1 A molecular phylogeny of Cochylina, with confirmation of its relationship to Euliina
2 (Lepidoptera: Tortricidae)
3
4 John W. Brown*1, Leif Aarvik2, Maria Heikkilä3, Richard Brown4, and Marko Mutanen5
5
6 1 National Museum of Natural History, Smithsonian Institution, Washington, DC, USA, e-mail:
8 2 Natural History Museum, University of Oslo, Norway, e-mail: [email protected]
9 3 Finnish Museum of Natural History, LUOMUS, University of Helsinki, Helsinki, 00014,
10 Finland, e-mail: [email protected]
11 4 Mississippi Entomological Museum, Mississippi State, MS 39762, USA, e-mail:
13 5 Ecology and Genetics Research Unit, PO Box 3000, 90014, University of Oulu, Finland, e-
14 mail: [email protected]
15 *corresponding author
16
17 This is the peer reviewed version of the following article: Brown, J.W., Aarvik, L., Heikkilä, M.,
18 Brown, R. and Mutanen, M. (2020), A molecular phylogeny of Cochylina, with confirmation of its
19 relationship to Euliina (Lepidoptera: Tortricidae). Syst Entomol, 45: 160-174., which has been
20 published in final form at https://doi.org/10.1111/syen.12385.
21
1
22 Abstract. We conducted a multiple-gene phylogenetic analysis of 70 species representing 24
23 genera of Cochylina and eight species representing eight genera of Euliina, and a maximum
24 likelihood analysis based on 293 barcodes representing over 220 species of Cochylina. The
25 results confirm the hypothesis that Cochylina is a monophyletic group embedded within a
26 paraphyletic Euliina. We recognize and define six major monophyletic lineages within
27 Cochylina: a Phtheochroa Group, a Henricus Group, an Aethes Group, a Saphenista Group, a
28 Phalonidia Group, and a Cochylis Group. We summarize the groups (including related genera
29 not included in our analysis), provide morphological characters that support the molecular data,
30 and compare our results to previous phylogenies of Cochylina. We propose the following
31 nomenclatural changes: Brevicornutia, revised status; Neocochylis, revised status; Paracochylis,
32 revised status; Pontoturania, revised status; and Platphalonidia, revised status.
33
34 Key Words: Phtheochroa, Saphenista, Henricus, Aethes, Cochylis, Phalonidia, monophyly,
35 Euliina, Cochylina
36
37 Introduction
38 As presently defined, the tortricid tribe Cochylini comprises two large subtribes - Cochylina
39 (with about 2,234 described species) and Euliina (with about 1,084 described species) (Gilligan
40 et al., 2018) - each of which historically was considered its own tribe (i.e., Cochylini and Euliini)
41 (Horak & Brown, 1991; Brown & Powell, 1991; Horak, 1999). As recent as 1983 (Powell,
42 1983), Cochylina was considered a distinct family (i.e., Cochylidae), even though Kuznetsov &
43 Stekolnikov (1977) had earlier proposed that Cochylini was a tribe within Tortricinae. A
44 molecular phylogeny of Tortricidae by Regier et al. (2012) recognized a monophyletic
2
45 Cochylina embedded within a paraphyletic Euliina. Unfortunately, that study included
46 exceedingly few taxa, i.e., four species/genera of Euliina and three species (two in genera that
47 are now considered synonyms) of Cochylina, so the results are not particularly robust.
48 There is no comprehensive phylogeny or classification of the genera of Cochylina
49 worldwide. Razowski (1970) presented an “intuitive” phylogeny for the Palearctic genera (n =
50 21) (Fig. 1), and Pogue & Mickevitch (1990) presented a parsimony analysis of North American
51 genera (n = 20) (Fig. 2). Hence, it is impossible to place many previously described and/or new
52 genera into a meaningful system or classification.
53 In this contribution we focus primarily on two issues: 1) we test the hypothesis that
54 Cochylina is a monophyletic group within a paraphyletic Euliina; and 2) we define generic
55 groups within Cochylina and propose a classification. We also propose taxonomic changes (i.e.,
56 the elevation of previously described subgenera to generic status) that result in more natural or
57 monophyletic genera.
58
59 Historical Background
60 Cochylina (as Cochylidi) was recognized by Guenée (1845) as one of 10 higher groups in
61 Tortricidae. Stephens (1852) considered Cochylina a subfamily of Tortricidae (i.e., Cochylinae),
62 and Morris (1898) elevated it to family rank (i.e., Phaloniidae). This concept was followed by
63 Stainton (1859), Meyrick (1882, 1895), Clarke (1963), Diakonoff (1961), and others for over 100
64 years. Razowski (1960) proposed the name Cochylidae for the group based on the oldest higher
65 taxon name used for it – Cochylidi (Guenée, 1845) – but in the years that followed, the group
66 was referred to inconsistently as either Cochylidae (e.g., Diakonoff, 1961; Falkovitsh, 1963;
67 Razowski, 1967a, 1967b, 1968a, b, c; Bradley et al., 1973) or Phaloniidae (e.g., Obraztsov, 1967;
3
68 Clarke, 1963, 1968). Based on larvae, Swatschek (1958) and MacKay (1959, 1962) continued
69 the exclusion of Cochylina from Tortricidae, although larval characters provided little evidence
70 for this.
71 In the 1970s, Kuznetzov & Stekolnikov (1973, 1977) and Razowski (1976) proposed
72 phylogenies of Tortricidae relying primarily on musculature of the male genitalia. Kuznetzov
73 and Stekolnikov (1973, 1977) were the first to return Cochylina to Tortricidae, treating it as the
74 supertribe Cochylidii (Tortricinae). They also proposed Euliae as a subtribe of Cochylidii.
75 Razowski (1976) likewise included Cochylina within the subfamily Tortricinae. However, these
76 results were not immediately accepted by all. For example, in the Check list of the Lepidoptera of
77 America north of Mexico (Powell, 1983), the group was retained as a distinct family (i.e.,
78 Cochylidae). However, by the late 1980s momentum was growing for recognition of the group as
79 a tribe (e.g., Gibeaux, 1985; Pogue & Friedlander, 1987; Pogue, 1988; Razowski, 1989), and by
80 the early 1990s there was general consensus that the group was best considered a tribe within the
81 subfamily Tortricinae (Horak & Brown, 1991). With few exceptions, it has been treated as such
82 in virtually all taxonomic revisions and systematic treatments since then (e.g., Razowski, 1990,
83 1992, 1993, 1995, 1997; Horak, 1999; Metzler, 2000; Pogue, 2001; Metzler & Sabourin, 2002;
84 Sabourin et al., 2002).
85
86 Relationship Between Euliina and Cochylina
87 Kuznetsov and Stekolnikov (1977) were the first to recognize Euliina (comprised of the two
88 genera Eulia Hübner and Pseudargyrotoza Obraztsov) as a higher category, placing it as a
89 subtribe of Cochylini. Prior to their work, Nearctic genera currently assigned to Euliina had been
90 placed in Cnephasiini by Powell (1964), and the Neotropical genera had been considered part of
4
91 Archipini by Razowski (e.g., 1987a, b, 1988). Powell (1986) elevated the concept of Euliini to
92 tribal level, into which he placed all the Neotropical genera from his previous concept of
93 Cnephasiini and Razowski’s group of Archipini. Brown & Powell (1991) reviewed the
94 Chrysoxena group of Euliini and proposed a preliminary, morphology-based phylogeny for the
95 tribe (including 23 genera), but the number of genera now associated with the group has
96 quadrupled. Brown (2005) retained Powell’s concept of Euliini in the world catalogue of the
97 family to include a single Palearctic genus (i.e., Eulia) and over 100 Neotropical genera.
98 Most recently, a molecular phylogenetic analysis of Tortricidae by Regier et al. (2012)
99 concluded that Cochylina is best treated as a subtribe within a broader, paraphyletic Euliina.
100 They recommended the retention of the two taxa as subtribes (Euliina and Cochylina) for
101 purposes of stability, even though Euliina were shown to be paraphyletic. Because Regier et al.
102 (2012) included only four species/genera of Euliina and three species/genera of Cochylina, their
103 results indicated that further investigation was necessary, stimulating the research presented
104 herein.
105
106 Materials and Methods
107
108 Taxon sampling and specimen acquisition
109 A primary objective was to maximize sampling at the genus level in the subtribes Cochylina
110 and Euliina, and multiple representatives of each genus were included when available. Because
111 dry specimens more than 10 years old are unlikely to provide data for nuclear genes, we limited
112 the sampling for full genetic analyses to taxa that we considered likely to be functional. We
113 sampled six species representing six genera of Euliina and 71 species representing 27 genera of
5
114 Cochylina for full genetic analyses. As outgroups, we included one species of Chlidanotinae
115 (Olindia schumacherana (Fabricius)), two species of Olethreutinae (Apotomis betuletana
116 (Haworth), Bactra lancealana Hübner), and seven species of tribes of Tortricinae (Archipini:
117 Zelotherses paleana (Hübner), Pandemis cinnamomeana (Treitschke), Thrincophora lignigerana
118 (Walker); Cnephasiini: Exapate congelatella (Clerck), Arotrophora arcuatalis (Walker);
119 Sparganothini: Sparganothis rubicundana (Herrich-Schäffer); Tortricini: Tortrix viridana
120 Linnaeus). The multi-gene dataset was supplement by sequences of the euliine genera Netechma
121 (CAD, COI), Pseudomeritastis (CAD, COI), and Bonagota (CAD) published in Regier et al.
122 (2012). For Lorita scarificata (Meyrick), we used COI data concatenated from BIOUG01827-
123 G12 (COI barcode region) and MH_USNM_18 (COI-end); and for Neocochylis molliculana
124 (Zeller) we used data concatenated from IT_3 (CAD, EF1a-begin and EF1a-end) and
125 TLMF_Lep_23781 (COI barcode region).
126 Because we were unable to obtain suitable material of many genera, we also conducted a
127 phylogenetic analysis of all species of Cochylina based on the mitochondrial COI gene data
128 (DNA barcode region) available to us in the BOLD database, which slightly increased generic-
129 level sampling and greatly increased species-level sampling. Additionally, if barcodes of a
130 species fell into more than one BIN (Barcode Index Number operational taxonomic unit, see
131 Ratnasingham & Hebert 2013), a representative of each BIN was included. In this analysis, 294
132 specimens plus Eulia ministrana as an outgroup were included, altogether representing 199
133 species and 279 BINs.
134
135 Molecular techniques
6
136 Legs of dry and pinned, but sometimes alcohol preserved, adult specimens were used for
137 extraction of genomic DNA with DNeasy Blood & Tissue Kit (Qiagen) at the University of
138 Oulu, Finland. One mitochondrial (cytochrome c oxidase subunit I, COI) and the following
139 seven nuclear regions were PCR amplified: carbamoylphosphate synthase domain protein
140 (CAD), elongation factor 1 alpha (EF1α), glyceraldhyde-3-phosphate dehydrogenase (GAPDH),
141 isocitrate dehydrogenase (IDH), cytosolic malate dehydrogenase (MDH), ribosomal protein S5
142 (RpS5), wingless (see Table S2). Altogether, the sequenced regions comprise 6211 base pairs.
143 This set of markers has become the standard in Lepidoptera phylogenetics and has been used in
144 numerous studies (e.g., Mutanen et al., 2010; Heikkilä et al., 2015). Sequencing pipeline largely
145 followed that of Wahlberg & Wheat (2008), except PCR clean-up was carried out with ExoSAP-
146 IT (Affymetrix) and Sephadex columns (Sigma-Aldrich), and sequencing that was done using an
147 ABI 3730 DNA Analyzer (Applied Biosystems). Sequences were manually aligned and edited
148 using BioEdit (Hall 1999). Sequencing of the COI barcode region was conducted primarily at the
149 Canadian Centre for DNA Barcoding (CCDB) following protocols explained in detail in
150 DeWaard et al. (2008). GenBank accession numbers for the sequences of full-gene analysis are
151 provided in Table S1 and sequence lengths in Table S2. Full collection and taxonomic data as
152 well as sequences and GenBank accession numbers for the specimens used in the COI analysis
153 can be retrieved from the public BOLD dataset DS-COCHY at DOI.XXXX.
154
155 Phylogenetic analyses
156 Maximum Likelihood (ML) analyses of the multiple-gene dataset were conducted using
157 RAxML (Stamatakis, 2014) in CIPRES (Miller et al., 2010) with GTR for nucleotide
158 substitution and gamma model for among-site rate heterogeneity. Support for nodes was
7
159 evaluated with 100–1000 bootstrap replicates. Separate analyses with the dataset partitioned by
160 gene, by codon position, and without the third codon position were performed. Resulting trees
161 were examined in FigTree 1.4.3.and rooted at Olindia schumacherana. Gene trees were also
162 generated following the same method with the exception of using the unpartitioned model and
163 rooting the RpS5 gene tree at Sparganothis rubicundana because we did not have the RpS5
164 sequence for Olindia schumacherana.
165 A phylogenetic analysis based on Bayesian inference was also performed with the program
166 MrBayes v3.2.6 (Ronquist & Huelsenbeck, 2003) in CIPRES (Miller et al., 2010). Molecular
167 data were partitioned by genes, and the number of substitution types set to nst = mixed with
168 among-site rate variation modelled by a gamma distribution (rates = gamma). The analyses were
169 run for 10 million generations and sampled every 1000 generations. Convergence was assessed
170 in Tracer v1.6 (Rambaut & Drummond, 2007). The first 25% of generations were discarded as
171 burn-in. FigTree 1.4.2 was used to view the tree.
172 A Maximum Likelihood analysis for COI barcode sequence data was performed using
173 RAxML-NG Black Box service available at https://raxml-ng.vital-it.ch/#/ (Kozlov et al., 2018)
174 under the following settings: GTR for nucleotide substitution with gamma model for among-site
175 rate heterogeneity, unpartitioned model, and automatic bootstrapping (bootstopping) with a value
176 of 0.03 for cutoff. The resulting tree was rooted with Eulia ministrana, and FigTree 1.4.2 was
177 used for the depiction of the tree.
178
179 Results and Discussion
180
8
181 The different partition strategies used in our phylogenetic analyses based on maximum
182 likelihood did not result in different topologies, and we choose to present the tree based on the
183 data partitioned by gene as our primary result (Fig. 3). The topology obtained in the Bayesian
184 inference analysis (see supplementary material: Fig. S1) is identical to that of the ML analysis
185 (apart from smlll differences in the placement of taxa within the outgroup and the placement of
186 species within a couple genera in the ingroup) and therefore in not discussed separately.
187 Our multi-gene analysis provides support for the conclusions of Regier et al. (2012). With
188 one notable exception (i.e., Hyptiharpa Razowski), all Euliina and Cochylina form a strongly
189 supported (i.e., 93) monophyletic lineage. Within this lineage, Bonagota+Apotomops are sister to
190 Eulia. The close relationship between Bonagota Razowski and Apotomops Powell & Obraztsov
191 has been recognized previously based on morphology (Brown & Powell, 1991; Brown &
192 Razowski, 2003). Cuproxena+Dorithia are sister to Pseudomeritastis Obraztsov, a relationship
193 supported by a similarly shaped signum in the female genitalia. Together with Anopina
194 Obraztsov and Netechma Razowski, all these euliine genera form a paraphyletic assemblage
195 surrounding a monophyletic Cochylina. Hence, our results corroborate and significantly expand
196 the findings of Regier et al. (2012), with the addition of four euliine genera and 22 cochyline
197 genera.
198 Hyptiharpa, an enigmatic monotypic genus (i.e., H. baboquavariana (Kearfott), with its
199 synonym H. hypostas (Razowski)) considered to belong to either Euliini or Cochylini by
200 Razowski (1992) and Brown (2016), did not cluster with other euliine genera. Instead, it found a
201 position closest to Eucosmini, and this result cannot be explained. To rule out the possibility of
202 contamination in the laboratory, we examined the gene trees of the taxa included in our study. In
203 each gene tree Hyptiharpa was grouped with the outgroup taxa. We also ran the barcode region
9
204 of our Hyptiharpa specimen through the specimen identification engine in BOLD and the closest
205 matches obtained were species of Eucosmini and another specimen of Hyptiharpa
206 baboquavariana (JWB-08-0210). GenBank’s BLAST tool also gave olethreutine species as
207 closest matches. These tests speak against the possibility of contamination as does the fact that
208 specimens belonging in the outgroup were not handled in the laboratory.
209
210 Generic Classification of Cochylina
211 The British genera of Cochylina were classified by Meyrick (1895) based primarily on wing
212 venation. Meyrick’s phylogenetic tree portrayed Hysterosia Stephens (currently treated as a
213 synonym of Phtheochroa Stephens) as the most primitive genus. Razowski (1970) presented a
214 classification of the Palearctic Cochylini, placing the 21 genera into three large groups (Fig. 1),
215 and his treatment remains the most comprehensive study on the taxon to date. In an unpublished
216 thesis on North American Cochylini, Pogue (1986) recognized 23 genera, 10 of which were new,
217 and Pogue & Mickevitch (1990) proposed a phylogeny for the North American members of the
218 tribe (Fig. 2) (20 genera) based on a parsimony (transformation series) analysis of morphological
219 characters.
220 Based on our multi-gene analysis, six monophyletic generic groups can be recognized
221 and defined (Fig. 3, Table 1): a Phtheochroa Group, a Henricus Group, an Aethes Group, a
222 Saphenista Group, a Phalonidia Group, and a Cochylis Group. Each group is detailed below.
223 As would be expected, the ML barcode analysis resulted in a tree with less compelling
224 structure than that of the multi-gene analysis (Figs. 4‒7). It successfully recovered the
225 monophyly of only three of the six groups recognized in the multi-gene tree, i.e., the Henricus
226 Group, the Saphenista Group (although embedded with the Aethes Group), and the Cochylis
10
227 Group. Nonetheless, the ML barcode tree provides evidence, although limited, of the
228 approximate position of some genera either not included, or represented by a single species, in
229 the multi-gene analysis. It also provides additional insight into genera that appear to be para- or
230 polyphyletic in the multi-gene analysis based on more thorough sampling; i.e., it provides
231 evidence of relationships among congeneric species, which is particularly helpful for untangling
232 the large, polyphyletic genus Cochylis. The ML barcode tree is divided by generic group and is
233 presented in Figs. 4‒7 as support for the narrative of each generic group below.
234
235 Phtheochroa Group
236 The Phtheochroa Group is our best sampled taxon, with 23 species representing nine genera.
237 Its monophyly is well supported (i.e., 94) (Fig. 3), and it is sister to the remaining generic groups
238 of Cochylina. Most of the genera included by Razowski (1970) in his three large lineages [i.e.,
239 Hysterosia, Phtheochroa (variously treated as Hysterosia, Phtheochroa, and Prohysterophora
240 Razowski), Hysterophora Obraztsov, Cochylimorpha Razowski (as Stenodes Guenée, 1845),
241 Ceratoxanthis Razowski, Fulvoclysia Obraztsov, Agapeta Hübner, and Phtheochroides
242 Obraztsov] are also present in our circumscription of the group. Differences between our tree and
243 that of Razowski (1970) include our placement of Phalonidia Le Marchand in the Phalonidia
244 Group (rather than in the Phtheochroa Group), and Razowski’s assignment of Eugnosta Hübner
245 and Eupoecilia Stephens to the Aethes Group, both of which we include in the Phtheochroa
246 Group.
247 Among the plesiomorphic features found in this group are the female frenulum with three
248 acanthi rather than two (present in Agapeta hamana (Linnaeus), Hysterophora, a few species of
249 Eugnosta, many species of Cochylimorpha, and most Phtheochroa, Phtheochroides,
11
250 Prochlidonia Razowski, and Commophila Hübner) (Monsalve et al., 2011); the presence of a
251 male forewing costal fold (found in many Phtheochroa); the presence of an uncus in the male
252 genitalia (in most Phtheochroa); and the distally united or fused arms of the vinculum (present in
253 Phtheochroa and Hysterophora). Also, the rigid, erect socii of many genera of the Phtheochroa
254 Group are found nowhere else in the Cochylina. Although these character states are not uniform
255 or consistent throughout the Phtheochroa Group, they are encountered rarely in other generic
256 groups of Cochylina.
257 Within the Phtheochroa Group (Fig. 3) the genus Phtheochroa is represented by a
258 monophyletic clade with Hysterophora embedded within, and this clade is sister to the remainder
259 of the genera. While this topology suggests that Hysterophora should be synonymized with
260 Phtheochroa, the combined morphological and molecular evidene is not altogether convincing.
261 Hence, for the interim, Hysterophora is retained as a valid genus, and it is considered the sister to
262 Phtheochroa. In the barcode tree (Fig. 4) there is a large, uniform cluster of Phtheochroa species
263 (32 OTUs) that is sister to Actihema Razowski plus two potentially misplaced species: Eugnosta
264 percnoptila (Meyrick) and Phtheochroa aarviki Razowski & Brown.
265 Eugnosta percnoptila is sister to Actihema in the multi-gene tree and sister to
266 Actihema+Phtheochroa aarviki in the barcode tree. Its genitalia retain many plesiomorphic
267 features, with the valvae, phallus, and transtilla similar to that of many species in the
268 Phtheochroa Group, and the rigid socii characteristic of Eugnosta, Actihema, and several other
269 genera. In Actihema the socii arise from a broad sclerotized plate, whereas in Eugnosta they arise
270 much closer together; however, there is considerable transition between the two states. In the
271 putative sister relationship between E. percnoptila and Actihema (Fig. 3) the broad base of the
272 socii in the former can be interpreted as homologous with that of Actihema. Also, the ring-like
12
273 sclerite in the corpus bursae of the female genitalia of E. percnoptila resembles that of Actihema.
274 Hence, E. percnoptila may represent an undescribed genus sister to Actihema.
275 There is also morphological support for the sister relationship between Actihema+E.
276 percnoptila and Phtheochroa aarviki (and its sister P. kenyana Aarvik) - the presence of a long,
277 rod-like, dorso-median process of the juxta. Otherwise, the genitalia of P. kenyana and P. aarviki
278 are characterized by plesiomorphies. The African genus Afropoecilia Aarvik also has a dorso-
279 median extension of the juxta, but it is broad and band-like. If the positions of E. percnoptila and
280 P. aarviki (plus P. kenyana) are correct, these species should be removed from Eugnosta and
281 Phtheochroa, respectively, and two new genera should be proposed. Alternatively, their potential
282 assignment to Actihema, which is very compact and distinct, is not a particularly compelling
283 resolution. In E. percnoptila the forewing has numerous tufts of raised scales. There is a
284 tendency for this in Actihema, but the tufts are not as pronounced as in E. percnoptila. This
285 feature may help distinguished E. percnoptila from other (“true”) Eugnosta, which lack raise
286 scales or tufts.
287 Our multi-gene phylogeny (Fig. 3) also shows a clade of Eugnosta (four species) as sister
288 to Cochylimorpha (two species), but again, with weak branch support. It also captures a
289 monophyletic Agapeta as sister to Ceratoxanthis+Fulvoclysia, similar to the hypothesis of
290 Razowski (1970) (Fig. 1). The adults of the last three genera all share a large forewing length
291 and bright yellow forewing maculation.
292 In our multi-gene tree (Fig. 3) Cochylimorpha occurs in two widely separated branches, with
293 one represented by the single species C. alternana (Curtis). The COI tree (Fig. 4) also shows two
294 clusters of Cochylimorpha species: one a monophyletic group (13 OTUs; support of 49) sister to
295 a large clade of genera (i.e., Eugnosta+Cryptocochylis+Fulvoclysia+Agapeta), and the second a
13
296 monophyletic group (5 OTUs; support of 22) embedded within the last-mentioned clade. Hence,
297 evidence from both the multi-gene and barcode trees suggests that Cochylimorpha may represent
298 two different genera. Within the smaller Cochylimorpha cluster, C. alternana is sister to C.
299 straminea (Haworth). Razowski (1960) treated these two species, along with C. chamomillana
300 (Herrich-Schäffer) (= decolorella (Zeller)) and C. santolinana (Staudinger), as a separate genus,
301 Euxanthoides Razowski, with C. straminea as the type species. Within Euxanthoides, he further
302 placed C. alternana in a separate subgenus (Bleszynskiella Razowski). Although these “species
303 groups” are fairly distinct morphologically, their circumscription into subgenera leaves a large
304 number of outliers and species clusters unaffiliated or unassigned to a genus or subgenus.
305 Consequently, Razowski (1970) subsequently gave up on the subgeneric division of
306 Cochylimorpha (as Stenodes), concluding that Cochylimorpha is a large and complicated genus.
307 Our analysis does not include enough representatives to provide compelling conclusions;
308 nonetheless, it suggests that Euxanthoides [to include C. straminea and C. alternana, and
309 perhaps C. perfuscana (Guenée), as well] may be a valid genus, possibly sister to Cryptocochylis
310 Razowski.
311 The most incongruent aspect of the COI tree (Fig. 4) is the presence of three separate
312 lineages of Eugnosta: one that represents a group of species formerly placed in Carolella Busck
313 (which is considered a synonym of Eugnosta); one that includes the single Palearctic species E.
314 magnificana (Rebel); and one that includes the remainder of the Eugnosta species (Nearctic and
315 Afrotropical representatives). The synonymy of Carolella and Eugnosta is supported by
316 morphological features, and this relationship is reflected in our multi-gene tree (Fig. 3).
317 However, the position of E. magnificana is inexplicable. The two African species of Eugnosta in
318 our sample cluster convincingly with North American species. These two (i.e., E. misella
14
319 Razowski and E. trimeni (Felder & Rogenhofer)) represent extremes in male genitalia: E. trimeni
320 has a massive process of the transtilla (similar to that of E. percnoptila) and weak, broad socii; E.
321 misella has a slender process of the transtilla and slender, rigid socii. These observations
322 exemplify one of the major difficulties in reconstructing the phylogeny of Cochylini, i.e., the
323 genitalia of species within a clade are sometimes more similar to those of species outside the
324 clade than they are to species within the same clade. Palearctic species of Eugnosta are mostly
325 characterized by a silvery forewing ground color, whereas virtually all New World and
326 Afrotropical species have a whitish or brownish ground color.
327 Genera not included in our multi-gene analysis (Fig. 3) that likely belong in the Phtheochroa
328 Group include the following: Acarolella Razowski & Becker, Afropoecilia, Anielia Razowski &
329 Becker, Combosclera Razowski, Commophila, Cryptocochylis (associated by barcodes),
330 Mimeugnosta Razowski, Oligobalia Diakonoff, Prochlidonia, Phtheochroides, and
331 Rigidsociaria Razowski. Among these, Commophila was placed by Razowski (1970) (Fig. 1) in
332 an unresolved trichotomy with Eugnosta and Eupoecilia, supporting its putative assignment to
333 the Phtheochroa Group. According to Razowski (2011), Eugnosta, Commophila, and
334 Prochlidonia form a distinct group characterized by long, erect socii, but the long rigid socii
335 occur in many Phtheochroa Group genera. Razowski (2011) also mentioned that the Neotropical
336 genera Anielia and Acarolella are closely related to genera that we include in the Phtheochroa
337 Group.
338 Afropoecilia likewise can be assigned to the Phtheochroa Group based on morphology.
339 According to Aarvik (2010), the structure of the socii of its male genitalia is the is the same as
340 that of Eupoecilia and Actihema, and the ring-shaped sclerite in corpus bursae in the female
341 genitalia is shared with Actihema.
15
342 Although the biology of the majority of the members of the Phtheochroa Group is unknown,
343 the existing records indicate a strong tendency for an internal/concealed feeding mode, primarily
344 in Asteraceae. For example, Razowski (2002) stated that the larvae of Phtheochroa feed in seeds,
345 fruit, stems, and roots of dicotyledonous plants in eight different families, but primarily
346 Asteraceae. Likewise, nearly all rearings of Cochylimorpha are from seeds, stems, roots, or galls
347 of Asteraceae; and nearly all known larvae of Eugnosta are gall-inducers in the stems of
348 Asteraceae (e.g., Razowski, 1970; Goeden & Riker, 1981; Vargas et al., 2015). Actihema was
349 recorded from the seeds/fruit of Asteraceae in Kenya (Brown et al., 2014). Eupoecilia and
350 Agapeta appear to have slightly broader host ranges, but both have been recorded from
351 Asteraceae, as well, and internal feeding is common in these two genera.
352
353 Henricus Group
354 In our multi-gene analysis (Fig. 3), the monophyly of the Henricus Group is strongly
355 supported (i.e., 100), but it is represented by only two genera that are almost certainly sisters
356 (Brown 2013): Henricus Busck, with 43 described species, and Eupinivora Brown, with eight
357 described species. The two genera share a number of features with many members of the
358 Phtheochroa group, including large forewing length, the presence of three acanthi in the female
359 frenulum (Monsalve et al., 2011), and the presence of an uncus (although small in the Henricus
360 group), all of which are almost certainly plesiomorphies. The similarities are so remarkable that
361 Razowski (1986) described two species of Eupinivora in Phtheochroa and one in Henricus.
362 None of the genera included in the Henricus Group was included in Razowski’s (1970)
363 phylogeny because they are found only in the New World, so there is no clue of where he might
364 have placed them. However, his original assignment of two species of Eupinivora to
16
365 Phtheochroa suggests he would have placed the Henricus Group within his Phtheochroa Group.
366 Similarly, Pogue & Mickevitch (1990) placed Henricus as sister to Eugnosta among North
367 American Cochylina.
368 In our barcode tree (Fig. 5), Henricus forms a large monophyletic group sister to the rest of
369 Cochylina, as in our multi-gene tree (Fig. 3), as in our multi-gene tree (Fig. 3), within which
370 Eupinivora is embedded, suggesting it is a synonym of Henricus. The Henricus Group is
371 exclusively New World and includes at least three other genera not included in our analysis:
372 Cartagogena Razowski, Imashpania Razowski & Wojtusiak, and Parirazona Razowski. The
373 first encompasses three high-elevation species from Costa Rica; Razowski (2011) suggested that
374 it may be synonymous with Henricus. The second is monotypic and likewise was considered
375 closely related to Henricus by Razowski & Wojtusiak (2008). The third is a small Neotropical
376 group (nine species) known only from Brazil; the name is derived from Irazona Razowski,
377 currently considered a synonym of Henricus. The type species of Irazona (i.e., comes
378 Walsingham) is convincingly embedded within Henricus (Fig. 5). Although barcodes place
379 Parirazona near Nycthia Pogue (Fig. 7), morphology argues for its placement in the Henricus
380 Group.
381 Different clades within the Henricus Group appear to have radiated on different host
382 families, but primarily on conifers. For example, all species of Eupinivora either have been
383 recorded from, or are suspected to feed within, the cones of Pinaceae (Brown, 2013), as does
384 Henricus fuscodorsanus (Kearfott) and H. melanoleucana (Clarke). Three species of Henricus,
385 H. infernalis (Heinrich), H. macrocarpana (Walsingham), and H. inchoatus Razowski & Becker,
386 are recorded only from Cupressaceae; and H. umbrabasana (Kearfott) shows a strong preference
387 for Quercus (Fagaceae).
17
388
389 Aethes Group
390 In the multi-gene tree (Fig. 3), the Aethes Group is comprised of a relatively well-supported
391 (i.e., 61), monophyletic group of Aethes Billberg, 1820 (11 OTUs) plus Rudenia Razowski (a
392 single example) as its sister; the latter is rather weakly associated with the Aethes Group. In the
393 barcode tree, Rudenia (at the bottom right side of Fig. 6) is sister to Spinipogon Razowski, and
394 the two are fairly distant from Aethes (the last represented by 54 OTUs). The female genitalia of
395 Spinipogon argue convincingly for its placement near Gynnidomorpha Turner and Phalonidia in
396 the Phalonidia Group. Hence, it is possible as well that Rudenia belongs in the Phalonidia
397 Group.
398 Razowski (1985a) originally hypothesized that Rudenia was closely related to Eugnosta (in
399 our Phtheochroa Group); he subsequently (Razowski 1994) placed the genus near Lorita Busck (in
400 our Phalonidia Group) based on the similarity of the female genitalia. Pogue & Mickevitch (1990)
401 likewise placed Rudenia near Lorita. In our barcode tree Rudenia (8 OTUs) forms a monophyletic
402 group near Gynnidomorpha (in our Phalonidia Group). Hence, the position of Rudenia in the
403 Aethes Group is likely incorrect. Rudenia is a small group (5 described species) ranging across the
404 southern portion of North America (from the Atlantic to the Pacific), south to Venezuela as a
405 complex of closely related species that are not easily separated by genital morphology, but can
406 be separated by barcodes; they feed exclusively on Fabaceae.
407 In the Aethes Group the most significant conflict between our multi-gene (Fig. 3) and
408 barcode trees (Fig. 6) is the presence of a branch that supports and a monophyletic Saphenista
409 Walsingham (5 OTUs) sister to Aphalonia Razowski+Aethes turialba (Busck) that is deeply
410 embedded within Aethes in the barcode tree (Fig. 6). This placement of Saphenista is
18
411 inexplicable and is almost certainly incorrect. Also, five species of Aethes fall outside the large
412 monophyletic group of this genus, somewhat scattered throughout the Phlaonidia group.
413 The monophyly of Aethes is strongly supported by a pair of unusual, sickle-shaped socii from
414 the dorsum of the tegumen in the male genitalia that are present in nearly all species. The genus
415 is widely distributed throughout Europe, Asia, North America, and South America (absent from
416 Australia and Africa), from boreal forests to rain forests, encompassing about 150 described
417 species that feed mostly on Asteraceae and Apiaceae, but with many exceptions. Among the
418 Neotropical species of Aethes is a small group of closely related species that are large and
419 superficially similar to Phtheochroa species (e.g., A. turialba, A. macasiana Razowski & Pelz, A.
420 chilesi Razowski & Wojtusiak), represented in our dataset by A. turialba. Species of Aphalonia
421 Razowski are also large and superficially similar to these large Aethes, and in his description of
422 Aphalonia, Razowski (1984) indicated that the type species (Aphalonia monstrata Razowski) is
423 similar to species of the Aethes agelasta Razowski species group, with similar wing venation.
424 Hence, the placement of Aphalonia within the Aethes Group is not surprising. The only other
425 genera that are assigned convincingly in the Aethes Group are Aethesoides Razowski, a small
426 Neotropical group that is almost certainly sister to Aethes, and Lincicochylis Razowski, which
427 may be a synonym of Aethesoides. According to Razowski (1994), Aethes and Aethesoides share
428 the presence of slender, curved socii (the processes mentioned above) from a broad, hairy base
429 and a rod-like sclerite coupling the tegumen with the valva. Aethesoides and Lincicochylis share
430 a pair of stout, sclerotized processes from the dorso-posterior end of the tegumen, reminiscent of
431 similar processes in Eugnosta; a valva that has a well-developed, rod-like sacculus and a short,
432 mostly atrophied remainder; and an unusual, semicircular, sometimes spiny flap, located
433 immediately anterior to the socii.
19
434
435 Saphenista Group
436 In our multi-gene tree (Fig. 3), the Saphenista Group is represented by a single Nearctic
437 species. However, in our barcode tree (Fig. 6), three Neotropical and two Nearctic species of
438 Saphenista form a monophyletic group sister to Aphalonia+Aethes turialba, and this cluster is
439 embedded within the main lineage of Aethes. Aphalonia and Aethes turialba are large,
440 superficially similar species that are quite distinct from Saphenista in facies and genital
441 morphology. Hence, the relationships of Saphenista to these groups in the barcode tree does not
442 seem particularly reliable. The monophyly of Saphenista is convincingly supported by the
443 broadly “Y”- or “T”-shaped distal expansion of the median process of the transtilla. The
444 monotypic Amallectis Meyrick, 1917, for which we have no molecular data, has a median
445 process of the transtilla that is very similar to that of Saphenista, and Razowski (2011) suggested
446 that based on wing venation and male genitalia, the two may be synonymous. Hence, its
447 inclusion in the group is supported by morphology.
448 Saphenista includes 97 described species that are restricted to the New World, occurring in
449 the Nearctic and Neotropical regions. Recorded hosts include Asteraceae, Garryaceae,
450 Rhamnaceae, and Rutaceae (Brown et al., 2018).
451
452 Phalonidia Group
453 The Phalonidia Group is a well-supported (i.e., 87) monophyletic unit comprised of
454 Gynnidomorpha Turner and Phalonidia Le Marchand (Fig. 3). The three species of
455 Gynnidomorpha included in our analysis form a monophyletic group, and Phalonidia is shown
456 as paraphyletic, with Gynnidomorpha embedded within. However, support for this arrangement
20
457 is not particular strong, so the paraphyly of Phalonidia is not altogether convincing. The close
458 relationship of Gynnidomorpha to Phalonidia is strongly supported by a unique circle of spines
459 in the corpus bursae of the female genitalia (Razowski 1985b), somewhat reminiscent of that of
460 Actihema (in our Phtheochroa Group). An extremely similar and probably homologous structure
461 in the corpus bursae of most Spinipogon Razowski (barcodes only) and Pogospinia Brown (not
462 included in the analysis) appears to link these genera, as well, although Spinipogon is sister to
463 Rudenia in our barcode tree (Fig. 6).
464 Gynnidomorpha includes 13 described species and has a broad geographic distribution (i.e.,
465 Europe, Asia, Australia, and North America); recorded larval hosts include numerous families of
466 dicotyledonous plants. Phalonidia, likewise, is nearly worldwide (Europe, Asia, North America,
467 and South America) and encompasses about 88 described species; larval hosts encompass several
468 plant families, but the majority of records are from Asteraceae.
469 Our circumscription of the Phalonidia Group closely parallels that of the Saphenista Group
470 of Razowski (1985b). Additional taxa assigned to the Saphenista Group by Razowski (1985b)
471 but not included in our analysis are the following Neotropical genera: Amallectis, Banhadoa
472 Razowski & Becker, Dinophalia Razowski & Becker, Juxtolena Razowski & Becker,
473 Lasiothyris Meyrick (barcode data), Macasinia Razowski & Pelz, Marylinka Razowski &
474 Becker, Mielkeana Razowski & Becker, Mourecochylis Razowski & Becker, Phaniola Razowski
475 & Becker, Spinipogon, and Thysanphalonia Razowski & Becker. Most of these genera have very
476 few species; seven are monotypic. As mentioned above, Amallectis is probably best assigned to
477 the Saphenista Group based on the shape of the distal expansion of the median process of the
478 transtilla. Although no synapomorphy has been identified to link the remaining genera, they
479 show many similarities in the male and female genitalia. Barcodes (Fig. 6) place Cirrothaumatia
21
480 Razowski & Becker and Honca Pogue in this group; the former likely belongs, but
481 morphological features of the latter argue for its placement elsewhere.
482 In a large number of genera in Cochylina, there is a unique, although variable, fascicle of male
483 secondary scales in a roll along the costa of the hindwing (Baixeras et al., personal communication).
484 Males of nearly all species in the Saphenista, Phalonidia, and Cochylis groups possess this feature.
485 Because it is absent (or different in its development) in the Phtheochroa and Henricus groups, it
486 may represent a synapomorphy for these three generic groups.
487
488 Cochylis Group
489 The Cochylis Group is our second most sampled generic group, with 13 genera and 22
490 species. The monophyly of the group is strongly supported (i.e., 84). Our Cochylis Group
491 includes many of the genera included by Razowski (1970, 1985b) in his Cochylis Group. Genera
492 in common between the two include Cochylis Treitschke, Diceratura Djakonov, Cochylidia
493 Obraztsov, and Falseuncaria Obraztsov & Swatschek. Platphalonidia Razowski, Atroposia
494 Pogue, and Nycthia Pogue are New World genera not included in Razowski’s (1970) study of the
495 Palearctic fauna.
496 Within the Cochylis Group, Platphalonidia, with its type species P. felix (Walsingham), is a
497 well-defined group with Lorita Busck embedded within it. This branch (e.g., Platphalonidia plus
498 Lorita) is sister to the remainder of the group (Fig. 3). Razowski (2011) synonymized
499 Platphalonidia with Phalonidia because he believed that the type species of the former (i.e., P.
500 felix) belonged in Phalonidia. Then he described Platphalonia Razowski to accommodate the
501 remaining species. However, our results strongly support Platphalonidia as distinct from
502 Phalonidia, and we return it to generic status (see Appendix S1). Although Lorita falls within
22
503 Platphalonidia, this placement is based only on COI data and is incorrect based on morphology.
504 Lorita includes four described species occurring in the Nearctic and Neotropical regions. Its
505 monophyly is convincingly supported by the unique ventral “scoop” of the male abdomen
506 (Pogue, 1988).
507 Within the Cochylis Group, females of nearly all species of Platphalonidia have three acanthi
508 in the frenulum, whereas females all of the remaining genera have two (Monsalve et al., 2011). It
509 is remarkable that while this character is highly variable within many species of Cochylina, it is
510 remarkably consistent within the Cochylis Group. Although our multi-gene tree (Fig. 3) places
511 Lorita withiin Platphalonidia, our barcode tree (Fig. 7) shows it as sister to Platphalonidia, the
512 latter of which is much more compelling based on morphology.
513 The monophyly of the lineage that is sister to Platphalonidia+Lorita is also well supported
514 (i.e., 91). Within this group, the three sampled species of Cochylidia form a monophyletic group
515 (support of 55) with Falseuncaria as its sister (support of 89). According to Razowski (2002,
516 2011), synapomorphies for Falseuncaria, Cochylidia, Diceratura, and Cochylis include the
517 presence of a cluster of minute spines in the vesica of the phallus in the male genitalia, and a
518 membranous sack anterior to the sterigma in the female genitalia. Furthermore, according to
519 Razowski (1987c), “Cochylidia is extremely similar to Diceratura and differs from it only by a
520 single supposed autapomorphy, viz. the presence of minute setae or spines at the end of the
521 costal arm of the valva.” The unique costal arm of the valva represents a convincing
522 morphological synapomorphy for the two genera, and the male genitalia of Falseuncaria are
523 quite different from those of Cochylidia and Diceratura. Hence, the sister relationship between
524 Cochylidia and Falseuncaria in our tree (Fig. 3) is not particularly compelling even though the
525 molecular support is strong (i.e., 89).
23
526 Two other species, Cochylidia rupicola (Curtis) and the Japanese C. contumescens
527 (Meyrick), are closely related other based on morphology of the male genitalia, and differ
528 considerably from other Cochylidia. Barcodes of C. rupicola indicate that there is considerable
529 genetic distance between this pair and “true” Cochylidia, and a new genus may be necessary to
530 accommodate these two species.
531 In our multi-gene analysis (Fig. 3) we recovered monophyly for Nycthia, Cochylidia, and
532 Diceratura. However, monophyly was enhanced considerably by the recognition of several
533 subgenera of Cochylis as detailed below.
534 Cochylis Treitschke has long been considered a polyphyletic assemblage of similar appearing
535 species, and our initial results confirm this. Cochylis was subdivided into several subgenera by
536 Razowski (1960), but less than a decade later, all were synonymized by Razowski (1970). More
537 recently, in a review of the Cochylis Group of genera, Razowski (1985b) stated “Almost all
538 subgenera [of Cochylis] require re-examination and probably some of them shall be treated as
539 valid genera.” By elevating some of the subgenera, several species groups are circumscribed that
540 result in more convincing monophyly within our multi-gene tree. Hence, we propose the
541 following modified concepts for the subgenera of Cochylis (see Appendix S2) that have been
542 incorporated in the multi-gene and barcode trees.
543 Cochylis (type species: C. roseana (Haworth)) includes C. roseana and C. flaviciliana
544 (Westwood), which cluster together in our COI tree (Fig. 7). The two species share a slender,
545 deeply notched valva; a slender phallus that lacks cornuti; and a telescopic ovipositor. Although
546 Razowski (1960) indicated that “Several species belong here,” we know of no other species that
547 can be convincingly associated with C. roseana and C. flaviciliana based on morphology. Hence,
548 we propose a significantly narrowed concept of Cochylis (sensu stricto).
24
549 Neocochylis Razowski (type species: C. molliculana (Zeller)), revised status, includes N.
550 molliculana, N. dubitana (Hübner), N. hybridella (Hübner), N. millierana Peyerimhoff) (syn.
551 sannitica Trematerra), and N. salebrana (Mann), which share a broad process of the transtilla
552 and a phallus that lacks cornuti but has spinules. In the multi-gene tree (Fig. 3), C. hybridella and
553 C. dubitana are sisters, and this pair is sister to C. molliculana. Likewise, in our COI tree (Fig.
554 7), C. molliculana, C. hybridella, C. dubitana, and C. millierana form a monophyletic group.
555 Paracochylis Razowski (type species: C. amoenana Kennel), revised status, is considered
556 monotypic, with the single species P. amoenana (syn. apricana Kennel) from Asia Minor. The
557 valva is simple, broad basally and gradually tapering distally, and the phallus has two groups of
558 cornuti plus a patch of spinules. We know of no other species that belong here.
559 Cochylichroa Swatschek (type species: C. atricapitana (Stephens)) was recently returned to
560 generic status by Brown (2019) and includes the following: C. hoffmanana (Kearfott), C. viscana
561 (Kearfott), C. avita (Razowski), C. hospes (Walsingham), and C. foxcana (Kearfott), all from
562 North America, and C. atricapitana from Europe. The species share extremely similar male
563 genitalia with a large rounded sacculus with a large field of spines, and a slender phallus with a
564 group of short spines. A second group of species with quite different genitalia are associated with
565 this species group by barcodes (Fig. 7), i.e., C. arthuri (Dang), C. aurorana (Kearfott), and C.
566 temerana (Busck), and these were also transferred to Cochylichroa by Brown (2019). Given their
567 different genital morphology, their assignment to Cochylichroa may be only an interim solution.
568 Brevicornutia Razowski (type species: C. pallidana Zeller), revised status, is considered to
569 include two species: B. pallidana and B. dolosana (Kennel) (the latter considered a subspecies of
570 pallidana by Brown (2005)). In these two species the cucullus is small and narrow, the sacculus
571 is stout with a heavily sclerotized ridge and teeth, and the phallus has a group of small cornuti.
25
572 Pontoturania Obraztsov (type species: C. defessana Mann), revised status, includes P.
573 defessana, P. maestana Kennel, P. piana (Kennel), P. faustana (Kennel), P. lutosa Razowski, P.
574 indica Razowski, P. aestiva (Walsingham), and P. posterana Zeller. Razowski (1970) included
575 only P. defessana in this subgenus, but in reference to P. posterana he commented that the “male
576 genital armature resembles C. refessana [sic.].” The male genitalia of these species share the
577 following features: 1) phallus with a long, free, terminal process; 2) vesica bearing a dense patch
578 of extremely long, slender, spinelike cornuti; and 3) apex of median process of transtilla with a
579 small patch of teeth.
580 Thyraylia Walsingham (type species: T. bunteana (Robinson)) [with its synonym Acornutia
581 Obraztsov) (type species C. nana (Haworth))] was treated as a valid genus (as its junior synonym
582 Acornutia) in the Phtheochroa Group by Razowski (1960). He (Razowski, 1970, 1997, 2011)
583 later considered Acornutia a synonym of Cochylis. However, Thyraylia has long been considered
584 a valid genus in North America (e.g., Powell, 1983; Metzler & Brown, 2014) with the following
585 species: bana (Kearfott), bunteana (Robinson), discana (Kearfott), gunniana (Busck),
586 hollandana (Kearfott), nana, and voxcana (Kearfott). The included species have an elongate,
587 somewhat upcurved, distally attenuate valva; a sacculus with a long, free distal tip; and a slender
588 phallus that lacks cornuti.
589 Rolandylis was described as monotypic by Gibeaux (1985) based on a single North American
590 species apparently introduced into Europe. Although it was considered a synonym of Cochylis by
591 Razowski (2011), Pogue (1986) recognized it as a distinct North American genus (under the
592 manuscript name “Clothoa”) in his unpublished Ph.D. dissertation, and he (Pogue, 2001) later
593 revised the genus, adding two more species. Metzler & Brown (2014) likewise considered the
594 genus as valid.
26
595 Cochylis epilinana Duponchel is an isolated species that cannot be assigned convincingly to
596 any of these genera, and may require a new genus. In the barcode tree (Fig. 7) it occupies a
597 position sister to Cochylis (sensu stricto). In addition, there are many species currently included
598 in Cochylis for which we have no convincing generic assignments, and hence for the time being
599 must remain in a large polyphyletic Cochylis (sensu lato).
600 The three most basal genera in the Cochylis Group (i.e., Platphalonidia, Atroposia, and
601 Nycthia) are restricted to the New World. Platphalonidia includes 21 species, with Asteraceae as
602 the most common foodplants. Atroposia is monotypic, feeding on Onagraceae. And Nycthia
603 includes two described species, both of which specialize on Agavaceae. Hence, the three genera
604 have little in common biologically.
605 Cochylidia is recorded from Europe and Asia, with one species (C. subroseana (Haworth))
606 either Holarctic or introduced to North America inadvertently. With very few exceptions, the
607 larvae of Cochylidia feed on Asteraceae. Falseuncaria includes seven species that occur in
608 Europe, Asia, and Africa; foodplants have been reported for only two species (i.e., F. ruficiliana
609 (Haworth) and F. degreyana (McLachlan)), encompassing Scrophulariaceae, Plantaginaceae,
610 Primulaceae, and Asteraceae.
611 Genera not included in our molecular analyses that likely belong in the Cochylis Group
612 include the following: Cochylidichnium Razowski, Coristaca Razowski, Geitocochylis
613 Razowski, Gryposcleroma Razowski, Mimcochylis Razowski, Monoceratuncus Razowski,
614 Revertuncaria Razowski, and Rolandylis.
615
616 Summary and Conclusions
617
27
618 Based on our multi-gene analysis, we recognize six major monophyletic lineages within
619 Cochylina: a Phtheochroa Group, a Henricus Group, an Aethes Group, a Saphenista Group, a
620 Phalonidia Group, and a Cochylis Group. Our ML analysis of DNA barcodes resulted in a tree
621 that is much less resolved, but provides insight into relationships among species and genera
622 within the major lineages identified in the multi-gene analysis, especially those in the Cochylis
623 Group.
624 The Phtheochroa Group is sister to the remaining Cochylina. Its members retain the most
625 plesiomorphic morphological characters of the subtribe, and most appear to be internal feeders
626 (crown borers, gall-inducers, fruit-feeders) as larvae, primarily in Asteraceae. The monotypic
627 Hysterophora is embedded within Phtheochroa, but this position is only weakly supported. The
628 relationship among Agapeta, Ceratoxanthis, and Fulvoclysia is well resolved, but Eugnosta and
629 Cochylimorpha are both represented by two widely separated branches, suggesting that neither is
630 monophyletic as currently defined. In the case of Eugnosta, the disjunct position of E.
631 magnificana is puzzling. In the case of Cochylimorpha, the clade represented by C. alternana
632 may be a distinct genus for which the name Euxanthoides is available.
633 The Henricus Group, with only two genera included in our analysis, is well supported and is
634 sister to the remaining generic groups (after the Phtheochroa Group). Most of the species for
635 which hosts are known feed in the seeds and cones of conifers, primarily Pinaceae and
636 Cupressaceae. The Aethes Group is comprised of a relatively well-supported, monophyletic
637 Aethes with Rudenia as its sister. However, the position of Rudenia is weakly supported and is
638 not particularly consistent with morphological evidence. Hence, it is possible that Rudenia is
639 incorrectly placed in the Aethes Group. The Saphenista Group is a small, monophyletic lineage
640 comprised of Saphenista and Amallectis, the latter included in the group based on morphology.
28
641 There are many inconsistencies between the multi-gene tree and the barcode tree, and the
642 position of the genus Saphenista embedded within the genus Aethes in the barcode tree is almost
643 certainly incorrect. In the barcode tree, Aethes turialba, which is a member of a small group of
644 exceptionally large and superficially similar species of Aethes, clusters with Aphalonia, and this
645 relationship is convincing based on facies and morphology of the male genitalia.
646 The Phalonidia Group is well supported, comprised of Gynnidomorpha and Phalonidia.
647 However, the three species of Gynnidomorpha included in our analysis form a monophyletic
648 group embedded within a paraphyletic Phalonidia. The association of Gynnidomorpha and
649 Phalonidia is strongly supported by morphological characters, especially features of the female
650 genitalia.
651 The Cochylis Group includes several genera that are convincingly supported as monophyletic
652 (e.g., Nycthia, Diceratura, Cochylidia), however, a few compelling relationships based on
653 morphology are not captured in our multi-gene tree. For example, Diceratura and Cochylidia are
654 assumed to be sisters based on synapomorphies of the male genitalia, but they are widely
655 separated in our tree. Platphalonidia is shown to be distinct from its putative senior synonym
656 Phalonidia, but Lorita falls between the two species groups of Platphalonidia (Appendix S1). To
657 no surprise, species assigned to the polyphyletic genus Cochylis were scattered nearly throughout
658 the Cochylis Group. However, by elevating many of the previously proposed subgenera of
659 Cochylis (e.g., Pontoturania, Neocochylis, Brevicornutia), we are able to define several
660 monophyletic groups, thus providing better resolution to relationships within the Cochylis
661 Group. Unfortunately, many species of Cochylis cannot be convincingly associated with one of
662 the subgenera, and additional phylogenetic studies are required to place these “leftover” species.
663
29
664 Author contributions
665
666 MM and MH designed the study and conducted the analyses, and JB and LA interpreted the
667 results and prepared the initial draft of the paper. MM and MH provided text on methods, and all
668 authors commented on various drafts of the manuscript. JB, LA, and RB provided specimens.
669 MH prepared the illustrations. The authors declare that there are no conflicting financial
670 interests.
671
672 Supporting Information
673
674 Additional supporting information may be found online in the Supporting Information section
675 oat the end of the article.
676
677 Appendix S1. Explanation of revised status for Platphalonidia
678 Appendix S2. Proposed taxonomic changes and arrangement of genera related to Cochylis.
679 Table S1. Taxa and genes used in phylogenetic analysis.
680 Table S2. Sequence lengths for genes used in phylogenetic analysis.
681 Figure S1. Bayesian analysis of Cochylini and out-groups.
682
683
684 Acknowledgements
685
30
686 We thank David Agassiz for specimens of Cochylini from Africa; Pasquale Trematerra and
687 Kai Berggren for filling gaps in our sample of European species; and John Phillips, who
688 provided the type species of Phtheochroa, P. rugosana. We also thank all of those who
689 contributed specimens of Cochylini to BOLD and generously shared barcode data with us,
690 especially Yves Bassett, Sophia Gripenberg, Jean-François Landry, Daniel Janzen, Winnie
691 Hallwachs, and Scott Miller. We also express our gratitude to Paul Hebert, Jeremy DeWaard,
692 Allison Brown, and other staff of the Biodiversity Institute of Ontario, University of Guelph,
693 who provided considerable assistance accessing barcode data. Laura Törmälä is thanked for
694 supervising the lab work, and Vlad Dinca for kindly lending a hand in the lab. Funding for this
695 study was provided by the Academy of Finland (MM). The following provided helpful reviews
696 that enhanced the clarity and quality of the paper: Joaquin Baixeras (University of Valencia),
697 Jason Dombroskie (Cornell University), and Józef Razowski (Polish Academy of Sciences).
698
699 Literature Cited
700
701 Aarvik, L. (2010) Review of East African Cochylini (Lepidoptera, Tortricidae) with description
702 of new species. Norwegian Journal of Entomology, 57, 81–108.
703
704 Bradley, J.D., Tremewan, W.G. and Smith, A. (1973) British tortricoid moths. Cochylidae and
705 Tortricidae: Tortricinae. The Ray Society, London. 251 pp.
706
707 Brown, J.W. (2005) World catalogue of insects. Volume 5: Tortricidae (Lepidoptera). Apollo
708 Books, Stenstrup. 741 pp.
31
709
710 Brown, J.W. (2013) A new genus of pine-feeding Cochylina from the western United States and
711 northern Mexico (Lepidoptera: Tortricidae: Euliini). Zootaxa, 3640, 270–283.
712
713 Brown, J.W. (2016) A new generic assignment for Tortrix baboquavariana Kearfott, 1907
714 (Lepidoptera: Tortricidae) with comments on its tribal assignment. Journal of the Lepidopterists’
715 Society, 70, 173–175.
716
717 Brown, J. W., Robinson, G. & Powell, J. A. (2018) Food plant database of the leafrollers of the
718 world (Lepidoptera: Tortricidae) (Version 1.0). http://www.tortricid.net/foodplants.asp.
719
720 Brown, J.W. (2019) New genera, new species, and new combinations in New World Cochylina
721 (Lepidoptera: Tortricidae: Tortricinae). Zootaxa, in press.
722
723 Brown, J.W., Copeland, R.S., Aarvik, L., Luke, Q., Miller, S. & Rosati, M. (2014) New host
724 records for fruit-feeding Afrotropical Tortricidae (Lepidoptera). African Entomology, 22, 343–
725 376.
726
727 Brown, J.W. & Powell, J.A. (1991) Systematics of the Chrysoxena group of genera
728 (Lepidoptera: Tortricidae: Euliini). University of California Publications in Entomology, 111, 1‒
729 87.
730
32
731 Brown, J.W. & Razowski, J. (2003) Description of Ptychocroca, a new genus from Chile and
732 Argentina, with comments on the Bonagota Razowski group of genera (Lepidoptera: Tortricidae:
733 Euliini). Zootaxa, 303, 1–31.
734
735 Brown, J.W., Robinson, G. & Powell, J.A. (2018) Food plant database of the leafrollers of the
736 world (Lepidoptera: Tortricidae) (Version 1.0). http://www.tortricid.net/foodplants.asp.
737
738 Clarke, J.F.G. (1963) Catalogue of the Type Specimens of Microlepidoptera in the British
739 Museum (Natural History) described by Edward Meyrick, Volume 4. Trustees of the British
740 Museum, London. 521 pp.
741
742 Clarke, J.F.G. (1968) Neotropical Microlepidoptera, XVII. Notes and new species of
743 Phaloniidae. Proceedings of the United States National Museum, 125 (3600), 1–58.
744
745 DeWaard, J.R., Ivanova, N.V., Hajibabaei, M. & Hebert, P.D.N. (2008) Assembling DNA
746 barcodes: analytical protocols. Methods in Molecular Biology: Environmental Genetics (ed. By
747 C. Martin), pp. 275–294. Humana Press, Totowaa, New Jersey,.
748
749 Diakonoff, A. (1961) Taxonomy of the higher groups of the Tortricoidea. XI Internationalen
750 Kongresses fur Entomologie: Wien, 17, bis 25, August 1960 (Volume 1) (ed. by H. Stouhal).
751 Verhandlungen, pp. 124–126.
752
33
753 Drummond A.J. & Rambaut, A. (2007) BEAST: Bayesian evolutionary analysis by sampling
754 trees. BMC Evolutionary Biology, 7, 214.
755
756 Falkovitsh, M.I. (1963) New species of the family Cochylidae (Lepidoptera) from Kazakhstan
757 and the Caucasus. Zoologicheskiĭ Zhurnal Moskva, 42, 697–703.
758
759 Gibeaux, C. (1985) Description d'un genre et de trois especes de tordeuses nouveaux pour la
760 France (Lep. Tortricidae, Cochylini). Entomologica Gallica, 1, 346–351.
761
762 Gilligan, T.M., Baixeras, J. & Brown, J.W. (2018) T@RTS: Online World Catalogue of the
763 Tortricidae (Ver.4.0). http://www.tortricid.net/catalogue.asp.
764
765 Guenée, M.A. (1845) Essai sur une nouvelle classification des Microlepidopteres et catalogue
766 des especes Europennes. Annales de la Société Entomologique de France, (2) 3, 297–344.
767
768 Hall, T. (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis
769 program for Windows 95/98/NT. Nucleic Acids Symposium Series, 41, 95–98.
770
771 Heikkilä, M., Mutanen, M., Wahlberg, N., Sihvonen, P. & Kaila, L. (2015) Elusive ditrysian
772 phylogeny: An account of combining systematized morphology with molecular data
773 (Lepidoptera). BMC Evolutionary Biology, 15 (260), 1–27. https://doi.org/10.1186/s12862-015-
774 0520-0
775
34
776 Horak, M. (1999) Tortricoidea. Lepidoptera, moths and butterflies. Volume 1: Evolution,
777 systematics, and biogeography. Handbook of Zoology 4 (35), Arthropoda: Insecta (ed. By N.
778 Kristensen), pp. 199–215. Walter de Gruyter, Berlin & New York,
779
780 Horak, M. & Brown, R.L. (1991) 1.2 Taxonomy and phylogeny. Tortricid Pests, Their Biology,
781 Natural Enemies and Control (ed. by L.P.S. van der Geest & H. H. Evenhuis), pp. 23–48.
782 Elsevier Science Publishers, Amsterdam.
783
784 Kozlov, A.M., Darriba, D., Flouri, T., Morel, B. & Stamatakis, A. (2018) RAxML-NG: A fast,
785 scalable, and user-friendly tool for maximum likelihood phylogenetic inference. bioRxiv.
786 doi:10.1101/447110
787
788 Kuznetsov, V.I. & Stekolnikov, A.A. (1973) Phylogenetic relationships in the family Tortricidae
789 (Lepidoptera) treated on the base of study of functional morphology of genital apparatus. Trudy
790 Russkago Entomologischeskago Obschchestva, 56, 18–43. [In Russian]
791
792 Kuznetsov, V I. & Stekolnikov, A.A. (1977) Functional morphology of the male genitalia and
793 phylogenetic relationships of some tribes in the family Tortricidae (Lepidoptera) fauna of the Far
794 East. Trudy Zoologicheskogo Instituta Leningrad, 70, 65–97.
795
796 MacKay, M.R. (1959) Larvae of the North American Olethreutidae (Lepidoptera). Canadian
797 Entomologist Supplement, 10, 1–338.
798
35
799 MacKay, M.R. (1962) Larvae of the North American Tortricinae (Lepidoptera: Tortricinae).
800 Canadian Entomologist Supplement, 28, 1–182.
801
802 Metzler, E. (2000) Two new species of Cochylini (Lepidoptera: Tortricidae: Tortricinae) from
803 the eastern United States. The Great Lakes Entomologist, 32 (1999), 185–197.
804
805 Metzler, E.H. & Brown, J.W. (2014) An updated check list the Cochylina (Tortricidae,
806 Tortricinae, Euliini) of North America north of Mexico including Greenland, with comments on
807 classification and identification. Journal of the Lepidopterists’ Society, 68, 274–282.
808
809 Metzler, E. & Sabourin, M. (2002) A new species of Spinipogon Razowski, 1967 from two
810 remnant prairies in Ohio (Lepidoptera: Tortricidae, Cochylini). Fabreries, 27, 69–76.
811
812 Meyrick, E. (1882) Descriptions of New Zealand Micro-lepidoptera. New Zealand Journal of
813 Science, 1, 159–162.
814
815 Meyrick, E. (1895) A handbook of British Lepidoptera. London: MacMillan, 843 pp.
816
817 Miller, M.A., Pfeiffer, W., & Schwartz, T. (2010) Creating the CIPRES Science Gateway for
818 inference of large phylogenetic trees, Proceedings of the Gateway Computing Environments
819 Workshop (GCE), 14 November 2010, New Orleans, Louisiana, pp. 1–8.
820
36
821 Monsalve, S., Dombroskie, J., Lam, W., Rota, J., & Brown, J.W. (2011) Variation in the female
822 frenulum in Tortricidae (Lepidoptera). Part 3. Tortricinae. Proceedings of the Entomological
823 Society of Washington, 113, 335–370.
824
825 Morris, F.O. (1898) A natural history of British moths, volume 3, 1–223. London.
826
827 Mutanen, M., Wahlberg, N. & Kaila, L. (2010) Comprehensive gene and taxon coverage
828 elucidates radiation patterns in moths and butterflies. Proceedings of the Royal Society B, 277
829 (1695), 2839–2848. https://doi.org/10.1098/rspb.2010.0392
830
831 Obraztsov, N. S. (1967) Genera Tortricoidarum. Check list of genera and subgenera belonging to
832 the families Tortricidae (Ceracidae, Chlidanotidae, Schoenotenidae and Olethreutidae included)
833 and Phaloniidae. Journal of the New York Entomological Society, 75, 2–11.
834
835 Pogue, M.G. (1986) A Generic Revision of the Cochylidae (Lepidoptera) of North America.
836 Ph.D. thesis, University of Minnesota.
837
838 Pogue, M.G. (1988) Revision of the genus Lorita Busck (Lepidoptera: Tortricidae: Cochylini),
839 with a description of a new species. Proceedings of the Entomological Society of Washington,
840 90, 440–457.
841
37
842 Pogue, M.G. (2001) Revised status of Rolandylis Gibeaux (Lepidoptera: Tortricidae: Cochylini),
843 with descriptions of two new species from North America. Proceedings of the Entomological
844 Society of Washington, 103, 788–796.
845
846 Pogue, M.G. & Friedlander, T.P. (1987) Cochylis caulocatax Razowski (Lepidoptera:
847 Tortricidae: Cochylini): a redescription of the male with descriptions of the female, larva, and
848 pupa. Proceedings of the Entomological Society of Washington, 95, 320–327.
849
850 Pogue, M.G. & Mickevitch, M.F. (1990) Character definitions and character state delineation:
851 the bete noire of phylogenetic inference. Cladistics, 6, 319–361.
852
853 Powell, J.A. 1964. Biological and taxonomic studies on tortricine moths, with reference to the
854 species in California. University of California Publications in Entomology, 32, 1–317.
855
856 Powell, J.A. (1983) Tortricoidea. Check list of the Lepidoptera of America north of Mexico (ed.
857 by R. W. Hodges), pp. 31–42. E.W. Classey, Ltd., and Wedge Entomological Research
858 Foundation, London.
859
860 Powell, J.A. (1986) Synopsis of the classification of Neotropical Tortricinae, with descriptions of
861 new genera and species (Lepidoptera: Tortricidae). Pan-Pacific Entomologist, 62, 372–398.
862
863 Ratnasingham, S. & Hebert, P.D.N (2013) A DNA-based registry for all animal species: The
864 Barcode Index Number (BIN) System. PLoS ONE, 8: e66213.
38
865
866 Razowski, J. (1960) Studies on the Cochylidae (Lepidoptera). Part II. The genera of the
867 Palearctic Cochylidae. Polskie Pismo Entomologiczne, 30, 281–356.
868
869 Razowski, J. (1967a) South American Cochylidae (Lepidoptera) from the collection of the
870 British Museum (Natural History). Acta Zoologica Cracoviensia, 12, 163–210.
871
872 Razowski, J. (1967b) Afghanische Wickler-Arten (Lepidoptera: Tortricidae und Cochylidae).
873 Beitrage zur Naturkundlichen Forschung in Sudwestdeutschland, 26, 91–108.
874
875 Razowski, J. (1968a) Revision of the generic group Agapeta Hubner (Lepidoptera, Cochylidae).
876 Acta Zoologica Cracoviensia, 13, 73–100.
877
878 Razowski, J. (1968b) Revision of the genus Eupoecilia Stephens (Lepidoptera, Cochylidae). Acta
879 Zoologica Cracoviensia, 13, 103–130.
880
881 Razowski, J. (1968c) Revision of the generic group Cochylis Treitschke (Lepidoptera,
882 Cochylidae). Acta Zoologica Cracoviensia, 13, 131–147.
883
884 Razowski, J. (1970) Cochylidae. Microlepidoptera Palaearctica, volume 3 (ed. by H.G. Amsel,
885 F. Gregor & H. Reiser). Verlag G. Fromme & Co., Wien, 528 pp. + xxx.
886
39
887 Razowski, J. (1976) Phylogeny and system of Tortricidae (Lepidoptera). Acta Zoologica
888 Cracoviensia, 21, 73–120.
889
890 Razowski, J. (1984) Descriptions of the Neotropical Cochylidii (Lepidoptera, Tortricidae).
891 Annales Zoologici, 38, 275–280.
892
893 Razowski, J. (1985a) Descriptions of Rudenia gen n. and its two new species (Lepidoptera,
894 Tortricidae). Polskie Pismo Entomologiczne, 55, 519–522.
895
896 Razowski, J. (1985b) On the generic groups Saphenista and Cochylis (Tortricidae). Nota
897 Lepidopterologica, 8, 55–60.
898
899 Razowski, J. (1986) New and little known Neotropical Cochylidii (Lepidoptera, Tortricidae).
900 Acta Zoologica Cracoviensia, 29, 373–396.
901
902 Razowski, J. (1987a) New taxa of Archipini (Lepidoptera: Tortricidae) from South America.
903 Tinea, 12 (supplement), 123–138.
904
905 Razowski, J. (1987b) Descriptions of new Neotropical genera of Archipini and rectification of
906 the Deltinea problem (Lepidoptera: Tortricidae). Sciences Nat, 52 (1986), 21–25.
907
908 Razowski, J. (1987c) The genera of Tortricidae (Lepidoptera). Part 1. Palearctic Chlidanotinae
909 and Tortricinae. Acta Zoologica Cracoviensia, 30, 141-355.
40
910
911 Razowski, J. (1988) New genera and species of the Neotropical Archipini (Lepidoptera:
912 Tortricidae). Acta Zoologica Cracoviensia, 31, 387–422.
913
914 Razowski, J. (1989) Descriptions of four Neotropical taxa of Cochylini (Lepidoptera:
915 Tortricidae). SHILAP Revista de Lepidopterologia, 17 (66), 205–208.
916
917 Razowski, J. (1990) Contribution to Cochylini fauna of Mexico (Lepidoptera: Tortricidae).
918 SHILAP Revista de Lepidopterologia, 18, 337–345.
919
920 Razowski, J. (1992) Descriptions of some Neotropical Euliini and Archipini (Lepidoptera,
921 Tortricidae). Miscellanea Zoologica, 14, 105–114.
922
923 Razowski, J. (1993) Cochylini (Lepidoptera, Tortricidae) from Peru and Bolivia. Acta Zoologica
924 Cracoviensia, 36, 161–181.
925
926 Razowski, J. (1994) Synopsis of the Neotropical Cochylini (Lepidoptera: Tortricidae). Acta
927 Zoologica Cracoviensia, 37, 121–320.
928
929 Razowski, J. (1995) Catalogue of the species of Tortricidae (Lepidoptera). Part III. Afrotropical
930 Chlidanotinae and Tortricinae: Phricanthini, Cochylini and Tortricini. Acta Zoologica
931 Cracoviensia, 38, 183–193.
932
41
933 Razowski, J. (1997) Cochylini (Lepidoptera: Tortricidae) of Canada. Acta Zoologica
934 Cracoviensia, 40, 107–163.
935
936 Razowski, J. (2002) Tortricidae of Europe. Part I. Tortricinae and Chlidanotinae. Frantisek
937 Slamka, Bratislava, 247 pp.
938
939 Razowski, J. (2011) Diagnoses and remarks on genera of Tortricidae, 2: Cochylini (Lepidoptera:
940 Tortricidae). SHILAP Revista de Lepidopterologia, 39, 397–414.
941
942 Razowski, J. & Becker, V.O. (1983) Brazilian Cochylidii (Lepidoptera: Tortricidae). Acta
943 Zoologica Cracoviensia, 26, 421–464.
944
945 Razowski, J. & Wojtusiak, J. (2008) Tortricidae from the mountains of Ecuador. Part III.
946 Western Cordillera. Genus, 19, 497–575.
947
948 Regier, J.C., Brown, J.W., Mitter, C., Baixeras, J., Cho, S., Cummings, M.P. & Zwick, A. (2012)
949 A molecular phylogeny for the leaf-roller moths (Lepidoptera: Tortricidae) and its implications
950 for classification and life history evolution. PLoS ONE, 7 (4): e35574.
951
952 Ronquist, F. & Huelsenbeck, J.P. (2003) MrBayes 3: Bayesian phylogenetic inference under
953 mixed models. Bioinformatics, 19, 1572–1574.
954
42
955 Sabourin, M., Miller, W.E., Metzler, E.H. & Vargo, J.T. (2002) Revised identities and new
956 species of Aethes from, midwestern North America. Journal of the Lepidopterists’ Society, 56,
957 216–233.
958
959 Stainton, H.T. (1859) A Manual of British Butterflies and Moths, Volume 2. London. 475 pp.
960
961 Stamatakis, A. (2014). RAxML Version 8: A tool for phylogenetic analysis and post-analysis of
962 large phylogenies. Bioinformatics, 30 (9), 1312–1313.
963
964 Stephens, J.F. (1852) List of the specimens of British animals in the collection of the British
965 Museum, 10 (Lepidoptera – continued), 1–120.
966
967 Swatschek, B. (1958) Die larval systematik der wickler (Tortricidae and Carposinidae) aus dem
968 zoologischen Institut der Universitat Erlangen. Akademie-Verlag, Berlin. 269 pp.
969 [Abhandlungen zur larvalsystematik Insekten 3.]
970
971 Wahlberg, N. & Wheat, C.W. (2008) Genomic outposts serve the phylogenomic pioneers:
972 designing novel nuclear markers for genomic DNA extractions of Lepidoptera. Systematic
973 Biology, 57, 231–242.
974
975
976
43
977 978 Explanation of Figures 979 980 Figure 1. Phylogenetic tree of Palearctic genera of Cochylini from Razowski (1970). 981 982 Figure 2. Phylogenetic tree of North American Cochylini from Pogue & Mickevitch (1990). 983 984 Figure 3. Multi-gene phylogenetic tree of Cochylini and out-groups. *Lorita scarifacata COI
985 data concatenated from BIOUG01827-G12 (COI barcode region) and MH_USNM_18 (COI-
986 end). **Neocochylis molliculana data concatenated from IT_3 (CAD, EF1a-begin and EF1a-
987 end), and TLMF_Lep_23781 (COI barcode region).
988 Figure 4. ML barcode tree of Phtheochroa Group species. 989 990 Figure 5. ML barcode tree of Henricus Group species. 991 992 Figure 6. ML barcode tree of Aethes, Saphenista, and Phalonidia Group species. 993 994 Figure 7. ML barcode tree of Cochylis Group species. 995
44
Fig. 1. Fig. 2.
Eugnosta Hysterosia Henricus Hysterophora Phalonidia Prohysterophora Aethes Phtheochroa Morta Stenodes Spinipogon Phalonidia Honca Ceratoxanthis Cagiva Fulvoclysia Clothoa Agapeta Poterioparvus Phtheochroides Cochylis Cochylidae Eugnosta Cochylidia Eupoecilia Atroposia Commophila Ceratunucus Prochlidonia Cybilla Aethes Nycthia Cochylidia Rudenia Diceratura Cochylichroa Cochylis Lorita Cryptocochylis Chlidanotini Falseuncaria Trachysmia Aprepodoxa Tortricini
Razowski (1970) Pogue & Mickevitch (1990)
Figure 1. Phylogenetic tree of Palearctic genera of Cochylini from Razowski (1970).
Figure 2. Phylogenetic tree of North American Cochylini from Pogue & Mickevitch (1990).
68 Phtheochroa aegrana_CGWC-3927 32 Phtheochroa inopiana_MM15670 4 Phtheochroa modestana_08BBLEP-04263 99 Phtheochroa waracana_08BBLEP-01868 Phtheochroa birdana_10BBCLP-2045 99 Phtheochroa riscana_BIOUG01602-A08 38 Phtheochroa vulneratana_MM04162 Phtheochroa vitellinana_NoA-08-150 99 Phtheochroa aureoalbida_08BBLEP-01889 99 82 Phtheochroa vitellinana_08BBLEP-04018 Phtheochroa birdana_DNA-ATBI-6188 99 10 Phtheochroa terminana_DNA-ATBI-6191 Phtheochroa_04HBL006520 Phtheochroa pulvillana_BC ZSM Lep 79080 Phtheochroa huachucana_CCDB-20277-C09 22 Phtheochroa schreibersiana_TLMF Lep 22918 Phtheochroa_TLMF Lep 25354 18 59 Phtheochroa cantabriana_TLMF Lep 08336 88 Phtheochroa_TLMF Lep 25353 Phtheochroa frigidana_TLMF Lep 08349 99 Phtheochroa drenowskyi_TLMF Lep 00934 Phtheochroa apenninana_TLMF Lep 03037 Phtheochroa schawerdae_TLMF Lep 05081 59 Phtheochroa duponchelana_NHMO Lep08295 98 Phtheochroa cypriana_IB139 10 39 55 Phtheochroa sodaliana_MM17752 Phtheochroa rugosana_TLMF Lep 03223 41 Phtheochroa syrtana_MM24104 Phtheochroa procerana_BCBZ 0480 33 50 Phtheochroa purana_TLMF Lep 23756 99 Phtheochroa unionana_BCBZ 0478 99 Phtheochroa unionana_BCBZ 0477 Eugnosta percnoptila_USNM ENT 00676743
53 Phtheochroa aarviki_USNM ENT 00676737 79 Actihema hemiacta_USNM ENT 00808473 99 Actihema msitumi_MM24042 96 Eugnosta bimaculana_MDOK-0922 99 Eugnosta sartana_06-FLOR-0761 1 93 Eugnosta sartana_06-JBTOR-0112 36 Eugnosta brownana_CCDB-20277-F10 Eugnosta_06-FLOR-0803 31 99 Eugnosta erigeronana_MDOK-2264 Eugnosta_CCDB-22959-B06 99 Eugnosta_CCDB-22959-B03 Cochylimorpha agenjoi_MM24103 94 Cochylimorpha hilarana_BC ZSM Lep 28683 Cochylimorpha erlebachi_TLMF Lep 04349 14 99 Cochylimorpha halophilana_TLMF Lep 23758 Cochylimorpha woliniana_TLMF Lep 18430 Cochylimorpha discopunctana_TLMF Lep 21182 1 49 99 Cochylimorpha fucatana_KLM Lep 06382 47 Cochylimorpha jucundana_TLMF Lep 23761 48 Cochylimorpha decolorella_TLMF Lep 25051 Cochylimorpha perturbatana_KLM Lep 06353 24 Cochylimorpha_MM25920 99 Cochylimorpha_MM25921 55 Cochylimorpha cultana_TLMF Lep 03177 99 Eugnosta n. sp._CCDB-28998-A07 61 Eugnosta n. sp._CCDB-20822-G12 1 75 Eugnosta deceptana_CCDB-20822-H03 71 Eugnosta mexicana_JWB-08-0214 13 Eugnosta beevorana_CCDB-28998-A12 99 Eugnosta busckana_06-JBTOR-0102 Eugnosta sp._KM023733 99 Cochylimorpha straminea_BC ZSM Lep 50992 36 Cochylimorpha straminea_TLMF Lep 03196 1 22 Cochylimorpha alternana_MM17866 4 Cochylimorpha_MM25898 99 Cochylimorpha perfusana_MM25903 Cryptocochylis conjunctana_CCDB-30812-E06 3 Cochylimorpha meridiana_TLMF Lep 23759 99 Eugnosta magnificana_MM24102 58 12 Eugnosta magnificana_KN00236 Fulvoclysia nerminae_KLM Lep 00329
16 Agapeta hamana_MM11780 99 Agapeta zoegana_BC ZSM Lep 23108 97 Agapeta zoegana_TLMF Lep 13440
99 Agapeta zoegana_HLC-23776 Agapeta zoegana_TLMF Lep 05267 63 Agapeta zoegana_MM12372 82 Agapeta zoegana_UKLB43C02 Eupoecilia acrographa_11ANIC-08971
42 Eupoecilia ambiguella_MM06468 68 Eupoecilia angustana_MM11846 52 Eupoecilia sanguisorbana_MM18869 77 Henricus fuscodorsana_10BBCLP-2190 55 Henricus phillipsEPR01_INB0004016107 26 Henricus phillipsEPR11_INB0003756515 57 Henricus infernalis_CCDB-20277-A06 99 Henricus macrocarpana_CCDB-20277-A11 Eupinivora ponderosae_JWB-08-0213 Henricus BioLep49_INB0003401687 22 Henricus comes_CCDB-20277-A03 40 Henricus platinus_INB0003339690 31 99 Henricus cognatus_CCDB-20275-H10 Henricus cognatus_CCDB-20275-H11 42 Henricus edwardsiana_MDOK-3878 74 Henricus umbrabasanus_CNCLEP00077906 99 Aethes citreoflavus_SWC-06-0255 98 Aethes kyrkii_MM10685 27 Aethes cnicana_MM01068 Aethes moribundana_TLMF Lep 01548 Aethes JB001_JWB-08-0174 5 99 Aethes_CNCLEP00007055 Aethes kindermanniana_MM13281 79 Aethes aurofasciana_TLMF Lep 00836 99 Aethes smeathmanniana_MM06319 42 Aethes decimana_TLMF Lep 00862 Aethes languidana_TLMF Lep 03199 0 Aethes rutilana_MM14588 24 Aethes margaritana_MM14117 23 Phalonidia lepidana_DNA-ATBI-0115 99 Aethes caucasica_TLMF Lep 11463 5 Cirrothaumatia vesta_CCDB-30812-E11 24 Aethes hartmanniana_MM14121 0 31 Lasiothyris luminosa_KU682789 Aethes tesserana_MM05753 18 Honca grandis_CCDB-20277-B05 Aethes triangulana_MM15676 Phalonidia curvistrigana_MM02414 Aethes_BC ZSM Lep 95109 69 33 Phalonidia gilvicomana_TLMF Lep 12054 50 0 7 Aethes deutschiana_MM00084 Phalonidia memoranda_jflandry0050 99 Aethes deutschiana_BIOUG06196-B09 Phalonidia albipalpana_TLMF Lep 05004 Aethes deutschiana_09PROBE-09448 15 99 82 Phalonidia affi nitana_TLMF Lep 23763 Aethes_10BBCLP-2345 49 97 Phalonidia manniana_MM14232 Aethes deutschiana_10BBCLP-2630 94 Phalonidia udana_RMNH.INS.550068 Aethes ardezana_TLMF Lep 01542 Phalonidia udana_MM10088 92 73 Aethes bilbaensis_TLMF Lep 03198 Phalonidia manniana_MM20726 59 20 Aethes bilbaensis_TLMF Lep 06438 Phalonidia manniana_TLMF Lep 05103 0 Aethes francillana_MM17590 88 75 Gynnidomorpha minimana_MM08602 98 Aethes fennicana_MM23220 97 Gynnidomorpha permixtana_MM02555 24 Aethes fennicana_MM23218 Gynnidomorpha sp._10BBCLP-3098 Aethes_MM24238 86 Gynnidomorpha luridana_MM17589 28 Aethes beatricella_MM24233 Gynnidomorpha mesoxutha_11ANIC-08975 64 Aethes dilucidana_BC ZSM Lep 38039 74 Gynnidomorpha alismana_TLMF Lep 08448 99 Aethes turialba_INB0003446750 15 98 Gynnidomorpha alismana_MM19265 6 Aethes turialba_INB0003520947 98 Gynnidomorpha vectisana_MM10086 Aphalonia praeposita_CCDB-30812-F01 0 Aethes margarotana_BC ZSM Lep 38033 0 Saphenista sp. JB-003_JWB-08-0204 99 Aethes williana_UKLB34E05 Saphenista n. sp._UBC-2007-0713 0 30 81 Cochylimorpha_KN00742 Saphenista_BIOUG25372-B03 83 11 Phalonidia contractana_TLMF Lep 03210 Saphenista phillipsEPR01_INB0003334513 96 Phtheochroa_CCDB-28565-E06 Saphenista sp. JB-001_JWB-08-0202 1 Spinipogon triangularis_SG-BCISP2337 Aethes monera_NoA-08-143 Rudenia leguminana_MDOK-1981 33 95 Aethes_BIOUG01451-C03 Rudenia leguminana_IAWAZ-0436 99 Aethes baloghi_MDOK-2942 Rudenia leguminana_08-JWB-0004 Aethes_CNCLEP00078339 73 30 25 89 Rudenia leguminana_08-JWB-0007 Aethes spartinana_CCDB-24281-B02 98 Rudenia leguminana_08-JWB-0013 Aethes bomonana_CCDB-24281-A08 23 50 Rudenia leguminana_BIOUG01928-A09 Aethes interruptofasciata_BIOUG07624-H04 49 35 Rudenia_JWB-08-0188 46 Aethes_BIOUG21145-C04 91 Rudenia leguminana_08-JWB-0003 41 Aethes promptana_MDOK-0932 96 Aethes floccosana_DNA-ATBI-6166 69 Aethes mymara_CNCLEP00041395 23 Aethes atomosana_CNCLEP00025154 Aethes razowskii_BIOUG03504-H09 40 99 Aethes promptana_JWB-05-0018 99 Aethes razowskii_CCDB-22958-A12 Aethes ringsi_CCDB-20822-C06 58 Aethes baloghi_CNCLEP00041396 24 99 Aethes seriatana_MDOK-4015 0 Aethes angustana_HLC-16811 99 Aethes_10BBCLP-2977 Aethes angulatana_SNS10IL-00892 61 Aethes rana_BIOUG35498-C04 59 Aethes_DNA-ATBI-2264 66 Aethes argentilimitana_10BBCLP-0769 99 Aethes mymara_BIOUG33271-B10 35 Aethes mymara_BIOUG08611-F06 45 Aethes biscana_2005-ONT-2080 Aethes sexdentata_BIOUG01690-A06 30 Aethes matheri_jflandry2160 57 Aethes_BIOUG08429-E11 55 76 Aethes angulatana_PPBP-0208 48 Aethes_BIOUG09769-F08 74 Aethes angulatana_BIOUG08751-D09 67 Aethes angulatana_BIOUG09769-G01 53 Platphalonidia felix_CNCLEP00042703 83 Platphalonidia sp. JB-1_BIOUG06941-F01 Platphalonidia felix_CNCLEP00056783 57 Platphalonidia_BIOUG09821-H07 96 Platphalonidia felix_CCDB-24281-C02 28 Platphalonidia_HLC-17151 Platphalonidia mystica_10ANIC-12256 Platphalonidia lavana_BIOUG08068-F11 12 35 Platphalonidia_BIOUG06722-H07 96 Platphalonidia lavana_CNCLEP00077887 Lorita baccharivora_CNCLEP00025827 99 Lorita scarificata_BIOUG01827-G12 99 Lorita scarificata_HLC-17161 98 Nycthia pimana_CCDB-20277-C06 62 Nycthia yuccatana_CCDB-20277-C01 99 Nycthia yuccatana_CCDB-20277-B12 2 24 Nycthia_CCDB-20277-C05 4 Cochylis caulocatax_CCDB-20277-G08 98 Phalonidia ontariana_CNCLEP00040354 Parirazona serena_CCDB-30812-E02 25 Cagiva cephalanthana_MDOK-3420 Cochylis yinyangana_CCDB-20277-G12 78 Cochylidia_KN00744 Pontoturania posterana_MM21169 12 23 Cochylis carmelana_JWB-08-0199 9 Cochylis parallelana_BIOUG01549-F11 Falseuncaria degreyana_MM19201 99 Falseuncaria ruficiliana_MM18278 80 Cochylis flaviciliana_MM06891 3 96 Cochylis flaviciliana_BC ZSM Lep 61566 Cochylis roseana_MM19611 96 98 Cochylis roseana_TLMF Lep 11967 Cochylis epilinana_TLMF Lep 03195 20 62 Cochylis epilinana_TLMF Lep 06466 Diceratura infantana_TLMF Lep 03208
46 Brevicornutia pallidana_TLMF Lep 01543 99 Brevicornutia pallidana_MM06762 96 Brevicornutia pallidana_MM06249 0 70 Cochylidia moguntiana_MM17263 36 Cochylidia moguntiana_TLMF Lep 18496 Cochylidia subroseana_MM14118 93 Cochylidia heydeniana_MM13298 56 Cochylidia implicitana_MM18279 7 Cochylidia richteriana_MM11012 Cochylidia rupicola_MM22758 97 Neocochylis dubitana_MM02551 25 Neocochylis hybridella_MM18829 71 Neocochylis sannitica_TLMF Lep 23778 75 Neocochylis molliculana_UKLB34E04 99 Neocochylis molliculana_TLMF Lep 23781 99 Atroposia oenotherana_10BBCLP-0736 16 Atroposia oenotherana_CNCLEP00056788 Cochylis bucera_MDOK-3023 0 96 Cochylichroa hoff manana_09BBELE-0123 28 Cochylichroa hoff manana_BIOUG24091-A09 Cochylichroa viscana_BIOUG34608-B09 13 99 Cochylichroa viscana_DNA-ATBI-6178 Cochylichroa temerana_2006-ONT-0375 29 Cochylichroa arthuri_CCDB-20277-G04 99 Cochylichroa_CNCLEP00040579 7 Cochylichroa atricapitana_MM19612 99 1 Cochylichroa atricapitana_RMNH.INS.538541 Cochylichroa atricapitana_MM19222 Cochylichroa punctadiscana_DNA-ATBI-6175
36 Cochylichroa sp. 3_BIOUG01602-G05 28 Cochylichroa avita_CNCLEP00041310 43 Cochylichroa sp. 4_MDH005817 84 Cochylichroa hospes_2005-ONT-1043 Cochylichroa hospes_CNCLEP00040582 99 Thyraylia_MDOK-4483 Thyraylia_MDOK-4496 12 91 Thyraylia nana_MM03201 99 Thyraylia nana_10BBCLP-2168
32 Thyraylia_MNBTT-1496 Thyraylia bunteana_10BBCLP-2863 99 Thyraylia_MDOK-3229 Thyraylia_MDOK-3476 49 27 Thyraylia_MDOK-3394 Thyraylia discana_MDOK-3272 Thyraylia bunteana_jflandry2730 74 33 Thyraylia hollandana_MDOK-3424 58 Thyraylia hollandana_MDOK-2938 Thyraylia_MDOK-3658 Thyraylia voxcana_DNA-ATBI-6179 31 Thyraylia_CNCLEP00056786 55 Thyraylia voxcana_CNCLEP00041307