating overdominance effects or, alternatively, selection against deleterious homozygotes.     

Multiple-Locus Heterozygosity and the Physiological Energetics of Growth in the Coot Clam, MULINIA LATERALIS, from a Natural Population

The relationship between individual energy budgets and multiple-locus heterozygosity at six polymorphic enzyme loci was examined in Mulinia lateralis. Energy budgets were determined by measuring growth rates, rates of oxygen consumption, ammonia excretion and clearance rates. Enzyme genotypes were determined using starch gel electrophoresis. Growth rate and net growth efficiency (the ratio of energy available for growth to total energy absorbed) increased with individual heterozygosity. The positive relationship between observed growth and multiple-locus heterozygosity was associated with a negative relationship between routine metabolic costs and increasing heterozygosity. Reduction in routine metabolic costs explained 60% of the observed increased growth of more heterozygous individuals. When routine metabolic costs were standardized for differences in feeding rates, these standard metabolic costs explained 97% of the differences in growth rate. Lower standard metabolic costs, associated with increasing heterozygosity, have been proposed as a physiological mechanism for the relationship between multiple-locus heterozygosity and growth rate that has been reported for a variety of organisms, ranging in diversity from aspens to humans. This study demonstrates that reduction of standard metabolic costs, at least in clams, accounts for virtually all of the differences in growth rate among individuals of differing heterozygosity.     

Does peripheral dislodgement contribute to heterozygote deficiencies in blue mussels?

Differential loss of heterozygous individuals that move to the periphery of mussel aggregations, where they are at greater risk for dislodgement, has been proposed as an explanation for observed heterozygote deficiencies in blue mussels. To test the dislodgement hypothesis, correlations between heterozygosity and mussel motility, as well as characteristics of byssogenesis and byssal thread attachment strengths, were determined in a wild and a farmed population of blue mussels (Mytilus edulis) from New Hampshire, USA. Although both populations exhibited a heterozygote deficit as measured by three microsatellite loci, no relationship was found between heterozygosity and increased motility in either population. Similarly, no relationship was found between heterozygosity and byssogenesis or attachment strength. Hence, differential dislodgement is highly unlikely as a possible contributor to the loss of heterozygous individuals.     

FLUCTUATING ASYMMETRY IN CENTRAL AND MARGINAL POPULATIONS OF LYCHNIS VISCARIA IN RELATION TO GENETIC AND ENVIRONMENTAL FACTORS

Developmental instability in the form of increased fluctuating asymmetry can be caused by either genetic or environmental stress. Because extinctions can be attributed broadly to these factors, fluctuating asymmetry may provide a sensitive tool for detecting such stresses. We studied the level of fluctuating asymmetry of flowers of a perennial outcrossing plant species, Lychnis viscaria, both in natural and common-garden populations. The degree of flower asymmetry was higher in small, isolated, and marginal populations of the species range. These marginal populations also were the most homozygous. In the core area of the species' range, flowers were more symmetrical The level of asymmetry was correlated with both population size and heterozygosity. However, a partial correlation analysis revealed that when the impact of population size was controlled for, there was a negative relationship between fluctuating asymmetry and heterozygosity, whereas when controlling for heterozygosity, no relationship between population size and fluctuating asymmetry was found. This indicates that genetic consequences of small population size probably underlie the relationship between the level of asymmetry and population size. Results from a transplantation experiment showed that individuals subjected to a higher environmental stress had an increased level of asymmetry compared to control plants. In the common-garden conditions the level of fluctuating asymmetry did not differ between the central and marginal populations. This suggests that presumably both genetic and environmental factors affected to the higher level of asymmetry among marginal populations compared to central ones. In all we conclude that even though fluctuating asymmetry seems to be a sensitive tool for detecting stresses, results from studies focusing on only one factor should be interpreted with caution.     

The heterozygosity for the biochemical and immunological markers of genes and the variability of morphophysiological traits in man

The relationship between heterozygosity of 9 biochemical and 5 blood group loci and variability of body length and age of menarche were studied in 467 women and 336 men from Moscow population. High and low levels of individual heterozygosity were shown in men to be associated with the maximal values of the coefficient of variation (CV) of body length, while in women strong positive correlation between the CV of body length and individual heterozygosity was demonstrated. The highest level of heterozygosity was revealed in medium-height men and low-height women. Highly heterozygous women were characterized either by early or by late age of menarche; early onset of menarche causes growth retardation. Positive correlation between the CV of body length and heterozygosity in women is due to the accumulation of low-height individuals having early age of menarche. The results are discussed in terms of Lerner's concept of genetic homeostasis. It is concluded that an average level of heterozygosity is optimal for a population.     

Heterozygosity–fitness correlations in a wild mammal population: accounting for parental and environmental effects

HFCs (heterozygosity–fitness correlations) measure the direct relationship between an individual's genetic diversity and fitness. The effects of parental heterozygosity and the environment on HFCs are currently under‐researched. We investigated these in a high‐density U.K. population of European badgers (Meles meles), using a multimodel capture–mark–recapture framework and 35 microsatellite loci. We detected interannual variation in first‐year, but not adult, survival probability. Adult females had higher annual survival probabilities than adult males. Cubs with more heterozygous fathers had higher first‐year survival, but only in wetter summers; there was no relationship with individual or maternal heterozygosity. Moist soil conditions enhance badger food supply (earthworms), improving survival. In dryer years, higher indiscriminate mortality rates appear to mask differential heterozygosity‐related survival effects. This paternal interaction was significant in the most supported model; however, the model‐averaged estimate had a relative importance of 0.50 and overlapped zero slightly. First‐year survival probabilities were not correlated with the inbreeding coefficient (f); however, small sample sizes limited the power to detect inbreeding depression. Correlations between individual heterozygosity and inbreeding were weak, in line with published meta‐analyses showing that HFCs tend to be weak. We found support for general rather than local heterozygosity effects on first‐year survival probability, and g2 indicated that our markers had power to detect inbreeding. We emphasize the importance of assessing how environmental stressors can influence the magnitude and direction of HFCs and of considering how parental genetic diversity can affect fitness‐related traits, which could play an important role in the evolution of mate choice.     

The strength of the association between heterozygosity and probability of interannual local recruitment increases with environmental harshness in blue tits

The extent of inbreeding depression and the magnitude of heterozygosity–fitness correlations (HFC) have been suggested to depend on the environmental context in which they are assayed, but little evidence is available for wild populations. We combine extensive molecular and capture–mark–recapture data from a blue tit (Cyanistes caeruleus) population to (1) analyze the relationship between heterozygosity and probability of interannual adult local recruitment and (2) test whether environmental stress imposed by physiologically suboptimal temperatures and rainfall influence the magnitude of HFC. To address these questions, we used two different arrays of microsatellite markers: 14 loci classified as neutral and 12 loci classified as putatively functional. We found significant relationships between heterozygosity and probability of interannual local recruitment that were most likely explained by variation in genomewide heterozygosity. The strength of the association between heterozygosity and probability of interannual local recruitment was positively associated with annual accumulated precipitation. Annual mean heterozygosity increased over time, which may have resulted from an overall positive selection on heterozygosity over the course of the study period. Finally, neutral and putatively functional loci showed similar trends, but the former had stronger effect sizes and seemed to better reflect genomewide heterozygosity. Overall, our results show that HFC can be context dependent, emphasizing the need to consider the role of environmental heterogeneity as a key factor when exploring the consequences of individual genetic diversity on fitness in natural populations.     

Heterosis × nutrition interaction in Drosophila melanogaster

Summary The relationship between heterozygosity and the expression of heterosis at two different nutrition levels was investigated using Drosophila melanogaster. Average daily egg production and egg hatchability were measured in two parental strains and in F₁, F₂ and reciprocal backcross generations. Heterosis was more pronounced in the poor nutritional conditions. Two electrophoretic markers used to estimate the level of heterozygosity in F₂ and backcrosses revealed an excess of heterozygous genotypes. Quantitative genetic effects (an additive line effect and individual and maternal heterosis) were estimated for both traits in the two environments. Although this model gave a reasonable fit in most cases, some epistatic interaction would have to be invoked in order to explain fully the results.     

Multilocus heterozygosity and inbreeding in the Siberian jay

Because of its common negative association with fitness, inbreeding is a major concern in conservation biology. Traditionally it has been measured as individual inbreeding coefficient calculated from the pedigree, but recently multilocus heterozygosity estimates have become commonly used as proxies. However, theoretical and simulation studies have cast doubt on the validity of these surrogates especially when they are based on only a few molecular markers. Yet, empirical studies reporting the correlation between multilocus heterozygosity and inbreeding coefficient are rare. We studied this relationship in a wild Siberian jay (Perisoreus infaustus) population subject to a long-term field study over 30 years. The correlations between inbreeding coefficient and the employed heterozygosity measures—standardized heterozygosity and internal relatedness—based on 21 microsatellite loci were weak. These results together with results from theoretical and simulation studies caution against use of multilocus heterozygosity estimates to study inbreeding in natural populations.     

GENETIC VARIATION IN BROMUS TECTORUM (POACEAE): POPULATION DIFFERENTIATION IN ITS NORTH AMERICAN RANGE

Allelic variation in seedlings from 60 North American populations of the alien annual grass Bromus tectorum was determined at 25 loci using starch gel electrophoresis. Populations were collected from four regions; east of the Rocky Mountains, Nevada and California, the Intermountain West, and British Columbia. Compared to other diploid seed plants, genetic variation within these populations of B. tectorum is low: 4.60% of loci are polymorphic per population, with an average of 1.05 alleles per locus and a mean expected heterozygosity of 0.012. Although 2,141 individuals were analyzed, no heterozygous individuals were detected, and consequently, mean observed heterozygosity is 0.000. Extensive deviations from Hardy-Weinberg expectations were observed at every polymorphic locus due to heterozygote deficiencies. The mean genetic identity (Nei's I) between population pairs was 0.980 and indicates a high level of overall genetic similarity among populations. The among-population component of the total gene diversity is high (G_ST ⁼ 0.478), indicating substantial genetic differentiation among populations. These results are consistent with previous reports for highly self-pollinating plants of low genetic variation and substantial genetic differentiation among populations. Despite the lack of genetic variation as measured by enzyme electrophoresis, this weedy grass has become exceedingly abundant in a diverse array of arid environments throughout much of western North America, perhaps due to phenotypic plasticity.     

Influence of mother's heterozygosity on the variation of anthropometric traits in newborns]

The relationship between mother's heterozygosity as revealed by 14 polymorphic loci and variation of the set of anthropometric traits were studied in normal singleton newborns (174 boys, 127 girls). Statistically significant negative correlation between mother's heterozygosity and the index of fluctuating asymmetry was found in girls. Variances of four traits--body weight, body length, circumference of head and breast measured by first principal component, and mean number of minor deviations from development (stigma) are shown to be minimal in the children born by mothers with the average level of heterozygosity. The groups of newborns with different level of mother's heterozygosity are characterized by different combinations of first principal component, stigma, and the index of fluctuating asymmetry. It is concluded from the whole set of data that children born by mothers with the average level of heterozygosity have the highest level of viability.     

Variation at isozyme loci inTriticeae

An electrophoretic comparison of variation at 16 presumptive isozyme gene loci was performed for 17 species from the tribeTriticeae. Included in the analysis were annuals and perennials, and self- and cross-pollinating species, representing the H, I, P, N, R, V, S, E, J, J₁J₂, A, B, and D genomes. Perennial species were found to contain a significantly (marginally, at the 5% level) higher proportion of polymorphic loci and level of heterozygosity, than annual species. There were no significant differences between self- and crosspollinating species. Across all species, mean heterozygosity levels ranged from 0–0.225 and the % polymorphic loci from 6.3–56.3%. Genetic distance estimates varied from 0.08–0.39 for congeneric species. Relationships were deduced between the 17 species using phenetic and cladistic analyses and compared with relationships inferred from other parameters such as morphology and nucleotide sequence data. In general, the trees derived from the various relationships were concordant; the evolutionary basis for minor discrepancies between trees is also discussed.     

Assessment of identity disequilibrium and its relation to empirical heterozygosity fitness correlations: a meta‐analysis

Heterozygosity fitness correlations (HFCs) have frequently been used to detect inbreeding depression, under the assumption that genome‐wide heterozygosity is a good proxy for inbreeding. However, meta‐analyses of the association between fitness measures and individual heterozygosity have shown that often either no correlations are observed or the effect sizes are small. One of the reasons for this may be the absence of variance in inbreeding, a requisite for generating general‐effect HFCs. Recent work has highlighted identity disequilibrium (ID) as a measure that may capture variance in the level of inbreeding within a population; however, no thorough assessment of ID in natural populations has been conducted. In this meta‐analysis, we assess the magnitude of ID (as measured by the g₂ statistic) from 50 previously published HFC studies and its relationship to the observed effect sizes of those studies. We then assess how much power the studies had to detect general‐effect HFCs, and the number of markers that would have been needed to generate a high expected correlation (r² = 0.9) between observed heterozygosity and inbreeding. Across the majority of studies, g₂ values were not significantly different than zero. Despite this, we found that the magnitude of g₂ was associated with the average effect sizes observed in a population, even when point estimates were nonsignificant. These low values of g₂ translated into low expected correlations between heterozygosity and inbreeding and suggest that many more markers than typically used are needed to robustly detect HFCs.     

ASSOCIATION BETWEEN QUANTITATIVE TRAITS AND ALLOZYME HETEROZYGOSITY IN A TETRASOMIC SPECIES: DACTYLIS GLOMERATA

Tetraploid individuals of orchardgrass (Dactylis glomerata L. subsp. hispanica Roth.) sampled from a natural population were used to evaluate the correlation between both single and multilocus heterozygosity at 7 enzyme loci, and several quantitative traits (plant size at time of collection, leaf weight, and panicle number in experimental trials). Four hundred and forty-eight plants were studied at the 7 loci and 288 of these individuals were scored for an additional eighth locus. Five genotype classes (monogenic, simplex, and duplex digenic, trigenic, and tetragenic) were distinguished according to their heterozygosity level. Multilocus heterozygosity showed a significant positive correlation with both leaf and panicle yield in experimental conditions, but not with original plant size, which was found to be markedly influenced by environmental microheterogeneity. Multilocus heterozygosity, estimated from both the number of heterozygous loci and the number of distinct alleles per locus, had a significant influence on plant performance. Individual locus effects were positive and significant at two loci (GOT1 and PX1). Panicle number increased regularly with heterozygosity level (from monogenics to tetragenics) at the GOT1 locus, as did leaf weight and panicle number at the PX1 locus. Such variation would be predicted by overdominance at these loci or at linked loci. Significant relationships between leaf yield and heterozygosity level at the GOT1 locus distinguished the homozygotes from the heterozygotes (of any class) and was thus more consistent with inbreeding effects. No significant differences were observed among the five genotype classes for any quantitative trait at the six remaining loci. At both the GOT1 and PX1 loci, heterozygosity had a significant independent effect on leaf weight and panicle number even when the correlation between these traits was removed by analysis of covariance.     

Heterozygosity at a single locus explains a large proportion of variation in two fitness‐related traits in great tits: a general or a local effect?

In natural populations, mating between relatives can have important fitness consequences due to the negative effects of reduced heterozygosity. Parental level of inbreeding or heterozygosity has been also found to influence the performance of offspring, via direct and indirect parental effects that are independent of the progeny own level of genetic diversity. In this study, we first analysed the effects of parental heterozygosity and relatedness (i.e. an estimate of offspring genetic diversity) on four traits related to offspring viability in great tits (Parus major) using 15 microsatellite markers. Second, we tested whether significant heterozygosity–fitness correlations (HFCs) were due to ‘local’ (i.e. linkage to genes influencing fitness) and/or ‘general’ (genome‐wide heterozygosity) effects. We found a significant negative relationship between parental genetic relatedness and hatching success, and maternal heterozygosity was positively associated with offspring body size. The characteristics of the studied populations (recent admixture, polygynous matings) together with the fact that we found evidence for identity disequilibrium across our set of neutral markers suggest that HFCs may have resulted from genome‐wide inbreeding depression. However, one locus (Ase18) had disproportionately large effects on the observed HFCs: heterozygosity at this locus had significant positive effects on hatching success and offspring size. It suggests that this marker may lie near to a functional locus under selection (i.e. a local effect) or, alternatively, heterozygosity at this locus might be correlated to heterozygosity across the genome due to the extensive ID found in our populations (i.e. a general effect). Collectively, our results lend support to both the general and local effect hypotheses and reinforce the view that HFCs lie on a continuum from inbreeding depression to those strictly due to linkage between marker loci and genes under selection.     

Association between host's genetic diversity and parasite burden in damselflies

Recent research indicates that low genetic variation in individuals can increase susceptibility to parasite infection, yet evidence from natural invertebrate populations remains scarce. Here, we studied the relationship between genetic heterozygosity, measured as AFLP‐based inbreeding coefficient f_AFLP, and gregarine parasite burden from eleven damselfly, Calopteryx splendens, populations. We found that in the studied populations, 5–92% of males were parasitized by endoparasitic gregarines (Apicomplexa: Actinocephalidae). Number of parasites ranged from none to 47 parasites per male, and parasites were highly aggregated in a few hosts. Mean individual f_AFLP did not differ between populations. Moreover, we found a positive association between individual's inbreeding coefficient and parasite burden. In other words, the more homozygous the individual, the more parasites it harbours. Thus, parasites are likely to pose strong selection pressure against inbreeding and homozygosity. Our results support the heterozygosity‐fitness correlation hypothesis, which suggests the importance of heterozygosity for an individual's pathogen resistance.