Cladistics Cladistics 30 (2014) 26–66 10.1111/cla.12024 A total-evidence phylogenetic analysis of Hormaphidinae (Hemiptera: Aphididae), with comments on the evolution of galls Jing Chena,b, Li-Yun Jianga and Ge-Xia Qiaoa,* aKey Laboratory of Zoological Systematics and Evolution, Institute of Zoology, Chinese Academy of Sciences, No. 1 Beichen West Road, Chaoyang District, Beijing, 100101, China; bCollege of Life Sciences, University of Chinese Academy of Sciences, No. 19 Yuquan Road, Shijingshan District, Beijing, 100049, China Accepted 20 February 2013 Abstract A phylogenetic analysis of Hormaphidinae is presented based on a total-evidence approach. Four genes (two mitochondrial, COI and CytB, and two nuclear, EF-1a and LWO) are combined with 65 morphological and seven biological characters. Sixty- three hormaphidine species representing three tribes and 36 genera as well as nine outgroups are included. Parsimony and model-based approaches are used, and several support values and implied weighting schemes are explored to assess clade stabil- ity. The monophyly of Hormaphidinae and Nipponaphidini is supported, but Cerataphidini and Hormaphidini are not recov- ered as monophyletic. Based on the parsimony hypothesis from the total-evidence analysis, the phylogenetic relationships within Hormaphidinae are discussed. Cerataphidini is re-delimited to exclude Doraphis and Tsugaphis, and Hormaphidini is redefined to include Doraphis. Ceratocallis Qiao & Zhang is established as a junior synonym of Ceratoglyphina van der Goot, syn. nov. Lithoaphis quercisucta Qiao, Guo & Zhang is transferred to the genus Neohormaphis Noordam as Neohormaphis quercisucta (Qiao, Guo & Zhang) comb. nov. Galls have evolved independently within three tribes of Hormaphidinae. In Cerataphidini, pseudogalls are ancestral, both single-cavity and multiple-cavity galls have evolved once, and galls appear to have evolved towards greater complexity. Galling on secondary hosts has evolved twice in hormaphidines. © The Willi Hennig Society 2013. Introduction and eastern North America (Heie, 1980; Ghosh, 1985, 1988; von Dohlen et al., 2002). The subfamily Hormaphidinae (Hemiptera: Aphidi- Hormaphidinae species have complex life cycles. dae) is a clade of extraordinary aphids characterized Many species are heteroecious, seasonally obligately by the possession of several intrinsically fascinating alternating between primary host plants where the sex- biological characteristics. It comprises more than 200 ual phase of the life cycle is completed and galls are species within 45 genera and three tribes worldwide. produced, and secondary host plants where only par- Many genera (17 of 45) are monotypic, whereas sev- thenogenetic generations occur (Ghosh, 1985, 1988; eral genera show great species diversity (e.g. Astegop- Moran, 1988, 1992). Also abundant within this sub- teryx Karsch contains 21 described species). The tribes family are non-alternating species, which are believed Cerataphidini and Nipponaphidini are restricted to to have descended from heteroecious ancestors by los- eastern and south-eastern Asia, whereas the Horma- ing one set of hosts (Moran, 1988, 1992; Dixon and phidini exhibits a widespread distribution in Europe as Kundu, 1994; Blackman and Eastop, 2000). Strong well as a disjunctive distribution between eastern Asia host specificity is well defined and represented in the Hormaphidinae, with different patterns of host associ- ation among tribes. The Cerataphidini is primarily associated with Styrax (Styracaceae), and the Horma- *Corresponding author: E-mail address: [email protected] phidini and Nipponaphidini occupy Hamamelis and © The Willi Hennig Society 2013 J. Chen et al. / Cladistics 30 (2014) 26–66 27 Distylium (Hamamelidaceae), respectively, as their pri- might have evolved towards a better ability to mary hosts. The secondary host association is more manipulate their host plants, thus achieving higher relaxed, with Cerataphidini on Compositae, Grami- reproductive success by enlarging gall volume, chang- neae, Loranthaceae, Palmaceae, and Zingiberaceae; ing galling sites, or forming complicated gall struc- Hormaphidini on Betula (Betulaceae) and Picea (Pina- tures. So far, however, the evolution of galls in ceae); and Nipponaphidini on Fagaceae, Lauraceae, Hormaphidinae has not yet been thoroughly studied. and Moraceae. Based on the basic structure and mode of gall for- A great many hormaphidine aphids are known to mation, Fukatsu et al. (1994) suggested that multi- induce galls on their primary host plants (Chen and ple-cavity galls appear to be apomorphic and have Qiao, 2009, 2012a; Aoki and Kurosu, 2010). The mor- evolved only once in Cerataphidini. Stern (1995) phology and ontogeny of galls have been documented confirmed the single origin of multiple-cavity galls in in detail for many species (Kurosu and Aoki, 1990, Cerataphidini using the mitochondrial cytochrome 1991a,b, 1994, 1997, 2001, 2003, 2009; Aoki and oxidase subunits I (COI) and II (COII) genes. It Kurosu, 1992, 2010; Aoki et al., 1995, 2001, 2002; So- would be interesting to investigate the evolutionary rin, 1996, 2001; Qiao and Zhang, 2004; Kurosu et al., history of gall morphology within Homaphidinae 2006, 2008). Galls provide abundant nutrition (Price and to test the evolutionary trend that has been et al., 1986, 1987; Inbar et al., 2004; Koyama et al., revealed in Eriosomatinae. 2004; Zhang and Qiao, 2007a,b), a favourable micro- Hormaphidinae was once classified within Erioso- environment (Felt, 1940; Price et al., 1986, 1987; matinae or Thelaxinae in early studies (Mordvilko, Miller et al., 2009), and protection against natural ene- 1908, 1948; Borner,€ 1930; Borner€ and Heinze, 1957). It mies (Cornell, 1983; Price et al., 1986, 1987) to the was first regarded as a subfamily of Aphididae by inducer and its offspring. They can also mitigate clonal Baker (1920). Subsequently, Hille Ris Lambers (1964) mixing and maintain genetic integrity (Foster and considered Hormaphidinae a distinct clade closely Northcott, 1994; Stern and Foster, 1996). In Hor- related to Eriosomatinae and Thelaxinae, based on maphidinae, galls are quite diverse in terms of type, retaining three-faceted eyes in nymphs, which was site, shape, and structure (Ghosh, 1985, 1988; Chen approved by many authors (Zhang and Zhong, 1983; and Qiao, 2009, 2012a; Fig. 1). A few species form Ghosh, 1985, 1988; Blackman and Eastop, 1994, 2000; pseudogalls that appear as leaf rolls, leaf curls, or leaf Remaudiere and Remaudiere, 1997). Present knowl- blisters on the hosts (e.g. Aleurodaphis stewartiae Sorin edge of Hormaphidinae has resulted largely from the & Miyazaki, see Fig. 4k in Sorin and Miyazaki, 2004). great contributions of van der Goot (1917) and Noor- The majority produce variously shaped and structured dam (1991) on the Javan fauna; Takahashi (1931, true galls at different sites on the host plants (Fig. 1). 1935, 1936, 1939, 1941, 1957, 1958a,b,c,d, 1959a,b, Gall morphology is also highly specific to a particular 1962) and Takahashi and Sorin (1958) on the Chinese, species (Wool, 2004; Chen and Qiao, 2009, 2012a). Japanese, Sumatran, and Thai fauna; Sorin (1979, For instance, Astegopteryx spinocephala Kurosu, 1987, 1996, 1999, 2001, 2006) on the Japanese fauna; Buranapanichpan & Aoki forms banana-bunch-shaped Raychaudhuri et al. (1980) and Ghosh (1988) on the galls composed of several subgalls on twigs of Styrax Indian fauna; and Qiao and Zhang (1998, 1999, 2001, benzoides (see Fig. 1 in Kurosu et al., 2006); Hama- 2003, 2004) on the Chinese fauna. melistes miyabei (Matsumura) induces spherical galls The first high-level molecular phylogenetic analysis of with many spine-like hairs on twigs of Hamamelis the Aphididae including hormaphidines was based on japonica (see Fig. 1a in Aoki et al., 2001); and Neotho- mitochondrial ribosomal DNA (partial 12S and 16S) racaphis yanonis (Matsumura) produces spherical galls and included only three hormaphidine representatives, with a pointed bottom that protrude from both sides wherein the monophyly of Hormaphidinae was not of the leaves on Distylium chinense and D. racemosum recovered (von Dohlen and Moran, 2000). More recent (Fig. 1e). Therefore galls are commonly regarded as molecular studies conducted by Ortiz-Rivas et al. the extended phenotype of aphids (Dawkins, 1982; (2004) and Ortiz-Rivas and Martınez-Torres (2010) Stern, 1995; Stone and Schonrogge,€ 2003; Wool, included one and 12 hormaphidines, respectively. Both 2004). They are very helpful for species identifications, studies revealed a close relationship among Hormaphid- especially for species that are difficult to distinguish inae, Anoeciinae, Eriosomatinae, Mindarinae (not morphologically (e.g. Sorin, 2003), and also useful in included in the former study), and Thelaxinae. In the phylogenetic studies of aphids. latter phylogenetic analysis, the monophyly of Hor- Many studies have been conducted on the evolu- maphidinae was retrieved, albeit with low statistical tion of gall morphology in another galling aphid supports, based on the nuclear elongation factor-1a group, Eriosomatinae (Inbar et al., 2004; Zhang and (EF-1a) gene and the mitochondrial cytochrome oxidase Qiao, 2007a,b, 2008; Sano and Akimoto, 2011). It subunit II (COII) gene (excluding the third codon posi- has been suggested that the Eriosomatinae aphids tions). The monophyly of this subfamily was not estab- 28 J. Chen et al. / Cladistics 30 (2014) 26–66 (a) (b) (c) (d) (e) (f) Fig. 1. Galls of Hormaphidinae. (a) A gall of Tuberaphis owadai Kurosu