Exceptional larval morphology of nine species of the Anastrepha mucronota species group (Diptera, Tephritidae) Author Rodriguez, Erick J. https://orcid.org/0000-0001-8132-0863 Department of Entomology and Nematology, University of Florida, Gainesville, FL, USA erick.rodriguez@ufl.edu Author Steck, Gary J. https://orcid.org/0000-0003-3714-0560 Florida Department of Agriculture and Consumer Services, Division of Plant Industry (FDACS / DPI), Gainesville, FL, USA Author Moore, Matthew R. https://orcid.org/0000-0002-6313-3690 Florida Department of Agriculture and Consumer Services, Division of Plant Industry (FDACS / DPI), Gainesville, FL, USA Author Norrbom, Allen L. https://orcid.org/0000-0002-5854-089X Systematic Entomology Laboratory, USDA, ARS, c / o Smithsonian Institution, Washington, DC, USA Author Diaz, Jessica https://orcid.org/0000-0001-7013-2349 Department of Entomology and Nematology, University of Florida, Gainesville, FL, USA Author Somma, Louis A. https://orcid.org/0000-0003-4023-0997 Florida Department of Agriculture and Consumer Services, Division of Plant Industry (FDACS / DPI), Gainesville, FL, USA Author Ruiz-Arce, Raul https://orcid.org/0000-0002-0790-0218 USDA APHIS PPQ S and T Insect Management and Molecular Diagnostic Laboratory, 22675 N. Moorefield Road, Edinburg, TX 78541, USA Author Sutton, Bruce D. https://orcid.org/0000-0001-7374-3778 Research Associate, Department of Entomology, Smithsonian Institution, USNM, Gainesville, FL, USA Author Nolazco, Norma Centro de Diagnostico de Sanidad Vegetal, Servicio Nacional de Sanidad Agraria, Av. La Molina 1915, La Molina, Peru Author Muller, Alies https://orcid.org/0000-0002-4782-3536 (retired) Ministry of Agriculture, Animal Husbandry and Fisheries, Paramaribo, Suriname Author Branham, Marc A. https://orcid.org/0000-0002-2187-4503 Department of Entomology and Nematology, University of Florida, Gainesville, FL, USA text ZooKeys 2022 2022-11-03 1127 155 215 http://dx.doi.org/10.3897/zookeys.1127.84628 journal article http://dx.doi.org/10.3897/zookeys.1127.84628 1313-2970-1127-155 8A484FF467F140E2BB0BBE756CF0883A 5E797D2798B25C2EBD6A6596A3CB817F Anastrepha nolazcoae Norrbom & Korytkowski, 2011 Figs 66-69 , 70-75 , 76-80 Material examined. Peru • 20 larvae ; Madre de Dios , Puerto Maldonado , Centro de Investigacion y Capacitacion Rio Los Amigos (CICRA), trail 21; 12.5722°S , 70.0885°W ; 233 m a.s.l. ; 1-5 Feb. 2014 ; E. J. Rodriguez and J. Caballero leg.; reared from fruit of Quararibea cordata ; FSCA (AP20180222.01-AP20180222.10, AP20180206.01-AP20180206.10) . Diagnosis. The larva of A. nolazcoae differs from those of all other species of Anastrepha that have been adequately described by the combination of having fringed posterior margins of the oral ridges and accessory plates, and the presence of 6-8 comb-like processes adjacent to the labium. The posterior margins of the oral ridges and accessory plates resemble those of A. crebra , A. haplacantha , Anastrepha sp. Peru-82, and Anastrepha sp. nr. protuberans , but those species lack the comb-like processes. In addition, A. nolazcoae resembles A. korytkowskii and Anastrepha sp. Sur-16 in the presence of comb-like processes, but A. nolazcoae can be distinguished from them by the fringed posterior margins of its oral ridges. Other characters such as the ventral surface of the mouthhook, number of tubules and apical width of the prothoracic spiracle, and dorsal spinules on thoracic segments further differentiate A. nolazcoae (see Tables 2 - 4 ). Anastrepha nolazcoae shares the same host plant, Quararibea cordata , with species within the Anastrepha fraterculus group ( A. fraterculus complex), Anastrepha mucronota group ( A. mucronota ), and Anastrepha striata group ( A. striata ). The larva of A. mucronota was described with limited data ( Steyskal 1977 ) but can be morphologically separated from A. nolazcoae by the lower number of oral ridges (13-15 vs. 16-19) and dorsal irregularly light brown plaques on the abdominal segments (present vs. absent). The description of A. mucronota lacks information for most of the characters of the pseudocepalon (Table 2 ) and most of the available data overlap with those of A. nolazcoae (Table 3 , 4 ). Anastrepha nolazcoae differs from five morphotypes within the A. fraterculus complex ( Canal et al. 2015 , 2018 ) and A. striata as follows: 1) greater number of oral ridges (16-19; see the dichotomous key in Steck et al. 1990 ), except unknown for Andean and Peruvian morphotypes of A. fraterculus complex; 2) posterior margin of oral ridges fringed in A. nolazcoae , irregularly serrate in A. fraterculus (Brazil-1 and Ecuadorian morphotypes), scalloped or emarginate in A. fraterculus (Mexican morphotype), entire or serrate in A. striata ; and 3) approximately 36 accessory plates with fringed posterior margins in A. nolazcoae , apparently seven plates and serrate in A. fraterculus (Ecuadorian morphotypes; see plate 4b in White and Elson-Harris 1992 ), eight plates and serrate in A. fraterculus (Mexican morphotype), 8-9 plates and entire in A. striata . Anastrepha nolazcoae differs further from the A. fraterculus complex in having a greater number of tubules on the prothoracic spiracle (18-21 vs. 9-18 in Anastrepha fraterculus complex, see Rodriguez et al. 2021 ), although in this character it overlaps with A. striata . Figures 66-69. Scanning electron photomicrographs of third instar of Anastrepha nolazcoae 66 pseudocephalon 67 oral ridges 68 comb-like processes 69 antenna and maxillary palp. Abbreviations: CLP, comb-like processes; URS, upper right section with right angle shape. Scale bars: 10 μm ( 69 ); 20 μm ( 67 ); 50 μm ( 66 ). Description. Habitus . Third instar elongate, cylindrical, tapered anteriorly and caudal end truncate; color creamy; amphipneustic. Length 5.33-11.76 mm and width 0.93-1.92 mm at the sixth abdominal segment. Pseudocephalon (Figs 66 - 70 ). Antenna and maxillary palp on moderately developed lobe. Antenna with cylindrical base and apical knob. Maxillary palp bearing three papilla sensilla, two knob sensilla; dorsolateral group of sensilla bearing two well-developed papilla sensilla, aligned perpendicular to palp and surrounded by collar. Facial mask partly globular in lateral view, upper right section lacking ridges and accessory plates and forming almost a right angle. Preoral organ bearing one unbranched peg sensillum, located apically on a small, elongate-rounded lobe directly anterior to mouthhook; adjacent medial preoral lobe separate, short-elongate, extending partially posterior to lobe bearing preoral organ. Oral ridges in 16-19 rows, 13-15 anterior ridges with fringed posterior margins, three or four posterior ridges entire, undulant; 6-8 comb-like processes adjacent to labium and posterior to oral ridges; approximately 36 accessory plates lateral to oral ridges covering a much smaller area than oral ridges, with fringed posterior margins as on oral ridges, in two series. Labium triangular, anterior surface knobby, ventrally with two visible sensilla. Figures 70-75. Optical photomicrographs and scanning electron photomicrographs of third instar of Anastrepha nolazcoae 70 preoral organ 71 ventral surface of mouthhook 72 cephaloskeleton, lateral view 73 cephaloskeleton, dorsal view 74 prothoracic spiracle, lateral view 75 prothoracic spiracle, dorsolateral view. Scale bars: 10 μm ( 70, 71 ); 50 μm ( 74, 75 ); 200 μm ( 72, 73 ). Figures 76-80. Scanning electron photomicrographs and optical photomicrographs of third instar of Anastrepha nolazcoae 76 caudal segment 77 anal lobe 78-80 posterior spiracles. Scale bars: 50 μm ( 78-80 ); 100 μm ( 77 ); 200 μm ( 76 ). Cephaloskeleton (Figs 71-73 ). Total length from tip of mouthhook to end of ventral cornu 0.69-1.10 mm. Mouthhook well sclerotized, black apically and basally; length a 0.20-0.23 mm; length b 0.12-0.15 mm; height c 0.14-0.17 mm; ratio a:b 1.5-1.7; ratio a:c 1.3-1.4. Tooth long, sharp, strongly curved, concave ventrally with weak medial carina, ventral surface smooth. Intermediate sclerite 0.16-0.20 mm long, 0.18-0.21 mm wide at ventral bridge. Epipharyngeal sclerite visible only in dorsal view, with medial lobe directed anteriorly. Labial sclerite robust, sclerotized, and triangular in dorsal view. Parastomal bar extending three-fourths length of intermediate sclerite. Dorsal arch 0.23-0.29 mm high. Dorsal cornu with well-defined sclerotized area adjacent to notch, 0.38-0.53 mm long. Dorsal bridge prominently projecting anteriorly from dorsal cornu and slightly sclerotized. Anterior sclerite irregularly shaped and sclerotized. Cornu notch (N) 0.25-0.34 mm long and cornu notch index (N/DC) 0.6-0.7. Ventral cornu with well-defined sclerotized area between notch and pharyngeal bar and grooves. Pharyngeal filter with weakly sclerotized anterior bar and seven ridges forming a series of grooves along length of ventral cornu. Ventral cornu 0.44-0.71 mm long from pharyngeal bar to posterior end of grooves. Ventral cornu 1.18-1.34 x as long as sclerotized area of dorsal cornu. Thoracic and abdominal segments . Thoracic segments with dorsal spinules conical, symmetrical to slightly curved posteriorly; dorsal spinule pattern as follows: T1 with 3-5 rows; T2 with 3-5 rows; T3 with one or two rows; ventral spinule pattern as follows: T1 with 8-11 rows; T2 with four or five rows; T3 with three or four rows. Abdominal segments (A1-A8) lacking dorsal spinules; ventral creeping welts present on all abdominal segments (A1-A8); ventral spinule pattern as follows: A1 with three or four rows; A2 with six or seven rows; A3-A6 with 6-8 rows; A7 with six or seven rows; A8 with 6-9 rows. Additional two or three irregular rows of spinules anteriorly and posteriorly to anal lobes, two rows laterally, spinules large, conical, pointing away from anal lobes. Prothoracic spiracle (Figs 74 , 75 ). Bilobed, bearing 18-21 tubules, distally rounded and arranged in a single sinuous row. Spiracle distal width 0.26-0.34 mm; basal width 0.12-0.17 mm at junction with trachea. Caudal segment (Figs 76 , 77 ). Dorsal (D1 and D2), intermediate (I1 and I2), lateral (L1), and ventral (V1 and V2) tubercles and sensilla weakly developed; D1 distinctly anterior to D2. Intermediate tubercles I1 and I2 more strongly developed, but associated sensilla weakly developed; I1 lateral and sometimes slightly ventral to I2. L1, V1, and V2 tubercles and associated sensilla weakly developed. Anal lobe entire and very protuberant. Posterior spiracle (Figs 76 , 78-80 ). Located above horizontal midline. Posterior spiracle openings with thick rimae and numerous trabeculae; 83-108 µm long; 27-32 µm wide; ratio length/width 3.0-3.4. Ecdysial scar apparent. Felt chamber oval, 187-210 µm in diameter at junction with trachea. Spiracular process SP-I comprising 8-11 trunks and 9-26 tips; ratio tips/trunks 1.1- 2.4; basal width 9-15 µm ; ratio basal width/length of spiracular opening 0.09-0.17. SP-II comprising 3-7 trunks and 6-14 tips. SP-III comprising 3-9 trunks and 5-20 tips. SP-IV comprising 4-12 trunks and 8-24 tips; ratio tips/trunks 2.0; basal width 7-12 µm ; ratio basal width/length of spiracular opening 0.08-0.12. Distribution. Anastrepha nolazcoae is known only from Peru (Amazonas, Cajamarca, Huanuco , San Martin ) ( Norrbom and Korytkowski 2011 ; Barr et al. 2017 ; Bartolini et al. 2020 ). Biology. We reared this species from fruit of Quararibea cordata , the only known host. It was previously reared from the same fruit in Peru: Huanuco : Tingo Maria ( Norrbom and Korytkowski 2011 ). The larvae feed only on the pulp of the fruit. Molecular identification. COI barcodes were generated for 29 larvae and five adults and submitted to GenBank (MH070234, MT643950-MT643954, MT654802-MT654827, MT884299, MT884396). These data further confirm the identity of the described larvae. K2P distances between A. nolazcoae larvae and the nine available adult sequences ranged from 0.0-1.1%. BLAST searches were consistent with our new data, yielding only four good matches: A. nolazcoae (99.21-100% sequence identity; KY428297, MN454445, MN454488, MF695205 [identified as A. kuhlmanni in GenBank, reported as A. nolazcoae in Barr et al. 2017 ]). Additionally, 27 larval barcodes returned consensus identifications of A. nolazcoae with either three or two votes, and two samples returned ambiguous identifications ( Moore et al. in press ).