Group I Introns and Independent Marker Genes Reveal the Existence of Three Sexual but Reproductively Isolated Biospecies in Trichia Varia (Myxomycetes)
Total Page：16
File Type：pdf, Size：1020Kb
Org Divers Evol (2015) 15:631–650 DOI 10.1007/s13127-015-0230-x ORIGINAL ARTICLE Sex or no sex? Group I introns and independent marker genes reveal the existence of three sexual but reproductively isolated biospecies in Trichia varia (Myxomycetes) Yun Feng 1 & Martin Schnittler1 Received: 17 April 2015 /Accepted: 28 July 2015 /Published online: 11 August 2015 # Gesellschaft für Biologische Systematik 2015 Abstract Plasmodial slime molds are members of the class pattern of three cryptic biological species that reproduce pre- Amoebozoa forming elaborate fruit bodies releasing airborne dominantly sexual but are reproductively isolated. The spores. Two species concepts have been developed indepen- pattern of group I introns and inserted homing endonucle- dently: a morphological relying on fruit body characters, and a ase genes mounts evidence that the Goddard-Burt intron life biological relying on crossing studies of a few cultivable spe- cycle model applies to naturally occurring myxomycete cies. In an attempt to reconcile both concepts, we obtained for populations. 198 specimens of the common species Trichia varia partial sequences of three independent markers (nuclear small- Keywords Extrachromosomal nuclear ribosomal DNA . subunit (SSU) ribosomal RNA gene, extrachromosomal; Group I intron . Homing endonuclease . Plasmodial slime elongation factor 1 alpha gene, chromosomal; cytochrome mold . Reproductive system . Speciation oxidase subunit 1 gene, mitochondrial). The resulting phylogeny revealed 21 three-marker genotypes clustering into three groups. Combinations of the single-marker genotypes Introduction occurred exclusively within these groups, called 1, 2a, and 2b. To examine the suitability of group I introns to monitor Plasmodial slime molds (Myxomycetes, Myxogastria), some- speciation events, complete SSU sequences were generated times called “sexual amoebae,” are the most species-rich for 66 specimens, which revealed six positions that can carry monophyletic group of Amoebozoa (Adl et al. 2012; Lahr group I introns. For each of the groups 1 and 2a, five of these et al. 2011a; Smirnov et al. 2011). These free-living amoebae positions were occupied by different intron genotypes; and no are abundant in soil (Urich et al. 2008) and multiply by binary genotype was shared by the two groups. Group 2b was devoid fission. According to the life cycle found in many textbooks, of introns. Putatively functional or degenerated homing endo- haploid amoebae or amoeboflagellates undergo syngamy, and nuclease genes were found at different positions in groups 1 zygotes form a diploid, syncytially organized plasmodium as and 2a. All observations (genotypic combinations of the three the second vegetative stage. Most of the plasmodial biomass markers, signs of recombination, intron patterns) fit well into a converts into sporocarpic fruit bodies which release airborne meiospores like fungi. Myxomycetes are one of the few groups of simple eukaryotes that appear to be macroscopic Electronic supplementary material The online version of this article in dimension, which has led to a considerable body of data (doi:10.1007/s13127-015-0230-x) contains supplementary material, which is available to authorized users. on distribution and ecology (Stephenson et al. 2008). Only a limited number of species, mostly of the order Physarales, * Martin Schnittler will complete their life cycle in culture, even if food [email protected] organisms are supplied (Clark and Haskins 2010). Well known are Physarum polycephalum and Didymium iridis 1 Institute of Botany and Landscape Ecology, Ernst Moritz Arndt as model organisms in cell physiology, developmental stud- University of Greifswald, Soldmannstr. 15, ies (Goodman 1980; Sauer 1982), and systems biology 17487 Greifswald, Germany (Rätzel et al. 2013). 632 Y. Feng, M. Schnittler Apart from the advanced knowledge we have about a few debris, since a single spore colonizing a new habitat island model species, myxomycetes in nature are poorly studied by can complete the entire life cycle and disperse anew by spores molecular methods. Only recently were first phylogenies (Schnittler and Tesmer 2008). A first study on a wild popula- published (Fiore-Donno et al. 2010, 2012, 2013), which tion of Lamproderma spp. (Fiore-Donno et al. 2011) sug- recovered two main clades: the bright-spored clade gested the existence of asexual populations, since specimens (spores lightly colored by various pigments; Trichia varia from geographical independent accessions were found to be as the target species of this study belongs to this clade), and the identical in three molecular markers: the first part of SSU, its dark-spored clade (spores dark colored by melanin, most of internal transcribed spacer 1 (ITS1), and the first part of the the cultivable species). Many taxa are in critical need of revi- elongation factor 1 alpha gene (EF1A). Therefore, sexual and sion (Schnittler et al. 2012). First studies employing partial asexual biospecies may coexist within one morphologically nuclear small-subunit (SSU) ribosomal RNA gene sequences circumscribed species. This was the second question for our as a barcode marker revealed that many morphospecies seem study: Can we find evidence for asexual strains in a natural to be highly diverse (Novozhilov et al. 2013b), mounting ev- population? idence that the “everything is everywhere” hypothesis In this study, we analyze one species for the first time with (Fenchel and Finlay 2004) being popular for microorganisms three independent marker genes: SSU (in myxomycetes locat- should be rejected, and cryptic speciation is likely to occur ed in multiple extrachromosomal DNA molecules; Torres- (Aguilar et al. 2013). Machorro et al. 2010), EF1A (chromosomal), and cytochrome Still, the only species concept applicable for the group as a oxidase subunit 1 gene (COI, mitochondrial). T. varia (Pers. whole is entirely based on the morphology of the sporocarps ex J.F. Gmel.) Pers. is a common myxomycete described al- (Schnittler and Mitchell 2000). However, culture experiments ready by Persoon (1794) based on the Linnean name carried out with members of the Physarales pose a challenge Stemonitis varia (Linne 1792). In contrast to all established for this morphospecies concept prevailing for nearly 200 years model species in myxomycetes, T. varia belongs to the bright- (Clark 2000). Most isolates of a morphospecies behave het- spored clade of myxomycetes (Fiore-Donno et al. 2013)andis erothallic: amoebal populations derived from a single spore easy to recognize by elaters within the spore mass that show will not sporulate if they do not come in contact with amoebae only two, not several, spiral bands like the other members of from a different clone (Collins 1981). Heterothallism is main- this genus. This might be the reason why the name remained tained via mating types which can be arranged in a single- valid until today, although the morphological variability of the locus multiallelic system (Didymium iridis; Betterley and species is reflected by a rather long list of heterotypic syno- Collins 1983;Clark1993), or in three loci with multiple al- nyms (Lado 2014), the last one dating from 1974. leles (Physarum polycephalum;ClarkandHaskins2010; Additionally, we studied group I introns in complete SSU Collins 1975; Moriyama and Kawano 2010). Often, some sequences. Perhaps caused by the lifestyle of plasmodia, but not all of the mating types in a morphologically defined which prey on various microbes (besides bacteria, also yeasts species are compatible. Groups of compatible strains can be and green algae may be utilized; Haskins and Wrigley de seen as biospecies (Clark 1995; Collins 1979), thus providing Basanta 2008;Lazo1961), myxomycete sequences are rich an alternative species concept. This was the first question of in introns (Johansen et al. 1992; Lundblad et al. 2004). As for the study: Can we detect evidence for coexisting biospecies in mating types, cultivable members of the Physarales were nature? established as a model system for the study of group I introns If we assume that myxomycetes reproduce sexually (as it (Nandipati et al. 2012; Wikmark et al. 2007a, b). Ten intron should be expected for Amoebozoa, Lahr et al. 2011b)with insertion positions are known for SSU of myxomycetes; four mating types ensuring outcrossing (Clark and Haskins 2010; of these (positions 516, 911, 959, and 1199) were reported to Collins 1979), asexual lineages can develop via clonal popu- occur in T. varia (Fiore-Donno et al. 2013). Two mechanisms lations of non-fruiting amoebae (Clark and Haskins 2011). for intron homing are known. As the most powerful mecha- However, in culture experiments involving multiple isolates, nism, intron homing via an intron-coded homing endonucle- some strains behave non-heterothallic and form fruit bodies in ase (HE) is assumed, but reverse splicing through an RNA single-spore cultures (Collins 1981). Automixis, i.e., incom- intermediate was invoked as well (Haugen et al. 2005a; plete meiosis and the formation of diploid instead of haploid Lambowitz and Belfort 1993). Although, in a strict sense, meiospores, was invoked as the most likely reproductive sys- every intron poses an additional burden to the DNA replica- tem for such non-heterothallic, presumably asexual isolates tion of its host and introns can thus be seen as parasitic genes, (Clark and Haskins 2013). Such diploid spores should give they are selectively neutral (Edgell et al. 2011;Haugenetal. rise to diploid amoebae