Botryosphaeriaceae species occurring on various woody hosts in Serbia

U zemljama Zapadnog Balkana zabeleženo je intenzivno sušenje brojnih drvenastih i žbunastih vrsta tokom poslednje decenije. Simptomi bolesti bili su tipični za one koje izazivaju gljivični patogeni iz familije Botryosphaeriaceae. Ciljevi ovog istraživanja bili su: 1. izolacija gljiva iz familije Botryosphaeriaceae iz obolelih stabala u Srbiji, Crnoj Gori i Bosni i Hercegovini i njihova karakterizacija, 2. da se utvrde domaćini i distribucija gljiva iz familije Botryosphaeriaceae u Srbiji, Crnoj Gori i Bosni i Hercegovini, u kontinentalnom i mediteranskom području, 3. da se utvrde domaćini gljive Diplodia sapinea u Srbiji i Crnoj Gori, 4. da se utvrdi genetički diverzitet i razmena genotipova između različitih populacija gljiva Neofusicoccum parvum i D. sapinea u Srbiji i Crnoj Gori. Karakterizacija izolata najpre je izvršena uz pomoć DNA sekvenci internog transkribovanog spejsera (ITS) rDNA i identifikovano je šest glavnih klada. Reprezentativni izolati svake klade su zatim dodatno okarakterisani uz pomoć DNA sekvenci translacionog elongacionog faktora 1-alpha (TEF 1-α), β-tubulin-2 (BT2) dela gena i velike podjedinice (LSU) rRNA, kao i morfoloških karakteristika aseksualnog izgleda ovih gljiva. D. sapinea-slični izolati su dodatno okarakterisani uz pomoć TEF 1-α dela gena. Genetički diverzitet i protok genotipova gljiva N. parvum i D. sapinea utvrđeni su uz pomoć DNA sekvenci ITS regiona, TEF 1-α, BT2 dela gena i mikrosatelitskih markera. Izolati gljive D. sapinea poreklom iz različitih domaćina proučavani su na nivou populacija poreklom iz Cedrus spp. i Pinus spp. Izolati gljiva D. sapinea i N. parvum koji potiču iz regiona sa različitim uslovima spoljašnje sredine proučavani su na nivou populacija iz kontinentalnog (CR) i Mediteranskog (MR) regiona. Identifikovano je deset vrsta gljiva iz familije Botryosphaeriaceae od kojih su osam i to Dothiorella sarmentorum, Neofusicoccum parvum, Botryosphaeria dothidea, Phaeobotryon cupressi, Sphaeropsis visci, Diplodia seriata, D. sapinea i Diplodia mutila bili nauci poznati taksoni. Preostale dve vrste mogle su biti identifikovane samo do nivoa roda i to kao Dothiorella spp. Takođe, opisan je Dichomera sin-aseksualni izgled gljiva D. sapinea, Dothiorella sp. 2 i B. dothidea, kao i jedinstvene morfološke karakteristike osam nauci poznatih vrsta...Extensive die-back and mortality of various trees and shrubs has been observed in parts of the Western Balkans region during the past decade. The disease symptoms have been typical of those caused by fungal pathogens residing in the Botryosphaeriaceae. The aims of this study were to 1. isolate and characterize Botryosphaeriaceae species associated with diseased trees in Serbia, Montenegro, Bosnia and Herzegovina, 2. consider the host range and distribution of Botryosphaeriaceae species, spanning a Mediterranean and a Continental region, 3. consider the host range of Diplodia sapinea in Serbia and Montenegro 4. determine genetic diversity and genotype flow between different populations of Neofusicoccum parvum and D. sapinea from Serbia and Montenegro. Isolates were initially characterized based on the DNA sequence data for the internal transcribed spacer (ITS) rDNA and six major clades were identified. Representative isolates from each clade were further characterized using DNA sequence data for the translation elongation factor 1-alpha (TEF 1-α), β-tubulin-2 (BT2) and large subunit (LSU) rRNA gene regions, as well as the morphology of the asexual morphs. D. sapinea-like isolates were additionally characterized using TEF 1- α. Genetic diversity and genotype flow of N. parvum and D. sapinea were determined using DNA sequence data of the ITS region, TEF 1-α, BT2 and microsatellite markers. The relationship between isolates of D. sapinea from different hosts was considered using populations from Cedrus spp. and Pinus spp. The relationships between isolates from environmentally different regions included populations of each species from the Continental (CR) and Mediterranean (MR) regions. Ten species of the Botryosphaeriaceae were identified of which eight, i.e. Dothiorella sarmentorum, Neofusicoccum parvum, Botryosphaeria dothidea, Phaeobotryon cupressi, Sphaeropsis visci, Diplodia seriata, D. sapinea and Diplodia mutila were known taxa. The remaining two species could be identified only as Dothiorella spp. Dichomera syn-asexual morphs of D. sapinea, Dothiorella sp. 2 and B. dothidea, as well as unique morphological characters for a number of the known species are described..

HOW DO TEACHERS IN SERBIA MANAGE THEIR CLASSROOMS?

The purpose of the current study was to first examine whether there is a statistically significant difference in the importance of the styles teachers use in managing their classrooms (interventionist, interactionist, and non-interventionist). Considering that these styles are also influenced by and involve teacher personality, the second aim of this study was to investigate whether there is a relationship between management styles, teachers' resilience and aspects of their emotional competence (perception and understanding of emotions, their expression and naming, and their regulation), but also with their gender and professional experience. The sample consisted of 100 elementary school teachers of both sexes (86 of them were women) with an average professional experience of 23.12 years. The following instruments were used: The Teacher Self-Assessment of Classroom Management Styles Inventory, the Connor-Davidson Resilience Scale, and the Emotional Competence Inventory. The results of the repeated measures ANOVA showed that there was a statistically significant difference in the expression of the different management styles. Further comparison showed that the interactionist style was the most pronounced, followed by the interventionist and non-interventionist styles. The results also showed that there was a positive, statistically significant correlation between the interventionist management style and resilience, and the same type of correlation was found between the interactionist management style, resilience, and all three aspects of emotional competence. No correlation was found with gender and work experience. Since resilience is positively related to both the interventionist and interactionist management styles, which imply some degree of control over the classroom situation, it can be considered a factor that enables the teacher to act in this manner. The interactionist style, which is the most pronounced, requires shared control, so the current results support the claim that teachers must have adequate emotional competencies

Pimpinella tragium Vill. (Apiaceae) – anatomy and essential oil composition

Pimpinella tragium Vill. is a perennial, herbaceous plant inhabiting dry grasslands, limestone rocky areas and screes of wide sense Mediterranean area, east European lowlands and Caucasus. This study explores the anatomical structure, content and composition of the essential oil of the vegetative organs and fruit of this species. The plant material was collected on two localities: the village Izvor (Bosilegrad, Serbia) and Mt. Galičica (FYR Macedonia). The anatomical studies were conducted on permanent slides obtained by standard method of preparation for viewing under a light microscope. The essential oils obtained by hydrodistillation from roots (collected during flowering and fruiting period), aerial parts with inflorescences, and fruits, were analysed by GC-FID and GC-MS. The anatomical analysis revealed a secondary structure of root and primary structure of stem with closed collateral vascular bundles. Leaves are isobilateral, amphistomatic; petiole is with arched vascular bundles and the fruit (mericarp) is semi-circular and lightly ribbed in cross section. Non-glandular unicellular and bicellular cuneate trichomes are sparsely distributed on the stem, leaves and petiole, but very dense on the fruit. Secretory channels are present in all organs: in the parenchyma of the root and stem cortex, stem pith, in the phloem of root and vascular bundles of stem and petiole, by the leaf vascular bundles and in the fruit pericarp. Essential oil yields from different parts of P. tragium ranged from 0.2-1.1% (v/w). The main compounds in the oils from roots (in both stages, from both localities), as well as from aerial parts and fruits from Mt. Galičica are C-12 norsesquiterpenes (trinorsesquiterpenes): pregeijerene (29.0-56.2%) and gejerene (14.1-22.9%), whereas those from aerial parts and fruits from village Izvor are β-bisabolene (19.1-57.2%) and a phenylpropanoid epoxy-pseudoisoeugenyl-2-metohybutyrate (17.4-22.2%). Trinorsesquiterpenes, which were also found to be characteristic constituents in some other Pimpinella species oils, are the most dominant constituents in all investigated oils, except in the oil of fruit from village Izvor. In addition, phenylpropanoids of pseudoisoeugenol type are present in all oils confi rming them as chemical markers of Pimpinella species analysed so far.7th Balkan Botanical Congress, 10-14th September 2018, Novi Sad, Serbi

Anatomy and chemical analysis of essential oil of Pimpinella saxifraga l. (Apiaceae)

Koren vrste Pimpinella saxifraga L. se tradicionalno koristi kao ekspektorans, bronhosekretolitik i antiflogistik. Cilj ovog rada je ispitivanje anatomske građe, sadržaja i sastava etarskog ulja vegetativnih organa i ploda P. saxifraga. Biljni materijal je sakupljan u jugoistočnoj Srbiji: Bojanine vode (Suva planina) i Ostrovica (Sićevačka klisura). Anatomska analiza izvršena je na trajnim preparatima dobijenim standardnom metodom pripreme za posmatranje pod svetlosnim mikroskopom. Etarska ulja su izolovana iz herbe u cvetu, ploda, korena u fazi cvetanja i plodonošenja, destilacijom vodenom parom i analizirana GC‐FID i GC‐MS metodama. Rezultati anatomske analize su pokazali da je koren sekundarne građe a stablo primarne sa zatvorenim kolateralnim sprovodnim snopićima. Listovi su dorziventralni, hipoamfistomatski, lisna drška je sa lučno raspređenim sprovodnim snopićima, a plodovi (merikarpi) su polukružnog oblika sa slabo izraženim rebrima, bez trihoma. Retke, nežlezdane, višećelijske, uniserijatne trihome sa oštrim vrhom su uočene na stablu, listovima i lisnoj dršci. Sekretorni kanali su brojni u svim organima. Najviši sadržaj etarskog ulja određen je u plodu (1,48‐1,52%), dok su herba (0,13‐0,21%) i koren (0,50‐0,53%) sadržali manju količinu ulja (v/m). Etarska ulja istih organa sa različitih lokaliteta su sličnog kvalitativnog sastava sa manjim kvantitativnim razlikama. U uljima herbe i ploda dominantna jedinjenja su β‐bisabolen (28,8‐76,0%) i epoksi‐pseudoizoeugenil‐2‐metilbutirat (7,7‐21,8 %). Najzastupljenija jedinjenja u uljima korena, u obe faze, su azuleni (31,4‐35,7%) i pregeijeren (13,7‐18,8%). U ispitivanim etarskim uljima različitih organa P. saxifraga utvrđeno je prisustvo trinorseskviterpena (azulena i geijerena) i fenilpropanoidnih jedinjenja pseudoizoeugenil tipa, koja su karakteristični hemijski markeri za etarska ulja biljaka ovog roda.Root of Pimpinella saxifraga L. is traditionally used as expectorant, bronchosecretolytic and antiphlogistic. The aim was investigation of anatomy, content and composition of the essential oils of vegetative organs and fruits. Plant material was collected in South‐Eastern Serbia: Bojanine vode (Mt. Suva planina) and Ostrovica (Sićevo gorge). The anatomical analysis was conducted on permanent slides obtained by standard method of preparation. The essential oils obtained by hydrodistillation from flowering aerial parts, fruits and roots (from flowering and fruiting period) were analysed by GC‐FID and GC‐MS. The anatomical analysis revealed a secondary structure of root and primary structure of stem with closed collateral vascular bundles. The leaves are dorsiventral, hypoamphistomatic and petiole is with arched vascular bundles. The fruit (mericarp) is semi‐circular and lightly ribbed in cross section without trichomes. Rare, nonglandular, multicellular, uniseriate, cuneate trihomes were noted on the stem, leaves and petiole. Secretory channels are numerous in all organs. The highest content of essential oil was determined in the fruits (1.48‐1.52%), while aerial parts (0.13‐0.21%) and roots (0.50‐0.53%) contained a lower amount of oil (v/w). The oils from the same organs originated from different localities were similar in qualitative composition with some quantitative differences. The most dominant constituents in the oils from aerial parts and fruits were β‐bisabolene (28.8‐76.0%) and epoxy‐pseudoisoeugenyl‐2‐methylbutyrate (7.7‐21.8 %), whereas those of the oils from roots, in both stages, were azulenes (31.4‐35,7%) and pregeijerene (13.7‐18.8%). Trinorsesquiterpenes (azulenes and geijerenes) and phenylpropanoid compounds of the pseudoisoegenyl type, which are characteristic chemical markers of the essential oils of the plants of this genus, were present in the all analyzed oils.VII Kongres farmaceuta Srbije sa međunarodnim učešćem: Zajedno stvaramo budućnost farmacije, Beograd, Srbija, 10-14. oktobar 2018

Host specificity of co-infecting Botryosphaeriaceae on ornamental and forest trees in the Western Balkans

The Botryosphaeriaceae is a diverse family of endophytes and fungal pathogens of mainly woody plants. We considered the host range and distribution of these fungi by sampling diseased ornamental and forest trees and shrubs in Serbia, Montenegro, Bosnia and Herzegovina, spanning a Mediterranean and a Continental climatic region. In total, ten Botryosphaeriaceae species were identified in the Western Balkans and with the exception of Sphaeropsis visci and Phaeobotryon cupressi, which occurred on one host, all the species had a broader host range. Phaeobotryon cupressi was found only in the Mediterranean region and S. visci, Dothiorella sp., Dothiorella sarmentorum and Diplodia seriata were present only in the Continental region. Pathogenicity tests were conducted on a variety of hosts from which the Botryosphaeriaceae species were isolated. These included leaves and/or stems of seedlings of 21 hosts, and cut leaves and/or branches of six hosts. Moreover, stems of seedlings of Chamaecyparis lawsoniana, Cedrus deodara, Picea omorika, Pinus patula and Eucalyptus grandis were inoculated as hosts from which some or all of the Botryosphaeriaceae species used for inoculation were not isolated. Inoculations showed that the majority of these fungi could also co-infect hosts other than those from which they were isolated. The results suggest that most of the species have broad host ranges and can potentially cause disease on a broad range of tree species under certain conditions.Members of Tree Protection Co-operative Programme (TPCP), the University of Pretoria, South Africa and the Ministry of Education, Science and Technological Development of the Republic of Serbia (TR37008 and III43007). The first author also wishes to acknowledge partial financial support from European Cooperation in Science and Technology (COST) Actions Pathway Evaluation in Pest Risk Management In Transport (PERMIT FP1002), ALIEN Challenge (TD1209) and A global network of nurseries as early warning system against alien tree pests (Global Warning FP1401).http://onlinelibrary.wiley.com/journal/10.1111/(ISSN)1439-03292019-04-01hj2018Forestry and Agricultural Biotechnology Institute (FABI)GeneticsMicrobiology and Plant Patholog

Chemical composition of volatile compounds of eight Geranium L. species from Vlasina plateau

The Vlasina plateau shows significant floristic diversity counting 956 species of which 91 may be categorized as endangered in the flora of Serbia. The genus Geranium is present in the flora of this region with few species and they are the subject of our research. The chemical composition of volatile compounds of aerial and underground parts of eight Geranium L. species (G. macrorrhizum L., G. phaeum L., G. sanguineum L., G. robertianum L., G. palustre L., G. pyrenaicum Burm. f., G. columbinum L. and G. lucidum L.) collected in flowering stage from Vlasina plateau (Serbia) was examined. The samples of volatile fraction were obtained by steam distillation and analyzed using GC-FID and GC-MS. The cluster analysis revealed the separation of volatile compounds of Geranium species to two clusters. The aerial and underground parts volatiles of G. macrorrhizum were separated from the other species, based on the high sequiterpenes content (92.3% and 94.6%), mostly germacrone (73.3%) in the aerial and δ-guaiene (76.8%) in the underground parts. The volatile fractions of other species were mainly composed of sesquiterpenes (10.8-61.8%), diterpenes (12.9-43.0%) and fatty acids and their derivatives (6.6- 21.6%). The exception was the fraction of the underground parts of G. phaeum which was predominantly made by fatty acids and their derivatives (76.6%). In the volatile fraction of G. palustre aerial parts β-selinene (18.6%) and (E)-caryophyllene (15.7%) prevailed. Hexadecanoic acid (15.4%), germacrene D (15.4%), (E)-caryophyllene (10.6%), phytol (10.4%) and caryophyllene oxide (10.2%) were the most abundant compounds in volatile fraction of G. pyrenaicum aerial parts, while phytol (13.2%) and (E)-caryophyllene (9.5%) in the volatile fraction of G. lucidum aerial parts. Hexadecanoic acid and phytol were dominant compounds in the volatile fraction of aerial parts of G. sanguineum (21.1% and 17.8%), G. robertianum (12.5% and 19.3%), G. phaeum (7.3% and 41.4%) and G. columbinum (14.5% and 29.5%). Volatiles of aerial parts of G. robertianum were also characterized by (E)-caryophylene (8.0%) and caryophyllene oxide (7.4%), while the volatiles of G. sanguineum contained β-bisabolene (5.2%). The volatiles of G. palustre aerial parts have been studied for the first time. The chemical characterization of volatile compounds of Geranium species may possibly have chemotaxonomic importance and is significant for further research of Vlasina plateau native flora.7th Balkan Botanical Congress, 10-14th September 2018, Novi Sad, Serbi

Botryosphaeriaceae associated with the die-back of ornamental trees in the Western Balkans

Extensive die-back andmortality of various ornamental trees and shrubs has been observed in parts of the Western Balkans region during the past decade. The disease symptomshave been typical of those caused by pathogens residing in the Botryosphaeriaceae. The aims of this study were to isolate and characterize Botryosphaeriaceae species associated with diseased ornamental trees in Serbia, Montenegro, Bosnia and Herzegovina. Isolates were initially characterized based on the DNA sequence data for the internal transcribed spacer rDNA and six major clades were identified. Representative isolates from each clade were further characterized using DNA sequence data for the translation elongation factor 1-alpha, b-tubulin-2 and large subunit rRNA gene regions, as well as the morphology of the asexualmorphs.Ten species of theBotryosphaeriaceae were identified of which eight, i.e., Dothiorella sarmentorum, Neofusicoccum parvum, Botryosphaeria dothidea, Phaeobotryon cupressi, Sphaeropsis visci, Diplodia seriata, D. sapinea and D. mutila were known taxa. The remaining two species could be identified only as Dothiorella spp. Dichomera syn-asexual morphs of D. sapinea, Dothiorella sp. 2 and B. dothidea, aswell as unique morphological characters for a number of the known species are described. Based on host plants and geographic distribution, the majority of Botryosphaeriaceae species found represent new records. The results of this study contribute to our knowledge of the distribution, host associations and impacts of these fungi on trees in urban environments.Tree Protection Co-operative Programme (TPCP), the University of Pretoria (South Africa) and the Ministry of Education and Science of the Republic of Serbia (TR37008).http://link.springer.com/journal/104822017-04-30hb2016Forestry and Agricultural Biotechnology Institute (FABI)GeneticsMicrobiology and Plant Patholog

Antimicrobial activity of eight Geranium L. species extracts

The antimicrobial activity of methanol extracts of aerial parts of 8 Geranium L. species (G. macrorrhizum L., G. phaeum L., G. sanguineum L., G. robertianum L., G. palustre L., G. pyrenaicum Burm. f., G. columbinum L. and G. lucidum L.) was tested against 8 standard strains (Staphylococcus aureus, Enterococcus faecalis, Bacillus subtilis, Escherichia coli, Klebsiella pneumaniae, Pseudomonas aeruginosa, Salmonella abony and Candida albicans) as well as 10 clinical isolates of E. coli. Also, total phenolics and tannins were quantified using spectrophotometric Folin- Ciocalteu method. The extracts exhibited antibacterial and anticandidal activity with minimal inhibitory concentrations (MICs) between 12.5 and 200 μg/ml. E. faecalis was most sensitive strain (MIC 12.5-50 μg/ml), especially to G. columbinum and G. phaeum extracts (MICs 12.5 μg/ml). The determined amounts of total phenolics and tannins were in the range of 170-534 and 38-386 mg GAE/g of dry extract, respectively, with the highest content in G. sanguineum extract. However, the antimicrobial activity was not related to total polyphenolics or tannin content and further investigation in regard with phenolic profile is needed

Trichoderma spp. from Pine Bark and Pine Bark Extracts: Potent Biocontrol Agents against Botryosphaeriaceae

Pinus sylvestris bark represents a rich source of active compounds with antifungal, antibacterial, and antioxidant properties. The current study aimed to evaluate the antifungal potential of P. sylvestris bark against Botryosphaeria dothidea, Dothiorella sarmentorum, and Neofusicoccum parvum (Botryosphaeriaceae) through its chemical (water extracts) and biological (Trichoderma spp. isolated from the bark) components. The water bark extracts were prepared at two temperatures (80 and 120 °C) and pH regimes (7 and 9). The presence of bark extracts (30%) caused inhibition of mycelial growth of B. dothidea and D. sarmentorum for 39 to 44% and 53 to 60%, respectively. Moreover, we studied the antagonistic effect of three Trichoderma isolates originating from the pine bark. Trichoderma spp. reduced growth of B. dothidea by 67%–85%, D. sarmentorum by 63%–75% and N. parvum by 55%–62%. Microscopic examination confirmed typical mycoparasitism manifestations (coiling, parallel growth, hook-like structures). The isolates produced cellulase, β-glucosidase and N-acetyl-β-glucosaminidase. The volatile blend detected the emission of several volatile compounds with antimicrobial activity, including nonanoic acid, cubenene, cis-α-bergamotene, hexanedioic acid, and verticillol. The present study confirmed in vitro potential of P. sylvestris bark extracts and Trichoderma spp. against the Botryosphaeriaceae. The study is an important step towards the use of environmentally friendly methods of Botryosphaeriaceae disease control

Assessment of polyphenol content, in vitro antioxidant, antimicrobial and toxic potentials of wild growing and cultured rue

 Ruta graveolens L. (rue) is edible medicinal plant that is traditionally used in various countries. This study aimed to investigate and compare the phenolic content, antioxidant capacity, antibacterial and cytotoxic activities of the methanolic and ethanolic extracts of wild growing and cultured rue. The total phenolic content of the tested extracts varied from 57.90 to 166.91 mg of catechin equivalent (CE)/g of extract and the total flavonoid content from 4.18 to 26.87 mg of rutin equivalent (Ru)/g of extract.  All the tested samples exhibited significant antioxidant potential in DPPH radicals scavenging and lipid peroxidation inhibition assays (comparable with activity of rutin in the same test systems), antimicrobial activity determined by microdilution method (particularly against Gram (+) bacteria strains) and ability to induce inhibition of HeLa cells growth and proliferation (up to 71.81%). In addition, rue-treated HeLa cells showed various morphological changes after 72 h of incubation with rue extracts. Extracts from wild growing rue with the highest polyphenol, tannin and flavonoid contents demonstrated the strongest activities in all tested systems. The present study also emphasized the fact that the rue leaves and herb should be harvested at the beginning of blossoming stage in order to achieve the maximal level of secondary metabolites and optimal pharmacological effects.