Dtsch. Entomol. Z. 71 (2) 2024, 241-254 | DO! 10.3897/dez.71.120389

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BERLIN

Phylogeny of the Chinese species groups of the subgenus Homoneura
Wulp, 1891 (Diptera, Lauxantidae, Homoneurinae) based on
morphological characters

Yao Yao', Chaoyang Kong", Pu Miao’, Shengjuan Zhao*, Wenliang Li!

1 College of Horticulture and Plant Protection, Henan University of Science and Technology, Luoyang 471023, China
2 Henan Tobacco Company Luoyang company, Luoyang 471000, China
3 College of Food & Bioengineering, Henan University of Science and Technology, Luoyang 471023, China

https://zoobank. org/243 FF CE92-70C3-43 1 B-93F8-BF&$2B2A 9823E

Corresponding author: Wenliang Li (wenliangli@haust.edu.cn)

Academic editor: A. Camargo # Received 7 February 2024 # Accepted 9 August 2024 Published 2 September 2024

Abstract

The subgenus Homoneura Wulp, 1891 (Diptera, Lauxaniidae, Homoneurinae) is highly diverse with more than 220 species known
from China, representing more than 80% of the Chinese genus Homoneura Wulp, 1891. These species were assigned into 21 species
groups in studies mainly focusing on the classification and description of species. The phylogenetic relationships of each subgenus
of Homoneura and the phylogenetic relationship of the species groups are still not well understood. We investigated the male mor-
phology to provide the basis to further revise the species groups of this subgenus. In this survey, 230 species were examined and 117
morphological characters obtained, a phylogenetic analysis was conducted using the maximum parsimony analysis with TNT and
WinClada. The analyses yielded 45 most parsimonious trees and one strict consensus tree. A phylogenetic hypothesis is proposed
dividing the subgenus Homoneura into 12 species groups: H. (H.) nigrifacies, H. (H.) pallida, H. (H.) patella, H. (H.) beckeri,
H. (H..) formosae, H. (H.) henanensis, H. (H.) nigra, H. (H.) notostigma, H. (H.) ornatifrons, and H. (H.) trispina, H. (H.) laticosta,
and H. (H.) quinquenotata. This research provides valuable contributions towards a better understanding of the phylogenetic relation-
ships within the subgenus Homoneura. However, the monophyly of the genus and subgenus was not supported.

Key Words

Maximum parsimony, monophyly, morphology, phylogenetic relationship, revision

Introduction

The genus Homoneura Wulp, 1891 (Diptera, Lauxanii-
dae, Homoneurinae) comprises more than 750 described
species in eight known subgenera distributed worldwide
(Chen 2022). Homoneura is widely distributed in all major
zoogeographical regions except for the Neotropical region
and plays a very important role in the ecological system
(Shi et al. 2017). The subgenus Homoneura Wulp, 1891
comprises more than 700 described species and has the
highest species richness of Lauxaniidae (You et al. 2023).
When the classification of this group entered the sec-
ond stage of prosperity and development, the number of

described species increased markedly, significantly reduc-
ing the efficiency of comparative morphological studies.
Therefore, some scholars began to study species groups
among the subgenera of Homoneura based on characters
such as wing spots and male genitalia. Miller (1977a,
1977b) made a comprehensive revision of 50 species, nine
groups and four subgenera of Homoneura distributed in
the Nearctic region in his monograph “Taxonomy and Bi-
ology of Homoneura in the Nearctic region’. In particu-
lar, the establishment of species groups was an important
contribution to the systematic study of Homoneura (Miller
1977a, 1977b). Later, Papp proposed two species groups
from the Palaearctic region and four species groups from

Copyright Yao Yao et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use,
distribution, and reproduction in any medium, provided the original author and source are credited.

242 Yao Yao et al.: Phylogeny of homologous subgenera populations in China based on morphological characters

the Oriental region (Papp 1978; Papp et al. 2006). Sasaka-
wa (1992) proposed a new species group from the Oriental
region. Kim (1994) distributed the Australian species of
Homoneura into 15 species groups, and established 20 spe-
cies groups of the global subgenera of Homoneura. Shi and
Yang (2014) divided the Chinese species of the Homoneu-
ra subgenus into 21 species groups, but some of the species
could not be classified into species groups. Up to now, only
five scholars have made a morphological classification of
Species groups, because of the loss or destruction of type
specimens. The descriptions were too simple to study care-
fully. The accuracy and availability of a species group sys-
tem 1s doubtful, and the phylogenetic relationships of exist-
ing species groups have not been comprehensively tested.

The phylogenetical approach is an important way to re-
veal the relationships between taxa; however, relationships
within Homoneura are relatively behind. Stuckenberg (1971)
hypothesized the relationships among 19 genera of Homo-
neurinae based on morphological characters, and divided the
19 genera into three generic groups, with Homoneura in the
first group; Sasakawa (1992) hypothesized the phylogenetic
relationships among five subgenera of Homoneura based on
six morphological characters, with the subgenera distributed
into two clades, one containing the subgenera Chaetohomo-
neura Malloch, 1927 and Neohomoneura Malloch, 1927,
and the other comprising three subgenera with Euhomoneu-
ra Malloch, 1927 and Homoneura presented as sister groups,
and the subgenus Minettioides Malloch, 1929 closely relat-
ed to these (Sasakawa 1992). In the only existing molecular
phylogenetic study on the generic relationships of Homo-
neura, Shi et al. (2017) explored the relationships based on
two mitochondrial and two nuclear genetic markers, and the
hypothesis of subgeneric monophyly was not corroborated.
Kong et al. (2022) presented the first morphological phylog-
eny of Homoneura, and the hypothetical monophyly of the
genus and subgenus was also not supported. Furthermore,
the phylogenetic relationships among other subgenera of
Homoneura were also discussed. The evolutionary relation-
ships among the Homoneura subgenera are not completely
clear, and phylogenetic relationships of some species groups
have not been studied yet, making systematic classification
Studies in this genus difficult. At present, no phylogenetic
studies of the Homoneura subgenera have been published
using morphological evidence. Despite its high diversity
and ecological significance, the phylogenetic relationships
in Homoneura and its subgenera remain to be studied.

Methods

All the specimens are deposited in College of Horticul-
ture and Plant Protection, Henan University of Science
and Technology, Henan, China (Suppl. material 1).

Morphological study and terminology

General terminology follows Cumming and Wood
(2017), and Gaimari and Miller (2021). The specimens

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were observed with a Motic SMZ-168 stereomicroscope
and the external morphological characters were exam-
ined. Genitalia preparations were made by removing and
macerating the apical portion of the abdomen in cold sat-
urated NaOH for six hours, then rinsing and neutralizing
them with glacial acetic acid for dissection and study.
After examination in glycerine, they were transferred to
fresh glycerine and stored in a microvial pinned below
the specimen or moved to an ethanol tube together with
the wet specimens. Most characters were illustrated using
photographs and line drawings. Photographs were taken
using a Canon EOS6D microscope (Canon, Tokyo, Ja-
pan) and stacked using Helicon Focus v7.0.2.0 (Helicon
Soft, Kharkiv, Ukraine). Line drawings were drawn with
Adobe Illustrator 2021 v25.2.1 (Adobe, San Jose, USA).

Specimen morphological characters

The morphological characters were numerically coded
(Suppl. material 2). Ninety-eight characters are binary
and 25 are multistate. Plesiomorphic states were coded
with (0), and apomorphic with (1,2,3), missing character
states were coded with (?), and inapplicable states were
scored as (—).

List of characters used in the cladistic analysis

The partial characters and all pictures were adapted from
Kong et al. (2022) and revised accordingly for the pur-
poses of deeper research.

Head:

1. Size of head, height of head/width of head in frontal
view and the eyes are included: (0) < 4/5 (Fig. 1A);
(1) > 4/5 (Fig. 1J).

2. Color of ocellar triangle: (0) black (Fig. 1A); (1)
brown to yellow (Fig. 1E).

3. Number of ocellar seta: (0) 2; (1) 3.

4. Length of ocellar seta/length of anterior fronto-or-
bital seta: (0)> 1 (Fig. 1F); (1)< 1 and= 1/2;(2)< 1/2
(Fig. 1E).

5. Length of anterior fronto-orbital seta/length of pos-
terior fronto-orbital seta: (0) < 1 (Fig. 1B); (1) > 1
(Fig. 1M).

6. Length between anterior fronto-orbital seta and
posterior fronto-orbital seta/length between poste-
rior fronto-orbital seta and inner vertical bristles:
(0) > 1/2 (Fig. 1G); (1) < 1/2.

7. Frons: (0) flat (Fig. 1F); (1) distinct uplifted; (2) dis-
tinct concave (Fig. 1J).

8. Length of frons/width of frons: (0) < 1 (Fig. 1A);
(1) > 1 (Fig. 1E).

9. Middle of frons: (0) without spot or stripe (Fig. 1L);
(1) with a dark median longitudinal stripe extending
from anterior margin to the ocellar triangle or the
top of the head (Fig. 1C).

Dtsch. Entomol. Z. 71 (2) 2024, 241-254

10.

Lil

Between the middle of frons and fronto-orbital seta:
(0) with two longitudinal stripes extending to both
sides of the ocellar triangle (Fig. 1A); (1) without
stripe (Fig. 1B).

The area around the fronto-orbital setae and the out-
er vertical bristles: (0) without spot or stripe (Fig.
1H); (1) with spot (Fig. 1A); (2) with stripe along
the bases (Fig. 1D).

. Presence of a spot at the costal margin of frons: (0)

absent (Fig. 11); (1) present.

. Facial angle of frons: (0) approach straight angle:

(1) approach right angle.

. Color of face: (0) black (Fig. 1L); (1) brown to yel-

low (Fig. 1D).

. Spot on face: (0) absent (Fig. 11); (1) irregular; (2)

round (Fig. 1M).

. Middle of face: (0) flat (Fig. 11); (1) distinct uplifted

(Fig. 1K).

23.

24.

243

. Ventral margin of face: (0) strumaes on both sides

(Fig. L); (1) lamellar processes in the middle (Fig.
1J); (2) flat (Fig. 1D).

. Width of ventral margin of face/height of gena:

(0) >3 (Fig. 11); (1) <3 (Fig. 1J).

. Spot on gena: (0) absent (Fig. IN); (1) present

(Fig. 10).

. Below eye on gena: (0) without strong seta (Fig.

1M); (1) with strong seta (Fig. 1H).

. Length of gena/length of eye: (0) <1/3 (Fig. 1N);

(1) > 1/3 (Fig. 15).

. Face and gena: (0) do not extend (Fig. 1L); (1) dis-

tinct extend ventrally (Fig. 1J).

Color of pedicel: (0) black (Fig. 1H); (1) brown to
yellow (Fig. 1G).

Number of scape long bristle that is not shorter than
the length of the first flagellomere: (0) 0 (Fig. 1G);
(1) 2 (Fig. 1J).

Figure 1. Head characters. Homoneura (Homoneura) picta (de Meijere, 1904) (A, N); Homoneura (Homoneura) flavida Shi &
Yang, 2009 (B, C, D, O); Homoneura (Minettioides) orientis (Hendel, 1908) (E); Homoneura (Euhomoneura) yangingensis Shi,
Gao & Li, 2017 (F, I); Homoneura (Neohomoneura) tricuspidata Shi & Yang, 2008 (G); Prosopophorella yoshiyasui Sasakawa,
2001 (H, J, K); Minettia (Frendelia) longipennis (Fabricius, 1794) (L, P); Pachycerina decemlineata de Meijere, 1914 (M).

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244

2p

26.

27.

28.

29.

30.

Sik

Yao Yao et al.: Phylogeny of homologous subgenera populations in China based on morphological characters

Color of flagellomere: (0) monochrome (Fig. 1P);
(1) bicolor (Fig. 1N).

Arista: (0) plumose (arista with longest setulae,
not less than the width of the first flagellomere)
(Fig. 1M); (1) pubescent (arista with longest setu-
lae shorter than the width of the first flagellomere)
(Fig. 1H).

Length of the first flagellomere/width of the first
flagellomere: (0) > 2 (Fig. 1M); (1) <2.

Spot between base of antennae and inner margin of
eye: (0) present (Fig. 1H); (1) absent (Fig. 1).
Color of proboscis: (0) black; (1) brown to yellow
(Fig. IN).

Color of palpus: (0) monochrome, black (Fig. 1P); (1)
monochrome, brown or yellow (Fig. 10); (2) bicolor.
Occiput: (0) without stripe (Fig. 1G); (1) with dis-
tinct narrow stripe (as wide as ocellar triangle);
(2) with distinct wide stripe (distinctly longer than
ocellar triangle) (Fig. 1C).

Thorax:

32

Siok

34.
aD:
36.
SHY

38.

39.

AQ.

4].

42.

43.

Leg:

44.
45.

46.

Mesonotum: (0) with distinct longitudinal stripe
(Fig. 2C); (1) without longitudinal stripe (Fig. 2E).
Base of dorsocentral seta and prescutellar acrosti-
chal seta: (0) without spot (Fig. 2E); (1) with spot
(Fig. 2B).

Pre-sutural dorsocentral seta: (0) absent (Fig. 2E);
(1) present (Fig. 2D).

Post-sutural dorsocentral seta: (0) 3 (Fig. 2A); (1)
2(Fig. 2D).

Rows of acrostichal seta: (0) seven or more rows
(Fig. 2E); (1) six or less rows (Fig. 2D).
Acrostichal seta: (0) weak, short hair (Fig. 2B); (1)
strong seta (Fig. 2D).

Mesonotum prescutellar acrostichal seta: (0) longer
than acrostichal seta with hair; (1) hair, as long as
acrostichal seta with hair.

Mesonotum scutellar suture: (0) without spot (Fig.
2D); (1) with spot.

Mesonotum pruinose stripe: (0) no pruinose stripe;
(1) a broad pruinose stripe from face, antenna,
frons, mesonotum to the end of scutellum.
Supra-alar seta: (0) one (Fig. 2F); (1) two, the latter
is about half as long as the former; (2) two, the latter
is about the same length as the former one.
Intra-alar seta: (0) one, strong (as long as supra-alar
seta); (1) one, weak (half the length of supra-alar
seta); (2) 0.

Katepisternal seta: (0) two (Fig. 2F); (1) one (Fig.
20:

Length of leg/length of body: (0) <1; (1) > 1.
Posterior ventral seta on fore femur: (0) five or
more (Fig. 2L); (1) four or less (Fig. 2G).
Ctenidium short seta on fore femur: (0) absent; (1)
ten or less; (2) eleven or more (Fig. 2G).

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47.

48.

49.

50.

oll,

Anterior seta on mid femur: (0) six or more; (1) five
or less (Fig. 21).

Posterior seta on mid tibia: (0) absent (Fig. 2J); (1)
present (Fig. 2K).

Apical ventral seta on mid tibia: (0) one; (1) two
(Fig. 2J); (2) three (Fig. 2K); (3) four.

Preapical anterior dorsal seta on hind femur: (0)
present (Fig. 2H); (1) absent.

Anteroventral seta on hind femur: (0) present; (1)
absent.

Wing:

52

a3:

54.

ao

56.

OT.

58.

59:

60.

OF

62,

63.

64.

Length of wing/width of wing: (0) < 2.7 (Fig. 3A);
(1) => 2.7 (Fig. 3D).

Wing: (0) most hyaline or pale yellow (five or less
spots and wing spots occupy a small area of the
wing) (Fig. 3E); (1) most brown or black (six or
more spots and wing spots occupy a large area of
the wing) (Fig. 3A).

Anterior margin of the wing (in front of R2+3): (0)
most hyaline or pale yellow (spots on the wings less
than half of the anterior margin of the wings) (Fig.
3F); (1) mostly brown or black (spots on the wings
not less than half of the wings, while still a little part
with no spots) (Fig. 3B); (2) all brown, fading from
costa vein to the middle; (3) all brown, no change in
color from costa vein to the middle.

Short black setae on costal margin of wing: (0) ex-
tend to between R,,, and R,,. (Fig. 3G); (1) extend
to R,,. (Fig. 3C).

Spot on r, cell: (0) absent (Fig. 3E); (1) present
(Fig. 3C).

Spot on R,,,: (Q) absent (Fig. 3G); (1) 1, a hyaline
space (Fig. 3H); (2) 2 (Fig. 3C); (3) = 3; (4) all
brown (Fig. 3A).

Tip of R,,.: (0) without spot (Fig. 3E); (1) with one
spot (irregular) (Fig. 3H); (2) with two spots (irregu-
lar), far away from crossvein r-m (Fig. 3D); (3) with
two spots (irregular), next to crossvein r-m (Fig.
3A); (4) with three spots (irregular) (Fig. 3C); (5)
many small spots, reticular connection (Fig. 3A); (6)
many sexangular spots (a small spot in the middle).
Crossvein r-m: (0) without spot (Fig. 3G); (1) only
strengthened or with a inconspicuous stripe around
(Fig. 3E); (2) distinct spots (Fig. 3H); (3) circular
spots (Fig. 3B).

Crossvein dm-cu: (0) without spot (Fig. 3G); (1)
only overstrike crossvein or an inconspicuous stripe
around (Fig. 3E); (2) distinct spots (Fig. 3F); (3) cir-
cular spots (Fig. 3D).

Tip of M,: (0) without spot (Fig. 3G); (1) one spot
(Fig. 3F); (2) two spots (Fig. 3H); (3) many small
spots, reticular connection (Fig. 3A).

Stripe on penultimate section of CuA1: (0) absent
(Fig. 3F); (1) present (Fig. 3D).

Spot on base of radial vein and medial vein: (0) ab-
sent (Fig. 3E); (1) present (Fig. 3B).

Anal vein: (0) normal (Fig. 3H); (1) lack (Fig. 3B).

Dtsch. Entomol. Z. 71 (2) 2024, 241-254 245

tie sca aaa
y too

~ — —— a

ar \

Figure 2. Thoracic and leg characters. Homoneura (Homoneura) flavida Shi & Yang, 2009 (A, G, H); Homoneura (Homoneura)
picta (de Meiere, 1904) (B); Pachycerina decemlineata de Meijere, 1914 (C); Homoneura (Euhomoneura) yangingensis Shi, Gao
& Li, 2017 (D, I, J); Homoneura (Neohomoneura) tricuspidata Shi & Yang, 2008 (E, kK); Homoneura (Minettioides) orientis (Hen-
del, 1908) (F); Minettia (Frendelia) longipennis (Fabricius, 1794) (L).

65. Anterior cubital cell: (0) without spot (Fig. 3C); (1) 67. 3™ (between R,,, and R,, .) section/4" (between R, , .

A+5

with spot (Fig. 3A). and M,) section: (0) > 1.5; (1) < 1.5.
66. 2nd (between R, and R,,,) section/3™ (between R,,, , 68. The end of R,,,: (0) not bend to costal margin (Fig.
and R,, .) section: (0) > 3; (1) <3. 3F); (1) bend to costal margin (Fig. 3C).

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246

Yao Yao et al.: Phylogeny of homologous subgenera populations in China based on morphological characters

60:0

60:2

6122

Figure 3. Wing characters. A. Homoneura (Homoneura) picta (de Meijere, 1904); B. Noonamyia umbrellata Shi & Yang, 2009;
C. Homoneura (Homoneura) posterotricuspis Gao, Shi & Han, 2016; D. Prosopophorella yoshiyasui Sasakawa, 2001; E. Homo-
neura (Homoneura) flavida Shi & Yang, 2009; F. Homoneura (Neohomoneura) zengae Shi & Yang, 2008; G. Minettia (Frendelia)
longipennis (Fabricius, 1794); H. Homoneura (Euhomoneura) yangingensis Shi, Gao & Li, 2017.

69:

70.

a

AZ:

Length of the ultimate section of M1/ length of the
penultimate section of M1: (0)> 1; (1) <1.

Length of the ultimate section of CuA1/length of the
penultimate section of CuA1: (0) < 1/5; (1) = 1/5.
Crossvein r-m: (0) before or in the middle of the
discal cell (Fig. 3F); (1) behind the middle of the
discal cell (Fig. 3D).

Color of knob part of haltere: (0) black; (1) brown
or yellow.

Abdomen:

fiom

Spot on middle of tergite 2: (0) absent (Fig. 4B); (1)
present (Fig. 4A).

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74.

TS:

LO:

Ts

78.

79:

80.

Spot on side of tergite 2: (0) absent (Fig. 4F); (1)
present (Fig. 4D).
Spot on middle of tergite 3: (0) absent (Fig. 4C); (1)
present (Fig. 4A).
Spot on side of tergite 3: (0) absent (Fig. 4E); (1)
present (Fig. 4D).
Spot on middle of tergite 4: (0) absent (Fig. 4C); (1)
present (Fig. 4A).
Spot on side of tergite 4: (0) absent (Fig. 4F); (1)
present (Fig. 4D).
Spot on middle of tergite 5: (0) absent (Fig. 4C); (1)
present (Fig. 4B).
Spot on side of tergite 5: (0) absent (Fig. 4F); (1) with
striped spot (Fig. 4D); (2) with circular spot (Fig. 4E).

Dtsch. Entomol. Z. 71 (2) 2024, 241-254

247

Figure 4. Abdomen characters. Homoneura (Homoneura) picta (de Meijere, 1904) (A, D); Homoneura (Neohomoneura) tricus-
pidata Shi & Yang, 2008 (B); Homoneura (Euhomoneura) yangingensis Shi, Gao & Li, 2017 (C, F); Homoneura (Minettioides)
orientis (Hendel, 1908) (E).

81.
82.
83.
84.

85.

Spot on middle of tergite 6: (0) absent; (1) present.
Spot on side of tergite 6: (0) absent; (1) present.
Posterior margin of tergite: (0) not change color
(Fig. 4E); (1) distinct dark brown stripe (2) distinct
light brown stripe (Fig. 4B).

Sternite 5: (0) not differentiated; (1) differentiated
into a digitiform process.

Short spine at posterior margin of sternite 5: (0) ab-
sent; (1) present.

Male genitalia:

86.

87.

88.

89.

Syntergosternite and epandrium: (0) not fused; (1)
fused.

Shape of syntergosternite: (0) semicircular (Fig.
5C); (1) circular, without ventral processes (Fig.
5A); (2) circular, with ventral processes (Fig. 5B).
Length of dorsal margin of syntergosternite/length
of posterior margin of syntergosternite: (0) < 2/3;
(1).2 2/3.

Dorsal margin of syntergosternite: (0) without short
hair (Fig. 5C); (1) with short hair (Fig. 5B).

le

92.

93°

94.

oS.

96.

oF}

98.

oo

100.

. Syntergosternite around the spiracle: (0) without

short hair (Fig. SC); (1) with short hair (Fig. 5A).
Costal margin of epandrium: (0) with sharp process
or concave; (1) without sharp process or without
concave.

Length of dorsal margin of epandrium/length of
ventral margin of epandrium: (0) > 1/2; (1) < 1/2.
Surstylus: (0) separated from epandrium (Fig. 6A);
(1) not separated from epandrium (Fig. 6D).
Length of the longest surstylus/height of epandri-
um: (0) > 1/2 (Fig. 6D); (1) < 1/2 (Fig. 6B).

Shape of the apex of surstylus: (0) sharp (Fig. 6A);
(1) blunt (Fig. 6C).

Shape of surstylus: (0) bent (Fig. 6A); (1) straight
(Fig. 6C).

Width of the middle of surstylus/length of surstylus:
(0) < 1/2 (Fig. 6B); (1) = 1/2 (Fig. 6A).

Fine teeth or terminal processes on surstylus: (0)
absent (Fig. 6D); (1) present (Fig. 6C).
Hypandrium: (0) present; (1) absent.

Middle of hypandrium: (0) connected; (1) uncon-
nected.

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248 Yao Yao et al.: Phylogeny of homologous subgenera populations in China based on morphological characters

101:1

POs
Figure 5. Syntergosternite and hypandrium characters. A. Homoneura (Homoneura) dorsacerba Gao, Shi & Han, 2016; B. Homo-
neura (Homoneura) posterotricuspis Gao, Shi & Han, 2016; C. Homoneura (Homoneura) procerula Gao & Yang, 2005; D. Cestro-
tus liui Shi, Yang & Gaimari, 2009.

101. Shape of hypandrium: (0) U-shaped (Fig. 7A); (1) 107. Seta on gonite: (0) present (Fig. 7D); (1) absent

Y-shaped (Fig. 5D); (2) H-shaped (Fig. 7D); (3) (Fig. 7A).
W-shaped (Fig. 7B). 108. Length of gonite/length of phallus: (0) > 1/2
102. Middle of anterior margin of hypandrium: (0) with- (Fig. 7D); (1) < 1/2 (Fig. 7A).
out inner processes (Fig. 7D); (1) with inner pro- 109. Tip of gonite: (0) sharp (Fig. 7D); (1) blunt
cesses (Fig. 5D). (Fig. 7A).
103. Both sides at anterior margin of hypandrium: (0) — 110. Phallus: (0) without thorn or sharp process (Fig. 7C);
without inner processes; (1) with inner processes. (1) with thorn or sharp process (Fig. 7B).
104. Middle of posterior margin of hypandrium: (0) — 111. Lateral view of the tip of phallus: (0) bent (Fig. 7C);
without ventral process (Fig. 7A); (1) with ventral (1) straight (Fig. 7A).
process (Fig. 7B). 112. Tip of phallus: (0) not inflated; (1) blunt round api-
105. Both sides at posterior margin of hypandrium: (0) cally.
without ventral process; (1) with ventral process. 113. Apex of phallus: (0) sharp (Fig. 7C); (1) not sharp
106. Gonite: (0) present (Fig. 7D); (1) absent (Fig. 7C). (Fig. 7B).

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Dtsch. Entomol. Z. 71 (2) 2024, 241-254

97:1

95:0;96:0

95:1;96:1

(Homoneura) posterotricuspis, D. Pachycerina decemlineata.

114. Phallus: (0) with distinct apical concavity; (1) with-
out distinct apical concavity (Fig. 7B).

115. Aedeagal apodeme: (0) present; (1) absent.

116. Length of aedeagal apodeme/length of phallus:
(0) < 1 (Fig. 7D); (1) = 1 (Fig. 7A).

117. Base of aedeagal apodeme: (0) separate and extend
(Fig. 7D); (2) not separate (Fig. 7C).

Phylogenetic analysis

In this research, two species of Lauxaniinae: Minet-
tia (Frendelia) longipennis (Fabricius, 1794) and

249

"98:0

Figure 6. Epandrium characters. A. Homoneura (Homoneura) trispina, B. Homoneura (Homoneura) dorsacerba, C. Homoneura

Pachycerina decemlineata Meijere, 1914 and representa-
tive species of all genera of Homoneurinae found in China
except Homoneura, Cestrotus liui Li et al., 2009, Dioides
incurvatus Shi et al., 2009, Noonamyia umbrellata Shi et
al., 2009, Phobeticomyia motuoensis Li et al., 2020. and
Prosopophorella yoshiyasui Sasakawa, 2001 were used
as an outgroup. Minettia (Frendelia) longipennis is the
first outgroup.

The phylogenetic construction was conducted using
maximum-parsimony analysis. The unambiguous char-
acters were mapped on the tree using WinClada version
v1.00.08 (Nixon 2002). The maximum-parsimony tree
is shown in Suppl. material 3, Bootstrap values (BS)

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250 Yao Yao et al.: Phylogeny of homologous subgenera populations in China based on morphological characters

\

110:1 113:1:114:1

116:0;117:0

|| 106:0;107:0

Figure 7. Aedeagal complex characters. A. Noonamyia umbrellata, B. Homoneura (Homoneura) beckeri; C. Pachycerina decem-
lineata, D. Homoneura (Homoneura) procerula.

and Bremer support (B) values are presented next to the 25 and 30 were used, maintaining the maximum num-
nodes. The analysis was performed in TNT (version 1.1; ber of trees (10000 trees). All analyses were performed
Goloboff et al. 2008) using implied weighting. For the — using traditional searches, 900 replicates holding up to
implied weighting analyses, K values of 2-5, 10, 15,20, 45 trees per replication. Branch support values were ver-

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Dtsch. Entomol. Z. 71 (2) 2024, 241-254 251

ified through bootstrap analyses on NONA 2.0 (Goloboff — ters, Fig. 2A—F), legs (8 characters, Fig. 2G—L), wings
1999). The Bremer support value or decay index was cal- (21 characters, Fig. 3), abdomen (13 characters, Fig. 4),
culated using TNT. and genitalia (32 characters, Fig. 5—7). Branch length is
the shortest and the topology structure no longer changes
when K is not less than 20. Forty-five maximum-parsimo-
nious trees were produced when K = 30. Different maxi-
mum-parsimonious trees mainly affect the monophyletic
and interspecific relationships of Homoneura (Homoneu-

ra) henanensis group, and the topology structure of each
On the basis of our research, 117 morphological charac- branch end. The resultant strict consensus tree was cal-

ters were obtained from different body parts of the adults; culated using TNT (Fig. 8) [branch length = 1413, con-
from the head (31 characters, Fig. 1), thorax (12 charac- sistency index (CI) = 0.11, retention index (RI) = 0.73].

Results

Phylogenetic analysis of Homoneura

é
s+
33 8
= = ga 3 g
S & = 3
EM en ee ORE A es Sey
3 Be Ste Sears
BR LER ETEERPRASRSES SFE
S BEE RES Sea tAZAK eS Sse E
5 = =) Pees 3
RE ewERRLLELLL SEES EFF rss iS2
3 2382 BRET EES = 3 eRe
S eh BSR at Feats is isigses se Fk
E% CELEEELIR EP PESEP PSPSPS sss Fs
Se etERPELet et Ei PF i ssss sss iss ss
*BREREEELELELE RES ES SEESSSS Sess es
SCeREERRESRERLERS ZELER EEE SEESSS
Sees he Seah bs ZEEE FERS ESF Pee FS8
@S%GhetSretezea sl tagi ei esSeigsssiss>
SESE EEES EER EE EEG ESLEGER SEES fess
@%eereueseraesEs Fgh SR Eistesgsssssse Fs
‘Se PSPELESen ds GESER ES SES EE EF 2 FF F
tne EZ Eee eles eh ee ees Sel ees Eg
4% EY 3% Yaeger eee ss
J %%ak e OBS = ,y id z=
SOLS iis | SEU py edd
OOOO AS Cae tA | fe} pa Od / SESE FS ESSE
Oy Ge, 4, eee eS ee wp Wot ff PEE EEE SEP OOM OS
a . % 7
Ag, Pag, ey My POA a vey Vv aa i ¥S & S og fg

Veep

5/78 ~—\

Homoneura
=c = Pachycerina decemlineata
Homoneura (Neohomoneuray tricuspidata 10/100 4 = Minettia (Frendelia) longipennis

Homoneura (Neohomoneura) indica ——_— et oe

Homoneura (Neohomoneura) dischida Homoneura (Homoneura) sp38

= = : Faones (Homoneura) sp37

ul
Homoneura (Nechomone oe : = =
—< 5 oneura (Homoneuray
HHomoneura (Net (Chactohomoneura) =P Vv y Homoneura (Hom, euaresta
Homoneura ( i) trisurstylata omonery ‘oneura) sp35
onewr® earth = :

&
S2e8an ==
SSP sePRESaeESSEETEEE
Ss s aoe R22Z3322 s2e5
SSEPSPFEZPPESESESLEERB ES
SSESSSESPEESSZ RR EEE Fy
$s FeF PPP PEPE ELS SER 225e% %
gers gs Fa 8 sssessad aoe Se
SSF FEC FIT FS SESEERRRRESTETR2A3 3 23%
¥ = &s sls F258 ESS EEDEFSTS3225% %. S %
Wo Fe FSF PESTS SSES TELL LSS Pb kkk h S385
FFs Fs Fs FFeFssrsssssSSsb gab Res Reyes
SF PF FFF FTTFFESSP STP PEPER RS SER =
Gy = 4 SS.
Ed PEGSE PUP PPLE SEL es soe ahat
sSFrss SEFTIUTESSaBeE Se
gs gst FFs sess TeEger se” 5
g 368 2 2. =.
fi ae *
= § Be 3
a) 8 § 2

Figure 8. Phylogenetic relationships of Homoneura. 1. H. (H.) ornatifrons group; UL. H. (H.) patella group; I. H. (H.) notostigma
group; IV. H. (#.) nigra group; V. H. (H.) nigrifacies group; VI. H. (H.) trispina group; VIL. H. (4.) beckeri group; VII. H. (#.)
formosae group; IX. H. (#.) laticosta group; X. H. (H.) pallida group; XI. H. (H.) henanensis group; XII. H. (H.) quinquenotata
group. Left, bootstrap values > 50%; right, Bremer support values.

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252 Yao Yao et al.: Phylogeny of homologous subgenera populations in China based on morphological characters

Relationships among the Homoneura
subgenera occurring in China

The monophyly of the subgenus Homoneura, Neohomo-
neura, Euhomoneura and Chaetohomoneura is not sup-
ported. The monophyly of the subgenus Minettioides

could not be verified due to the limited taxa. One species
of the subgenus Chaetohomoneura appears at an end node
within the subgenus Neohomoneura, while the subgenus
Euhomoneura, Neohomoneura, Minettioides and Chaeto-
homoneura also appear mosaically distributed within the
subgenus Homoneura.

Key to the species groups of the Homoneura subgenus based on the maximum-parsimony tree

1 Wine without spots oronly with qpalé SOOlS. OM chOSSVEINS CFIS. Sie Gite: hr mee Beem er RA Nee 2 ae ire ns ee ae 2
- VV GBs MELE They HULSE CUIOROWAT.SOOLSQCh ee Son: ff ee eta ee hte tastes dei sate see dee el a ee iets es Fe oer kL 10
2 BOCWVEIOWGOr FOWh (eles 2U PDR EP ak, ocvyeens coverennasesnteaecuyesueree agmneidbawieagens ceed ehaen ele anes Seow eee bend seei¥a dh muyeny ecauiednaaen S
- BOC yrblaG kK Cie O25 x) heel Puts eit tit as eee Aes Amo Ll a Boe Rae oe eects Me, Oa ead eb teed! 6
3 Two stripes along orbital bristles (Fig. 1A), a triangular brown stripe in the middle of frons (Fig. 1C); hypandrium

A esd ge 181210). ah 3 elias i POR Ric Ses Se PT oe oe a Re A eee pee eee CP ee eee eek ie rere H. (H.) beckerl group
~ FronsSwitheuhstriper(rig.. LBs nypancrium-mormw-SNaoeGhs -jsn20 Sh snp end lets. Me class us reams ea bedee tha Sebaehadadiee Means ud tade 4
4 Post pedicel bicolor (Fig. 1N); mesonotum scutoscutellar suture with black spots (Fig. 1H)..... H. (H.) notostigma group
- Flagellomere yellow. (Fig. 1P);mesonetum scutellar-suture without spot (Fig: 2D). ccs..2.t2..ccchciss bec seethtedeteete he cestaacetiete 6
5 Length of ocellar seta is shorter than the length of anterior fronto-orbital seta (Fig. 1E)................ H. (H.) patella group
- Length of ocellar seta is longer than the length of anterior fronto-orbital seta (Fig. 1F) .............. H. (H.) laticosta group
6 Prescutellaracrostichal-seta weak, Same as acrostichal Sela «...5555;, 0010) gaye seeseeeneyyygodeten treedeeses H. (H.) ornatifrons group
_ Prescutellar acrostichal seta strong, longer than acrostichal Se@ta................cccccecceceeeneeeseeeeeeeeeeceeceessecseeeseeceeserteeeereneens 7
7 With a broad white pruinose stripe from face to the end of scutellum (Fig. 4B) ..........cceeecc cece eeee enna es H. (H.) nigra group
~ WiLL: PRULN@ Sess tal DS Om haCere.SC US| [LIM dlal ard cake, buat. Mes a0 0,9 noe wrest erler UV ee he te) ee Rosebc tae e 8
Sf = TESS Sawith Salt Ol DlACK SOUS CI: ALB ey cecvescet saaee tees au roc ceva te banure poe yicey cevernsee-cbeeeenimencones H. (H.) formosae group
- EF eICeco-WTOU SOO TS Rts ie pee aa as health Re eRe ce Oe ieee Pb R ae le os a rina Pee Na, | a eee Pe ee ns a esta en oes 2]

9 Base of fronto-orbital seta with a stripe (Fig. 1D); stripe on the middle of frons extending to ocellar triangle (Fig. 1C)..
Jes Se casa caper Re wey eae En od eainchee RD pails 9 see Seine oe talents a gs a Vee ahem an ager out a MRM el Tasceee cu PEE, CAN an ce ton ace H. (H.) nigrifacies group

- Base of fronto-orbital setae without a stripe; only costal margin of frons yellow (Fig. 1E).............. H. (H.) trispina group
TS VIS MTR OUESOOUS OM EGhOSSVOIN dette ae SON) aSaase che Seaqs dep tao spent Dietmar gpk vai PanaNeee ore eaaas tisk paaenm hs Bicaawepeh ete vaRaRENE arr tas Abell
- WiINBEWITT; SOGLS ON rCrOSS Vel MCh SSE). once. nt oie Sec eee leiede cas cynecteiiceesss Miboidareetee sti es tebhboes H. (H.) quinquenotata group
1d SBi oe without spotsbetweens-iandrapical: Spot (Fig. SE). asdF/) ses. has acedcssefasts il ndeaglel: ecaaseess H. (H.) pallida group

- R,,. with spots between r-m and apical spot (Fig. 3D)

Discussion

Our study proposes a phylogenetic relationship hypoth-
esis for the genus and subgenus Homoneura using mor-
phological data. In this study, five genera of Homoneuri-
nae except for Homoneura were included for the purpose
of serving as an outgroup. Previously, Kong et al. (2022)
published the phylogeny of the Chinese subgenera of
Homoneura based on morphological characters, and ob-
tained the conclusion that the monophyly of the genus
and subgenus Homoneura is not supported, and this con-
clusion is further verified in this study. Some relationships
among genera are in close agreement with the literature,
suggesting that traditional taxonomic characters are more
reliable at the generic level. Although the non-monophy-
ly of Homoneura was strongly supported, the position of
the species groups across the subgenus Homoneura was
less consistent.

Based on the increment of available morphological
characters and on the result of our analyses, we pro-
pose to divide the subgenus Homoneura. This 1s dif-
ferent from the the method of dividing the subgenus

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REET. Ff Sade, SRI OE es Be fe eT H. (H.) henanensis group

Homoneura solely based on spots on the wing. Based
on the resultant strict consensus tree we propose to re-
duce the 21 species groups into 12 species groups by
establishing three new species groups: H. (H.) nigrifa-
cies, H. (H.) pallida, and H. (H.) patella, keeping sev-
en species groups: H. (H.) beckeri, H. (H.) formosae,
H. (H.) henanensis, H. (H.) nigra, H. (H.) notostigma,
H. (#.) ornatifrons, and H. (H.) trispina; and combin-
ing the remaining species groups into two new species
groups: H. (H.) laticosta, and H. (H.) quinquenotata.
However, we advise for caution since these possible
placements are tentative at best and further studies are
needed to verify these observations.

Previously, the only existing molecular phylogenet-
ic study of the generic level relationships of Lauxani-
idae, Shi et al. (2017) explored the phylogeny of the
subgenus Homoneura based on two mitochondrial and
two nuclear markers. But the results did not support the
published taxonomy of the species groups of the sub-
genus Homoneura in China (Shi and Yang 2014). All
of these indicates that there are many problems in the
division of species group of the subgenus Homoneura.

Dtsch. Entomol. Z. 71 (2) 2024, 241-254

In our research, we provide solid evidence that the cur-
rently recognized genus and subgenus Homoneura are
not monophyletic. Thus, our results indicate that the
current classification of the Homoneura does not accu-
rately reflect the evolutionary history of the group. This
result is consistent with the results obtained by Shi and
Yang (2014). We do not seek to formally propose one
of these changes here because the resolution of these
clades is poor and because our taxon sampling is limited
(only Chinese species are involved). We can only try
to separate species groups of the subgenus Homoneura.
Furthermore, we need to take a closer inspection of the
morphology, focusing on an alternative level of analy-
sis. Therefore, in order to enable such taxonomic deci-
sions, we urge for a complete phylogeny of the Homo-
neura with datasets including more morphological and
molecular data.

Conclusions

Our research results show that the monophyly of the
genus and subgenus Homoneura is not supported,
and the confusion of the subgeneric division 1s one of
the main problems of this genus, which makes some
scholars ignore the subgeneric rank in their works, and
there are often taxonomists who misclassify subgen-
era when describing species. The main reason is that
the key characteristics and validity of subgeneric level
classification are not clear. Although not the primary
focus of this study, our results also provide a prelim-
inary glimpse of potential major relationships within
the genus Homoneura. Nevertheless, we emphasize
that this study was focused on testing the monophyly
and the relationships of the species groups within the
subgenus Homoneura. Thus, our findings regarding the
major relationships within the entire genus should be
considered, at best, preliminary. And although these
conclusions are based on a small sample size compared
with the diversity of the genus as a whole, our results
indicate that the classification of the genus at the sub-
genus and species group levels will require a thorough
reformulation that reflects the family’s evolutionary
history. In these cases, we should re-analyze the mor-
phological data, trying to look for misinterpretations of
morphological characters.

As a next step for the phylogeny and classification
of the genus and the subgenus, we will try to find syn-
apomorphies of early diverging lineages, whose phylo-
genetic placements are not well supported in our study.
Additionally, the phylogeny of each subgenus of Homo-
neura is still unclear, and the monophyly and phyloge-
netic relationship of species groups is still blank, and
none of them have been examined using molecular data.
The availability of molecular data for species of Homo-
neura has increased during recent years, and we encour-
age the phylogenetic analysis among the species groups
at the subgeneric level using a combination of morpho-

253

logical and molecular data. Furthermore, we will review
specimens from other zoogeographical regions and re-
vise the species groups of the Homoneura subgenera by
using geographic distribution information to reconstruct
the phylogeny of the genus Homoneura making taxo-
nomic revisions.

Acknowledgements

We express our sincere thanks to Xulong Chen for identi-
fication of the specimens of Homoneura.

References

Chen XL (2022) Taxonomy of Homoneura in China. D. M.S. Henan
University of Science and Technology, 247 pp.

Cumming JM, Wood DM (2017) Adult morphology and terminology.
In: Kirk-Spriggs AH, Sinclair BJ(Eds) Manual of Afrotropical Dip-
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Africa Vol. 1, 89-133.

Gaimari SD, Miller, RM (2021) Lauxaniidae (Lauxaniid flies). In Man-
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South African National Biodiversity Institute: Pretoria, South Africa
Volume 3: 1757-1781.

Goloboff PA (1999) NONA (NO NAME) ver. 2.0. Z. Published by the
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Goloboff PA, Farris JS, Nixon KC (2008) TNT, a free program for phy-
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Kong CY, Feng KL, Zhao SJ, Li WL, Li XK (2022) Phylogeny of the
Chinese Subgenera of the Genus Homoneura (Diptera, Lauxaniidae,
Homoneurinae) Based on Morphological Characters. Insects 13:
665. https://doi.org/10.3390/insects 13080665

Miller RM (1977a) Taxonomy and biology of the Nearctic species of
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Nixon, KC (2002) WinClada; ver. 1.00.08; Nixon, K.C.: Ithaca, NY,
USA.

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Sasakawa M (1992) Lauxaniidae (Diptera) of Malaysia (part 2): a re-
vision of Homoneura van der Wulp. Insecta Matsumurana 46:
133-210.

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254 Yao Yao et al.: Phylogeny of homologous subgenera populations in China based on morphological characters

Shi L, Yang D (2014) Supplements to species groups of the subgenus
Homoneura in China (Diptera: Lauxaniidae: Homoneura), with de-
scriptions of twenty new species. Zootaxa 3890.1: 1-117. https://
doi.org/10.11646/zootaxa.3890.1.1

Shi L, Zhang MJ, Shen RR, Li SD, Bai J, Wang YJ, He GW, Cui Y
(2017) Preliminary study on phylogeny of species groups in subge-
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org/10.11646/zootaxa.5257.1.11

Supplementary material |

The species studied

Author: Yao Yao

Data type: pdf

Copyright notice: This dataset is made available under
the Open Database License (http://opendatacommons.
org/licenses/odbl/1.0). The Open Database License
(ODbL) is a license agreement intended to allow us-
ers to freely share, modify, and use this Dataset while
maintaining this same freedom for others, provided
that the original source and author(s) are credited.

Link: https://do1.org/10.3897/dez.71.120389.suppl1

dez.pensoft.net

Supplementary material 2

Morphological dataset used for the analysis
of the phylogeny

Author: Yao Yao

Data type: pdf

Copyright notice: This dataset is made available under
the Open Database License (http://opendatacommons.
org/licenses/odbl/1.0). The Open Database License
(ODbL) is a license agreement intended to allow us-
ers to freely share, modify, and use this Dataset while
maintaining this same freedom for others, provided
that the original source and author(s) are credited.

Link: https://doi.org/10.3897/dez.71.120389.suppl2

Supplementary material 3

Maximum parsimony tree

Authors: Yao Yao, Chaoyang Kong, Pu Miao, Shengjuan
Zhao, Wenliang Li

Data type: zip

Copyright notice: This dataset is made available under
the Open Database License (http://opendatacommons.
org/licenses/odbl/1.0). The Open Database License
(ODbL) is a license agreement intended to allow us-
ers to freely share, modify, and use this Dataset while
maintaining this same freedom for others, provided
that the original source and author(s) are credited.

Link: https://doi.org/10.3897/dez.71.120389.suppl3