Phylogenetic relationships of Hydropsychidae ...

Proceedings of the 11th International Symposium on Trichoptera (2003, Osaka) pages 131-136, © 2005 Tokai University Press, Kanagawa. eds. K. Tanida and A. Rossiter

Phylogenetic relationships of Hydropsychidae subfamilies based on morphology and DNA sequence data

Christy J. GERACI1, Kari M. KJER2, John C. MORSE1 and Roger J. BLAHNIK3 1

Department of Entomology, Soils and Plant Sciences, Clemson University, Clemson, South Carolina 29634-0315, USA; 2Department of Ecology, Evolution and Natural Resources, Cook College, Rutgers University, New Brunswick, New Jersey 08901, USA; 3Entomology Department, University of Minnesota, St. Paul, Minnesota 55108, USA Abstract. This study presents preliminary data regarding subfamilial and generic relationships of the family Hydropsychidae based on morphological, mitochondrial COI gene fragment, nuclear ribosomal RNA and EF-1 alpha sequence data from Kjer et al. (2001, 2002). Specimens from twenty genera of Hydropsychidae were examined for morphological characters, reinterpreted from Frania and Wiggins (1997), Ross (1944), and Schefter (1996). Phylogenies obtained from morphological, molecular, and combined data sets yielded similar trees with respect to subfamilial relationships; however, support values for subfamilial relationships were weak. Combined data support the monophyly of the Arctopsychinae, Smicrideinae and Macronematinae, but that of the Hydropsychinae and Diplectroninae remains unresolved.

Introduction

study Hydropsychinae phylogeny, and outlined their interpretation and hypotheses about the groundplan for the genitalic structure of the Hydropsychinae. Kjer et al. (2001, 2002) used molecular evidence to revise the higher phylogeny of the Trichoptera. Using COI, ribosomal and EF-1 alpha gene sequence data for ten hydropsychid species, Kjer et al. (2001, 2002) also provided preliminary information regarding hydropsychid subfamily relationships. Those relationships are still tentative and need corroboration from additional species and from morphological data. Our study presents preliminary data regarding subfamilial and generic relationships of Hydropsychidae based on morphological data from Frania and Wiggins (1997), Ross (1944) and Schefter (1996), and on mitochondrial COI gene fragment, nuclear ribosomal RNA and EF-1 alpha sequence data from Kjer et al. (2001, 2002).

The family Hydropsychidae (Insecta: Trichoptera) is a diverse, cosmopolitan group of caddisflies that plays an important role in stream ecosystems. Globally, 1,462 species in 50 recognized extant genera constitute five subfamilies of Hydropsychidae: Arctopsychinae, Diplectroninae, Hydropsychinae, Macronematinae and Smicrideinae (Morse 2001). Hydropsychid larvae display a wide range of tolerance values and are used in biomonitoring programs throughout the world. For example, North Carolina Biotic Index (NCBI) tolerance values for hydropsychids range from 0.0 for Hydropsyche carolina Banks to 8.8 for Hydropsyche betteni Ross (on a scale of 0-10, with 0 = least tolerant of pollution) (Lenat 1993). Despite the environmental importance of the family, a combined analysis of the phylogeny of world Hydropsychidae subfamilies, tribes and genera using both morphological and molecular data has never been attempted. The phylogenetic relationships among the five subfamilies of Hydropsychidae have been debated (Flint 1961, 1974; Schefter 1996) using various morphological characters from larvae, pupae, and adult males and females. Schefter (1996) addressed the subfamily relationships in Hydropsychidae using larval, pupal and adult synapomorphies. Schefter (1996) and Flint (1974) focused on adult wing and body characters in their phylogenetic studies. Ross and Unzicker (1977) used male genitalic characters to

Methods Specimens from 20 genera of Hydropsychidae were examined with an Olympus SZ60 dissecting microscope with up to 63X magnification. Morphological characters presented by Frania and Wiggins (1997) (Character 1), Ross (1944) (Character 2), and Schefter (1996) (Characters 3-30) were reinterpreted for these genera using binary coding (Table 1). Synapomorphies were coded as “1” and plesiomorphies as “0”.

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C.J. GERACI, K.M. KJER, J.C. MORSE1 and R.J. BLAHNIK

Table 1: Characters examined in morphological study (adapted from Schefter 1996). (0 = plesiomorphic; 1 = apomorphic; ? = missing data). Taxa Ancestor Arctopsyche Parapsyche Diplectrona Homoplectra Sciadorus Cheumatopsyche Hydromanicus Hydronema Ceratopsyche Hydropsyche Potamyia Abacaria Aoteapsyche Calosopsyche Streptopsyche Macrostemum Leptonema Macronema Pseudomacronema Centromacronema Smicridea (Rhyacophylax) Smicridea (Smicridea) Smicrophylax Asmicridea

Morphological Characters 1 2 3 4 5 6 7 8 9 10 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 0 1 0 1 1 1 1 1 0 0 0 1 0 1 1 1 1 1 0 1 0 1 0 0 1 1 1 1 0 1 0 1 0 0 1 1 ? ? ? ? ? ? ? ? ? 1 0 1 1 1 1 0 1 0 1 1 0 1 1 1 0 0 1 0 1 1 0 1 1 1 1 0 1 1 1 1 0 1 1 1 1 0 1 0 1 1 0 1 1 1 1 0 1 1 1 1 0 1 1 1 1 0 1 1 1 1 ? ? ? ? ? ? ? ? ? 1 0 1 1 1 1 0 1 1 1 1 0 1 1 1 0 0 0 0 1 1 0 1 1 1 0 0 1 0 1 1 0 1 1 1 0 0 0 0 1 1 0 1 1 1 0 0 0 0 1 1 0 1 1 1 0 0 0 0 1 1 ? ? ? ? ? ? ? ? ? 1 ? ? ? ? ? ? ? ? ? 1 0 1 1 1 0 0 0 1 0 1 0 1 1 1 0 0 0 1 0 1 0 1 1 1 0 0 0 1 1 1 0 1 1 1 0 0 0 1 1

11 0 0 0 0 0 ? 1 0 1 0 1 1 ? 1 0 0 0 0 0 ? ? 0 0 0 0

12 0 1 1 0 0 ? 0 0 0 0 0 0 ? 0 0 0 0 0 0 ? ? 0 0 0 0

13 0 1 1 1 1 ? 1 1 1 1 1 1 ? 1 1 1 1 1 1 ? ? 1 1 1 1

14 0 0 0 1 1 ? 1 1 1 1 1 1 ? 1 1 1 1 1 1 ? ? 1 1 0 1

15 0 0 0 0 0 ? 1 1 1 1 1 1 ? 1 1 0 1 1 1 ? ? 0 0 0 0

16 0 1 1 1 1 ? 1 1 1 1 1 1 ? 0 0 0 0 0 0 ? ? 0 0 ? 0

17 0 0 0 0 0 ? 0 0 0 0 0 0 ? 0 1 1 1 1 0 ? ? 0 0 ? 1

18 0 0 0 1 1 ? 1 1 1 1 1 1 ? 1 ? ? 1 1 1 ? ? 1 1 ? 1

19 0 0 0 0 0 0 0 0 1 1 1 1 1 1 0 0 0 0 0 ? ? 0 0 0 0

20 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1

21 0 0 0 0 0 0 1 0 1 0 0 1 0 0 0 0 0 0 0 0 0 1 1 1 1

22 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 1

23 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0

24 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 0 0 0 0

25 26 27 0 0 0 1 0 0 1 0 0 1 0 0 1 10 0 1 1 0 1 1 0 1 1 0 1 1 0 1 1 0 1 1 0 1 1 0 1 1 1 1 1 0 1 1 0 1 1 0 1 1 0 1 1 0 1 1 0 1 0 0 1 1 0 1 1 1 1 1 1 1 1 0 1 1 0

28 0 1 1 0 ? 0 1 1 1 0 0 1 1 1 0 0 1 0 0 ? ? 0 1 0 0

29 0 0 0 1 0 ? 0 0 0 0 0 0 ? 0 ? ? 0 ? ? ? ? 0 1 1 0

30 0 0 0 0 0 ? 1 0 0 0 0 0 ? 0 0 0 0 0 0 ? ? 1 1 1 1

Character 1: 0 = Anterior tentorial pits in contact with anterior arms of dorsal ecdysial line; 1 = Anterior tentorial pits arising on dorsal apotome some distance mesad of cleavage line. Character 2: 0 = Frontoclypeal suture of larvae ogival; 1 = Frontoclypeal suture of larvae lyre-shaped. Character 3: 0 = Posterior ventral apotome absent; 1 = Posterior ventral apotome present. Character 4: 0 = Posterior ventral apotome long; 1=Posterior ventral apotome minute. Character 5: 0=Ventral apotome single; 1=Anterior and posterior ventral apotomes fused. Character 6: 0 = Larval mentum subrectangular; 1 = Larval mentum cleft anteriorly. Character 7: 0 = Larval mesonotal and metanotal sclerotized plates present and either undivided or divided mesally; 1 = Larval mesonotal and metanotal plates with transverse ecdysial line. Character 8: 0 = Larval foretrochantin lacking well developed dorsal ramus; 1 = Foretrochantin with dorsal ramus. Character 9: 0 = Scale hairs absent from larval abdominal terga; 1 = Scale hairs present on abdominal terga. Character 10: 0 = Hair-like setae absent from larval abdominal terga; 1 = Hair-like setae present on larval abdominal terga. Character 11: 0 = Dorsum of larval abdomen lacking minute spines; 1 = Dorsum of abdomen with minute spines. Character 12: 0 = Dorsum of larval abdomen lacking minute denticles; 1 = Dorsum of abdomen with minute denticles. Character 13: 0 = Larval abdominal gills without apical filaments; 1 = Larval abdominal gills with apical filaments. Character 14: 0 = Larval abdominal gills without subapical filaments; 1 = Larval abdominal gills with subapical filaments. Character 15: 0 = Venter of larval abdominal segment VII with primary setae 10; 1 = Venter of larval abdominal segment VII without primary setae 10. Character 16: 0 = Pupal anal processes uniramous or absent; 1 = Pupal anal processes apically divided. Character 17: 0 = Posterior hookplate present on pupal abdominal segment IV; 1 = Posterior hookplate absent on pupal abdominal segment IV. Character 18: 0 = Posterior hookplate present on pupal abdominal segment V; 1 = Posterior hookplate absent on pupal abdominal segment V. Character 19: 0 = Lateral protarsal setal brush absent from adult male; 1 = Lateral protarsal setal brush present on adult male. Character 20: 0 = Hindwing and forewing fork 2(R4+R5) sessile, contiguous with discoidal cell; 1 = Fork 2 on stalk, dividing distad of discoidal cell. Character 21: 0 = Forewing crossveins m-cu and cu separated by more than twice the length of m-cu; 1 = m-cu and cu in line, separated by less than twice the length of m-cu. Character 22: 0 = Microspines evenly arrayed between PC and A1 on forewing; 1 = Microspines concentrated in linear or irregular patch between PC and A1. Character 23: 0 = Forewing A1 without short row of recurved setae; 1 = Forewing A1 with short row of recurved setae. Character 24: 0 = Forewing A1 without file-and-groove structure; 1 = Forewing A1 part of file-and-groove structure. Character 25: 0 = Forewing median and thyridial cells without overlap; 1 = Forewing median and thyridial cells with overlap. Character 26: 0 = Forewing median and thyridial cells with overlap less than length of m-cu; 1 = Forewing median and thyridial cells with overlap at least length of m-cu. Character 27: 0 = Hindwing fork 1 present; 1 = Hindwing fork 1 absent. Character 28: 0 = Filament-like lobes present on adult abdominal segment V; 1 = Filament-like lobes absent on adult abdominal segment V (low protuberances may be present). Character 29: 0 = Reticulate sacs absent from adult abdominal segments VI and VII; 1 = Reticulate sacs present on adult abdominal segments VI and VII. Character 30: 0 = Ventral plates mesally conjoined or fused with adult female sternite VIII; 1 = Ventral plates entirely separated mesally.

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50% Majority Rule consensus of 16 trees Ancestor Arctopsyche

Arctopsychinae

Parapsyche Diplectrona

1,3,13,25

Diplectroninae

Homoplectra 27

5

Smicridea (R) Smicridea (S)

21

Smicrideinae

Smicrophylax Asmicridea

26

Cheumatopsyche Hydronema Potamyia 11 6

Aoteapsyche Hydropsyche

Hydropsychinae

Ceratopsyche 8 4

Hydromanicus

23

Streptopsyche Calosopsyche Macrostemum Leptonema

24

Macronematinae

Macronema

Fig. 1: Fifty-percent majority consensus tree of 16 most parsimonions trees based on morphological data (see Table 1) (TL= 54). Synapomorphic characters are included on their appropriate nodes.

and 50 repetitions. Abacaria, Sciadorus, Pseudomacronema and Centromacronema were omitted from the morphological analysis because of the large amounts of missing data. A consensus tree (50% majority rule) was generated for the morphological data, and bootstrap analyses were performed on the molecular and combined trees.

Adult heads were cleared in 10% KOH to examine the tentorium. Larvae, pupae and adults of as many genera as possible were examined. If all life stages were not available from the Clemson University Arthropod Museum, the literature was consulted. If the character still could not be determined, it was coded as “?” = missing data. Several specimens of three other genera of Annulipalpia (Polycentropodidae: Polycentropus, Dipseudopsidae: Phylocentropus, and Psychomyiidae: Lype) were examined as the outgroup taxa and labeled “Ancestor” in the morphological tree, following Schefter (1996). Additional DNA sequences for the COI mitochondrial gene fragment (441 nts), nuclear ribosomal RNA (1078 nts) and EF-1alpha (1098 nts) were added to the dataset of Kjer et al. (2001, 2002): for sequencing procedures. All data were analyzed in PAUP (Swofford 1999) using maximum parsimony with a heuristic search routine

Results Phylogenies obtained from morphological (Fig. 1, Table 1), molecular (Fig. 2) and combined data sets (Fig. 3) yielded similar trees with respect to the subfamily relationships. These relationships, however, have weak support values. In all three trees, the Arctopsychinae, Macronematinae and Smicrideinae are monophyletic. Arctopsychinae are basal to other hydropsychids, and Smicrideinae are basal to the (Macronematinae + Hydropsychinae).

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Wormaldia gabriella Polycentropus interruptus

96

Ecnomus tenellus

99

Xiphocentron n. sp. Parapsyche elsis

99

Arctopsyche grandis 78

Arctopsychinae

Diplectrona modesta

55

Homoplectra doringa

Diplectroninae

Sciadorus acutus

36

Asmicridea edwardsi

43

Smicridea talamanca (R)

58

21

Smicridea talamanca (S)

97

Smicrideinae

Smicridea bivittata (S) 9

Leptonema salvini

85

Leptonema crassum 98

Centromacronema excsisum

99

Centromacronema apicali

86 27

42

Macronematinae

Macrostemum zebratum Pseudomacronema vivittatum

46

Synoestropsis punctipennis Weakly supported

12

Plectropsyche hoogstraali

45

Calosopsyche continentalis

99

Streptopsyche parander Cheumatopsyche oxa

66

Hydropsychinae

Potamyia flava

38

Hydropsyche occidentalis 100

Ceratopsyche bronta

97

Aoteapsyche colonica

Fig. 2: Generic relationships of Hydropsychidae based on DNA sequences for the COI mitochondrial gene fragment (441 nts), nuclear ribosomal RNA (1078 nts) and EF-1alpha (1098 nts) (Kjer et al. 2001, 2002). Bootstrap values are provided at each node.

monophyly of the Hydrospychinae. The molecular and combined data sets do not support the placement of Calosopsyche, Plectropsyche, and Streptopsyche within Hydropsychinae (Fig. 3), but the morphological data show the two genera to be basal hydropsychines based on synapomorphic Character 23 (from Schefter 1996), the presence of a row of recurved setae on A1 of the forewing (Fig. 1). Again, the bootstrap values are so weak that we cannot make any conclusions regarding the placement of these genera. The monophyly of the Diplectroninae also remains unresolved by either morphological or DNA sequence data, although the subfamily appears to be paraphyletic (Fig. 1, 2). This preliminary study illustrates the need for a comprehensive world revision of the family Hydropsychidae. More support is needed to

The monophyly of Diplectroninae is not supported by any tree, but neither is it strongly refuted. Discussion Trees obtained from these preliminary results cannot resolve the subfamilial relationships among the Hydropsychinae. Bootstrap values from the molecular and combined data set analyses (Figs. 2 and 3) do show strong support for the monophyly of Arctopsychinae, Smicrideinae and Macronematinae, and the placement of Arctopsychinae as a basal lineage. This supports the opinions of Fischer (1963), Schmid (1968) and Levanidova (1982) that the Arctopsychidae are a separate family. One main discrepancy among the trees is the

134


Wormaldia gabriella Xiphocentron n. sp.

63 48

Polycentropus interruptus Ecnomus tenellus Parapsyche elsis

100

Arctopsychinae

Arctopsyche grandis 98

Homoplectra doringa Diplectrona modesta

62

Diplectroninae

Sciadorus acutus 33

Asmicridea edwardsi

81

Smicridea talamanca

82 44

87

Smicridea turrialbana

Smicrideinae

Smicridea bivittata Cheumatopsyche oxa

37 29

Potamyia flava

79

Abacaria sp. 50

Hydromanicus 36

Hydropsyche occidentalis 35

Ceratopsyche bronta

31 33

24

Herbertorossia sp. Aoteapsyche colonica Plectropsyche hoogstraali

40

Streptopsyche parander

96 51 Weakly supported

Hydropsychinae

Calosopsyche continentalis Calosopsyche dominicensus

26

Leptonema salvini

96

Leptonema crassum 100

Macrostemum zebratum 71

Macronema varia

56 64

Centromacronema excsisum

Macronematinae

Centromacronema apicali

26

Pseudomacronema vivittatum

17 31

Synoestropsis punctipennis Oestropsyche vitrina

Figure 3: Generic relationships of Hydropsychidae based on a combined analysis of molecular and morphological data. Bootstrap values are provided at each node.

KMK acknowledges support from the New Jersey Agricultural Experiment Station, and NSF grants DEB9796097 and DEB9974081. CJG, KMK, and JCM are also grateful for the support of NSF grant DEB0316504. Thanks to R. Holzenthal (University of Minnesota) and O. Flint (Smithsonian Institution) for specimens and helpful discussions. This is Technical Contribution No. No. 4934 of the South Carolina Agriculture and Forestry Research System, Clemson University.

resolve the arrangement of the subfamilies and genera. Future research will focus on incorporating both morphological data (i.e., identifying additional synapomorphies) and molecular data (i.e., sequencing more genera) to produce a more conclusive combined phylogeny of the Hydropsychidae. Acknowledgements CJG acknowledges support from the Clemson E.W. King Endowed Memorial Grant Fund.

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References

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