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Molecular identification of Fusarium graminearum pathogen of small grains and maize in Serbia
No full textFuarium held blight (FHB) ranka among the most pcrvaive and economically damaging diseasee
afl'O(ting =eats glob.Uy. The intens.i.&d occurrence offusariosis on wheat spikes has resulted in
signifieant economic loaaes worldwide. Beyond economic impam, FHB is also lilllred to
mycotoxin eontamination in maize bmela. The primary causltive agent of FHB is F'UllQl'hlln
gramineantm Sdlwabe, initially recopized 111 a linale species but now clauified wichin the
F'fllaTirtm gramiManlm apecies complex (FGSC). The FGSC it genetically diverse, comprising at
lellt 15 phylogeneticllly distinct apecies, includina 1he tnlditionaJ. F. graminemvm. The aim of
1hi1 study was to make a molecular idc!d:ifiad:ion of F. grammeanan species. In this n:&ean:h, 8
isolates were selected from the collection of funai aria;inating from grains of wheat, barley and
maize from differmt localitif:ll in Serbia. Total DNA W81 ����� from mycelium collected from
7-day4d coloaia of sillglHpOR ilOl.llel arown on PDA using the DNeasy Pl.mt Mini Kit
(Qiagen. Hilden, Gcrmmy). I.clem:ifiGatia proc:edwes involved anplttntion aod bidirectiooal
sequenc:ing of tmlacription elongation factor (TEF-JaJ, hiatone H3, md p..tubWn lllillg the
primen efl/cf2, H3-la/H3-1b, md Tl/1'22, rmpec:tivdy. The obtained &eqUCDCCS lW:rC depollited
in OmBd Ullder ICCCllion lllllllben MF9744�����MF974408 (TEF-Ja.), MG063784-MG063792
(p..tubWn), llld MF999140-MF999148 (b.istone H3). Sequeru:e malyii.s was conducted using
BLAST, while genetic 1imilmity wat11S1C111Cd usms MEGA 6.0 softwan:. Temd isolates shared
99-100% nucleotide idemitywith TEF-la, hilltone H3 llld P-tubulin compered to most iaolates of
F. ����� availlble in 1he gene bank
Specific detection of Waitea circinata var. zeae using conventional and real‑time PCR
No full textWaitea circinata var. zeae, a pathogen with a relatively narrow host range, has recently been detected in cabbage and oilseed
rape in Europe and worldwide. In this study, we developed specific conventional and real-time PCR protocols for direct
detection of W. circinata var. zeae from mycelium and diseased plant tissue. The newly developed primer pair zeaefor1/
zeaerew1, used in PCR protocols, specifically amplified only target isolates of W. circinata var. zeae when tested against
isolates of 11 different binucleate and multinucleate anastomosis groups of Rhizoctonia spp. including AG-A, AG-G, AG-F,
AG-U, AG-2-1, AG-2-2, AG-3, AG-4 HGI, AG-4 HGII, AG-4 HGIII, and AG-6 and common soil-borne pathogens. Total
of nine previously published primer pairs designed for the detection of various Rhizoctonia spp. were also tested and did
not amplify target isolates of W. circinata var. zeae. The detection limit of conventional and real-time PCR protocols was
10 –2 and 10 –5 (with starting concentration 9.5 ng/μl), respectively, and both methods are the first available tools for direct
detection and identification of W. circinata var. zeae from mycelium and diseased oilseed rape seedlings. Both conventional
and SYBR-Green-based real-time PCR protocols are cost-effective and provide a solid basis for further investigations of W.
circinata var. zeae, particularly in relation to distribution, host range, and epidemiology
Analysis of yield components of tomatoes grown in intercropping with marigold
No full textLekovite biljke poput nevena mogu uticati na povećanje prinosa i
kvaliteta plodova povrtarskih vrsta, kao i produţiti njihov rok trajanja posle
berbe. Cilj istraţivanja je bio da se procene komponente prinosa paradajza
gajenog u zdruţenom usevu sa nevenom. Eksperiment je sproveden tokom
2022. i 2023. godine u poljskim uslovima korišćenjem SP-109 genotipa
paradajza, koji je selekcionisan u Institutu za povrtarstvo Smederevska
Palanka. U toku vegetacije biljke su prihranjivane i navodnjavane.
Eksperiment je postavljen po sluĉajnom blok sistemu u ĉetiri ponavljanja.
Primenjena su tri tretmana: T1 - ĉist usev paradajza (razmak izmeĊu redova
50 cm i izmeĊu biljaka 25 cm), T2 - zdruţen usev paradajza i nevena (sa
istom gustinom kao u T1 tretmanu) i T3 - zdruţen usev paradajza i nevena
(sa redukovanim razmakom izmeĊu redova na 25 cm). Mereni su broj
plodova i masa plodova po biljci, kao glavne komponente prinosa. Najveći
broj plodova, kao i masa plodova po biljci je zabeleţen u tretmanu T2 u obe godine istraţivanja. U 2022. godini povećanje broja plodova po biljci
je bilo 12% i 36,3% a povećanje mase plodova iznosilo je 41,9% i 66,1%,
dok je u 2023. godini povećanje broja plodova bilo 77% i 27,9% a
povećanje mase plodova iznosilo 64,7% i 47,8% u odnosu na tretmane T1
i T3. Na osnovu dobijenih rezultata moţe se zakljuĉiti da su faktori godina
i razliĉiti tretmani zdruţivanja useva paradajza i nevena imali statistiĉki
vrlo znaĉajan uticaj, kao i njihova meĊusobna interakcija na ispitivane
parametre.Medicinal plants such as marigold can increase the yield and quality of
fruits of vegetable species, as well as extend their shelf life after harvest.
The aim of the research was to evaluate of yield components of tomatoes
grown in intercropping with marigold. The experiment was conducted
during 2022 and 2023 in field conditions using the SP-109 tomato
genotype, which was selected at the Institute for Vegetable Crops
Smederevska Palanka. During the growing season, the plants were fed and
irrigated. The experiment was set up in a random block system with four
replications. Three treatments were applied: T1 - tomato crop (distance
between rows 50 cm and between plants 25 cm), T2 - intercropping of
tomato and marigold (with the same density as in the T1 treatment) and T3
- intercropping of tomato and marigold (with reduced spacing between
rows at 25 cm). The number of fruits and fruit weight per plant were
measured as the main components of the yield. The highest number of fruits, as well as the fruit weight per plant, was recorded in treatment T2 in
both years of the study. In 2022, the increase in the number of fruits per
plant was 12% and 36.3%, and the increase in fruit weight was 41.9% and
66.1%, while in 2023, the increase in the number of fruits was 77% and
27.9% and the increase in fruit weight was 64.7% and 47.8% compared to
treatments T1 and T3. Based on the obtained results, it can be concluded
that the factors of year and different treatments of intercropping tomato and
marigold had a statistically significant influence, as well as their interaction
on the examined parameters
In vitro and in vivo testing of Steinernema feltiae (Filipjev) populations to control Lycoriella ingenua (Dufour) and the impact on mushroom yield
No full textIn most cases, insect pest control in cultivated mushroom production of Agaricus bisporus
is based on chemical insecticides. Due to the environmental side effects and often negative
effect on mushroom yield and non-target species, the world's trend is replacing chemical control
measures with sustainable alternatives. The mushroom fly, Lycorie/la ingenua (Dufour)
(Sciaridae: Diptera) is the major mushroom pest worldwide. Conversely, entomopathogenic
nematodes (EPN) are one of the most potent and frequently used biocontrol agents in control of
the harmful mushroom flies. Therefore, the study aimed to test virulence and control potential of
different populations of 5teinernema fe/tiae (Filipjev) against the mushroom fly, and their
influence on mushroom yield.
Two experiments were carried out. The first, in vitro experiment, aimed at testing the
virulence of EPN, was carried out in plastic Petri dishes (35 mm diameter) with the bottom
covered with filter paper. Commercial and three native popu lations of 5. feltiae were used. In
each Petri dish 0.5 ml of a nematode suspension containing 0, 50, 100, 200, 500 and 1000
infective juveniles (IJ) was applied to filter paper, after which 10 fourth instar larvae of L. ingenua
were added to each Petri dish. The entire experiment was carried out in 10 replicates. The
mortality of mushroom fly larvae was recorded three and seven days after exposure to the
nematodes. The second, in vivo experiment was conducted in a climatecontrolled mushroom
growth chamber. Commercial and two of the most virulent native populations of 5. feltiae for
mushroom sciarid fly and their impact on mushroom yield was evaluated. Commercially
produced phase Ill mushroom substrates were placed into 24 plastic containers (w 31 x d 21.5 x
h 13 cm), and inoculated with 20 fourth instar larvae of L. ingen ue each. Nem atode suspension of
each of the tested EPN strains containing 50000 IJ was added on the substrate surface per
container at the casing time and seven days after casing. Containers were held in the insectrearing
cages and a number of L. ingenua adults were observed weekly using yellow sticky traps
inside each cage.
The results of the in vitro experiment showed the highest virulence of the commercial
EPN strain for the mushroom fly larvae, followed by the native populations. Log-probit
regression analyses of the concentration-mortality data showed that all tested populations of 5.
feltiae caused significant mortality of L. ingenua larvae, with estimated LC�����o values (median
Lethal Concentrations) above 11 IJ/cmi of surface for all populations. The LCw values for the
native populations tested corresponded to the highest IJ concentration used (approximately 100
IJ/cmi), while for the commercial population a concentration of 84 IJ/cmi caused mortality of 90
per cent of the exposed sciarid larvae. The results of an in vivo experiment showed the highest
efficacy in controlling L. ingenua larvae by the commercial EPN population, followed by two
native populations. No statistically significant differences in mushroom yield were observed
between the control and the treatments
Adsorption of carbamate pesticide desmedipham from aqueous solution using reactivated active carbon spheres
No full textHuman activities such as agricultural usage of herbicides, desiccants, and pesticides involving crop
farming and industrial processes such as petroleum refining and combustion of fossil fuels, are
contributing to the growing pollution of ground and surface water in amounts higher than the ecosystem
can receive [1]. It has been proven that exposure of the organism to pesticides and their decomposition
products can have carcinogenic and mutagenic effects, and the toxicity of pesticides certainly causes
cell damage and the creation of metabolites harmful to the organism [2].
Desmedipham belongs to the broad-spectrum carbamate pesticides with neurotoxic influence, used to
manipulate or control undesirable vegetation. However, extensive pesticide use in agricultural
practices can lead to environmental pollution and human health issues [3]. Many studies have proven
that lethal and sub-lethal effects have been described following desmedipham exposure in aquatic and
soil organisms, even in more complex organisms such as rodents, amphipods, and fish. The high
toxicity of the carbamate pesticide desmedipham places it in the group of easily accessible chemical
compounds that can be used in crises such as terrorist acts or war for water pollution, which is an
essential resource for survival in addition to food [4]. Adsorption was widely investigated as a process
for dissolved or dispersed pollutants in untreated effluent. The effectiveness of the adsorbent depends
on the porosity, surface area, number of available sites, and possible interactions with the target
contaminant. The various possible adsorbents were used for wastewater treatment [4].
This study investigated the adsorption of desmedipham using the carbon (RAC) microspheres
obtained from Saratoga™ filtering protective suiteactivated at 650℃ and with the density of used
activated carbon between 180 and 220 g m-2. These suits provide a minimum of one-day protection
and more than 40 days of wear under battlefield conditions, including several washings cycles [5]. The
dependence of pesticide removal efficiency of different initial parameters such as pesticide and
adsorbent particles concentration, pH, and the temperature was studied. The kinetic study includes
the pseudo-first, pseudo-second (PSO), and second-order kinetic models. Desmedipham removal
using reactivated carbon was followed by ultraviolet-visible (UV-Vis) spectroscopy. The kinetics
parameters of pesticide adsorption were monitored from 20 to 180 minutes. After 3 hours of the
pesticide removal, equilibrium was established. According to the obtained results the kinetics for all
adsorbents are best described using a second order kinetics model. At optimal conditions (CRAC= 1
g/L, CDES=2.33×10–5, and pH 5), the adsorption capacity was qe = 29.649 mol g-1 × 10-8 and the
adsorption rate was k = 4.3×10–4g mmol-1 min-1, and activation energy was Ea=14.935KJ mol-
Plectosphaerella cucumerina ̶ new and emerging pathogen of lettuce in Balkan region
No full textIn March 2021, unusual plant stunting, collar and root rot of lettuce during rosette stage were
observed in two commercial fields in Serbia. Initial above-ground symptoms were yellowing
and wilting of leaves, while below-ground symptoms were collar and root rot. Eventually,
whole plants wilted, collapsed and died. In total, twenty Plectosphaerella spp. isolates
obtained from 35 symptomatic lettuce plants were identified on the basis of morphology and
evaluated for their pathogenicity. To confirm the species identity, internal transcribed spacer
(ITS) region and the D1/D2 region, of a selected representative isolate 13-3-c, were amplified
and sequenced by using primer pairs ITS1/ITS4 and N1/N2, respectively. Sequence analysis
of both regions revealed 100% nucleotide identity with Plectosphaerella cucumerina isolates
from different countries deposited in the NCBI GenBank. Neighbour-joining analysis was
conducted based on combined ITS and D1/D2 regions, and the tree was constructed with the
substitution models. Combined phylogeny confirmed that the sequences shared a common
clade with P. cucumerina. To our knowledge, this is the first molecular identification of P.
cucumerina on lettuce or any other crops in Serbia and Balkan region. P. cucumerina has
already been known as a pathogen of lettuce and other hosts grown in many countries
worldwide, as well as in some European countries. This emerging pathogen may cause
significant economic losses in lettuce production in Serbia and in the other Balkan countries.
Our results may help to develop efficient monitoring and detection of P. cucumerina and
effectively manage the spread of the disease
Trichothecene chemotypes of Fusarium graminearum species complex in Serbia
No full textThe species Fusarium graminearum represents one of the most economically important factor that
threaten agricultural production in the world. In addition to affecting yield reduction and grain quality,
this species is of great importance due to its ability to synthesize mycotoxins. Knowing the toxicological
profile of F. graminearum species is essential for agriculture and food industry, because trichothecene
chemotypes (NIV, 3ADON, 15ADON) have different toxicological properties that directly affect
human and animal health. In this study, 20 isolates of F. graminearum originating from maize and wheat
were examined. DNA isolation was performed from the mycelium of the fungus using the commercial
kit Dneasy Plant Mini Kit according to the manufacturer’s instructions. Chemotype detection was
performed using two sets of specific primers (TRI3 and TRI12) designed for trichothecenes by Ward et
al. in 2002. After the polymerase chain reaction was completed, the products were electrophoretically
separated on a 1% agarose gel, and visualization was performed by staining with ethidium bromide and
observing under UV light. Amplified fragments were read visually, and a positive reaction indicated
the presence of DNA fragment of the expected size. All isolates of F. graminearum examined in this
study had products of size 610bp at TRI3 and 670bp at TRI12, which indicates that isolates originating
from Serbia belonged to 15ADON chemotype
Divergence within the Taxon ‘Candidatus Phytoplasma asteris’ Confirmed by Comparative Genome Analysis of Carrot Strains
No full textPhytoplasmas are linked to diseases in hundreds of economically important crops, including carrots. In carrots, phytoplasmosis is associated with leaf chlorosis and necrosis, coupled with inhibited root system development, ultimately leading to significant economic losses. During a field study conducted in Baden-Württemberg (Germany), two strains of the provisional taxon ‘Candidatus Phytoplasma asteris’ were identified within a carrot plot. For further analysis, strains M8 and M33 underwent shotgun sequencing, utilising single-molecule-real-time (SMRT) long-read sequencing and sequencing-by-synthesis (SBS) paired-end short-read sequencing techniques. Hybrid assemblies resulted in complete de novo assemblies of two genomes harboring circular chromosomes and two plasmids. Analyses, including average nucleotide identity and sequence comparisons of established marker genes, confirmed the phylogenetic divergence of ‘Ca. P. asteris’ and a different assignment of strains to the 16S rRNA subgroup I-A for M33 and I-B for M8. These groups exhibited unique features, encompassing virulence factors and genes, associated with the mobilome. In contrast, pan-genome analysis revealed a highly conserved gene set related to metabolism across these strains. This analysis of the Aster Yellows (AY) group reaffirms the perception of phytoplasmas as bacteria that have undergone extensive genome reduction during their co-evolution with the host and an increase of genome size by mobilome
Virulence of native populations of Steinernema feltiae (Filipjev) against Lycoriella ingenua (Dufour)
No full textPest and disease control in cultivated mushroom production in Serbia is mainly based on chemical
pesticides that are toxic to non-target organisms, humans and the environment. The European and
national policies, as well as the United Nations Sustainable Development Goals, strongly
encourage the use of low-risk active ingredients and sustainable alternatives to chemical
pesticides in mushroom disease and pest control. The mushroom fly, Lycoriella ingenua (Dufour)
(Sciaridae: Diptera) is the most damaging mushroom pest in Serbia and globally. The aim of the
study was to evaluate the virulence of two locally isolated populations of Steinernema feltiae
(Filipjev) (strains P9 and K2) against the mushroom fly.
Nematodes were reared on grater wax moth last instar larvae, and infective juveniles (IJ) not older
than 15 days were used in the experiments. Experiments were conducted in plastic Petri dishes
(35 mm in diameter) with the bottom surface covered with filter paper. Volume of 0.5 ml of
nematode suspension containing 0, 50, 100, 200, 500, and 1000 IJ were applied to filter paper and
10 fourth larval instar of L. ingenua were added subsequently in each Petri dish. Every
concentration of both nematode populations were replicated five times and whole experiment was
repeated twice. The mortality of mushroom fly larvae was recorded three and seven days after
exposure to nematodes. Log-probit regression analyses of the concentration-mortality data
showed that both local populations of S. feltiae tested caused significant mortality of the fourth
instar larvae of L. ingenua. In the dose-response bioassays, the following LC50s (median Lethal
Concentrations) were estimated: 15.7718 and 18.2742 nematodes/larva (i.e. 16 and 18 IJ/cm2),
for populations P9 and K2, respectively. Values LC90 for both populations corresponded with
the highest IJ concentration used, around 100 IJ/cm2. A lethal dose ratio test showed that the local
population P9 was significantly more virulent toward the mushroom fly larvae. Experiments
regarding in vivo testing of efficacy of local populations of S. feltiae against L. ingenua are
ongoin
Diversity of Coccinellidae in colonies of soft scales (Hemiptera: Coccidae)
No full textThe ladybird beetles (Coleoptera: Coccinellidae) is one of the most important predatory groups
in biological control of soft scales, which are significant pests of many horticultural and
agricultural crops. Coccinellidae in nature can provide successful control of these pests,
especially in environments where chemical control measures are not applied. This study was
carried out to determine the Coccinellidae species and their preys (Hemiptera: Coccidae) in
Serbia. The samples were collected from perennial plants, including fruit trees, vines, forest trees,
shrubs, and ornamental woody plants. Predator larvae collected with scales were reared
individually in petri dishes in order to prevent cannibalism. Eclosed ladybugs adults were
mounted on cards, and identified using the Bieńkowski (2018) key. Total of 163 individuals were
collected and 8 species belonging to 7 genera were identified. The most common species of
predator were Exochomus quadripustulatus L. that was reared from 16 species of soft scale
species and Harmonia axyridis Pallas, which has been identified on nine Coccidae species. Also,
these two species were the most numerous in the soft scale colonies. A total of 117 individuals
E. quadripustulatus and 22 individuals of H. axyridis were reared. Other species, Chilocorus
bipustulatus (L.), Chilocorus renipustulatus (Scriba), Coccinella septempunctata L.,
Hippodamia variegata (Goeze), Propylea quatuordecimpunctata (L.) and Scymnus abietis
Paykull, were represented individually in soft scale colonies. Ladybirds were present in Coccidae
colonies throughout the year, with young larvae feeding on eggs or first instar nymphs of
Coccidae, and older larvae and adults feeding on all developmental stages of the host. The future
research should be focused on the study of individual Coccinellidae species and their role in the
reduction of harmful Coccidae