Molecular diversity of Merodon aureus species group (Diptera:Syrphidae)

Род Merodon Meigen (Diptera: Syrphidae) се одликује великм бројем врста које имају  функцију у опрашивању  биљака. У оквиру овог рода својом разноврсношћу издваја се Merodon aureus  група  врста коју поред фенотипски различитих одликује и присуство већег броја криптичних  врста. Због морфолошке сличности криптичне врсте представљају изазов за таксономе, те су молекуларне методе од посебног значаја. У том светлу примарни циљ овог истраживања је био утврђивање молекуларног диверзитета групе и могућности његове примене у таксономији. Истраживање је било засновано на анализи варијабилности секвенци  COI  и  28S  рРНК гена за 718 јединки 41 описане или новоткривене врсте сирфида. Додатно, тестирана је примењивост мултилокусних  ISSR маркера у раздвајању врста  M.  luteomaculatus  комплекса. Узорци су тестирани и на присуство Wolbachia с обзиром да она може утицати на еволуцију митохондријалних гена. Добијени резултати су указали на високу варијабилност секвенци  COI  гена  који се показао корисним у  утврђивању граница  криптичних  врста у анализираним комплексима. Секвенце 28Ѕ рРНК гена у већини случајева нису биле од већег значаја за раздвајање врста, али би могле имати потенцијал у  раздвајању комплекса  или подгрупа  врста као допуна анализи  секвенци  COI  гена. Додатно,  ISSR  маркери су показали потенцијал за примену у молекуларној таксономији. Све анализиране врсте изузев М. balkanicus су биле заражене  Wolbachia, али није утврђена јасна веза између варијабилности  секвенци  митохондријалног  COI гена и инфекционог статуса врста. Утврђен образац генетичке варијабилности је вероватно  обликован драстичним климатским променама  током Плеистоцена и фрагментацијом хабитата. Да би се донели крајњи закључци о таксономском статусу предложених врста потребно је добијене резултате интегрисати са подацима других релевантних  научних дисциплина.Rod Merodon Meigen (Diptera: Syrphidae) se odlikuje velikm brojem vrsta koje imaju  funkciju u oprašivanju  biljaka. U okviru ovog roda svojom raznovrsnošću izdvaja se Merodon aureus  grupa  vrsta koju pored fenotipski različitih odlikuje i prisustvo većeg broja kriptičnih  vrsta. Zbog morfološke sličnosti kriptične vrste predstavljaju izazov za taksonome, te su molekularne metode od posebnog značaja. U tom svetlu primarni cilj ovog istraživanja je bio utvrđivanje molekularnog diverziteta grupe i mogućnosti njegove primene u taksonomiji. Istraživanje je bilo zasnovano na analizi varijabilnosti sekvenci  COI  i  28S  rRNK gena za 718 jedinki 41 opisane ili novotkrivene vrste sirfida. Dodatno, testirana je primenjivost multilokusnih  ISSR markera u razdvajanju vrsta  M.  luteomaculatus  kompleksa. Uzorci su testirani i na prisustvo Wolbachia s obzirom da ona može uticati na evoluciju mitohondrijalnih gena. Dobijeni rezultati su ukazali na visoku varijabilnost sekvenci  COI  gena  koji se pokazao korisnim u  utvrđivanju granica  kriptičnih  vrsta u analiziranim kompleksima. Sekvence 28Ѕ rRNK gena u većini slučajeva nisu bile od većeg značaja za razdvajanje vrsta, ali bi mogle imati potencijal u  razdvajanju kompleksa  ili podgrupa  vrsta kao dopuna analizi  sekvenci  COI  gena. Dodatno,  ISSR  markeri su pokazali potencijal za primenu u molekularnoj taksonomiji. Sve analizirane vrste izuzev M. balkanicus su bile zaražene  Wolbachia, ali nije utvrđena jasna veza između varijabilnosti  sekvenci  mitohondrijalnog  COI gena i infekcionog statusa vrsta. Utvrđen obrazac genetičke varijabilnosti je verovatno  oblikovan drastičnim klimatskim promenama  tokom Pleistocena i fragmentacijom habitata. Da bi se doneli krajnji zaključci o taksonomskom statusu predloženih vrsta potrebno je dobijene rezultate integrisati sa podacima drugih relevantnih  naučnih disciplina.Genus  Merodon  Meigen  (Diptera: Syrphidae)  is characterized by a large number of species which have a function in plants pollination. Within this genus, Merodon aureus species group is distinguished by its diversity and it is characterized not only by phenotypically divergent species but also with a large number of cryptic species. Because of the high morphological similarity, cryptic species are a challenge for taxonomist, thus molecular methods are of special importance. The primary goal of this research was to determine the molecular diversity of the group and the possibility of its application in taxonomy. The study was based on the sequences variability analysis of COI and 28S rRNA genes for 718 individuals belonging to 41 described or newly discovered hoverfly species. Additionally, the applicability in species delimitation of multilocus marker ISSR was tested on  M. luteomaculatus  species complex. Specimens have also been tested for Wolbachia since  it may affect the evolution of mitochondrial genes. The obtained results indicated a high variability of the COI gene  sequences that proved useful for determining the  cryptic  species boundaries in the analyzed complexes. Sequences of the 28S rRNA gene in   most cases were not of much significance for the species delimitation, but they could have the potential to separate species complexes  оr subgroups  as a complement to the analysis of the COI gene  sequences. Additionally, the ISSR markers showed potential for application in molecular taxonomy. All analyzed species  except  M. balkanicus  were infected with  Wolbachia, but no clear  relation  was established between the sequence variability of the mitochondrial COI gene and  the infectious status of species. The established pattern of genetic variability is probably shaped  by drastic climatic changes during Pleistocene and habitats fragmentation. In order to  achieve  the final conclusions on the taxonomic status of the proposed species, it is necessary to integrate the obtained results with the data of other  relevant scientific disciplines

The genus Xanthogramma Schiner, 1861 (Diptera: Syrphidae) in southeastern Europe, with descriptions of two new species

Examination of 122 specimens of Xanthogramma Schiner, 1861 (Diptera: Syrphidae) from varied localities in Europe (+Turkey) resulted in the description of two new species (X. aeginae Ricarte, Nedeljković, and Vujić new species and X. pilosum Nedeljković, Ricarte, and Vujić new species), as well as new data on six other species. Most of the examined material originated from the Balkan Peninsula and Greek islands. New species concepts were supported by morphological and molecular evidence. Relationships among the eight studied species were analysed and discussed based on the data of nuclear (ITS2) and mitochondrial (COI) genes sequences. An identification key to the European species of Xanthogramma is provided. Lectotypes are designated for Doros decoratum Zetterstedt, 1843, Lasiophthicus novus Rondani, 1857, Syrphus laetus Fabricius, 1794, Syrphus ornatus Meigen, 1822, and Xanthogramma nobilitatum Frey, 1946.Financial support was provided by the Serbian Ministry of Education and Science (projects OI173002 and III43002), the Provincial Secretariat for Science and Technological Development (project “Genetic resources of agro-ecosystems in Vojvodina and sustainable agriculture”), the FP7 EU project, Innosense, and the H2020 project “ANTARES” (664387), Antonio Ricarte’s position at the University of Alicante is funded by the Vicerrectorado de Investigación y Transferencia de Conocimiento)

Seed quality of the facelia-variety NS Priora grown in Serbia

Phacelia has been used for seed production and as a forage crops, either on its own or in a mix with peas or vetch to provide forage and honey production as a source of high quality nectar and pollen. The experiment was carried out in 2016, in the fields of the Institute of Field and Vegetable Crops in Bački Petrovac, in Serbia, with variety NS Priora. NS Priora had plant flowering continualy over 60 days and had high, good quality grain yield. NS Priora variety had average nitrogen content is 3.21%, protein content was 20.06% and the average thousand seeds weight was 1.42 g. Phacelia is presently very intensively used in organic agriculture and for sowing of arable land temporarily excluded from production which achieves high yields

Review of the Merodon natans group with description of a new species, a key to the adults of known species of the natans lineage and first descriptions of some preimaginal stages

Merodon natans group (Diptera, Syrphidae) taxa are reviewed using an integrative taxonomic approach combining morphological, morphometric and molecular techniques. The approach substantiates recognition of the three species: M. calcaratus (Fabricius, 1794), M. natans (Fabricius, 1794) and M. pulveris Vujić & Radenković in Radenković et al. 2011, and reveals the existence of a new species, M. makrisi Vujić, Radenković & Tot sp. nov., which is described. It also highlights the existence of a series of natans group populations, especially on some of the Mediterranean islands, in the Levant and in the Afrotropical Region, for which more comprehensive data are required to clarify their status. A key is provided to the natans lineage species currently recognised, and preimaginal stages of some natans-group species are described for the first time. Redescriptions for M. calcaratus and M. natans are provided. A neotype is selected for M. natans. Lectotypes are designated for M. annulatus (Fabricius, 1794) and M. melancholicus (Fabricius, 1794). Merodon annulatus is recognised as a synonym of M. natans.The authors acknowledge financial support of the Ministry of Education, Science and Technological Development of the Republic of Serbia (Grant No. 451-03-9/2021-14/200125 and Grant No. 451-03-9/2021-14/200358). André van Eck was financially supported for research in Cyprus by the Dutch Uyttenboogaart-Eliasen Foundation (UES), with grant nrs. SUB.2016.12.12 and SUB.2018.12.03

The puzzling mitochondrial phylogeography of the black soldier fly (Hermetia illucens), the commercially most important insect protein species

Abstract Background The black soldier fly (Diptera: Stratiomyidae, Hermetia illucens) is renowned for its bioconversion ability of organic matter, and is the worldwide most widely used source of insect protein. Despite varying extensively in morphology, it is widely assumed that all black soldier flies belong to the same species, Hermetia illucens. We here screened about 600 field-collected and cultured flies from 39 countries and six biogeographic regions to test this assumption based on data for three genes (mitochondrial COI, nuclear ITS2 & 28S rDNA) and in order to gain insights into the phylogeography of the species. Results Our study reveals a surprisingly high level of intraspecific genetic diversity for the mitochondrial barcoding gene COI (divergences up to 4.9%). This level of variability is often associated with the presence of multiple species, but tested nuclear markers (ITS2 and 28S rDNA) were invariant and fly strain hybridization experiments under laboratory conditions revealed reproductive compatibility. COI haplotype diversity is not only very high in all biogeographic regions (56 distinct haplotypes in total), but also in breeding facilities and research centers from six continents (10 haplotypes: divergences up to 4.3%). The high genetic diversity in fly-breeding facilities is mostly likely due to many independent acquisitions of cultures via sharing and/or establishing new colonies from field-collected flies. However, explaining some of the observed diversity in several biogeographic regions is difficult given that the origin of the species is considered to be New World (32 distinct haplotypes) and one would expect severely reduced genetic diversity in the putatively non-native populations in the remaining biogeographic regions. However, distinct, private haplotypes are known from the Australasian (N = 1), Oriental (N = 4), and the Eastern Palearctic (N = 4) populations. We reviewed museum specimen records and conclude that the evidence for introductions is strong for the Western Palearctic and Afrotropical regions which lack distinct, private haplotypes. Conclusions Based on the results of this paper, we urge the black soldier fly community to apply molecular characterization (genotyping) of the fly strains used in artificial fly-breeding and share these data in research publications as well as when sharing cultures. In addition, fast-evolving nuclear markers should be used to reconstruct the recent invasion history of the species

Molecular Approaches for Detection of <i>Trichoderma</i> Green Mold Disease in Edible Mushroom Production

Due to the evident aggressive nature of green mold and the consequently huge economic damage it causes for producers of edible mushrooms, there is an urgent need for prevention and infection control measures, which should be based on the early detection of various Trichoderma spp. as green mold causative agents. The most promising current diagnostic tools are based on molecular methods, although additional optimization for real-time, in-field detection is still required. In the first part of this review, we briefly discuss cultivation-based methods and continue with the secondary metabolite-based methods. Furthermore, we present an overview of the commonly used molecular methods for Trichoderma species/strain detection. Additionally, we also comment on the potential of genomic approaches for green mold detection. In the last part, we discuss fast screening molecular methods for the early detection of Trichoderma infestation with the potential for in-field, point-of-need (PON) application, focusing on isothermal amplification methods. Finally, current challenges and future perspectives in Trichoderma diagnostics are summarized in the conclusions

Loop-mediated isothermal amplification (LAMP) assay coupled with gold nanoparticles for colorimetric detection of Trichoderma spp. in Agaricus bisporus cultivation substrates

Abstract One of the significant challenges in organic cultivation of edible mushrooms is the control of invasive Trichoderma species that can hinder the mushroom production and lead to economic losses. Here, we present a novel loop-mediated isothermal amplification (LAMP) assay coupled with gold nanoparticles (AuNPs) for rapid colorimetric detection of Trichoderma spp. The specificity of LAMP primers designed on the tef1 gene was validated in silico and through gel-electrophoresis on Trichoderma harzianum and non-target soil-borne fungal and bacterial strains. LAMP amplification of genomic DNA templates was performed at 65 °C for only 30 min. The results were rapidly visualized in a microplate format within less than 5 min. The assay is based on salt-induced aggregation of AuNPs that is being prevented by the amplicons produced in case of positive LAMP reaction. As the solution color changes from red to violet upon nanoparticle aggregation can be observed with the naked eye, the developed LAMP-AuNPs assay can be easily operated to provide a simple initial screening for the rapid detection of Trichoderma in button mushroom cultivation substrate

Merodon vittatus Vujic & Likov 2021, sp. nov.

Merodon vittatus Vuji&cacute; & Likov sp. nov. urn:lsid:zoobank.org:act: 591CEDD4-80E4-4978-B13E-3F66511C9026 Figs 1, 4C, 5C, 6C, 7D–F, 8B, 9C, 10C, 11C, 12C, 18C, 21E Diagnosis Brown, large (12 mm), striped species, with golden yellow pile on the tip of abdomen (Fig. 5C); frons and postpronotum dark brown; scutum with five pollinose vittae (Fig. 10C); metafemur with distinct apical thorn on triangular process (Figs 6C, 8B); terga 3 and 4 with broad medial pollinose fasciae (Fig. 5C). Similar to Merodon multifasciatus from which differs only in male genitalia with narrow, S-shaped posterior lobe of surstyle, gradually narrowing toward tip (Fig. 7D: pl) (in M. multifasciatus broad, slightly curved, with almost parallel sides and rounded apex (Fig. 13D: pl). Etymology The specific epithet ‘ vittatus ’ ‘wearing or carrying a vitta’, an adjective, refering to the broad golden gray fasciae on the terga 2–4. Material examined Holotype GHANA • 1 &male;; Transvolta Togoland, Amedzofe; 6.853° N, 0.433° E; 2 Nov. 1959; N.D. Jago leg.; FSUNS ID 04085; NBCN. Description Length: large species, body 12 mm, wing 8.5 mm (n = 1). Male HEAD (Figs 4C, 9C, 11C, 12C). Antenna (Fig. 4C) reddish-yellow; pedicel elongated, approximately as long as basoflagellomere (relation scape:pedicel: basoflagellomere =1.0: 2.5: 2.5); basoflagellomere concave dorsally, with acute apex; arista yellow to reddish, thickened basally, 1.7 times as long as basoflagellomere. Face dark brown, covered with gray pollinosity and whitish pile, except on bare medial vitta that occupies ¼ width of face. Oral margin protruded (Fig. 9C). Frons dark brown, inflated, covered with gray pollinosity and whitish pile. Vertical triangle isosceles (Fig. 12C), reddish, covered with a long, whitish pile. Ocellar triangle equilateral. Eye pile dense, gray, slightly longer than scape. Eye contiguity about 15 ommatidia long. Occiput reddish-brown, pollinose, covered with a whitish pile. THORAX (Figs 6C, 8B, 10C). Mesonotum black, except lateral side of scutum including postpronotum, postalar callus, and posterior margin of scutellum red-brown; covered with grayish-yellow pile; scutum with five pollinose longitudinal vittae (Fig. 10C). Pleuron black to dark brown, covered with gray pollinosity and the following parts with whitish to yellow pile: anterior part of proepimeron, posterior part of anterior anepisternum, most of the posterior anepisternum except anterior end, antero-ventral and postero-dorsal part of katepisternum and anepimeron. Wing hyaline, with dense microtrichia and yellow to brown veins. Calypter pale yellow. Halter with pale yellow pedicel and capitulum. All three femora dark brown; tibiae mostly reddish, medially dark brown; tarsi reddish (Figs 6C, 8B). Metatrochanter without calcar. Metafemur moderately thickened, with less developed apicoventral triangular lamina, more or less dentate, the apical dens is distinct (Figs 6C, 8B). Pile on legs predominantly whitish to yellow. ABDOMEN (Fig. 5C). Black to dark brown. Tergum 1 with gray pollinosity, covered with black basal sockets of pile and with short pale yellow pile; tergum 2 with pollinose posterior margin and narrow medial fascia, interrupted in the middle, covered with pale yellow pile; terga 3 and 4 with pollinose posterior margin and broad medial fasciae (Fig. 5C); tergum 3 covered with pale yellow pile except anterior half with few short black pile; tergum 4 covered with long golden-yellow pilosity (Fig. 5C). Sterna reddish-brown, covered with pale yellow pile. GENITALIA (Figs 7D–F, 18C). Posterior lobe of surstyle S-shaped, gradually narrowing toward tip (Fig. 7D: pl); anterior margin of surstyle angular (Fig. 7D: ams); cercus elongated (Fig. 7D: c). Hypandrium with theca medially distinctly narrowed (Fig. 7F). Lateral sclerite of aedeagus small (Fig. 18C: s). Female Unknown. Period of flight and distribution (Fig. 1) Only the holotype is known, which originated from Ghana and was collected in November. It was found in a deciduous forest-woodland savanna type of vegetation.Published as part of Vujić, Ante, Radenković, Snežana, Zorić, Ljiljana Šašić, Likov, Laura, Tot, Tamara, Veselić, Sanja & Djan, Mihajla, 2021, Revision of the Merodon bombiformis group (Diptera: Syrphidae) - rare and endemic African hoverflies, pp. 88-135 in European Journal of Taxonomy 755 (1) on pages 115-117, DOI: 10.5852/ejt.2021.755.1401, http://zenodo.org/record/501300

Revision of the Merodon bombiformis group (Diptera: Syrphidae) – rare and endemic African hoverflies

In the present work, the Afrotropical species of the bombiformis species group, part of the aureus lineage, are revised. Six species are recognized, based on a combination of morphological and genetic features. Three of these species are new to science: Merodon lotus Vujić & Radenković sp. nov., M. vittatus Vujić & Likov sp. nov., and M. zebra Vujić & Radenković sp. nov. Redescriptions are provided for the other three species: M. bombiformis Hull, 1944, M. multifasciatus Curran, 1939, and M. nasicus Bezzi, 1915. The female of M. bombiformis is described. The name Merodon edentulus Macquart, 1855 is considered here as a nomen dubium. One new synonymy is proposed: M. apimima Hull, 1944 syn. nov. (junior synonym of M. multifasciatus). The distribution of the bombiformis species group is discussed. The larval host plant of M. multifasciatus is identified as Gladiolus. A key to the identification of both males and females of the bombiformis group is provided

Merodon Meigen 1803

Genus Merodon Meigen, 1803 Diagnosis of the Merodon aureus lineage Mid coxa with long pile posteriorly (Fig. 2A); anterior anepisternum below postpronotum with many long pile (Fig. 2B); species with stocky abdomen (Fig. 2C); lateral sclerites of the aedeagus very small or absent (Fig. 3A: marked with arrow). Diagnosis of the Merodon bombiformis group Pedicel elongated, approximately as long as basoflagellomere (Fig. 4); abdomen broad (Fig. 5); metafemur with less serrated apicoventral triangular lamina, usually only the apical dens is distinct (Fig. 6); metatrochanter of males smooth, without calcar (Fig. 6); male genitalia with posterior surstyle lobe usually bent (as on Fig. 7A, D: pl), and hypandrium narrowed medially (as on Fig. 7C: marked with arrow); distribution: Afrotropical Region (Fig. 1). The Merodon bombiformis group and M. funestus (Fabricius, 1794) differ from other species and groups among the aureus lineage by an elongated pedicel, approximately as long or even longer than basoflagellomere (Fig. 4) and small lateral sclerite of the aedeagus (as on Fig. 3B, D: s) (absent in other species and groups of the aureus lineage, on Fig. 3A: marked with arrow). Morphologically, the M. bombiformis group can be distinguished from M. funestus by the absence of a calcar on metatrochanter in males (present in M. funestus, on Fig. 8A), less dentate apicoventral triangular lamina on the metafemur, but usually with a distinct apical dens (Fig. 8B) (clearly dentate in M. funestus, on Fig. 8A) and by the shape of the posterior lobe of the surstylus: tip rounded in bombiformis group (as on Fig. 13A, D: pl), but tapering in M. funestus (Fig. 19A: pl).Published as part of Vujić, Ante, Radenković, Snežana, Zorić, Ljiljana Šašić, Likov, Laura, Tot, Tamara, Veselić, Sanja & Djan, Mihajla, 2021, Revision of the Merodon bombiformis group (Diptera: Syrphidae) - rare and endemic African hoverflies, pp. 88-135 in European Journal of Taxonomy 755 (1) on page 93, DOI: 10.5852/ejt.2021.755.1401, http://zenodo.org/record/501300