On four species of the genus Argiope Audouin, 1826 (Araneae, Araneidae) from China

Abstract Based on morphological and molecular evidence, Argiope macrochoera Thorell, 1891 from China is found to be the unknown female of A. cameloides Zhu & Song, 1994, the known male of A. perforata Schenkel, 1963 is mismatched and provisionally suggested to be the male of A. boesenbergi Levi, 1983, and the true male of A. perforata Schenkel, 1963 is described for the first time. Argiope abramovi Logunov & Jäger, 2015 is suggested to be a synonym of A. perforata Schenkel, 1963. Argiope chloreides Chrysanthus, 1961 and A. vietnamensis Ono, 2010 are newly recorded from China. The unknown male of A. vietnamensis Ono, 2010 is described for the first time.


Introduction
Argiope Audouin, 1826 is nested in the subfamily Argiopinae Simon, 1890 together with the genera Gea C.L. Koch, 1843 and Neogea Levi, 1983, and it comprises sexually dimorphic species well known for their showy, colorful females and their unique web stabilimenta (Levi 1983;Tan 2018). Argiope has 89 species worldwide, and it is the most diverse genus in Southeast Asia, including New Guinea and adjacent islands (Jäger 2012;WSC 2020). The genus is rather well studied on account of a series of revisions and reviews by Levi (1983Levi ( , 2004, Bjørn (1997), Yin et al. (1997), Tanikawa (2009), andJäger (2012). However, more than one-third of its species (31) are known only from a single sex: four by males and 27 by females. Two species lack diagnostic illustrations and cannot be confidently identified. Thus, Argiope remains inadequately known. To date, 19 species have been recorded from China, of which three are endemic.
While examining Argiope specimens from southwest China, A. chloreides Chrysanthus, 1961, A. vietnamensis Ono, 2010, and the previously unknown male of the latter were recognized. An extended study of morphological and molecular evidence has revealed that A. macrochoera Thorell, 1891 from China is misidentified and conspecific with A. cameloides Zhu & Song, 1994, and that the male of A. perforata Schenkel, 1963 is mismatched and may be A. boesenbergi Levi, 1983. Moreover, the true male of A. perforata is revealed for the first time, and A. abramovi Logunov & Jäger, 2015is synonymized with A. perforata Schenkel, 1963. The goals of the present paper are to revise and describe the misidentifications, mismatches, and the unknown sexes of A. cameloides, A. perforata, and A. vietnamensis, as well as to provide a distributional map of those species.

Materials and methods
All specimens were collected by beating shrubs or hand collecting and were preserved in 75% ethanol, except for the specimens for DNA extraction which were preserved in absolute ethyl alcohol. All specimens were deposited in the Museum of Tongren University, China (TRU).
The specimens were examined with an Olympus SZ51 stereomicroscope. After dissection, the epigyna were cleared in a trypsin enzyme solution before examination and imaging. Left male palps were used for the descriptions and illustrations. Photographs of the copulatory organs and habitus were taken with a Kuy Nice CCD or an Olympus C7070 camera mounted on an Olympus BX51 compound microscope. Compound focus images were generated using Helicon Focus v. 6.7.1.
All measurements are given in millimeters. Leg measurements are given as total length (femur, patella + tibia, metatarsus, tarsus). Abbreviations used in the text and figures are as follows: The total genomic DNA from spider legs was extracted using the Animal Genomic DNA Isolation Kit (Kangwei Biotech, Beijing, China) following the manufacturer's protocols. The primer pair LCO1490/HCO2198 (Folmer et al. 1994) was used to amplify cytochrome c oxidase subunit I (COI) under the following PCR reaction protocol: initial denaturation at 95 °C for 5 min; 35 cycles of denaturation at 95 °C for 1 min, annealing at 40 °C for 1 min, and elongation at 72 °C for 30 s; and a final extension at 72 °C for 7 min. The 25 μl PCR reactions consisted of 12.5 μl of 2×Taq MasterMix or 2×Es Taq MasterMix (KangWei Biotech, Beijing, China), 1 μl of each forward and reverse 10 μM primer, 1 μl of genomic DNA, and 9.5 μl of double-distilled H 2 O. All PCR products were purified and sequenced at Tsingke Biotechnology Company (Chengdu, China).   Jäger & Praxaysombath, 2009 in having a similarly shaped median apophysis and broad, flat conductor, but it differs in: 1) the embolus is almost directed towards 6 o'clock apically in prolateral view ( Fig. 2A), versus almost 9 o'clock in A. dang (Jäger and Praxaysombath 2009: fig. 38); 2) the distal end of the embolus is not expanded ( Fig. 2A), versus expanded in A. dang  (Jäger and Praxaysombath 2009: fig. 39); 3) the embolus has a distinct lamellar pendant ( Fig. 2A), versus absent in A. dang (Jäger and Praxaysombath 2009: fig. 38). The female of the species resembles A. macrochoera Thorell, 1891 in having the broad epigynal scape incrassated on the posterior-lateral margin but it differs in: 1) the epigynal scape is distinctly longer than wide in ventral view (Fig. 3A), versus almost as long as wide in A. macrochoera (Levi 1983: fig. 19); 2) the septum is narrowest anteriorly in the posterior view (Fig. 3C), versus narrowest medially in A. macrochoera (Levi 1983: fig. 20). Description. Male (TRU-Araneidae-37). Total length 2.62. Carapace 1.46 long, 1.34 wide; abdomen 1.38 long, 1.02 wide. Eye sizes and interdistances: AME 0.10, ALE 0.06, PME 0.11, PLE 0.09, AME-AME 0.13, AME-ALE 0.05, PME-PME   Fig. 1D, E) pale and flat, acutely narrowed anteriorly and followed by rounded thorax area, with a longitudinal brown stripe medially. Fovea linear. Chelicerae ( Fig. 1F) pale, mingled with green. Endites (Fig. 1F) brown laterally and white on the inner side. Labium ( Fig. 1F) pale, hairy. Sternum (Fig. 1F) heart-shaped, pale medially and greenbrown laterally. Legs (Fig. 1D) pale to yellow, more or less mingled with green, armed with macrosetae. Abdomen ( Fig. 1D-F) oval, dorsum with a longitudinal, branched pale-brown patch, and silver spots gradually smaller from the anterior margin; venter greyish-white, with marked silver spots median-laterally. Spinnerets brown, hairy. Palp ( Fig. 2A-D): patella with a long bristle; tibia swollen; paracymbium curved medially, with a blunt tip directed towards the bulb in retrolateral view; median apophysis looks like a swan, with two wide spurs pointed apically, and a small, slender, tapered, and curved spur; conductor broad and flat, slightly curled; embolus tapered, spiraled proximally, and curved posteriorly, with a transparent short pendant directed towards the conductor.
Comments. Argiope cameloides is known only from the descriptions of the holotype from Hainan, China (see Zhu et al. 1994: 33, fig. 8A-C). Because the pairing has been supported by the result of DNA barcoding, and the collection site is geographically near the type locality, we identified these specimens from Guangxi, China, as belonging to A. cameloides. Moreover, the female of A. macrochoera from China differs from the holotype in some details (the differences have also been noted by Yin et al. (1997)) and is almost identical to these A. cameloides specimens, and, consequently, it is proposed as the female of A. cameloides.   Diagnosis. The male of this species resembles A. aetheroides Yin, Wang, Zhang, Peng & Chen, 1989 in the general shape of the palp, but it differs in the distal end of embolus is slender (Fig. 6A) versus flattened in A. aetheroides (Yin et al. 1997: fig. 13h), and the median apophysis is bifurcated at its distal end in posterior view (Fig. 6D) versus bifurcated at the base in A. aetheroides (Yin et al. 1997: fig. 13i). The female of the species closely resembles A. anasuja Thorell, 1887 in having the epigynal flange and broad posterior plate, but it differs in the median septum being less than 1/3 of the posterior plate width in ventral view (Fig. 7A-C), versus more than 1/3 of the posterior plate width in A. anasuja (Levi 1983: fig. 167) and the dorsal abdomen possesses white patches laterally (Fig. 5A, B), versus has three wide, transverse white patches in A. anasuja (Levi 1983: fig. 170).
Palp (Fig. 6A-D): patella with a long bristle; tibia swollen; paracymbium curved medially, blunt at tip; median apophysis bifurcated, with a short terminal spur; conductor flat, slightly curled; embolus enlarged at proximal end, curved almost 90° medially in prolateral view and with a lance-like tip in apical view.
Comments. Argiope perforata was originally described based on the female holotype from Sichuan, China; the illustrations are poor. It was redescribed and better illustrated by Levi (1983) who based on a specimen from Guangdong, China. Herein, the opposite sexes of the new specimens were collected from the same localities, and their pairing has been supported by the result of DNA barcoding. Additionally, the male of A. perforata, as described by Wang (1988), is considered to be mismatched and herein proposed to be the male of A. boesenbergi Levi, 1983 due to the close resemblances in palp structure. Besides, A. abramovi Logunov & Jäger, 2015 Ono (2010). The male of this species resembles A.minuta Karsch, 1879 in having the elongated median apophysis terminally divided into two branches, of which the inner branch bears a short spur. It differs in having the embolus being completely visible in prolateral view (Fig. 9A), versus partly hidden by the conductor in A. minuta (Levi 1983: fig. 210), and in the median apophysis lacking a proximal protrusion (Fig. 9A), versus having a distinctly proximal protrusion in A. minuta (Levi 1983: fig. 210).
Distribution. China (Guizhou, Guangxi); Vietnam. Comments. A male and a subadult female were collected on the same web from Shiwandashan National Forest Park during the night. The embolus of the male is structurally consistent with the broken embolus found in an A. vietnamensis female epigyne. Moreover, other male and females were also collected from Maolan National Nature Reserve. Based on these points of evidence, we propose the males as A. vietnamensis.