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Botryosphaeriaceae kao patogeni jabuke – identifikacija i osetljivost na fungicide i etarska ulja in vitro
No full textApple production suffers significant economic losses and fruit quality reduction
due to fungal pathogens, particularly ones that cause postharvest fruit rot, such as
Botryosphaeriaceae fungi. Isolates used in this study were obtained from symptomatic
apples and, based on morphological characteristics and sequence analysis of two genes
(EF 1-α and β-tubulin), they were identified as Diplodia seriata and Botryosphaeria dothidea.
Pathogenicity tests on healthy apple fruits revealed that D. seriata was more aggressive
than B. dothidea, with significantly higher average values of lesion diameter and depth.
Fungicide sensitivity tests showed that D. seriata was more sensitive to the combination
fluopyram + tebuconazole (EC50=0.00023 μg a.i. ml-1), while B. dothidea exhibited higher
sensitivity to pyraclostrobin (EC50=0.025 μg a.i. ml-1). With 98.44% and 97.56% percent
growth inhibition (PGI) rate of D. seriata and B. dothidea (respectively) at 10 μg a.i. ml-1, the
tested combination of fungicides surpassed pyraclostrobin in inhibition potential. Four
essential oils (thyme, rosemary, lavender and lemongrass) were also tested for antifungal
activity using the fumigant macrodilution method. Thyme oil demonstrated the highest
antifungal potential, completely inhibiting the mycelial growth of both species at
0.05 μl ml–¹ of air. Strong inhibition potential was also shown by lemongrass oil with
100% inhibition of D. seriata and B. dothidea mycelial growth at 0.07 and 0.09 μl ml–¹ of air,
respectively. Rosemary oil showed a moderate inhibition potential, while lavender oil was
the least effective. These findings highlight the inhibiting potential of fungicides against
D. seriata and B. dothidea, but they also indicate that thyme and lemongrass essential
oils could be used as viable alternatives. Further research is needed to determine their
effectiveness in in vivo assays and potential impact on fruit quality and the environment.Proizvodnja jabuke suočava se sa značajnim ekonomskim gubicima usled smanjenog
kvaliteta plodova kao posledice prisustva velikog broja fitopatogenih gljiva, naročito
prouzrokovača truleži plodova nakon berbe kao što su gljive iz familije Botryosphaeriaceae.
Izolati korišćeni u ovom istraživanju prikupljeni su sa zaraženih plodova jabuka i na osnovu
morfoloških karakteristika i analize sekvenci dva genska regiona (EF 1-α and β-tubulin)
identifikovani kao pripadnici vrsta Diplodia seriata i Botryosphaeria dothidea. Na zdravim
plodovima jabuka sproveden je test patogenosti kojim je na osnovu merenja prečnika i
dubine truleži utvrđen značajno viši stepen agresivnosti izolata D. seriata. Veća osetljivost na
kombinaciju fluopiram + tebukonazol uočena je kod izolata D. seriata (EC50=0,00023 μg a.s./ml),
dok je piraklostrobin jače inhibitorno delovanje ispoljio na izolat B. dothidea (EC50=0,025 μg a. s./ml).
Ispitivana kombinacija fluopiram + tebukonazol pokazala je veći inhibitorni potencijal u
poređenju sa piraklostrobinom, sa inhibicijom porasta micelije od 98,44% za D. seriata i
97,56% za B. dothidea pri koncentraciji od 10 μg a.s./ml. Antifungalni efekat četiri etarska
ulja (majčine dušice, ruzmarina, lavande i limunske trave) ispitan je primenom fumigantne
makrodilucione metode. Ulje majčine dušice ispoljilo je najizraženije delovanje, potpuno
inhibirajući porast micelije izolata obe vrste pri koncentraciji 0,05 μl/ml vazdušne faze. Jako
inhibitorno delovanje pokazalo je i ulje limunske trave sa kompletnom inhibicijom porasta
micelije pri koncentraciji 0,07 μl/ml vazdušne faze (D. seriata), odnosno 0,09 μl/ml vazdušne
faze (B. dothidea). Umeren inhibitorni potencijal zabeležen je kod ulja ruzmarina, dok je ulje
lavande ispoljilo najniži antifungalni efekat. Rezultati prikazani u ovom radu ukazuju na visok
stepen osetljivosti izolata D. seriata i B. dothidea na ispitivane fungicide, ali i ulja majčine dušice
i limunske trave kao njihove ekološki prihvatljivije alternative. Dalja istraživanja su potrebna
kako bi se ispitala efikasnost ovih ulja u uslovima in vivo, kao i potencijalni uticaj na kvalitet
plodova i životnu sredinu
Compatibility of biological control agents in the Agaricus bisporus production system
No full textUnderstanding ecological interactions among cohabiting microorganisms after their introduction in the mushroom cultivation cycle is essential for improving both the quantity and quality of products. In addition, pest and disease management using biological control agents (BCAs) is compatible with organic agriculture and represents an environmentally safe alternative that can reduce the use of chemical pesticides. As of late, compost green mould, caused by Trichoderma aggressivum, has become a major fungal disease of button mushrooms. Relationships (synergistic/antagonistic/additive) between the following BCAs: bacterium Bacillus amyloliquefaciens B-241, actinobacterium Streptomyces flavovirens A06, and a commercial population of entomopathogenic nematode Steinernema feltiae in the control of T. aggressivum were evaluated in vivo. Their impact on mushroom yield was also assessed. The synergy factor (SF) was calculated as a ratio between the observed and expected impact on yield or the efficacy in pathogen suppression. The experiment was conducted under conditions of artificial inoculation with T. aggressivum after co-inoculation of these microorganisms. Satisfactory control of T. aggressivum was achieved by all tested microorganisms. The study revealed an additive interaction after simultaneous application of the bacterium B. amyloliquefaciens B-241 and actinobacterium S. flavovirens A06 in their efficacy against the pathogen. Similarly, additive relationships in efficacy against T. aggressivum were confirmed between the EPN population and each of the strains of the beneficial bacterium or actinobacterium. Regarding the impact on yield, synergistic interaction was detected between the two microorganisms, as well as between each beneficial microorganism and EPN S. feltiae. Since the results revealed compatibility among the tested BCAs in the mushroom cropping cycle, their synchronous application is recommendable
Diversity and patulin production of Penicillium spp. associated with apple blue mold in Serbia
No full textApple blue mold, caused by the Penicillium species, is a significant postharvest
disease, leading to food loss and impacting food safety due to mycotoxin contamination.
This study aimed to identify the Penicillium species associated with apple blue mold in
Serbia, assess their pathogenicity, and evaluate their patulin production potential. A total of
70 Penicillium isolates were collected from symptomatic apple fruit and identified as P.
expansum (92.9%), P. crustosum (4.3%), P. solitum (1.4%), and P. chrysogenum (1.4%). The
pathogenicity assay revealed P. expansum strains as the most virulent. Molecular detection of
msas gene and HPLC analysis confirmed patulin production exclusively in P. expansum
isolates. Principal Component Analysis (PCA) grouped P. expansum strains in two distinctive
clusters, while P. crustosum strains clustered separately with P. solitum and P. chrysogenum,
yet in distinct positions. This is the first report of P. solitum and P. chrysogenum as causal
agents of apple blue mold in Serbia. The results of the study provide insights that might be
useful in the development of effective control strategies for apple blue mold, ensuring
consumption of healthy and safe apple fruit and apple-based products
Biološko suzbijanje šampinjonske mušice Lycoriella ingenua (Diptera: Sciaridae) primenom domaćih populacija entomopatogene nematode Steinernema feltiae (Nematoda: Steinernematidae)
No full textEntomopatogene nematode (EPN) dobijaju sve veći značaj kao biološki kontrolni agensi u situaciji kada se sintetski hemijski insekticidi za zaštitu šampinjona od štetočina povlače sa tržišta. Korišćenje bioloških agenasa kao ekološki zasnovanih inputa za održivu proizvodnju šampinjona bez štetnih ostataka sintetskih insekticida, doprinosi zaštiti zdravlja ljudi, životne sredine i neciljanih organizama. U Novom tehničkom rešenju utvrđena je zadovoljavajuća virulentnost (u laboratorijskim uslovima) i efikasnost (u oglednom gajilištu) dve domaće (K2, R9), i jedne komercijalne populacije nematode Steinernema feltiae (Nematoda: Steinernematidae), u biološkom suzbijanju šampinjonske mušice, Lycoriella ingenua (Diptera: Sciaridae), kao i odsustvo negativnih efekata njihove primene na prinos šampinjona. Efikasnost domaćih populacija S. feltiae u suzbijanju larvi šampinjonske mušice (oko 77%), nije se statistički značajno razlikovala od efikasnosti koju je postigla komercijalna populacija ove vrste (oko 87%), ukazujući na značajan biološki potencijal domaćih populacija u suzbijanju šampinjonske mušice, ekonomski najznačajnije štetočine šampinjona. U Novom tehničkom rešenju se za optimalne efekte zaštite šampinjona od šampinjonske mušice preporučuje dvokratna primena ispitivanih populacija S. feltiae, u koncentraciji 1,5×106 IJ (infektivnih juvenila) po m2 pokrivke, sa prvim tretmanom 0,75×106 IJ u 1 l vode po m2 odmah po postavljanju pokrivke, i drugim 0,75×106 IJ u 1 l vode po m2 nakon sedam dana. Takođe, preporučuje se preventivno tretiranje komposta u kompostarama pre zasejavanja micelije šampinjona populacijom entomopatogenih nematoda, u koncentraciji 0,75×106 IJ u 2 l vode na 1000 kg komposta
Morphological, Pathogenic and Molecular Characterization of Sclerotinia sclerotiorum, the Causal Agent of White Rot of Cabbage (Brassica oleracea var. capitata), in Serbia
No full textSclerotinia sclerotiorum is a globally distributed necrotrophic pathogen with a broad host
range, including many Brassicaceae crops. In 2021, white rot symptoms on cabbage were
observed in 12 commercial fields in the northern part of Serbia. Twelve representative
isolates of S. sclerotiorum, forming white colonies and black sclerotia, were selected for
characterization and comparison with an isolate from sunflower, as the most important
host plant in Serbia. The isolates showed significant variation in growth rate and sclerotia
production, while ITS sequence analysis revealed the complete nucleotide identity and all
isolates grouped within the major phylogenetic clade of S. sclerotiorum. Despite the low
diversity of the global population of S. sclerotiorum, forty-four haplotypes were identified,
with one predominant haplotype encompassing all Serbian isolates. When six Brassicaceae
species (cabbage, cauliflower, broccoli, kale, mustard, and oilseed rape) and sunflower
were inoculated, sunflower was found to be the most and cauliflower the least susceptible,
while isolates from cabbage were generally more aggressive than those derived from
sunflower. This work represents the first detailed characterization of S. sclerotiorum infecting
cabbage in Serbia and highlights its genetic uniformity and differential pathogenic potential,
which are critical factors for integrated disease management and crop rotation planning in
Brassicaceae agroecosystems
Discovering Ecological Interactions Between Biocontrol Bacterial Strains and Entomopathogenic Nematodes in Button Mushroom Production
No full textThe substrate for button mushroom (Agaricus bisporus) cultivation includes
a highly complex microbiome. The aim of the study was to evaluate ecological inter-
actions (synergistic, antagonistic, or additive) between a commercial population of the
entomopathogenic nematode Steinernema feltiae (EPN) and beneficial microorganisms, bac-
terium Bacillus amyloliquefaciens B-241 (BA) or actinobacterium Streptomyces flavovirens
A06 (SF). Their relationships were evaluated in efficacy against the pathogenic fungus
Trichoderma aggressivum and the fungus gnat Lycoriella ingenua. Moreover, their impact on
mushroom yield was estimated. The synergy factor was calculated as the ratio of observed
to expected values regarding their efficacy against T. aggressivum/L. ingenua and influence
on mushroom production. Additive relationships in efficacy against T. aggressivum were
observed between EPN and BA or SF. As for the impact on yield, synergistic interactions
were indicated between each beneficial microorganism and EPN. Considering suppression
of L. ingenua, a mild antagonistic reaction between EPN and each beneficial microorganism
was observed in plots without T. aggressivum and additive in plots inoculated with the
pathogenic fungus, although high efficacy was achieved in all combinations (>80%). Tested
native strains of both beneficial microorganisms could be combined with the commercial
EPN strain for successful biological pest and disease control in mushroom productio
Antifungal activity of peppermint and spearmint essential oils against Trichoderma spp. green mold disease agents of oyster mushroom and shiitake
No full textProduction of oyster mushroom (Pleurotus spp.) and shiitake (Lentinula edodes) is seriously affected by various
Trichoderma species causing green mold disease. The purpose of the study was to determine the antifungal activity
of essential oils peppermint (Mentha piperita) and spearmint (Mentha spicata), both obtained from plants from
Serbia, against Trichoderma strains isolated from Pleurotus ostreatus oyster mushroom (Trichoderma pleuroti KM11
and Trichoderma pleuroticola KM12, 2018, North Macedonia) and shiitake substrates (Trichoderma guizhouense
T59, 2018, Serbia). Air-dried plant material was subjected to hydro-distillation in a Clevenger type apparatus. The
obtained essential oils were dried over anhydrous sodium sulphate. Antifungal activity of the oils was tested using
in vitro methods: contact microdilution by using a pathogen spore suspension, and fumigant macrodilution by
applying either pathogen mycelial inoculum or spore suspension. Both oils were lethal for all tested species when
the contact microdilution method was used at 12.5 μl mL-1, while no fungicidal effect of either oil was found on any
tested strain using the fumigant method. In the contact microdilution bio-assay, both oils inhibited T. guizhouense
T59 at 3.12 μl mL-1; peppermint oil more strongly suppresed T. pleuroti KM11 (3.12 μl mL-1) than T. pleuroticola KM12
(6.25 μl mL-1), while spearment oil had an opposite effect. Using fumigation, both oils inhibitied mycelial growth of
the tested species at 0.16 μl mL-1, with an exception of spearmint oil, which inhibited T. guizhouense T59 at 0.32 μl
mL-1 of air phase. Inhibition of spore germination of T. pleuroti KM11 by fumigation was detected at 0.08 (pepermint
oil) or 0.016 μl mL-1 (spearmint oil), T. pleuroticola KM12 at 0.016 μl mL-1 (both oils), and T. ghuizouense T59 at 0.32
μl mL-1 (peppermint oil) or inhibition was missing (spearmint oil). The most resistant taxon to both oils was T.
guizhoense from shiitake. Both oils showed similar contact effect, while peppermint oil exhibited a slightly stronger
fumigant activity than spearmint oil on the tested strains. Peppermint and spearmint oils could be recommended
for further in vivo investigation
The effect of Bacillus spp. isolates and calendula extract on tomato yield
No full textXIII International Symposium on Agricultural Sciences AgroReS 2024
BOOK OF ABSTRACTS
91
P1_50
The effect of Bacillus spp. isolates and calendula extract on tomato yield
Slađana Savić1 , Marina Dervišević2, Biljana Šević3, Jelena Maksimović4 ,
Magdalena Knežević4, Zoran Dinić4, Ivana Radović5
1 Institute for Plant Protection and Environment, Serbia
2 Institute of Pesticides and Environmental Protection, Serbia
3 Institute for Vegetable Crops Smederevska Palanka, Serbia
4 Institute of Soil Science, Serbia
5 University of Belgrade, Faculty of Agriculture, Serbia
Corresponding author: Slađana Savić, [email protected]
Abstract
In recent years, in order to protect the environment and food safety, intensive work has been done
on the development of biological fungicides, nematicides and herbicides, as well as products
based on biological agents and medicinal herbs for plant protection. The aim of this research was
to evaluate the effect of the application of Bacillus spp. and calendula extract on tomato yield.
The experiment was carried out during the 2023 growing season in the field conditions using SP
- 109 genotype of tomato, selected at the Institute for Vegetable Crops Smederevska Palanka.
Tomato seedlings were produced in a greenhouse and were planted in the field at the end of May.
The experiment was laid out in a random block system, with four replications. The number of
plants per main plot was 36. The distance between the rows was 50 cm and between the plants
in the row 25 cm. Five treatments were applied three times during the vegetable season (T1 -
Bacillus spp. isolate 1, T2 - Bacillus spp. isolate 2, T3 - Calendula extract, T4 - Fungicide, T5 -
Control). The number of fruits per plant in treatments T1 and T4 was 29. It was statistically
significantly higher compared to the number of fruits per plant in treatments T2, T3 and T5 (20,
21, 20). Also, the fresh weight of fruits per plant was significantly higher in treatments T1 and
T4 (847.7 and 868.5) compared to treatments T2, T3 and T5 (686.7, 680.4 and 634.4). Results
of this investigation indicate that the application of biological agents, specifically selected
Bacillus spp. isolate 1 (T1), can have effects comparable to fungicides, as evidenced by the
achieved tomato yield. The future research will include investigations of the impact of Bacillus
spp. isolates and calendula extract on the nutritional characteristics of tomato fruit
Diversity and interactions of the parasitoids (Hymenoptera: Chalcidoidea) of soft scales from Serbia
No full textThis research aimed to study the diversity of soft scale parasitoids and examine their potential in the biological control of harmful Coccidae on cultivated and ornamental plants. Forty species of parasitoid wasps from four families (Hymenoptera: Chalcidoidea) were reared and determined from 16 species of soft scales (Hemiptera: Coccidae). A total of 6,249 individuals of parasitoid wasps were collected from 35 localities in Serbia from 2014 to 2017. The most numerous genera are Metaphycus with 10, Coccophagus with 7, and Blastothrix with five reared species. The most abundant species is C. lycimnia, reared from second-instar larvae of all 16 species of scale insects and from females of 11 soft scale species, with a total of 1,053 individuals. Of the 40 identified species, 15 are new to the fauna of Serbia: Coccophagus piceae, Coccophagus proximus, Coccophagus silvestrii, Encarsia gigas (Aphelinidae), Adelencyrtus intersectus, Anagyrus schoenherri, Blastothrix brittanica, Blastothrix erythrostetha, Blastothrix hedqvisti, Encyrtus infidus, Metaphycus hageni, Metaphycus stanleyi, Metaphycus unicolor, Microterys hortulanus (Encyrtidae) and Eunotus obscurus (Pteromalidae). Ninety-nine (99) hymenopteran/coccoid associations were identified in this study, of which 16 associations are new
Selection of lettuce rhyzobacteria antagonistic to Sclerotinia spp. in vitro
No full textLettuce (Lactuca sativa L.) is the world's most popular and widely grown leafy vegetable. Lettuce drop, caused by Sclerotinia spp., is one of the major threats for growers that can cause significant losses under favourable conditions. In Serbia, two species, Sclerotinia sclerotiorum and Sclerotinia minor were detected as economically important pathogens of lettuce. Available control measures are either ineffective or environmentally unacceptable. Biological control using beneficial microorganisms could be a convenient and feasible solution. Endospore-producing bacterial species of the genus Bacillus are the most promising biocontrol tools for a variety of plant pathogens. The aim of this study was to test a hypothesis that a rhizosphere of healthy and vigorous lettuce plants originating from Sclerotinia-infested plots contains antagonistic Bacillus spp. strains that could effectively suppress Sclerotinia spp.
Isolation of endospore-forming bacterial strains with antagonistic potential to Sclerotinia spp. was performed from soil samples, taken from the rhizosphere of healthy lettuce plants that were grown in close proximity to diseased plants, naturally infected by Sclerotinia spp. Ten grams of each soil sample was suspended in 100 ml of water, mixed on a magnetic stirrer for 20 min and incubated at 80°C for 10 min. After the incubation period, serial dilutions of the samples were seeded on NA (nutrient agar) and PDA (potato dextrose agar) media and incubated at 28°C for 48h.
Fifty two gram-positive and catalase-positive bacterial isolates, forming whitish, convex, Bacillus-like colonies were derived from the samples. Based on the results of a confrontation test, performed with 24h-old bacterial colonies grown on NA medium and 3-day old colonies of S. sclerotiorum and S. minor isolates from PDA plates, 12 bacterial strains were selected as promising biocontrol agents against Sclerotinia spp. Their antagonistic effect on Sclerotinia spp. was further tested by using a Double-Layer Agar (DLA) method. The first layer was 20 ml of 2% PDA, while the second layer consisted of 10 ml of the suspension of fungal pathogens in 1.2% PDA, distributed homogeneously over the first layer. Bacterial suspensions were prepared by inoculation of a single bacterial colony into NB (nutrient broth) medium and incubated for 72h at 28ºC on a rotary shaker (240 rpm). Wells (10 mm in diameter) were cut in the central part of the DLA plates and 100 μl of the bacterial suspension was added. Sterile distilled water was used as a negative control. The experiment was performed twice in four replicates. Antagonistic activity of the bacterial strains was evaluated by measuring the growth inhibition zone of the fungal mycelium (mm) formed around the well after 48h at 25 ºC.
Among 12 selected bacterial strains, the strain B 7/16 showed the highest antagonistic effect on S. sclerotiorum, forming 64.2 mm-wide inhibition zone, while the strain B 7/6 showed the best result against S. minor with 46.8 mm-wide zone. This work demonstrates that a rhizosphere of healthy and vigorous plants contains promissing antagonistic bacterial strains that could be used as an alternative to synthetic pesticides. Our future work will be focused on examining in vivo efficacy of our strains compared to commercially available biofungicides