Biocontrol of Seedborne Fungi on Small-Grained Cereals Using Bacillus halotolerans Strain B33

The development of biological pesticides is rapidly becoming an integral aspect of pest management in sustainable agriculture. This study was conducted to evaluate the effectiveness of Bacillus halotolerans strain B33 against three common seedborne fungal pathogens—Fusarium graminearum, Alternaria alternata, and Aspergillus flavus. B33 strain identity was determined using the 16S rRNA and tuf gene sequences. Commercial wheat, barley, oat, and rye seeds were artificially infected by fungal isolates and then treated with B33 overnight culture in Nutrient Broth. The obtained results indicate high efficacy against F. graminearum (83.55–94.38%) and A. alternata (85.05–96.70%), whereby the highest efficacy was noted on wheat seed and the lowest was detected on rye seed. On the other hand, B33 achieved 100% efficacy against A. flavus on barley, rye, and oat seeds, while being 96.24% effective against this pathogen on wheat. Principal component analysis indicated the highest treatment influence on A. flavus. The effect of all tested treatments on seed germination was statistically significant compared to the controls, whereby the number of germinated seeds declined as the seed infection rate increased. B. halotolerans strain B33 effectively managed seedborne fungal pathogens, thereby enhancing seed germination

The effect of biopesticides on tomato yield and quality

Biopesticides are increasingly being used for plant protection as natural and environmentally friendly alternatives to synthetic pesticides. The aim of the research was to examine the impact of the Bacillus spp. and calendula extract on tomato yield (fresh weight of fruits) and fruit quality (content of phenolic compounds, vitamin C, lycopene and carotenoids). The experiment was carried out during the 2024 growing season in the field conditions using tomato cultivar for processing (SP-109). The experiment was laid out in a random block system, with four replications. 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). Results showed that T1 treatment significantly increased yield per plant (593 g) compared to treatments T2, T3, T4 and T5 (466.5 g, 509.4 g, 420.3 g and 525.2 g). Also, the T1 treatment had a significant impact on fruit quality, since vitamin C content in the fruits was increased (from 33.5% to 213.6%) compared to others treatments. The content of phenolic compounds was significantly reduced in all treatments compared to control treatment, while the lycopene and carotenoids were similar in all treatments. Results of this investigation indicate that the application of biological agents, specifically selected Bacillus spp. isolate 1 (T1), can have effects comparable to fungicides (even better), as evidenced by the achieved tomato yield and fruit quality, while calendula extract had no effect on tomato yield and quality

First report of Cucumber mosaic virus infecting Petroselinum crispum in Serbia

The Apiaceae family, a large and complex family with about 3820 species distributed throughout the world, includes important leaf and root vegetables such as carrot, parsley, celery, and parsnip. A number of species are cultivated as aromatic herbs and spices, while certain species have long been used in medicine. It is well-known that this family has been threatened by numerous pathogens, of which more than 30 widespread viruses have been described. Cucumber mosaic virus (CMV; Cucumovirus CMV, Cucumovirus, Bromoviridae) is one of the most widespread viruses, causing significant agronomic losses in many crops, including over 1,300 plant species from more than 500 genera and 100 families, with a high potential for rapid spread to new hosts. In autumn 2023, during a survey to determine the presence of Apiaceous viruses in Serbia, symptomatic parsley plants with chlorotic spots and line patterns on the leaf surface were observed in the Medvedja locality (district of Rasina). The collected samples were analyzed by double-antibody sandwich (DAS)-ELISA and CMV was detected in 7 out of 9 tested samples. The presence of CMV in parsley was also confirmed by RT-PCR with specific primers. The amplified product of isolate 4/23 was sequenced using Sanger sequencing method and analyzed by MEGAX software. BLAST search result revealed a 99,52% identity with Polish CMV isolate from host Lonicera caprifolium (EU 191027). The constructed phylogenetic tree showed that the Serbian parsley isolate belongs to CMV subgroup II. In Yugoslavia's former geographical region, CMV's presence was suspected based on symptoms observed on celery plants in the fields. To our knowledge, this is the first report of CMV on parsley in Serbia

First report of Pseudomonas avellanae causing hazelnut decline in Serbia

Grapevine trunk diseases (GTDs) are a group of diseases caused by a large number of fungal phytopathogens that infect and colonize the woody tissue of grapevine, causing chronic or apoplectic decline of plants, and as such posing a serious threat to viticulture. Among the most virulent pathogens are Neofusicoccum parvum, the causal agent of Botryosphaeria dieback, and Fomitiporia mediterranea considered as the main causal agent of grapevine rot within the Esca disease complex. This study aimed to isolate and select potent indigenous bacteria collected from Montenegrin vineyards to serve in the control of GTDs. Isolations were performed from soil samples taken in a zone of grapevine root (Montenegro) on NA after soil was dissolved in water and heating at 80°C for 20 min. A total of 262 isolates were tested for antagonistic activity in a direct in vitro test on PDA. The appearance of a clear zone was checked after fungi development. Results showed that 32 Montenegrin isolates inhibited the growth of N. parvum by 34.0-65.0%, and F. mediterranea by 34.8-71.0%. Neighbourjoining phylogenetic analysis based on the 16S rRNA (primers P0/P6) sequences differentiated those isolates into six clusters but lacked the resolution to differentiate between several species. Six groups were as follows: I (21 isolates: Bacillus amyloliquefaciens/B. velezensis/B. siamensis/B. subtilis), II (2: B. subtilis/B. velezensis/B. tequilensis), III (5: B. halotolerans/B. mojavensis/B. subtilis/B. velezensis/B. tequilensis/B. spizizenii), IV (1: B. licheniformis/B. paralicheniformis), V (2: B. cereus/B. thuringiensis/B. albus/B. anthracis), VI (1: Paenibacillus peoriae/P. polymyxa). Sequencing of housekeeping genes would provide more accurate identification

Storage of buckwheat products as an factor affecting colour condition and implications on meat products

Buckwheat is one of the traditionally high-quality crops recognized for its high nutritional and functional value. Buckwheat products (grain and flour) stored for 0, 3, 6, and 9 months and emulsion-type chicken sausages modified by adding those buckwheat flours (FB0, FB3, FB6, and FB9) were analyzed for instrumental colour parameters (L*, a*, b*, C*, h, S, ΔE). One- (P < 0.05) and two-way (P < 0.001) ANOVA tests were used to analyze the data, followed by Tukey’s HSD post-hoc test (P < 0.05). In terms of storage, a slight decrease in hue angle was observed in grains, with the lowest value of 57.48 (6 months). In flour, trends of decreasing L* and increasing a*, b*, C*, and S values were observed to the end of storage. The values of hue angle and total colour difference decreased, with the lowest point at 73.92 for h and the largest significant change at 4.73 for ΔE in 6 months. In terms of product type, compared to grains, flour was lighter and yellowish on average 2.3 (L*) and 1.3 (h) times. Grains consistently demonstrated significantly higher a*, C*, and S values. In composite products, added flour slightly affects L*, and significantly a* and b*, where FB0 was lighter, redder and yellower than the other combined products. A significant decrease in C* and increase in h values caused lower chromatic purity and a more yellow nuance in FB9 (20.66, 45.38). A decrease in S resulted in the greatest attenuation in overall colour perception at FB6, and FB9. An increase trend of ΔE indicating the differentiation of sausages with respect to effect on total colour (ΔE ≤ 5.10). The results could indicate the potential utilization of long-term stored buckwheat grains (flour) as a functional ingredient and valuable plant-based raw material in the production of enriched, composite meat products

80 ГОДИНА РАДА ИНСТИТУТА ЗА ЗАШТИТУ БИЉА И ЖИВОТНУ СРЕДИНУ

Институт се од оснивања до данас стално развијао и унапређивао основну делатност, научноистраживачки рад из области пољопривреде - заштите биља. Основан је 29. септембра 1945. године, у време када није било ни средстава, ни кадрова за научни рад, па је рађено на постепеном опремању лабораторија, оспособљавању истраживача и истовременом решавању бројних актуелних проблема у заштити биља. Руководство и истраживачи су увек били фокусирани на изврсност и релевантност научноистраживачких резултата са јасно зацртаним циљевима: стварање квалитетног истраживачког кадра, конкурентности истраживања и иновативним решавањем важних питања у пољопривреди и заштити биља. Данас је рад истраживача у Институту усмерен на решавање присуства штетних организама у биљној производњи на мултидисциплинаран и интердисциплинаран начин, обједињавањем основних (природно-математичких наука - биолошких наука) и примењених истраживања (биотехничких наука - биотехнологије и пољопривреде). У складу са тим је и висок ниво постигнутих резултата из фитопатологије, хербологије, ентомологије, нематологије, примене средстава за заштиту биља, са проценом утицаја на животну средину. Институт се увек успешно носио са бројним тешкоћама које су се периодично јављале током његовог вишедеценијског рада, уз ентузијазам и жељу запослених да се постављени циљеви остваре и да Институт остане на путу водеће институције у области заштите биља у Републици Србији. У лабораторијама Института, поред стално запослених, велики број колега из других институција, пољопривредних станица и производних организација обавио је усавршавања из области заштите биља. Многи истакнути научни радници и професори универзитета започели су професионалну каријеру у Институту и стекли основна знања из области заштите биља.Urednici: Šikuljak Danijela i Savić Slađan

Application of Microscopy in Identification of Water Pollutants

Lake Ada Ciganlija is located on the southern bank of the Sava River, near the confluence of the Sava and Danube, 4 km from the center of Belgrade (Serbia). In summer, the lake and the surrounding peninsula become a cultural and entertainment center, attracting many visitors every day. Covered with dense deciduous forests and dotted with glades and meadows, Ada is the largest and most visited recreation and swimming spot for Belgrade residents. Another name for this popular site is „The Belgrade Sea“. A few years ago, something resembling bird feathers was spotted floating on the surface of the water. Pigeons and gulls live in the area, so the presence of feathers in the water was not surprising. Most visitors did not notice it, and some assumed it was of natural origin. As it appeared every day, the author decided to take some samples and collect them in 1.5 ml molecular tubes. The aim of the study was to examine this material under a microscope. Microphotographs were made using an Olympus BH-2 microscope with Nomarsky contrast and bright-field microscopy, a digital camera and Motic Images Plus 2.0 ML software. The results were surprising, revealing transparent, plastic-like hollow microtubes or microfibers that were clearly not of natural origin. The microfibers varied in length (100-3,000 μm) and width (5-20 μm). They were connected to each other in pairs or multiple pairs, sometimes fused together in clumps, and adhered very well to human tissue. Many questions arose: Where did it come from? What is the purpose of these microtubes? Who made it? A literature survey of microfibers in fresh waters in foreign countries showed that their microfibers originate mainly from wastewater. But it was not the case here. The Belgrade sea i.e. lake is fed directly by rivers, and the microfibers are so tiny that they float for a few seconds and sink to the bottom when the water moves. They must first fly through the air and then settle on the surface until they reach the bottom or some organism. In both cases, the microfibers are water pollutants, since they are not natural and were not present in the lake before. Government authorities should pay more attention to such pollutants, since they can cause disruptions in the aquatic ecosystem and become a serious environmental hazard. In addition, microfibers can have adverse effects on the lungs, gastrointestinal tract and many other physiological systems of organisms. In this sense, citizen science will be important in future, to alert for unusual artifacts and processes in our environment. But first, the public needs to be informed

In vitro effectiveness of commercial biofungicides in controlling Botrytis cinerea on strawberrirs and raspberries

Gray mold on strawberries and raspberries is caused by the fungus Botrytis cinerea. Infection occurs during the flowering period, and symptoms appear in the form of fruit brown rotting spots and velvety gray coating. It represents a major problem in all production regions in Montenegro, causing fruit deterioration during production and after harvest in the storage. Disease control is mainly carried out by applying synthetic fungicides, but one of the problems is the increasing occurrence of pathogen resistance. Also, great public concern has been expressed about the residual effect of synthetic fungicides, which is why more environmentally friendly solutions are necessary. One of the solutions is the potential application of biofungicides. During field activities in 2024, samples of strawberry and raspberry diseased fruits were collected from several locations in Montenegro. The pathogen Botrytis cinerea was isolated on PDA in the phytopathology laboratory of Biotechnical Faculty in Podgorica. The obtained isolates were used in directly in vitro tests to examine the efficacy of three commercial biofungicides, Vintec® (Trichoderma atroviride SC 1), Taegro® (Bacillus amyloliquefacies strain FZB24) and Serenade ASO (Bacillus subtilis strain QST 713). The tested biofungicides reduced the growth of isolate Botrytis cinerea from strawberries by 54,55% for Vintec®, 53,59% for Taegro® and 66,67% for Serenade ASO, respectively, compared to the control. Similar results of reduction were achieved by the tested biofungicides against Botrytis cinerea isolate from raspberries; Vintec® (49,69%), Taegro® (53,23%) and Serenade ASO (65,30%). Statistical analysis of the data confirmed the effectiveness of the tested biofungicides against Botrytis cinerea isolates from strawberries and raspberries in regard to untreated control, but that there was no statistically significant difference in their effectiveness between them. The results obtained indicate the potential possibility of including the biofungicides Vintec®, Taegro® and Serenade ASO in strawberry and raspberry protection programs against Botrytis cinerea

Erwinia amylovora specific bacteriophages distribution during the fire blight epidemic in central Serbia

Since first detection of Erwinia amylovora on apple and pear in Serbia, in 1989, the fire blight (FB) disease occurred periodically depending mostly on weather conditions. Rainy weather during blooming period induced severe symptom development, resulting in reduced yield, plant damage and eradication. During the spring of 2024, fruit growers reported frequent occurrence of apple, pear and quince FB in several localities of Central Serbia. We assume that unusual high average monthly temperatures for February, March and April significantly contributed to a high disease incidence. Typical symptoms included blackening along the leaf midrib and veins or full necrosis of leaves as if scorched by fire, shepherd's crook and occasional occurrence of bacterial exudate on infected tissue. The etiology of the symptoms was confirmed by isolation of bacteria from the diseased tissue. The isolates showed growth and biochemical characteristics typical for E. amylovora. Pathogenicity of the isolates was confirmed by prick-inoculation of young pear fruits. After 48 h at high humidity, the tissue necrosis developed around the inoculation site followed by oozing of bacterial exudate one day later. The identity of the isolates with E. amylovora was confirmed by real-time PCR analysis using Ams116F/Ams189R primers and Ams141T hydrolysis probe and by conventional PCR. Due to lack of efficient bactericides, control of fire blight is one of major challenges in pome fruit production. Being widespread natural bacterial enemies, simple for cultivation, host- specific, suitable for integration with other control practices, human and environment friendly, bacteriophages became a promising alternative for FB control. Samples for bacteriophage isolation were collected from different localities in Serbia, primarily from the regions where FB presence was observed, during 2024 and 2025. Isolation was attempted from leaves of host plants with or without FB symptoms (apple, pear, quince and medlar), rhizosphere soil, and from irrigation water by standard procedure for phage isolation from environmental samples. Presence of phages in samples was detected by “spot test”, pipetting 10 μl of suspensions onto NYA medium surface, previously inoculated with E. amylovora strains. Following purification, preliminary screening for phage specificity was performed to 35 E. amylovora strains originating from Serbia. Bacteriophages specific to E. amylovora were isolated from 8 out of 154 samples. The low percentage of successful phage isolation may be due to high temperature and UV index, and long dry periods during the season. Six samples originated from plant material with characteristic symptoms of FB, while two positive samples originated from rhizosphere soil. All phages were specific to all E. amylovora strains but some differences in their virulence were revealed. Our results showed that bacteriophages specific to E. amylovora were isolated more frequently from diseased plant material than from rhizosphere soil. Since they lysed E. amylovora strains originating from different hosts and localities, they represent good candidates for future use in biological control of FB