Hierarchical Population Structure Analysis of the Milkweed Beetle, Tetraopes Tetraophthalmus (Forster)
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Heredity 58 (1987) 193—201 The Genetical Society of Great Britain Received 7 April 1986 Hierarchical population structure analysis of the milkweed beetle, Tetraopes tetraophthalmus (Forster) David E. McCauley and Department of General Biology, Vanderbilt Walter F. Eanes University, Nashville, TN 37235, U.S.A., and Department of Ecology and Evolution, SUNY at Stony Brook, Stony Brook, NY 11794, U.S.A. The population structure of the milkweed beetle Tetraopes tetraophthalmus (Forster) is tied intimately to the biology of its one host plant, Asciepias syriaca. The patchy distribution of the host plant and the limited dispersal of the beetle combine to organise the herbivore into numerous very localised populations. An analysis of hierarchical patterns of spatial variation in allozyme frequencies by means of Wright's F-statistics reveals that in some parts of the beetle's range this population structure can result in moderate genetic differentiation (F =0.03—0.06)among beetle populations inhabiting milkweed patches separated by only a few kilometres. Substantial differentiation occurs across the range of the species (F =0.154).A review of similar studies shows the values of F calculated for T. tetraophthalmus to be among the highest found for a flying insect. It is argued that the local differentiation results, in part, from a reduction in effective population sizes associated with the ephemeral nature of milkweed patches and that this population structure is favourable for the operation of Wright's Shifting Balance mode of evolution. INTRODUCTION though rarely are both approaches applied together. From recent reviews that have compiled Thespatial organisation of local populations and F-statistic data for numerous animal species (i.e., concomitant patterns of gene flow are important Eanes and Koehn, 1978; Wright, 1978; Ayala, determinants of the potential for a species to 1982; Barrowclough, 1983) it can be seen that, not become genetically differentiated over its geo- surprisingly, there is a general agreement between graphic range. This is true whether the agent of the relative degree of population structure as differentiation is spatial variation in selection measured by F(S x T) and the perceived vagility pressures or the chance effects of genetic drift. The of the species in question. For example, among study of population structure and associated gene animal species, sedentary groups such as land flow in natural populations can take two snails and salamanders tend to have relatively high approaches, one ecological and one genetic (Slat- values of F(S x T), while most species of flying kin, 1985a). Ecological studies of the dispersal of birds and insects have rather low values. individuals or their gametes use various mark and A further refinement of the application of F- recapture schemes to attempt to measure gene flow statistics to the analysis of population structure directly, and can be used to make predictions about involves hierarchical analysis. Here the total geo- the population structure of a species. In contrast, graphic component of genetic variance is parti- surveys of geographic patterns of gene frequency tioned into subcomponents due to (a) variance variation that utilise electrophoretic markers can among closely spaced local populations likely to be used to make inferences about ecological par- be exchanging migrants and (b) variance among ameters such as average local population size and groups of populations defined on a greater geo- rates of migration (e.g., Larson eta!., 1984; Slatkin, graphic scale. In one such analysis Selander and 1985b). The genetic data is often summarised by Whitham (1983) argue that localised measures of the use of F-statistics as suggested by Wright F(S x T) are most likely to reflect the effects of (1978) for the analysis of population structure. population structure and gene flow. Hierarchical Both approaches have been applied to the study F-statistic analysis was developed by Wright as of the population structure in a diversity of species, part of his Shifting Balance theory of evolution in 194 D. E. McCAULEY AND W. F. EANES which some evolutionary transitions due to the Adults can be found on the foliage of the host joint effects of selection, drift, and migration are plant from June until August, depending on loca- possible in highly structured populations but can tion. Adult beetles were collected from individual not occur in large panmictic units (Wright, 1977; milkweed patches according to a hierarchical col- 1978). lecting scheme. Milkweed patches were defined as The milkweed beetle Tetraopes tetraophthalmus nearly discrete aggregations of milkweed stalks (Forster) (Coleoptera: Cerambycidae) has been occupying from about 025 to 10 ha. All beetles the focus of several ecological studies of dispersal collected from the same patch are considered to and gene flow (McCauley et a!., 1981; McCauley, belong to the same population or random breeding 1983). These studies have concluded that the unit as evidenced by dispersal studies (McCauley, limited dispersal of the insect, combined with the 1983). Some number of populations (4-19) were patchy, island-like distribution of its host plant, sampled from each of 5 localities called regions. the common milkweed Asciepias syriaca, organise Each region consisted of an area of less than 30 x the species into thousands of local populations 30 km. Regions were defined in southern New across its range in eastern North America. Thus, Hampshire (NH), southwest Virginia (VA), middle the species would appear to have the potential for Tennessee (TN), and northern Illinois (ILL). Col- a high degree of genetic differentiation across its lections were made in New Hampshire in 1983, geographic range and should exhibit genetic Illinois in 1984, and Tennessee in 1985. Most Vir- differences between populations separated by as ginia populations were sampled in 1984, though a little as a few kilometres. The one limited genetic few localities were sampled in 1983 or 1985. All study of this species (Eanes et aL, 1977) used data presented for a particular population are polymorphic allozyme markers to document allele derived from a single generation. Because of the frequency heterogeneity among seven collections univoltine nature of this species only two gener- of T tetraophthalmus taken from individual milk- ations separate the 1983 and 1985 collections. In weed patches located in Suffolk Co., N.Y. addition, data taken from collections made on Here, we report on gene frequency variation at Long Island, New York (NY) in 1975 and three polymorphic loci derived from a more sys- described by Eanes et aL (1977) were used to tematic and hierarchical collection of beetles taken constitute a fifth region. Fig. 1 presents these across a broad geographic area. The collections regional locations against the range of the host were designed to examine whether the degree of plant as described by Chemsak (1963). local population structure documented by Eanes Beetles were stored at —70°C until they could eta!. (1977) in New York is representative of other be prepared for starch gel electrophoresis. Eanes localised regions across the species' range, and et a!. (1977) present data for leucine aminopep- also to estimate the total geographic component tidase (LAP) and phosphoglucomutase (PGM), of genetic variance. Groups of milkweed patches the only polymorphic loci they found in their were sampled on an individual patch basis within original survey. In that survey amylase, glucose-6- four 30 x 30 km regions, the regions being spaced phosphate dehydrogenase, glutamate oxaloacetate across a major portion of the range of the species. transaminase, isocitrate dehydrogenase, lactate The data, when combined with that reported by dehydrogenase, peptidase and phosphogluco- Eanes et aL (1977) are used in a hierarchical F- isomerase were resolved but found to be monomor- statistic analysis that partitions the total geo- phic (Eanes, unpubi.). Similar techniques were graphic component of genetic variance into com- followed for the resolution of LAP and PGM in ponents due to among patch variance within the the more recent samples with the following excep- local regions and a component among regions. The tions. Both enzymes could be resolved with an results are discussed in the context of the known electrode buffer described as "salamander B" by ecology of the beetle and its host plant, as well as Werth (1985) and a gel buffer consisting of a 1:9 in the context of a survey of F-statistics calculated dilution of the electrode buffer. B-mercaptoethanol for other species of insects. was added to the grinding buffer to stabilise the PGM electromorphs. A more recent enzyme survey was able to resolve two additional enzymes, malic MATERIALSAND METHODS enzyme and hexokinase. Hexokinase (HK) proved to be polymorphic when resolved with a tris-citrate Tetraopestetraophthalmus is a univoltine beetle electrode buffer (pH 8.0). The discovery of the HK whose adults and larvae feed almost exclusively polymorphism came after the on set of the Virginia on the common milkweed, Asc!epias syriaca. collecting. As a consequence not all Virginia popu- POPULATION STRUCTURE OF MILKWEED BEETLE 195 Figure1 The location of five regions from which adult Tetraopes retraophthalmus were collected. Numbers refer to the number of individual milkweed patches sampled in each region. Stippled area defines the range of the host plant, Asciepias syriaca as described by Chemsak (1963). lations were assayed for HK. Samples from various RESULTS regions were occasionally run side by side to stan- dardise allelic nomenclature. Resultsare presented as allele frequencies and Gene frequency variation was quantified by sample sizes for LAP, PGM and HK in tables 1-3 hierarchical F-statistics as suggested for the study respectively. All populations sampled expressed of population structure by Wright (1978). See Weir two electrophoretic alleles for LAP (named F and and Cockerham (1984) and Slatkin (1985a) for S), though there was considerable variation in recent discussions of the application of F-statistics frequency from region to region.