﻿Two new species and records of Neoperla (Plecoptera, Perlidae) from Yunnan, China

﻿Abstract Two new species of the stonefly genus Neoperla, N.gaoligongshanasp. nov. and N.hajekisp. nov. are described from Yunnan Province of southwestern China based on the morphological and distributional data, and the new species are compared with two congeners, N.wuzhishana Chen & Du, 2016 and N.orissa Stark & Sivec, 2015. Neoperlahubleyi Stark & Sivec, 2008 is recorded from Yunnan Province for the first time.

We recently examined a collection of stonefly specimens received from the National Museum Prague of the Czech Republic, and some of the results of our investigations into this material have been already published (Mo et al. , 2020b(Mo et al. , 2021a(Mo et al. , 2021bLi and Kong 2020;. Herein, we report our results on the Neoperla from Yunnan Province in this collection, including two new species and one new record from the province.

Materials and methods
Types are kept in the Insect Collection of Henan Institute of Science and Technology (HIST), Xinxiang of China and the National Museum Prague (NMP) of the Czech Republic, and the Collection of Smaller Insect Orders, Department of Zoology, Hungarian Natural History Museum (HNHM), Budapest, as indicated in the text. Specimens used in this study were collected using a light trap and stored in 75% ethanol. Specimens were examined with the aid of a Leica M420 dissecting microscope and the color photographs were taken with a Leica S8APO stereo microscope. Aedeagi were everted using the cold-maceration technique of Zwick (1983). The morphological terminology follows that of Stark and Sivec (2008). The map ( Fig. 7) was prepared using a base map of Yunnan Province downloaded from DataV.GeoAtlas (Alibaba, China) and ACME Mapper 2.2 (http://datav.aliyun.com/tools/atlas/index.html; http://mapper.acme.com/). Diagnosis. Males of this species are characterized by having the tergum 7 with a triangular process with sparse spines apically. The aedeagal tube is straight, and aedeagal sac is armed with spines.

Results and discussion
Description. Adult habitus (Fig. 1). Body color brown. Head mostly yellowish brown, with a black marking covering ocellar triangle, the marking extended forward to pale M-line and getting brown, and a narrow triangular marking occurring forward of M-line. Head approximately as wide as the pronotum; compound eyes black; antenna and palpi yellow. Distance between ocelli slightly wider than the diameter of an ocellus. Pronotum disc brown, midline darker, margins pale ( Fig. 1B, C). Wings subhyaline, veins brown; legs pale to yellowish brown, femorotibial joint dark brown. Abdomen brownish, cerci brownish ( Fig. 1B, C).
Aedeagus (Fig. 2). Aedeagal tube weakly sclerotized ventrally and dorsally sclerotized strongly, apex with two ventral spinous lobes ( Fig. 2A, B). Sac nearly straight, ca 2× as long as the tube. Spinose apex of sac slender, slightly ventrally curved, with an apical dorsolateral patch of black spines and a subapical ventral patch of spines ( Fig. 2A, C); two wide rows of numerous smaller spines covering most of the dorsal surface of the sac, basal half of spinous rows interrupted medially ( Fig. 2A, C).
Female. Unknown. Etymology. The specific name refers to the Gaoligongshan National Nature Reserve in Yunnan Province, where the type locality is situated.
Distribution. China (Yunnan). Ecology. Gaoligongshan National Nature Reserve is located in northwestern Yunnan Province and is the largest nature reserve of the province. At the same locality, accompanying stoneflies were Neoperla hajeki sp. nov. and Tyloperla illiesi Stark & Sivec, 2005. Remarks. The new species is a member of the N. montivaga group. The aedeagal tube and terga 7-10 of the new species are similar to those of Neoperla wuzhishana Chen & Du, 2016, but N. wuzhishana can be distinguished from the new species primarily by the shape of the aedeagal sac and aedeagal armatures (Fig. 8C). In N. wuzhishana, the aedeagal sac is distinctly curved and expanded apically (Fig. 8C, present study) (which is obscure in the original drawing because of an apical damage in the type) and has at least four large dorsal spines subapically (see figs 7, 8 in Chen, Du 2016b); however, the aedeagal sac of N. gaoligongshana is nearly straight and the apex is constricted and has only small spines subapically on its dorsal side. In addition, the aedeagal sac of N. wuzhishana bears a dozen moderately long spines at mid-length which are absent in N. gaoligongshana. Besides, the color pattern including that of the head and legs of N. wuzhishana and N. gaoligongshana is different: N. wuzhishana has brown legs and the head is pale with a small dark spot between the posterior ocelli, while in N. gaoligongshana, the legs are pale to yellowish brown, the femorotibial joint is dark brown, and the head has a large, black marking covering the ocellar triangle, with this marking extending anterolaterally under the pale M-line (Fig. 1B). We examined specimens of N. wuzhishana from Yinggeling, Hainan Province, and found slight intraspecific variations of head pattern and armatures of the aedeagal sac: the dark area between ocelli in males was slightly smaller than in females, which in both sexes are quite small (Fig. 8A, B); four large spines in types (both in the original illustrations and descriptions) (see figs 7, 8 in Chen, Du 2016b), which can number seven to nine in our specimens (Fig. 8C, D). Therefore, the absence of large aedeagal spines in N. gaoligongshana is regarded as a distinguishing character, separating it from N. wuzhishana. Diagnosis. This species is characterized by a small dark marking over the ocellar area and a dark brown stigma before the M-line. The male of new species is characterized by an S-shaped aedeagal tube and by a sac bearing a subapical triangular patch of spinules in dorsal aspect.

Neoperla hajeki
Description. Adult habitus (Figs 3, 5). General body color brown. Head general pale brown, a small dark marking covers ocellar area, with a dark brown stigma before M-line. Head slightly wider than pronotum; compound eyes blackish, antenna dark brown except the basal segment yellowish (Figs 3A, 5A), palpi pale brown. Distance between ocelli narrower than the diameter of ocellus and a small marking between ocelli dark brown. Pronotum yellow, with rugosities and a strip-like midline (Figs 3A, 5A). Wings hyaline, veins brown; legs brown, basal part of femur dark brown, with wider yellow bands in mid-and hind legs (Figs 3B, 5B). Cerci yellowish (Figs 3D, 5B).
Etymology. The species is named after Dr Jiri Hájek for collecting the specimens.

Remarks.
The new species is a member of the N. clymene group. Color pattern, pronotum, and male genital features are similar to Neoperla orissa Stark & Sivec, 2015 from India. The new species can be easily separated from N. orissa by the projection of tergum 7 and detail of the aedeagal sac armature. In N. hajeki, the projection of tergum 7 is pointed in dorsal aspect, and the dorsal spines of the aedeagal sac are triangularly arranged. In N. orissa, the slender and median projection of tergum 7 appears truncate in dorsal aspect, and the spines of the sac are arranged in several close-set rows.
Remarks. Neoperla lihuae Li & Murányi, 2014 was originally described by Li et al. (2014a) from Xishuangbanna in Yunnan Province. Our specimens from Yingjiang County agree well with original description of the head pattern and terminalia, the aedeagal tube and sac.

Neoperla hubleyi Stark & Sivec, 2008
Neoperla hubleyi Stark and Sivec 2008: 30 (original   Remarks. Stark and Sivec (2008) originally described this species from Vietnam, and Mo et al. (2020a) described a new record in Guangxi Zhuang Autonomous Region. The terminalia and aedeagus of our specimen fit the original description of Stark and Sivec (2008).

Concluding remarks
Neoperla was divided into the Neoperla clymene group and the Neoperla montivaga group by Zwick (1983). The N. clymene species group currently includes more than 150 species, and the N. montivaga species group includes over 123 species worldwide (Huo et al. 2021;Mo et al. 2021a).
So far 11 species of Neoeprla have been recorded from Yunnan Province, including the two new species and one species newly recorded in this paper. Among the six  endemic species of Neoperla, including the two new species listed below-N. lihuae, N. yanlii, N. obscurofulva, N. yunnana, N. hájeki, and N. gaoligongshana-three of them are distributed in Baoshan City. Five species are widely distributed: N. cavaleriei, N. diehli, N. hubleyi, N. limbatella, and N. lui. Neoperla binodosa Wu, 1973 was transferred to the genus Phanoperla and is placed as a synonym of P. pallipennis Banks, 1938 by Mo et al. (2021c). Most species are distributed in western and southeastern Yunnan Province and central and northern Yunnan still needs to be surveyed (Fig. 7).