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

Anatomical characteristics of two Ornithogalum L. (Hyacinthaceae) taxa from Serbia and Hungary and their taxonomic implication

Anatomical characters of two morphologically similar Ornithogalum taxa, O. umbellatum and O. divergens, were investigated. An analysis of leaf and scapus cross-sections was performed on plants from ten populations from Serbia and Hungary, using light microscopy. The aim of this research was to give data about the qualitative and quantitative anatomical characteristics of these taxa, in order to evaluate their taxonomic significance and single out distinctive anatomical features, as well as to contribute to the knowledge of the genus Ornithogalum in the studied region. On the basis of the variability of anatomical characters, similar populations formed two clusters, joining the plants previously determined as O. divergens and O. umbellatum. The two taxa significantly differed for most of the quantitative leaf and scapus characters. Since only quantitative differences were recorded in this research, anatomical characters could not be solely used to separate these two taxa. However, the results of anatomical investigations are consistent with the results of previous morphological and genetic analyses; therefore anatomical parameters could be useful as additional taxonomic characters

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)

Insights from the preimaginal morphology of the constans species‑group, to reveal novel morphological patterns of the Merodon albifrons‑evolutionary lineage (Diptera, Syrphidae)

Merodon triangulum Vujić, Radenković & Hurkmans, 2020 is a European endemic hoverfly species belonging to Merodon constans species-group, inside albifrons-lineage. The distribution of this species is known to be mostly central Europe and Balkan peninsula and it has been categorized as Near Threatened in the European IUCN red list of hoverflies; this paper cites the species for the first time in Ukraine (western Ukraine, specifically). In the present study, the preimaginal stages of this species are described and figured using Scanning Electron Microscopy. The material used for the descriptions were larvae collected in Ukraine and Serbia feeding inside underground storage organs of the spring snowflake Leucojum vernum L., 1753. This morphological description constitutes the first one inside the constans species-group, and the sixth description of the albifrons-lineage, in which there is only one species-group left to have at least one species of the preimaginal stages described (i.e., ruficornis species-group). The descriptions were compared with the rest available of the genus, stating the diagnostical characters of the present species and the shared characters inside the lineage. The novel information provided on the trophic interaction between M. triangulum larvae and Leucojum bulbs is stated for the first time and further supports the association of the constans species-group with the underground storage organs of snowflakes and snowdrops (Galantheae) in their role as host plants.Partial financial support was received from the research department of the University of Alicante in the frame work of a predoctoral grant (UAFPU2019-03). In addition, the study has been partially supported by the Ministry of Science, Technological Development and Innovation of the Republic of Serbia (Grant No. 451-03-47/2023-01/200125 and Grant No. 451-03-47/2023-01/200358)

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

Diversity and genetic structure of Ornithogalum L. (Hyacinthaceae) populations as revealed by RAPD-PCR markers

Random amplified polymorphic DNA (RAPD) PCR method was used to assess the level of diversity and genetic structure in Ornithogalum L. populations from Serbia and Hungary with the main goal of improving the knowledge of this genus in the given region. The material was collected from 19 populations and identified as two morphologically similar and phylogenetically close taxa: O. umbellatum L. 1753 and O. divergens Boreau 1887. All ten RAPD primers used for the analysis gave PCR products, with length between 3000bp and 300bp. There were 101 amplified fragments in total; number of polymorphic bands per primer varied between seven and 13. Percentage of polymorphic loci was 96% in total and 12% in average in each population. Genetic variation statistics for all loci also showed that genetic diversity for all populations was 0.29 and Shannon index 0.45, while mean values for these parameters calculated for each population were 0.04 and 0.06, respectively. Analysis of molecular variance demonstrated high population genetic differentiation; however Mantel test showed no significant correlation between geographic distances of populations and genetic distances expressed through population pairwise FST. UPGMA dendrogram based on Jaccard genetic similarity coefficients showed subclustering and principal coordinate analysis based on Nei and Li coefficients of genetic distances indicated grouping. Analysis of populations genetic structure was in accordance with these results and clearly separated populations of O. umbellatum from O. divergens. RAPDs proved to be a reliable and rapid method suitable for distinguishing genetic differentiation in Ornithogalum, thus could be applied as a useful additional tool in resolving taxonomic problems. [Projekat Ministarstva nauke Republike Srbije, br. 173002

Stability and changes in the distribution of Pipiza hoverflies (Diptera, Syrphidae) in Europe under projected future climate conditions.

Climate change is now considered a significant threat to terrestrial biodiversity. Species distribution models (SDMs) are among the modern tools currently used to assess the potential impacts of climate change on species. Pipiza Fallén, 1810 is a well known aphidophagous hoverfly genus (Diptera, Syrphidae) at the European level, for which sampling has been conducted across the region, and long-term databases and geo-referenced datasets have been established. Therefore, in this work, we investigated the potential current distributions of the European species of this genus and their response to future climate change scenarios, as well as evaluated stability in their ranges and potential changes in species-richness patterns. We applied three climate models (BCC_CSM1.1, CCSM4, HadGEM2-ES) to four representative concentration pathways (RCP 2.6, RCP 4.5, RCP 6.0, RCP 8.5) for two time frames (2050 and 2070). Our results show that the distribution of most Pipiza species may slightly differ under different climate models. Most Pipiza species were predicted not to be greatly affected by climate change, maintaining their current extent. Percentages of stable areas will remain high (above 50%) for the majority of studied species. According to the predicted turnover of species, northern Europe, could become the richest in terms of species diversity, thus replacing Central Europe as the current hot spot

FIGURE 11 in Two new enigmatic species of the genus Merodon Meigen (Diptera: Syrphidae) from the north-eastern Middle East

FIGURE 11. Metatrochanter and metafemur of Merodon male, anterior view. A Merodon mixtum sp. n., B Merodon geniculatus Strobl, C Merodon trochantericus, D Merodon trebevicensis Strobl, E Merodon caucasicus. t—thorn on trochanter, bb—basal bulge, cb—central bulge. 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 199, DOI: 10.11646/zootaxa.4555.2.2, <a href="http://zenodo.org/record/10073372">http://zenodo.org/record/10073372</a&gt

Assessment of genetic diversity within the Merodon ruficornis species group (Diptera: Syrphidae) by RAPD analysis

As one of the most distinct groups in the hoverfly genus Merodon, the monophyletic ruficornis species group has been the focus of several studies using different approaches. Molecular methods have shown incongruences between morphological and molecular data. In the present study, we investigated four species of the Merodon ruficornis group (i.e. M. loewi, M. armipes, M. papillus and M. hoplitis) with the aim of detecting intra- and interspecific genetic diversity, and we examined the usefulness of random amplified polymorphic DNA (RAPD) in an integrative taxonomic approach to species delimitation. Analysis of Nei’s genetic variation over all loci showed that genetic diversity for the analyzed Merodon species was h=0.24. Based on UPGMA, PCoA and Bayesian clustering analyses, our results clearly differentiated four groups that correspond to the four morphologically-defined Merodon species. Among the analyzed species, M. armipes and M. hoplitis showed the lowest level of genetic divergence; M. loewi was clearly separated from both M. armipes and M. papillus. Based on our data, we propose the use of RAPD-PCR as an additional tool for resolving taxonomic problems within Merodon. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no.173002