A new species of Anagyrus (Hymenoptera, Encyrtidae) from Malaysia, parasitoid of Lanceacoccus sp. (Hemiptera, Pseudococcidae)

A new species of Anagyrus Howard, 1896 (Hymenoptera: Encyrtidae), A. minipedicellus Zu & Zhang sp. n., is described from Borneo of Malaysia as a parasitoid of Lanceacoccus sp. (Hemiptera: Pseudococcidae), and DNA barcoding of the new species is given.


Introduction
Anagyrus Howard, 1896 is one of the most diverse genus of Encyrtidae, containing 282 species worldwide (Noyes 2017), which are mainly endoparasitoids of mealybugs (Hemiptera: Pseudococcidae), but some Australian species have been recorded as parasitoids of coccinellid (Coleoptera: Coccinellidae) (Noyes andHayat 1994, Noyes 2000).The members of the genus are very important as biological control agents and 19 species have been used in biological control (Noyes 2000).
In the present paper, A. minipedicellus Zu & Zhang, sp.n. is described as new to science, and a key to Malaysian species of Anagyrus is provided.

Material and methods
All the specimens in the present study were collected from Mt. Trus Madi by rearing, then dissected and mounted in Canada balsam on slides following the method described by Noyes (1982).Morphological terminology and abbreviations follow those of Noyes and Hayat (1994) and Noyes (2000) with some modifications.Photographs were taken with a digital CCD camera attached to an Olympus BX51 compound microscope, and most measurements were made from slide-mounted specimens using an eye-piece reticle, except body length, AOL, OCL, OD, OOL, POL.

Molecular studies
Eight individuals of A. minipedicellus were used to obtain their DNA barcodes.Total genomic DNA was extracted using the DNeasy Blood & Tissue Kit (Qiagen) following the manufacturer's protocols.Polymerase chain reactions (PCR) were carried out in 50 μL reaction volumes using TaKaRa ExTaq Polymerase kit.Final volumes included 5 μL of 10×Buffer, 25 mM MgCl2, 2.5 mM dNTP mixture, 10 pmol of each primer, 1U of ExTaq and 5 μL of genomic DNA.The COI gene fragment was amplified using the primers FWPTF1 (Li et al. 2010) and Lep-R1 (Hebert et al. 2004).The PCR cycle program for COI followed Hebert et al. (2003).Sequencing was performed directly from positive products in both directions using BigDye v3.1 on an ABI 3730xl DNA Analyzer (Applied Biosystems).
The following abbreviations are used in the text: Diagnosis.Female.Length, excluding ovipositor, 1.38-1.83mm.Body stout (Fig. 2); head generally orange with interantennal prominence and gena below mid eye level dark brown; scape dark brown, with a white subapical band; pedicel and funicle generally dark brown; clava white; mesosoma and metasoma mostly dark brown, except mesopleuron orange; frontovertex nearly 0.4× head width; clypeal margin slightly emarginate; antennal scape strongly broadened, about twice as long as broad; pedicel shorter and narrower than F1; all funicular segments at least 1.5× as long as broad; fore wing completely hyaline, 2.15× as long as broad; costal cell broad, with four complete lines of setae ventrally; ovipositor about 0.7× as long as mid tibia.
Male (length, 1.22-1.33mm): Color similar to female (Fig. 3), except head completely black, flagellum black, gradually becoming yellowish distad; antenna with scape about twice as long as broad, all funicular segments much longer than broad, F1 the longest; scale-like sensilla present only on F6; frontovertex nearly half head width; fore wing 2.08× as long as broad, costal cell ventrally with four lines of setae; metasoma shorter than mesosoma; phallobase without digiti; aedeagus about 1.6× as long as mid tibia.
Description.Female.Holotype.Length, 1.83 mm (excluding ovipositor).Head generally orange, interantennal prominence dark brown, gena dark brown to mid eye level, occiput mostly dark brown.Antenna with radicle dark brown; scape dark brown with a white subapical band; pedicel and funicle black, funicle gradually becoming paler distad and ventrally paler, clava white.Mesosoma dark brown; mesopleuron orange.Wings hyaline.Legs generally yellowish white, except dorsal and ventral margins of fore coxa, fore and mid femora, hind tibia dark brown, mid coxa mostly dark brown, hind coxa brown, all tarsi yellowish brown.Metasoma dark brown.
Variation.Length of female, excluding ovipositor, varies from 1.38-1.83mm, scape in smaller species as in Fig. 6B, colour of F6 in some specimens is paler, ventral 1/3 dark brown, dorsal 2/3 nearly white mixed with brown, otherwise very little in material available.
Male.Length 1.22-1.33mm.Color is similar to that of female except head completely black, flagellum gradually going from dark brown to yellowish.Antenna (Fig. 14) with scape about 2.02× as long as broad, all funicular segments much longer than broad, F1 the longest; longest flagellar setae about 1.5× as long as diameter of corresponding segments; scale-like sensilla present only on F6; antennal torulus with lower margin slightly below lower margin of eye and separated from mouth margin by about 1.46× its own height; frontovertex nearly (Fig. 12) 0.49× head width.Fore wing (Fig. 15) 2.08× as long as broad, costal cell ventrally with four lines of setae.Metasoma shorter than mesosoma; phallobase (Fig. 18) without digiti; aedeagus about 1.57× as long as mid tibia.Otherwise similar to female.
Etymology.The specific name refers to the small pedicel of female antennae.
Comments.This species is similar to A. alami Hayat, 1970 in the short and narrow pedicel.However, it can be easily separated from A. alami by its dark brown body (generally yellowish in alami), F1 1.33× as long as pedicel (twice in A. alami), MT 1.42× as long as OL (1.05× in borneensis).It is also probably close to A. subtilis Noyes &Hayat, 1994 andA. aceris Noyes &Hayat, 1994, but differs in the short pedicel, which is 0.75× as long as F1 (at least longer than F1 in A. subtilis and A. aceris), normal F1, which is similar to other funicular segments (F1 much shorter than any other funicular segments in A. subtilis), dark brown radicle (orange in A. subtilis), subparallel eye margins (diverging ventrally in A. aceris) and generally orange mesosoma (dorsum of mesosoma dark brown in A. aceris).

Molecular studies
The COI sequences the eight individuals of A. minipedicellus were successfully generated with high quality.All sequences have been deposited in GenBank (Accession Numbers: MH587108-MH587115).Only one base pair changes was detected between the eight individuals sequenced for COI.Blasting the COI sequences gave no close matches (over 90%) both on BOLD and GenBank.In GenBank, a COI sequence of Anagyrus (Anagyrus nr.pseudococci PLCO02, Accession No. KU499515) differs from those of the new species by about 14%.