Disclisioproctaedmondsii (Butler, 1882) comb. nov. (Lepidoptera, Geometridae, Larentiinae)

Abstract Background The generic assignment of the geometrid moth Xanthorhoeedmondsii (Butler, 1882) (Lepidoptera, Geometridae, Larentiinae), originally described under Hypochroma Guenée, [1858], a junior homonym of Hypochroma Herrich-Schäffer, [1855] (Geometridae, Ennominae), is assessed using genitalia morphology and analysis of mitochondrial DNA sequences. New information Morphological characters revealed closeness to the type species of Disclisioprocta Wallengren, 1861 (Larentiinae). In agreement with morphology, the molecular analysis clustered X.edmondsii with species of Disclisioprocta in a well-supported monophyletic group distantly related to members of Xanthorhoe Hübner, [1825]. Accordingly, Disclisioproctaedmondsii (Butler, 1882) comb. nov. is proposed.


Introduction
Natural environments of South America harbour a high diversity of geometrid moths (Lepidoptera, Geometridae), whose taxonomy remains insufficiently studied (Hausmann and Parra 2009, Brehm et al. 2016, Murillo-Ramos et al. 2021. Besides the frequent discovery of new species (Brehm 2018, Ramos-González et al. 2019, Moraes et al. 2021, Vargas 2021, generic assignments of most of the already described species deserve assessment, as suggested by new combinations arising in taxonomic revisions (Parra 2018) and molecular phylogenetic analyses ).
The geometrid moth Xanthorhoe edmondsii (Butler, 1882) (Geometridae, Larentiinae) is known from Chile and Argentina (Butler 1882, Chalup 2014). It was originally described under Hypochroma Guenée, [1858], a junior homonym of Hypochroma Herrich-Schäffer, [1855] (Geometridae, Ennominae) (Parsons et al. 1999). Individuals from northern Chile reared from larvae collected on the ornamental plant Bougainvillea glabra Choisy (Nyctaginaceae) were misidentified as Chrismopteryx undularia (Blanchard, 1852), based on comparison with material from central Chile deposited in the Museo Nacional de Historia Natural de Santiago (Vargas et al. 2010). Subsequent comparison with a photo of the type material deposited in the Natural History Museum, London, UK, allowed concluding that the specimens examined by Vargas et al. (2010) belong to X. edmondsii.
Morphological characters of the genitalia of geometrid moths are extremely useful in generic assignments (Pitkin 2002, Parra 2018, Viidalepp and Lindt 2019, which can be reinforced by phylogenetic analysis of DNA sequences , Matson 2022, Wanke et al. 2022). An examination of the genitalia of X. edmondsii revealed remarkable morphological differences with Xanthorhoe montanata ([Denis & Schiffermüller], 1775), the type species of Xanthorhoe Hübner, [1825], suggesting instead closeness with Disclisioprocta stellata (Guenée, [1858]), the type species of Disclisioprocta Wallengren, 1861. The aim of this study is to propose a new generic assignment for X. edmondsii, based on genitalia morphology and analysis of mitochondrial DNA sequences.

Materials and methods
Specimens examined in this study were collected at light or reared from larvae collected on B. glabra in the Azapa Valley (18°31'16''S, 70°10'42''W), Arica Province, northern Chile. Photos of the genitalia were taken with a Leica Flexacam C1 digital camera attached to a Leica M125 stereomicroscope. Each image was constructed with about 5-10 photos assembled with the software Helicon Focus 8. The specimens of X. edmondsii are deposited in the "Colección Entomológica de la Universidad de Tarapacá" (IDEA), Arica, Chile. The specimens of D. stellata are deposited in the "Coleção Pe. Jesus de Santiago Moure (DZUP)", Universidade Federal do Paraná, Curitiba, Paraná, Brazil.
Genomic DNA was extracted from two legs of a male adult using the QIAamp Fast DNA Tissue Kit, following the manufacturer's instructions. DNA purification, PCR amplification and sequencing of the barcode region (Hebert et al. 2003) with the primers LCO1490 and HCO2198 (Folmer et al. 1994) were performed at Macrogen Inc. (Seoul, South Korea). The PCR programme was 5 min at 94°C, 35 cycles of 30 s at 94°C, 30 s at 47°C, 1 min at 72°C and a final elongation step of 10 min at 72°C. Analysis of this mitochondrial marker represents a helpful tool in generic assignments of Lepidoptera, including Geometridae (Wanke et al. 2019, Wanke et al. 2022, Vargas et al. 2022. The sequence of X. edmondsii was submitted to a Maximum Likelihood (ML) analysis with additional representatives of Larentiinae downloaded from BOLD (Ratnasingham and Hebert 2007), following the classification provided by  in which Disclisioprocta belongs to an unnamed clade sister to Euphyiini. The alignment included sequences of Euphyia Hübner, [1825] and Oligopleura Herrich-Schäffer, [1855] as representatives of this tribe, sequences of Xanthorhoe due to the current generic adscription of X. edmondsii (Parsons et al. 1999) and sequences of Scotopteryx Hübner, [1825] as outgroups. The software MEGA11 (Tamura et al. 2021) was used to perform sequence alignment with the ClustalW method and to determine genetic distance using the Kimura 2-Parameter (K2P) method. Before the ML analysis, the substitution saturation of the alignment was assessed with the Xia test, using the software DAMBE7 (Xia 2018). The ML analysis was performed using the software IQTREE 1.6.12 (Nguyen et al. 2014) in the web interface W-IQ-TREE (Trifinopoulos et al. 2016) with data partitioned to codon position. ModelFinder (Kalyaanamoorthy et al. 2017) selected TNe+I, F81+F and TN+F+G4 as the best fit models for 1st, 2nd and 3rd partitions, respectively. Branch support was assessed with 1000 replications of the Shimodaira-Hasegawa-like approximate likelihood ratio test (SH-aLRT, Guindon et al. (2010)) and ultrafast bootstrap (UFBoot, Hoang et al. (2017)). The unrooted tree was visualised in FigTree (Rambaut 2014) to root on Scotopteryx.

Description
Male habitus in Fig. 1. Although the male abdominal segments VII and VIII are not part of the genitalia, these are described here and illustrated because the morphology of the sclerites of these segments can be modified in different groups of Larentiinae (Viidalepp 2011). Male abdominal segments VII and VIII (Fig. 2). Segment VII mostly membranous; tergum a transverse stripe strongly posteriorly folded in the middle; sternum a transverse stripe; pleura with pair of coremata. Segment VIII mostly membranous; tergum an anterior transverse stripe with semicircular expansion on tips, connected by a short longitudinal stripe with a posterior rectangular transverse plate; sternum an anterior transverse stripe posteriorly curved in the middle, projected as a narrow longitudinal stripe posteriorly bifid, triangular expansion near tip of the anterior transverse stripe.
Male genitalia (Fig. 3). Uncus bifid with broad posterior concavity in the middle, truncate points slightly down-curved. Saccus with small rounded anterior projection. Subscaphium slightly sclerotised. Labides with lobe-like tip bearing setae. Manica heavily spinose. Juxta trapezoidal, ventral half of lateral margin broadly concave, ventral margin broadly concave. Valva elongated; costal sclerotised band not reaching apex; cucullus mostly membranous on distal half with abundant setae; sacculus broad, well-sclerotised; sacculus projection stout, apex almost reaches that of the distal margin of the cucullus, with a broader, dorsally projected basal process. Phallus cylindrical, anterior half straight, posterior half curved, with a small spine-like projection ventrally on posterior tip; vesica mostly membranous with a plate-like cornutus.

Molecular analysis
Genetic distance of D. edmondsii (BOLD accession GEONC001-22) was 10.3-10.5% (K2P) with D. natalata and 11.0-11.5% with D. stellata, while the distance between the latter two was 6.2-7.1%. The alignment was suitable for phylogenetic analysis, as no evidence of stop codons was detected and the index of substitution saturation was smaller than the critical value (ISS < ISS.C; p < 0.001) in the Xia test. The ML analysis ( Fig. 4) clustered (D. edmondsii (D. natalata + D. stellata)) with high support. Although each genus had reasonable statistical support in the ML analysis, relationships between genera were not resolved.

Discussion
Although the identification of synapomorphies for Disclisioprocta deserves further assessments based on better knowledge of the morphology of related genera, the morphological similarities between the genitalia of D. edmondsii (Fig. 3) and D. stellata (Fig. 5) provide support to consider them as congenerics. The two species have uncus bifid, costal sclerotised band not reaching apex of the cucullus, a stout sacculus projection and a plate-like cornutus in the male and a flattened elongated antrum and a cluster of elongated spine-like signa arising from a plate on the corpus bursae in the female. The same morphological characters support the removal of D. edmondsii from Xanthorhoe. The type species of this genus has an elongated rod-like uncus, costa as a sclerotised free arm extending beyond the cucullus apex, sacculus lacking a projection and numerous spinelike cornuti in the male and a short antrum and two elongated plate-like signa with small spines in the female (Expósito-Hermosa and Viidalepp 2011).
The result of the ML analysis is congruent with the genitalia morphology, since D. edmondsii clustered with two other representatives of Disclisioprocta in a well-supported clade, distantly related to members of Xanthorhoe (Fig. 4) (Parsons et al. 1999). Accordingly, it is highly probable that, with an improvement in the knowledge of the taxonomic diversity of Disclisioprocta, the genetic distance of D. edmondsii to other congenerics would be smaller. Alternatively, further studies could reveal D. edmondsii as a member of another, either described or undescribed, lineage of generic level, sister to Disclisioprocta. In the meantime, its placement in this genus seems a better solution than its previous adscription to Xanthorhoe, in spite the deep genetic distance with congenerics.