Perlidae (Plecoptera) from the Paranapiacaba Mountains, Atlantic Forest, Brazil: Diversity and implications of the integrative approach and teneral specimens on taxonomy

Abstract

The study of complementary sources of biological variation (e.g. morphological, molecular) has allowed a better understanding of biodiversity through the construction of an integrative taxonomy. Using this approach, specimens from the Paranapiacaba Mountains, southeastern Brazil, were studied to update the knowledge on the stonefly family Perlidae from the region, characterize the species, and make associations between nymphs and adults using a fragment of the mitochondrial cytochrome c oxidase subunit I (COI) gene. The study also discusses the implications of integrative taxonomy and teneral specimens for the study of South American Perlidae. The molecular data were analyzed using Bayesian inference, Neighbor-joining, and delimiting species methods. Our results revealed that, in general, there was a morphological and molecular congruence between species. In the Paranapiacaba Mountains, three genera and 15 species were recorded: Anacroneuria boraceiensis Froehlich 2004, A. debilis (Pictet 1841) (new record), A. fiorentini De Ribeiro and Froehlich 2007 (new record), A. flintorum Froehlich 2002, A. iporanga Bispo and Froehlich 2004, A. itajaimirim Bispo and Froehlich 2004, A. polita (Burmeister 1913), A. subcostalis Klapálek 1921, A. tupi Bispo and Froehlich 2004 (with a description of the nymph), Kempnyia auberti Froehlich 1996, K. colossica (Navás 1934), K. flava Klapálek 1916, K. neotropica (Jacobson and Bianchi 1905) (including its new junior synonym K. petersorum Froehlich 1996), Kempnyia sp., and Macrogynoplax veneranda Froehlich 1984. COI sequences were obtained for 11 species, five of which had nymphs associated with adults. Among the five associated nymphs, the nymph of A. tupi is described here. The results of this study indicate that the color of adult teneral specimens differs from that of mature specimens. Given this, the synonym of K. neotropica and K. petersorum was proposed since these species have high morphological and molecular similarities and differ only in color patterns. In addition, the previous record of A. petersi Froehlich 2002 from the Paranapiacaba Mountains was invalidated since it was considered a teneral specimen of A. flintorum. These results suggest that the development of an integrative taxonomy is essential to continue advancing the study of Perlidae diversity in South America.

Fig 1. Paranapiacaba Mountains.

Map of Paranapiacaba Forest Continuum and the sampled sites. EEX: Ecological Station of Xitué; PECB: Carlos Botelho State Park; PEI: Intervales State Park; PENP: Nascentes do Paranapanema State Park; and PETAR: Alto Ribeira Tourist State Park, São Paulo State, Brazil.

Fig 2. Molecular analysis.

Bayesian inference tree obtained using mitochondrial cytochrome c oxidase subunit I (COI) sequences (636 bp) from specimens from the Paranapiacaba Mountains (São Paulo State, Brazil) and related stoneflies from Paraná State (red rectangle) and Santa Catarina State (blue rectangle), Brazil. Numbers are clusters support (bootstrap) recovered. The tree was rooted using a sequence of Gripopterygidae. Delimitation methods: 1) morphology (Morph); 2) Automatic Barcode Gap Discovery (ABGD) considering primary partition (ABGDp); 3) ABGD considering recursive partition (ABGDr); and 4) Poisson Tree Processes (PTP) and Bayesian implementation of the Poisson Tree Processes (bPTP).

Fig 3. Intra- and interspecific divergences comparisons.

Percentages of pairwise intra- and interspecific divergences obtained by Kimura-2-parameter (K2P) showing the barcode gap in Anacroneuria (A) and Kempnyia (B).

Fig 4. Teneral pattern of Anacroneuria species.

Head and pronotum of A. debilis adult male (A) and teneral adult males (B and C); A. itajaimirim adult male (D) and teneral adult males (E and F); A. tupi adult female (G) and teneral adult males (H and I); A. polita adult female (J) and teneral adult male (K). Scale: 1 mm.

Fig 5. Morphological variation within cluster Kempnyia neotropica + K. petersorum.

Head and pronotum of adult females (A and C) and teneral adult female (B) of K. neotropica. Head and pronotum of adult males (D–F) of K. petersorum according to description by Froehlich [70]. Scale: 1 mm.

Fig 6. Anacroneuria species.

Head and pronotum of A. boraceiensis adult male (A), A. iporanga adult male (B), A. itajaimirim adult male (C), A. polita adult male (D), A. subcostalis adult female (E), and A. tupi adult male (F). Scale: 1 mm.

Fig 7. Anacroneuria debilis.

Adult male, head and pronotum (A). Tergum 9 with hammer (B). Penial armature in ventral (C), dorsal (D), and lateral (E) views. Scale: 1 mm.

Fig 8. Anacroneuria fiorentini.

Teneral male, head and pronotum (A). Tergum 9 with hammer (B). Penial armature in ventral (C), dorsal (D), and lateral (E) views. Scale: 1 mm.

Fig 9. Anacroneuria tupi nymph.

Head and pronotum (A), mesonotum and metanotum (B), abdomen (C), labium (D), right mandible (E), right maxilla (F), foreleg (G), cercus in proximal, medial and distal views (H). Scales: 1 mm for A–G and 0.5 mm for H.

Fig 10. Kempnyia and Macrogynoplax species.

Head and pronotum of a K. colossica adult male from the Paranapiacaba Mountains (A), PR (B) and SC (C), a K. flava adult female (D), a K. sp. adult female (E) and an M. veneranda adult male (F). Scale: 1 mm.