Analysis of the variability of the Ornithogalum umbellatum L. 1753 (Hyacinthaceae) using molecular markers and morpho-anatomical characters

Predmet ovog istraživanja je varijabilnost taksona  Ornithogalum umbellatum  L. 1753. (Hyacinthaceae) na području Srbije i Mađarske. Ovaj najpoznatiji i najrasprostranjeniji predstavnik roda  Ornithogalum  L. izučava se dugi niz godina sa različitih aspekata: kao gajena baštenska vrsta i kao invazivni korov; kao otrovna biljka i potencijalno lekovita. Sa ekološkog aspekta  ove biljke su značajne kao  domaćini insektima polinatorima koji su višestruko i u različitim fazama razvića vezani za ove lukovičaste geofite. Veliki areal  i prilagođenost različitim tipovima staništa uzrokuju značajnu varijabilnost, koja rezultuje taksonomskom konfuzijom.  Jedan od razloga za  istorijski prepoznatljive taksonomski i filogenetski nerazjašnjene relacije  su  područja sa prirodnim staništima populacija  O.  umbellatum  čijem izučavanju nije posvećeno dovoljno pažnje. Jedno od takvih je i ovde istraživano područje. Kako je u Flori Srbije kao podvrsta  O. umbellatum  opisan i  O. umbellatum  subsp.  divergens  (Boreau), analizama su obuhvaćene i populacije ovog taksona. RAPD-PCR metodom ustanovljen je viši nivo genetičkog diverziteta između nego u okviru populacija, bez jasnog geografskog trenda dispozicije ove varijabilnosti. Ovi su rezultati očekivani za poliploidne taksone sa velikim udelom vegetativnog razmnožavanja u reproduktivnoj strategiji biljke, koji su pritom široko rasprostranjeni a u istraživanom području predstavljeni uglavnom relativno udaljenim te izolovanim populacijama.  Determinacija genetičke struktuiranosti među ispitivanim  genotipovima ukazala je na razdvajanje populacija dva taksona u dva klastera. Rezultate u saglasnosti sa ovim dala  je i analiza parametara poprečnih preseka lista, skapusa i plodnika, a ustanovljeni su i diskriminativni anatomski karakteri koji su razdvojili populacije dva taksona. Izdvojeni su pojedini kvantitativni karakteri, kako anatomski tako i morfološki, koji su  najviše doprineli varijabilnosti na interpopulacionom nivou. Pokazana je velika raznolikost morfoloških karaktera i heterogenost populacija  O. umbellatum, što je u skladu sa podacima u postojećoj litaraturi. Poznato je i da  habitus ovih  biljaka  veoma  zavisi  od  uslova  sredine,  kao  i  stadijuma  razvića  biljke,  dok  na molekularne  markere  ovi  faktori  ne  utiču.  Sa  druge  strane,  iako  primenjena  RAPD metoda  nije  dovoljna  za  determinaciju  filogenetski  blisko  srodnih  taksona  poput  ovih, pokazala  se  pouzdanom  i  efikasnom  za  ispitivanje  diverziteta  na  populaciono-genetičkom  nivou.  Stoga  je  u  cilju  što  sveobuhvatnije  procene  stanja  korišćena kombinacija  različitih  pristupa,  koji  su  dali  usaglašene  rezultate,  istovremeno  dajući uvid  u  nivo  polimorfnosti  i  struktuiranosti  populacija  i  ukazujući  na  jedinstvene  oblike varijabilnosti morfo-anatomskih karakteristika Sa  jedne  strane  ovo  je  važno  zbog  razjašnjenja  komplikovane  taksonomije,  a  sa  druge zbog  značaja  ovih  biljaka  sa  aspekta  potencijalne  koristi  odnosno  štete  za  čoveka,  ali  i iz  ugla  zaštite  biodiverziteta.  Uvid  u  nivo  varijabilnosti  populacija  doprineće adekvatnim  strategijama  konzervacije  i  menadžmenta  kako  ovih  biljnih  vrsta,  tako  i insekatskih sa kojima su povezane.Variability of  Ornithogalum umbellatum  L. 1753. (Hyacinthaceae) in  Serbia and Hungary  was analyzed in this research. This most famous and the most  widespread taxon in genus Ornithogalum L. has been a research subject for many years for various reasons: as garden plant and invasive weed; for its toxic and pharmaceutical properties. From an ecological point of view those bulbous geophytes  are the important host-plants for insect pollinators associated with them in different life stages.  Large distribution area and adaptation to diverse habitat types result in significant variability of these plants which is followed by  taxonomic confusion. Persisting taxonomic problems arise also from the fact that there are under-investigated regions with native populations of O. umbellatum, such as  localities  in Serbia and neighbouring countries. Since published floras of these countries  include  O. umbellatum  subsp.  divergens (Boreau), analyses involve populations of this taxon as  well. RAPD-PCR method revealed higher levels of genetic diversity among than within populations, without distinct geographical trend in the disposition of the variability. These results have been expected for polyploid taxa with high levels of vegetative reproduction, widespread and represented mostly with relatively distant and isolated populations in the given region. Genetic structure analyses have separated  populations of the two investigated taxa into two clusters. Analyses of the anatomical characteristics of the leaf, scapus and ovary cross-sections have shown results in accordance with those. Discriminative characters which divided populations of the two  taxa were determined. Distinctive quantitative characters, both anatomical and morphological, which have contributed the most to the interpopulation variability, were  singled out. Great diversity of morphological characters and population heterogeneity  of  O.  umbellatum  has been  shown, which  was  already described in literature dealing with the subject. Additionally, the high diversity is explained by the fact that habitus of those plants with very plastic phenotypes is influenced by environmental conditions and life stage of the plant. Those factors do not affect molecular markers; however techniques like RAPD-PCR could not be used independently for  characterization  of phylogenetically close taxa such as these. Nevertheless, RAPDs proved to be a reliable and efficient method suitable for distinguishing genetic differentiation on population level. Therefore, combination of different approaches was used in order to give comprehensive assessment of the situation. The results were consistent, revealing the levels of populations’  polymorphism and structure, as well as certain variations of morpho-anatomical features. This is important in the light of complicated  taxonomy clarification, and on the other hand from the aspect of potential value or harm of these plants to humans. In terms of  biodiversity conservation, insight into population variability levels might contribute to adequate management strategies for both plants and insects related to them

Analysis of the variability of the Ornithogalum umbellatum L. 1753 (Hyacinthaceae) using molecular markers and morpho-anatomical characters

Predmet ovog istraživanja je varijabilnost taksona  Ornithogalum umbellatum  L. 1753. (Hyacinthaceae) na području Srbije i Mađarske. Ovaj najpoznatiji i najrasprostranjeniji predstavnik roda  Ornithogalum  L. izučava se dugi niz godina sa različitih aspekata: kao gajena baštenska vrsta i kao invazivni korov; kao otrovna biljka i potencijalno lekovita. Sa ekološkog aspekta  ove biljke su značajne kao  domaćini insektima polinatorima koji su višestruko i u različitim fazama razvića vezani za ove lukovičaste geofite. Veliki areal  i prilagođenost različitim tipovima staništa uzrokuju značajnu varijabilnost, koja rezultuje taksonomskom konfuzijom.  Jedan od razloga za  istorijski prepoznatljive taksonomski i filogenetski nerazjašnjene relacije  su  područja sa prirodnim staništima populacija  O.  umbellatum  čijem izučavanju nije posvećeno dovoljno pažnje. Jedno od takvih je i ovde istraživano područje. Kako je u Flori Srbije kao podvrsta  O. umbellatum  opisan i  O. umbellatum  subsp.  divergens  (Boreau), analizama su obuhvaćene i populacije ovog taksona. RAPD-PCR metodom ustanovljen je viši nivo genetičkog diverziteta između nego u okviru populacija, bez jasnog geografskog trenda dispozicije ove varijabilnosti. Ovi su rezultati očekivani za poliploidne taksone sa velikim udelom vegetativnog razmnožavanja u reproduktivnoj strategiji biljke, koji su pritom široko rasprostranjeni a u istraživanom području predstavljeni uglavnom relativno udaljenim te izolovanim populacijama.  Determinacija genetičke struktuiranosti među ispitivanim  genotipovima ukazala je na razdvajanje populacija dva taksona u dva klastera. Rezultate u saglasnosti sa ovim dala  je i analiza parametara poprečnih preseka lista, skapusa i plodnika, a ustanovljeni su i diskriminativni anatomski karakteri koji su razdvojili populacije dva taksona. Izdvojeni su pojedini kvantitativni karakteri, kako anatomski tako i morfološki, koji su  najviše doprineli varijabilnosti na interpopulacionom nivou. Pokazana je velika raznolikost morfoloških karaktera i heterogenost populacija  O. umbellatum, što je u skladu sa podacima u postojećoj litaraturi. Poznato je i da  habitus ovih  biljaka  veoma  zavisi  od  uslova  sredine,  kao  i  stadijuma  razvića  biljke,  dok  na molekularne  markere  ovi  faktori  ne  utiču.  Sa  druge  strane,  iako  primenjena  RAPD metoda  nije  dovoljna  za  determinaciju  filogenetski  blisko  srodnih  taksona  poput  ovih, pokazala  se  pouzdanom  i  efikasnom  za  ispitivanje  diverziteta  na  populaciono-genetičkom  nivou.  Stoga  je  u  cilju  što  sveobuhvatnije  procene  stanja  korišćena kombinacija  različitih  pristupa,  koji  su  dali  usaglašene  rezultate,  istovremeno  dajući uvid  u  nivo  polimorfnosti  i  struktuiranosti  populacija  i  ukazujući  na  jedinstvene  oblike varijabilnosti morfo-anatomskih karakteristika Sa  jedne  strane  ovo  je  važno  zbog  razjašnjenja  komplikovane  taksonomije,  a  sa  druge zbog  značaja  ovih  biljaka  sa  aspekta  potencijalne  koristi  odnosno  štete  za  čoveka,  ali  i iz  ugla  zaštite  biodiverziteta.  Uvid  u  nivo  varijabilnosti  populacija  doprineće adekvatnim  strategijama  konzervacije  i  menadžmenta  kako  ovih  biljnih  vrsta,  tako  i insekatskih sa kojima su povezane.Variability of  Ornithogalum umbellatum  L. 1753. (Hyacinthaceae) in  Serbia and Hungary  was analyzed in this research. This most famous and the most  widespread taxon in genus Ornithogalum L. has been a research subject for many years for various reasons: as garden plant and invasive weed; for its toxic and pharmaceutical properties. From an ecological point of view those bulbous geophytes  are the important host-plants for insect pollinators associated with them in different life stages.  Large distribution area and adaptation to diverse habitat types result in significant variability of these plants which is followed by  taxonomic confusion. Persisting taxonomic problems arise also from the fact that there are under-investigated regions with native populations of O. umbellatum, such as  localities  in Serbia and neighbouring countries. Since published floras of these countries  include  O. umbellatum  subsp.  divergens (Boreau), analyses involve populations of this taxon as  well. RAPD-PCR method revealed higher levels of genetic diversity among than within populations, without distinct geographical trend in the disposition of the variability. These results have been expected for polyploid taxa with high levels of vegetative reproduction, widespread and represented mostly with relatively distant and isolated populations in the given region. Genetic structure analyses have separated  populations of the two investigated taxa into two clusters. Analyses of the anatomical characteristics of the leaf, scapus and ovary cross-sections have shown results in accordance with those. Discriminative characters which divided populations of the two  taxa were determined. Distinctive quantitative characters, both anatomical and morphological, which have contributed the most to the interpopulation variability, were  singled out. Great diversity of morphological characters and population heterogeneity  of  O.  umbellatum  has been  shown, which  was  already described in literature dealing with the subject. Additionally, the high diversity is explained by the fact that habitus of those plants with very plastic phenotypes is influenced by environmental conditions and life stage of the plant. Those factors do not affect molecular markers; however techniques like RAPD-PCR could not be used independently for  characterization  of phylogenetically close taxa such as these. Nevertheless, RAPDs proved to be a reliable and efficient method suitable for distinguishing genetic differentiation on population level. Therefore, combination of different approaches was used in order to give comprehensive assessment of the situation. The results were consistent, revealing the levels of populations’  polymorphism and structure, as well as certain variations of morpho-anatomical features. This is important in the light of complicated  taxonomy clarification, and on the other hand from the aspect of potential value or harm of these plants to humans. In terms of  biodiversity conservation, insight into population variability levels might contribute to adequate management strategies for both plants and insects related to them

DNA barcoding applied: identifying the larva of Merodon avidus (Diptera: Syrphidae)

The immature stages of phytophagous hoverflies and their plant hosts are mostly unknown. This paper describes the larva of Merodon avidus Rossi, 1790 (Diptera: Syrphidae), based on material collected from Đerdap National Park in Serbia. Larvae were found in the bulbs of Ornithogalum L. (Hyacinthaceae) and in the surrounding soil. DNA barcoding was used to identify the species and scanning electron microscopy was used to describe the morphological characteristics of Merodon avidus. Reared material previously identified as Merodon constans (Rossi, 1794) is shown to belong to M. hurkmansi Marcos-García, Vujić & Mengual, 2007.This study was supported by the Ministry of Education, Science and Technological Development, Republic of Serbia, Grant No. 173002 and Grant No. 43002, the Provincial Secretariat for Science and Technological Development, Grant No. 114-457-2173/2011-01 and by the project Status and Trends of European Pollinators, FP7-ENV-2009-1-244090 (STEP)

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

FIGURE 3 in Two new enigmatic species of the genus Merodon Meigen (Diptera: Syrphidae) from the north-eastern Middle East

FIGURE 3. Male of Merodon eumerusi sp. n.. lateral view. Scale bar, 1mm.Published as part of <i>Vujić, Ante, Radenković, Snežana, Likov, Laura, Andrić, Andrijana, Gilasian, Ebrahim & Barkalov, Anatolij, 2019, Two new enigmatic species of the genus Merodon Meigen (Diptera: Syrphidae) from the north-eastern Middle East, pp. 187-208 in Zootaxa 4555 (2)</i> on page 192, DOI: 10.11646/zootaxa.4555.2.2, <a href="http://zenodo.org/record/10073372">http://zenodo.org/record/10073372</a&gt

FIGURE 1. Merodon antenna, lateral view. A in Two new enigmatic species of the genus Merodon Meigen (Diptera: Syrphidae) from the north-eastern Middle East

FIGURE 1. Merodon antenna, lateral view. A Merodon eumerusi sp. n. male, inner side, B Merodon eumerusi sp. n. male, outer side, C Merodon eumerusi sp. n. female, inner side, D Merodon eumerusi sp. n. female, outer side, E Merodon pumilus Macquart male, outer side, F Eumerus armatus Ricarte et Rotheray male, outer side, G Eumerus truncatus Rondani male, outer side. f—fossette. Scale bar, 1 mm.Published as part of <i>Vujić, Ante, Radenković, Snežana, Likov, Laura, Andrić, Andrijana, Gilasian, Ebrahim & Barkalov, Anatolij, 2019, Two new enigmatic species of the genus Merodon Meigen (Diptera: Syrphidae) from the north-eastern Middle East, pp. 187-208 in Zootaxa 4555 (2)</i> on page 190, DOI: 10.11646/zootaxa.4555.2.2, <a href="http://zenodo.org/record/10073372">http://zenodo.org/record/10073372</a&gt

FIGURE 13 in Two new enigmatic species of the genus Merodon Meigen (Diptera: Syrphidae) from the north-eastern Middle East

FIGURE 13. Male genitalia of Merodon mixtum sp. n.. A, B epandrium, C hypandrium. A, C lateral view, B ventral view. al— anterior surstyle lobe, pl—posterior surstyle lobe, c—cercus, s—lateral sclerite of aedeagus. Scale bar, 0.5 mm.Published as part of <i>Vujić, Ante, Radenković, Snežana, Likov, Laura, Andrić, Andrijana, Gilasian, Ebrahim & Barkalov, Anatolij, 2019, Two new enigmatic species of the genus Merodon Meigen (Diptera: Syrphidae) from the north-eastern Middle East, pp. 187-208 in Zootaxa 4555 (2)</i> on page 201, DOI: 10.11646/zootaxa.4555.2.2, <a href="http://zenodo.org/record/10073372">http://zenodo.org/record/10073372</a&gt

FIGURE 4 in Two new enigmatic species of the genus Merodon Meigen (Diptera: Syrphidae) from the north-eastern Middle East

FIGURE 4. Metafemur of Merodon male. A, B Merodon eumerusi sp. n., C Merodon segetum (Fabricius), D Eumerus truncatus. A, anterior view, B, C, D antero-ventral view. it—row of inner thorns. Scale bar, 1 mm.Published as part of <i>Vujić, Ante, Radenković, Snežana, Likov, Laura, Andrić, Andrijana, Gilasian, Ebrahim & Barkalov, Anatolij, 2019, Two new enigmatic species of the genus Merodon Meigen (Diptera: Syrphidae) from the north-eastern Middle East, pp. 187-208 in Zootaxa 4555 (2)</i> on page 193, DOI: 10.11646/zootaxa.4555.2.2, <a href="http://zenodo.org/record/10073372">http://zenodo.org/record/10073372</a&gt