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Antagonistic activity of Bacillus spp. strains against Monilinia spp.
No full textBrown rot caused by Monilinia spp. is one of the major factors limiting the pre- and post-harvest
period of stone and pome fruits. Traditionally, synthetic fungicides have been used for managing
brown rot during bloom and preharvest periods, while the postharvest use of fungicides is restricted
in EU countries, as well as in Serbia. Because of the growing public concern over the health and
environmental hazards associated with high levels of pesticide use, as well as restricted usage of
fungicides in postharvest period, the use of environmentally friendly approaches, such as biological
control agents, has increased in recent years.
The aim of this study was to test the antagonistic activity of Bacillus spp. strains from Serbia
against three main pathogens of genus Monilinia: Monilinia laxa, Monilinia fructigena and Monilinia
fructicola, and to test efficacy of the most promising strains in apple protection against these brown
fruit rot pathogens.
We conducted in vitro screening of 33 Bacillus spp. strains originating from different substrates
for their antifungal properties. In vitro antagonistic activity was assessed using direct confrontation
test and well diffusion method. Based on antagonistic effects in vitro, three of 33 isolates were chosen
for further studies and identified to the species level. Additionally, bioactivity of the most promising
antagonistic strains was tested in vivo on apple fruits cv. Idared.
The results showed that the investigated bacterial isolates exhibited antifungal properties
against all three Monilinia species in vitro, and Bacillus amyloliquefaciens B-241, and Bacillus subtilis
B-313 and B-358 were selected for further testing in vivo. The experiment on wound-inoculated
apple fruits showed that tested isolates were effective and that their effectiveness remained stable
for at least 24 hours. On inoculated apple fruits, B. amyloliquefaciens B-241 was the most effective,
achieving 87.1 to 93.7% efficacy compared to the control. On the other hand, B. subtilis B-313 reached
59.1 to 76.4% efficacy, while B. subtilis B-358 was the least effective achieving 11.6 to 37.5 % efficacy,
depending on Monilinia isolates used in the study. B. amyloliquefaciens B-241 arose as a promising
candidate for developing a shelf-stable formulated product and for testing its activity spectrum and
efficacy under field conditions, as well as to estimate economic aspects of its use.VIII CONGRESS ON PLANT PROTECTION: Integrated Plant Protection for Sustainable Crop Production and Forestry ;
VIII КОНГРЕСС ПО ЗАЩИТЕ РАСТЕНИЙ: Интегрированная защита растений для устойчивого растительного
производства и лесного хозяйства ; Book of Аbstracts / Сборник тезисов ; November 25-29, 2019, Zlatibor, Serbia
25-29 ноября 2019 года, Златибор, Серби
Antifungal effect of Bacillus subtilis B6 strain on Monilinia fructicola
No full textBrown rot caused by Monilinia spp. is one of the major factors limiting the storage period and market life of stone and pome fruits. Control of postharvest decay is currently based on the use of synthetic fungicides during bloom and preharvest periods, while postharvest use of fungicides is restricted in EU countries, as well as in Serbia. In recent years, the use of environmentally friendly approaches, such as biological control agents, has increased continuously due to public concerns over the risks of pesticide residues remaining in food and their negative impact on the environment. One of the most significant practical constraints in the use of biocontrol products is the required development of their shelf-stable formulations.
The aim of this study was to test antifungal properties of numerous bacterial strains from Serbia against Monilinia fructicola, to develop a suspo-emulsion (SE) formulation, and to test its efficacy in apple protection against one of the most damaging brown fruit rot pathogens. We conducted in vitro screening of many bacterial isolates originating from different substrates for their antifungal properties against a M. fructicola isolate as a model organism. In vitro antagonistic activity assays were performed in 90 mm Petri plates using the wells technique. Based on antagonistic effects in vitro, seven of 108 isolates were chosen for further studies. The isolates were identified to the species level based on morphological and molecular characteristics. One the most promising antagonistic strains was then selected to develop a SE biopesticide formulation and test its bioactivity in vivo on apple fruits cv. Golden Delicious. The results showed that the investigated bacteria exhibited antifungal properties against M. fructicola in vitro, and that the B6 strain of Bacillus subtilis qualifies to be selected as the best for further testing in vivo. The experiment on wound-inoculated apple fruits showed that the SE formulation of B6 strain was as effective as a reference synthetic fungicide and that its effectiveness remained stable for at least 24 hours. On inoculated apple fruits, the 10% concentration of SE formulation was the most effective, achieving 80% efficacy compared to the control. This work confirmed a great potential of the isolated bacterial strain and its formulation for biological control of brown rot in apple fruit.https://events.uliege.be/postharvest2019/wp-content/uploads/sites/24/2019/06/Book-of-Abstract-ISPP2019-Lie%CC%80ge.pdfFrom Consumer to Laboratory: Sustainable Approaches to Managing Postharvest Pathogens, 19 – 24 May, 2019, Liège, Belgiu
Different aspects of non-standard foliar fertilizers based on amino acids, phytohormones and plant extracts
No full textWe studied different aspects of application of foliar non-standard fertilizers based on amino
acids, phytohormones and plant extracts. The trials were carried out at the level of the
seedlings, individual plants grown in semi-controled conditions and plants grown in the field.
Various energetic and thermodynamic parameters were analyzed, then the chemical
composition (mineral elements, different sugars, secondary metabolites, etc.), as well as
parameters of plant growth and their yield, in order to better assess the impact of these
fertilizers on crops. We found that on maize seedlings it works by changing the content of
various elements, then the polyphenol profiles, as well as thermodynamic parameters, where
this effect does not only depend on the dosage of the fertilizers, but also on the corn genotype.
We also found that the fertilizers affect the energetic and thermodynamic parameters of
individual maize plants, as well as the parameters of plant growth. The most significant and
most diverse results were obtained by analyzing the yield and components of the yield of
many different crops (crop, fruit, vegetable), as well as their chemical composition (mineral
elements, different sugars, secondary metabolites, etc.) in terms of improving nutritive
quality. It was noticed that these fertilizers greatly affect the content of microelements, starch
and crude proteins in maize and barley, sugar and polyphenol content in various fruit trees, as
well as in soybeans, in which we note that in certain agroecological situations these fertilizers
have led to spectacular magnification yields of different crops, but there were also situations
when they did not have any positive effect on crop yields. Overall, the early treatment of
cultivated plants with non-standard fertilizers greatly affects the vigor of seedlings of these
plants, which is of great importance for crop yield. These fertilizers also significantly affect
the quality of crop yield in terms of improved chemical composition of edible parts of these
plants
The activity of native Bacillus subtilis strains in control of green mould disease of oyster mushroom (Pleurotus spp.)
No full textCilj ovog rada je bio pronalaženje potencijalnih agenasa biološke kontrole, izolovanih iz supstrata za gajenje šampinjona, koji bi predstavljali alternativu toksičnim hemikalijama koje se koriste u suzbijanju prouzrokovača bolesti u proizvodnji gajenih gljiva. Ispitivan je antagonistički potencijal deset domaćih sojeva Bacillus subtilis protiv prouzrokovača zelene plesni bukovače, Trichoderma pleuroti i Trichoderma pleuroticola. Korišćen je in vitro metod za kvantifikaciju antagonističkog potencijala sojeva Bacillus spp. zasnovan na dvojnoj kultivaciji s patogenom. Inhibicija porasta T. pleuroti varirala je između 54,44% i 62,22% i nisu utvrđene statistički značajne razlike između antagonističke aktivnosti sojeva B. subtilis testiranih u radu. Inhibicija T. pleuroticola je bila neznatno veća, (55,56% do 69,62%), a soj B. subtilis B-358 je ispoljio najveći procenat inhibicije. Ovo istraživanje je potvrdilo da je supstrat za gajenje gljiva dobar izvor antagonističkih mikroorganizama sa potencijalom korišćenja u biološkom suzbijanju zelene plesni bukovače.The study aimed to isolate potential biocontrol agents from mushroom substrate that could serve as an alternative to toxic chemicals commonly used for disease control in mushroom production. The antagonistic potential of ten native Bacillus subtilis strains against the causal agents of green mould disease of oyster mushroom, Trichoderma pleuroti and Trichoderma pleuroticola, was evaluated. The antagonistic potential of Bacillus spp. strains was quantified in vitro based on dual cultivation with the pathogen. Growth inhibition of T. pleuroti ranged from 54.44% to 62.22% and no significant differences in antagonistic activity were found between the tested B. subtilis strains. Inhibition of T. pleuroticola was slightly higher, ranging from 55.56% to 69.62% and B. subtilis strain B-358 induced the highest growth inhibition. This research confirmed mushroom substrate to be a good source of antagonistic microorganisms with potentials for use in biological control of green mould in oyster mushroom production
Photocatalytic degradation of carbamate insecticides: Effect of different parameters
No full textProučavana je fotokatalitička degradacija model jedinjenja karbamatnog insekticida karbofurana u vodi, u prisustvu polihromatske svetlosti i katalizatora ZnO i TiO2. Ispitivan je uticaj reakcionih parametara, kao što su vreme reakcije i početna koncentracija karbofurana, na fotokatalitičku degradaciju. Ustanovljeno je da je reakcija bila pseudo-prvog reda i primenom optimalnih uslova skoro potpuno uklanjanje 88,4 mg L-1 karbofurana dogodilo se u toku 2 sata. Reakcije su praćene primenom UV spektroskopije i tečne hromatografije visokih performansi (HPLC). Naknadno, upoređena je fotokatalitička efikasnost katalizatora ZnO i TiO2 primenom identičnih reakcionih uslova.Photocatalytic degradation of a model compound of the carbamate insecticide carbofuran in water was studied using polychromatic light and ZnO and TiO2 catalysts. The influence of operational parameters, such as reaction time and initial carbofuran concentration, on photocatalytic degradation was studied. A pseudo-first-order kinetic model was established and an almost complete removal of 88.4 mg L-1 of carbofuran occurred within 2 h under optimized conditions. The reactions were examined by UV spectroscopy and high performance liquid chromatography (HPLC). In addition, the photocatalytic efficiencies of ZnO and TiO2 were compared under the same reaction conditions
Residual efficacy of deltamethrin against Sitotroga cerealella (Olivier) (Lepidoptera: Gelechiidae) in treated maize grain
No full textpests of stored maize in Serbia and worldwide. Contact insecticides continue to be the most significant
tool for residual control of stored-product insect pests. The present study therefore focused on
tests of the residual efficacy of an EC formulation of deltamethrin, which was applied at the recommended
rate (0.25 mg AI/kg of maize grain) against S. cerealella, after different periods of contact
with treated maize grain.
The residual efficacy of the insecticide containing 25 g/L AI deltamethrin and 225 g/L PBO (piperonyl
butoxide synergist), against a lab population of S. cerealella was investigated according to PP
1/203 (1) and PP 1/204 (1) OEPP/EPPO methods. The maize variety ’Pioneer P0725’ with 13,2±0,6%
moisture content in maize grain was used in the experiment. Two standard solutions (1 L each) were
prepared by adding 10 mL (0.25 mg a.i./kg) of deltamethrin product to 990 mL of water. A 50 kg maize
grain lot was poured into a concrete mixer for a 10 min round of mixing and treating with 50 mL water
solution of insecticide. The procedure was run two times to forming 2 lots of 50 kg of treated maize.
The same procedure, only using water alone, was used for control maize. After mixing, the treated
maize was stored in bags at room temperature of 24-26°C and 50±5% r.h. Residual efficacy was tested
in separate bioassays on 0, 7, 14 and 30 days old deposits. Three samples of 50 g were taken from each
lot and placed into plastic vessels and 10 unsexed ≤1 day old imagoes of S. cerealella were added into
each vessel, topped with cotton cloth and fixed with a rubber band. For each bioassay vessel placed
in the laboratory under the same (described) conditions, mortality was estimated after 2, 7 and 14
days of exposure to treated maize grain. In addition, S. cerealella progeny production/reduction (PR,
%) was recorded after a total of 8 weeks for each deposit age and after 14 days of insect exposure.
After 2 and 7 days of exposure to deposits of all ages, deltamethrin caused a wide range of S.
cerealella mortality, 47-87% and 70-100%, respectively, while mortality after 14 days of exposure was
100%. Lepidoptern S. cerealella had no progeny on any deltamethrin deposit age in maize grain (PR,
100%). The results showed that the synergized deltamethrin, applied at 0.25 mg/kg in maize grain, is
a highly effective insecticide for stored-product insect pest control, such as S. cerealella.VIII КОНГРЕСС ПО ЗАЩИТЕ РАСТЕНИЙ: Интегрированная защита растений для устойчивого растительного производства и лесного хозяйства - Сборник тезисов, 25-29 ноября 2019 года, Златибор, Серби
Primena Bacillus subtilis B6 u zaštiti uskladištenih plodova jabuke
No full textJabuka je jedna od najznačajnijih vrsta voćaka u Srbiji, koja se prema podacima iz 2014. godine gaji na površini od 23.737 ha (Republički zavod za statistiku Srbije, 2014). S obzirom da je tržište jabuke izuzetno zahtevno a konkurencija veoma velika, veliki je i izazov pred proizvođačima kako bi proizveli zdravstveno bezbedan plod dobrog kvaliteta i odgovarajuće krupnoće. Jedan od najznačajnijih uzročnika smanjenja kvaliteta plodova i mogućih gubitaka jesu bolesti prouzrokovane fitopatogenim gljivama. Osim patogena koji prouzrokuju značajne gubitke tokom vegetacije, mnoge fitopatogene gljive izazivaju ekonomski značajne gubitke posle berbe koji vrlo često premašuju gubitke u samom procesu proizvodnje (Grahovac i sar., 2011). Čak i prosečni potrošač, koji kupuje kvalitetne plodove u hipermarketima, uočava i prepoznaje značaj i trajnost problema, jer je često prinuđen da ubrzo nakon kupovine propale plodove baci. Literaturni podaci pokazuju da oboljenja uskladištenog voća predstavljaju najznačajniji faktor gubitaka u proizvodnji (Tian, 2007), sa gubicima od 5 do 25% u razvijenim, odnosno do 50% u zemljama u razvoju (Ewekeye et al., 2013; Marković et al., 2011)
Biological control of green mould and dry bubble diseases of cultivated mushroom (Agaricus bisporus L.) by Bacillus spp.
No full textCompost green mould and dry bubble are the most devastating diseases of the cultivated white button mushroom. In search of efficient disease control, previously and newly isolated Bacillus spp. strains obtained from mushroom substrates were examined for antagonistic activity against Trichoderma aggressivum f. europaeum T77, Trichoderma horzianum T54 and Lecanicillium fungicola var. fungicola Sa(2)V(6) in vitro. All 33 tested Bacillus spp. strains inhibited the growth of pathogens. The strains with the strongest antagonistic activity were selected for in vivo experiments in which the efficacy of strains in pathogen suppression and their effect on mushroom yield were evaluated, in comparison with a commercial biofungicide based on Bacillus velezensis QST 713 and a fungicide based on prochloraz-manganese. In mushroom growing room trials with T. aggressivum f. europaeurn, the most effective in pathogen suppression were (in decreasing order) prochloraz-Mn, Bacillus amyloliquefaciens B-241, Bacillus velezensis QST 713, Bacillus pumilus B-138 and B. subtilis B-233. The commercial prochloraz-Mn fungicide was the best at suppression of L. fungicola var. fungicola, while B. amyloliquefaciens B-241 was the most successful out of the three tested Bacillus spp. strains. In all in vivo experiments, B. amyloliquefaciens B-241 showed similar results in suppression of pathogens as the commercial biofungicide B. velezensis QST 713. Regarding the impact on mushroom productivity in trials with both pathogens, no statistically significant differences were noted when the treatments were compared with inoculated and uninoculated controls. Strain B. amyloliquefaciens B-241 showed the greatest potential for biocontrol of both compost green mould and dry bubble disease, which makes it a good candidate for further trials at commercial scale
High genetic diversity of ‘Candidatus phytoplasma solani’ infecting pepper in Serbia
No full textSurveys for the presence of symptoms typical for the “stolbur” disease were conducted during 2015 and 2016 in pepper growing areas of Serbia. ‘Candidatus Phytoplasma solani’ detection was performed by Stol11 primer amplification. Characterisation was done by multi-locus typing (MLST) of tuf, secY, vmp1 and stamp genes. The disease was observed in pepper in all the surveyed locations except in south Serbia, whith a low incidence (3-5%). Overall, 13 MLST genotypes were distinguished, and the most common were Rqg31/Rqg50, Rpm35/M5 and STOL
Botrytis squamosa - prouzrokovač lisne pegavosti luka u Bosni i Herzegovini
No full textOver the past several decades, necrotic spots, lesions and blight symptoms have been observed on onion leaves in several locations in Bosnia and Herzegovina, where the crop is grown intensively. The type of symptoms indicated a possible infection with Botrytis squamosa, a widespread pathogen of onion. As symptoms of leaf spots and necrotic lesions can also be caused by some other biotic and abiotic factors, our research focused on identifying the causal agent of the observed symptoms. The pathogen was isolated from diseased tissue using standard phytopathological procedure and identified based on pathogenic and morphological features. Identification was confirmed by amplification and sequencing of the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA). The influence of temperature and growth medium on mycelial growth rate of the isolates was also studied.U poslednjih nekoliko godina, na nekoliko lokaliteta u području intenzivne proizvodnje crnog luka u Bosni i Hercegovini uočena je pojava nekrotičnih pega i sušenja lišća. Simptomi oboljenja ukazali su na moguće prisustvo široko rasprostranjenog patogena luka, vrste Botrytis squamosa. S obzirom da pegavost i nekrotične lezije na listu luka mogu biti prouzrokovane različitim biotičkim i abiotičkim faktorima, cilj ovog istraživanja bio je da se precizno identifikuje prouzrokovač oboljenja. Patogen je izolovan primenom standardnih fitopatoloških metoda i identifikovan na osnovu proučenih patogenih i morfoloških karakteristika dobijenih izolata. Identifikacija je potvrđena amplifikacijom i sekvenciranjem ITS rDNA genomnog regiona. Takođe, proučen je uticaj temperature i hranljive podloge na porast izolata