Molecular Phylogenies of Fig Wasps: Partial Cocladogenesis of Pollinators and Parasites Carlos Lopez-Vaamonde,* Jean Yves Rasplus,† George D
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Molecular Phylogenetics and Evolution Vol. 21, No. 1, October, pp. 55–71, 2001 doi:10.1006/mpev.2001.0993, available online at http://www.idealibrary.com on Molecular Phylogenies of Fig Wasps: Partial Cocladogenesis of Pollinators and Parasites Carlos Lopez-Vaamonde,* Jean Yves Rasplus,† George D. Weiblen,‡ and James M. Cook*,1 *Department of Biology & NERC Centre for Population Biology, Imperial College, Silwood Park, Ascot, Berkshire SL5 7PY, United Kingdom; †INRA, Centre de Biologie et de Gestion des Populations, Campus International de Baillarguet, CS 30016, 34988 Montferrier-sur-Lez, France; and ‡Department of Zoology, 203 Natural Science Building, Michigan State University, East Lansing, Michigan 48824 Received October 24, 2000; revised April 13, 2001 derpins the idea (Fahrenholz’s rule) that host and par- Figs (Ficus spp., Moraceae) and their pollinating asite phylogenies should be mirror images (Lyal, 1986). wasps form an obligate mutualism, which has long Although cospeciation is not a new idea, the opportu- been considered a classic case of coevolution and co- nity offered by molecular techniques to test it has led to speciation. Figs are also exploited by several clades of a burst of interest and activity in this area (Hafner et nonpollinating wasps, which are parasites of the mu- al., 1994; Hafner and Page, 1995; Moran et al., 1995; tualism and whose patterns of speciation have re- Page, 1996a; Farrell and Mitter, 1998; Peek et al., ceived little attention. We used data from nuclear and 1998; Clark et al., 2000). Molecular markers may per- mitochondrial DNA regions to estimate the phylog- mit the estimation of phylogenies for parasites and enies of 20 species of Pleistodontes pollinating wasps their hosts by use of genes that (unlike some morpho- and 16 species of Sycoscapter nonpollinating wasps associated with Ficus species in the section Mal- logical traits) are not coadapted to the species interac- vanthera. We compare the phylogenies of 15 matched tion. Further, if cospeciation is inferred, molecular Pleistodontes/Sycoscapter species pairs and show that data can be used to compare rates of evolution in host the level of cospeciation is significantly greater than and parasite lineages (Hafner and Nadler, 1990; that expected by chance. Our estimates of the maxi- Hafner et al., 1994; Hafner and Page, 1995; Moran et mum level of cospeciation (50 to 64% of nodes) are very al., 1995; Page, 1996a; Paterson et al., 2000). similar to those obtained in other recent studies of To date few cospeciation studies have produced mo- coevolved parasitic and mutualistic associations. lecular phylogenies for both sets of taxa and insights However, we also show that there is not perfect con- are limited to a few systems. One interaction in par- gruence of pollinator and parasite phylogenies (for ticular—that between pocket gophers (geomyid ro- any substantial clade) and argue that host plant dents) and their chewing lice—has dominated both switching is likely to be less constrained for Sycoscap- methodological (Page, 1990, 1993a,b, 1994) and biolog- ter parasites than for Pleistodontes pollinators. There ical (Nadler et al., 1990; Demastes et al., 1998) debates is perfect correspondence between two terminal about cospeciation. The pioneering studies based on clades of two sister species in the respective phylog- enies, and rates of molecular evolution in these pairs allozyme (Hafner and Nadler, 1988) and DNA (Hafner et al., 1994) data provided evidence for a high degree of are similar. © 2001 Academic Press Key Words: Agaonidae; Pleistodontes; Sycoscapter; cospeciation in the gopher–louse assemblage. How- Ficus; fig wasps; Malvanthera; coevolution; cospecia- ever, both studies are based on small phylogenies and tion; 28S; ITS2; cytochrome b. may not be typical of mammals and lice, or other ecto- parasites, in general (Taylor and Purvis, 2001). A re- cent study of pinworms and primates (Hugo, 1999) INTRODUCTION suggests that at least some endoparasites have under- gone considerable cospeciation with their hosts. More The speciation of parasites is often thought to be predictably, high cospeciation levels have been in- driven by the speciation of their hosts, such that, when ferred between invertebrates and endosymbiotic algae an ancestral host species splits into two daughter spe- or bacteria that contribute significantly to host metab- cies, its parasite also speciates. This assumption un- olism (Peek et al., 1998; Funk et al., 2000). Finally, whereas endogenous retroviruses generally track host 1 To whom correspondence and reprint requests should be ad- phylogeny, occasional host shifts do occur, even be- dressed. Fax (ϩ44) 207 594 2339. E-mail: [email protected]. tween mammals and birds (Martin et al., 1999). 55 1055-7903/01 $35.00 Copyright © 2001 by Academic Press All rights of reproduction in any form reserved. 56 LOPEZ-VAAMONDE ET AL. Lice, endosymbiotic bacteria, retroviruses, and fig logenetic studies of Panamanian pollinating and asso- wasps are not expected to have the same levels of ciated nonpollinating wasps (Machado et al., 1996) and cospeciation as they encompass a range of interactions, of figs and pollinators in general (Herre et al., 1996), with different transmission mechanisms. However, the were consistent with high levels of cospeciation. same methods can be used to analyze these different Australasia harbors great Ficus diversity with at systems, and, as case studies accumulate, it should least 130 species in New Guinea alone (Wiebes, 1982a). become possible to identify general features of interac- The section Malvanthera Corner comprises 22 species, tions (e.g., transmission mode) that promote or limit 14 of which are endemic to Australia and 8 to New cospeciation (Clayton et al., 2001). Guinea (Corner, 1965). There are two distinct groups: a Phytophagous insects can be regarded as parasites of lineage of stranglers and free-standing trees found in their host plants (Price, 1977) and may be generalists rainforests of eastern Australia and New Guinea and a or specialists. Some specialists attack just a single clade of lithophytic shrubs and trees found in more arid plant species, whereas many others feed on a few parts of Australia. Malvanthera species are all polli- closely related species. Interaction and coevolution nated by wasps in the genus Pleistodontes and are have probably been important during the radiations of exploited by a range of nonpollinating wasps (Boucek, plant lineages and their insect herbivores, with plant 1988; Cook and Power, 1996), of which Sycoscapter chemistry playing a key role (Ehrlich and Raven 1964); species are the most prevalent. Sycoscapter larvae are however, strict sense cospeciation seems generally un- thought to be either true parasitoids or phytophagous likely (Farrell and Mitter, 1990, 1998). More specific kleptoparasites of the pollinators (Boucek, 1993; interactions occur between certain groups of plants Compton et al., 1994). Although both are referred to as and their pollinators and perhaps the two best-known fig wasps, Pleistodontes and Sycoscapter are distantly cases are figs and fig wasps (Herre, 1996; Anstett et al., related genera from different families of chalcid wasps 1997) and yuccas and yucca moths (Pellmyr and Lee- (Rasplus et al., 1998). bens-Mack, 1999). Both of these systems are mutual- In this paper, we use nuclear and mitochondrial isms but also involve conflicts of interest, principally DNA sequences to infer phylogenies for 20 Pleistodon- over whether female flowers nourish developing seeds tes and 16 Sycoscapter species from Malvanthera figs. or insect larvae. We test two null hypotheses of host–parasite associa- Figs (Ficus spp.) and their pollinating wasps (family tions (Fig. 1; see Huelsenbeck et al., 1997, 2000; Clark Agaonidae sensu Rasplus et al., 1998) provide a classic et al., 2000). H1 is that host and parasite phylogenies example of an obligate mutualism. Most Ficus species have been produced by independent random-branching have their own unique pollinating wasp species, whose processes. Acceptance of H1 infers that cospeciation larvae feed on the fig seeds. Each partner relies on the has been rare or absent, whereas rejection of H1 sug- other to complete its life cycle and many morphological gests that cospeciation (or another process imparting characters of the partners are thought to be coadapted congruence) has occurred. In the latter case we can (Herre, 1989; Van Noort and Compton, 1996). The bi- then test H2 that Pleistodontes and Sycoscapter data ology of figs and their pollinators has led to a general sets are consistent with the same underlying tree to- acceptance of the idea that they have coevolved and pology (Peek et al., 1998; Clark et al., 2000; Huelsen- cospeciated extensively. However, rigorous testing of beck et al., 2000, 2001; Johnson et al., 2001). Accep- this idea has been hindered by the fact that the polli- tance of H2 would provide stronger evidence for strict nator taxonomy has developed under the influence of cospeciation and permit a test of H3 that the rates of existing fig taxonomy and a strong cospeciation as- molecular evolution and speciation times are the same sumption (Ramirez, 1974, 1977; Wiebes, 1982b). in the two clades. Fig-based invertebrate communities lend them- selves to cospeciation studies because, in addition to pollinating wasps, there are diverse nonpollinating MATERIAL AND METHODS wasps, mites, drosophilids (Harry et al., 1996), and Taxon Sampling parasitic nematodes (Poinar and Herre, 1991). Many (perhaps