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Effective plant pathogen control presents an important challenge. Pesticide resistance and associated health and environmental problems demonstrate the need for novel, safe active ingredients for plant protection. Since both coumarins and 1,2,4-triazoles show pesticidal activity, in this study, we evaluated the antimicrobial activity of coumarin-1,2,4-triazole hybrids against plant pathogenic fungi (Fusarium oxysporum, Fusarium culmorum, Macrophomina phaseolina and Sclerotinia sclerotiorum), bacterial plant pathogens (Pseudomonas syringae and Rhodococcus fascians), and beneficial bacteria (Bacillus mycoides and Bradyrhizobium japonicum). Coumarin-1,2,4-triazoles inhibited the growth of S. sclerotiorum and F. oxysporum, while no antibacterial effect on either pathogens or soil-beneficial bacteria was observed. A quantitative structure–activity relationship models for antifungal activities on S. sclerotiorum and F. oxysporum, developed using Dragon descriptors, can explain 79% and 77% of the compounds inhibitory activity, respectively. According to molecular docking, title compounds are potential sterol 14α-demethylase inhibitors, with 7-((5-mercapto-4-(p-tolyl)-4H-1,2,4-triazol-3-yl)methoxy)-4-methyl-2H-chromen-2-one as the promising candidate for further research in plant protection.

During 2021 and 2022, a field investigation was conducted in Istria, Croatia, searching for trees exhibiting signs of Botryosphaeria dieback. Samples of symptomatic trees were collected from 26 different locations and analysed. Isolates that morphologically corresponded to species from the Botryosphaeriaceae family were selected, and detailed morphological characterisation and molecular identification of the isolates were conducted. Based on morphological characteristics and phylogenetic analysis using the internal transcribed spacer (ITS), beta-tubulin (TUB2), and translation elongation factor 1-alpha (TEF1-α) regions, six species of fungi from the Botryosphaeriaceae family were identified: Botryosphaeria dothidea (Moug. ex Fr.) Ces. & De Not.; Diplodia mutila (Fr.) Fr.; Diplodia seriata De Not.; Dothiorella iberica A.J.L. Phillips, J. Luque & A. Alves; Dothiorella sarmentorum (Fr.) A.J.L. Phillips, Alves & Luque; and Neofusicoccum parvum (Pennycook & Samuels) Crous, Slippers & A.J.L. Phillips. This is the first report of D. mutila, Do. sarmentorum, and Do. iberica causing Botryosphaeria dieback on olive trees in Croatia, and the first study investigating the resistance of Croatian olive varieties to species from the Botryosphaeriaceae family. Pathogenicity testing of selected isolates and assessment of variety resistance were conducted on four different olive varieties, namely Buža, Istarska bjelica, Leccino, and Rosinjola, using representative isolates of the mentioned species. The most aggressive species was found to be N. parvum. Olive varieties exhibited differences in susceptibility depending on the fungus they were infected with.

This paper describes the mycotoxins produced by fungi from the genera Fusarium, Penicillium, Aspergillus, Claviceps and other types of molds, their characteristics and importance. Mycotoxins are secondary fungi metabolites that serve as a defense mechanism in stressful conditions. Several hundred mycotoxins have been identified so far, and the most significant in terms of danger to human health and animals are aflatoxins, ochratoxin A, patulin, fumonisin, zearalenone and nivalenol/deoxynivalenol produced by toxigenic fungi from the genera Penicillium, Aspergillus, Claviceps, Stachybotris and Fusarium. Moisture and temperature are two factors that have a crucial influence on the development of the fungus and the synthesis of toxins. It is estimated that approximately 25% of agricultural crops are infected with fungi. Consequently, foods can also be contaminated with mycotoxins. By treating the stored grain with various chemicals, and in recent years, using environmentally friendly fungicides, the synthesis of mycotoxins is being prevented and the development of mycotoxicogenic fungi is being controlled. The mycotoxicosis can occur directly through the consumption of infected food, inhalation and skin contact, or indirectly through animals that eat contaminated feed. Mycotoxins have a pathological effect primarily on liver. Some mycotoxins also interfere with cellular protein synthesis, causing hypersensitivity and extreme immunodeficiency. Despite mycotoxins being resilient and remaining stable while undergoing chemical and thermal food processing, methods such as applying good hygiene and production practices from field to table can reduce their occurence.

A field investigation of olive trees in Istria, Croatia, revealed branch dieback and cracked bark. Samples of diseased branches were collected from eight different locations and analysed. Additionally, meteorological data from two locations were analysed to determine if there was a connection between climatic changes and the appearance of pathogens in the region. Pathogenicity tests were conducted on olive seedlings. This study provides a description of Biscogniauxia and Sordaria species’ morphology and elucidates their phylogeny based on the internal transcribed spacer (ITS), beta-tubulin (TUB2) and translation elongation factor 1-alpha (TEF1- α) regions. This research represents the first documented occurrence of Biscogniauxia mediterranea causing charcoal disease in olive trees in Croatia. Additionally, it is the first report of Biscogniauxia nummularia (Bull.) Kuntze and Sordaria fimicola causing diseases in olive trees anywhere in the world. Furthermore, this study marks one of the initial forays into molecular investigations of these species isolated from olive trees. Considering the potential threat posed by the inherent aggressiveness of Biscogniauxia species, further research is deemed necessary to curb the development of charcoal disease.

Grapevine is one of the most important culture grown in the world. It is attacked by a large number of phytopathogenic fungi, of which species of the family Botryosphaeriaceae pose one of the most significant threats. Symptoms occur in the form of necrotic lesions, rot of fruits and decline of branches. The disease is more often observed in older vineyards. The most important preventive measures include planting a healthy planting material and the choice of a suitable location for cultivation, and the most important protection measures include the removal of infected remains from the vineyards and their destruction, the application of fungicides, the coating of wounds after pruning, and the application of biological preparations for plant protection.

Vinova loza ubraja se u najvažnije kulture koje se uzgajaju u svijetu. Napada ju veliki broj fitopatogenih gljiva, među kojima se ističu i sve aktualnije postaju vrste iz porodice Botryosphaeriaceae. Simptomi se javljaju u obliku nekrotičnih lezija unutar drva, sušenja grozdova i sušenja mladica, krakova ili čitavih čokota. Bolest se češće uočava u starijim vinogradima. Najvažnije preventivne mjere podrazumijevaju sadnju zdravog sadnog materijala i izbor pogodne lokacije za uzgoj, a najvažnije izravne mjere zaštite podrazumijevanju uklanjanje zaraženih ostataka iz vinograda i njihovo uništavanje, primjenu fungicida i premazivanje rana nakon rezidbe te primjenu bioloških pripravaka za zaštitu bilja.

Dosadašnji načini kontrole uzročnika bolesti koji, među ostalim, podrazumijevaju uporabu fungicida prema novim zakonodavnim okvirima i novim strategijama u biljnom zdravstvu nailaze na sve veće restrikcije. Pronalaženje nekemijskih sredstava za zaštitu usjeva od biološki preparat, Fusarium graminearum, pšenica, zrnobolesti je od velikog interesa za poljoprivrednu proizvodnju, stoga je u našem istraživanju ispitan učinak novog biološkog preparata (BP) za kontrolu gljive Fusarium graminearum na klijancima pšenice. Biološki preparat primijenjen je na tri različita načina: inokulacijom zrna pšenice, zalijevanjem, te inokulacijom zrna i zalijevanjem. Utvrđeno je kako se intenzitet bolesti primjenom BP smanjuje, a najbolji rezultati dobiveni su inokulacijom zrna prije sjetve dok je zalijevanje površine BP polučilo slabije rezultate. Također, ukoliko je zrno već tretirano biološkim preparatom dodatna aplikacija zalijevanjem nije bila opravdana ni s ekološkog ni s ekonomskog aspekta.

Current methods of controlling pathogens, which include the use of fungicides under the new legislative framework and new strategies in plant health are facing increasing restrictions. For this reason, finding non-chemical products to protect crops from diseases is of great interest. Our study examined the effect of a new biological preparation (BP) for the control of Fusarium graminearum on wheat seedlings. The biological preparation was applied by wheat grain inoculation, watering, grain inoculation and watering. It was found that the disease intensity is reduced by the use of BP and the best results were obtained by inoculation of grains before sowing. Watering the BP on the surface achieved poorer results. Also, if the grain has already been treated with a biological preparation, additional application by watering was not justified from either an ecological or economic point of view.

Leaf spot symptoms were spotted in two olive orchards in Istria and in Kvarner Gulf, Croatia. Fungal species from three representative isolates (P13 LECIII, R18 BI, JA20 NP) have been morphologically characterized based on the colony and conidial characteristics. Several techniques were performed for inducing the sporulation of the JA20 NP isolate. Only PDA + banana medium was successful. PCR was conducted for ITS, TUB, and EF1α gene regions. Phylogenetic analyses were performed using internal transcribed spacer, beta-tubulin, and translation elongation factor 1-alpha sequence data. Three types of tests were conducted: a pathogenicity test on detached leaves, on detached and scratched leaves, and on olive seedlings. Ultimately, from the morphological characterizations, DNA sequence analysis of ITS, TUB, and EF1α gene regions, and phylogenetic analysis, these species were identified as Nigrospora gorlenkoana Novobr., Nigrospora osmanthi Mei Wang & L. Cai, and Nigrospora philosophiae-doctoris M. Raza, Qian Chen & L. Cai. This is the first report of Nigrospora species causing leaf spot on olive trees and the first report of Nigrospora philosophiae-doctoris as a plant pathogen. Fungal leaf diseases in conditions that are favorable for infection and disease development can lead to a decrease in the yield and olive oil quality. Therefore, it is necessary to conduct further research and the monitoring of fungal leaf diseases.

Olive (Olea europaea L.) is a very important crop grown in the Mediterranean part of Croatia. Olive branch and fruit dieback symptoms were observed in two olive orchards in Istria, Croatia. The samples from symptomatic trees were collected and brought to the laboratory for analysis. Based on their morphological characterization, isolated fungi were identified as Cytospora sp. Two representative isolates (one per orchard) were taken for molecular analysis, and based on DNA sequence data of the ITS and TUB gene regions, and phylogenetic analysis of the sequences, the isolates were identified as Cytospora pruinosa Défago. To determine pathogenicity, pathogenicity tests were conducted on detached olive branches and two-year-old olive trees in the greenhouse. This is the first report of C. pruinosa causing olive branch and fruit dieback in Croatia.

Coumarins are secondary plant metabolites widely distributed in higher plants, bacteria, fungi, and sponges. This great structural diversity of these natural compounds and their synthesized derivatives enables their wide range of pharmacological activities, such as antioxidant; antibacterial; antifungal; anti-human immunodeficiency infection; anti-tubercular; and anti-cancer activities, which were very well reviewed previously. There are also many reports about their effectiveness against plant pathogenic pests (phytopathogenic fungi, bacteria, nematodes, and insects). These secondary metabolites protect environmental enemies and competing plants. However, there is still limited literature on coumarins’ practical applications in agriculture, as well as their effects on beneficial populations of soil organisms. This review summarizes recent knowledge about the effects of natural and synthesized coumarins on phytopathogens and beneficial populations of soil organisms. A separate section is dedicated to poorly reviewed computer-aided molecular design (CAMD) methods of agrochemicals. It also reviewed CAMD techniques to develop low-toxicity and environmentally safe pesticides. Despite the many positive effects of coumarins related to plant protection, they do possess properties harmful to the environment and health. These properties are described in the last section. Prediction of coumarin hazardous properties using a quantitative structure-activity relationship (QSAR) approach was also reviewed.

Fluoro-substituted pyrazoles have a wide range of biological activities, such as antibacterial, antiviral, and antifungal activities. The aim of this study was to evaluate the antifungal activities of fluorinated 4,5-dihydro-1H-pyrazole derivatives on four phytopathogenic fungi: Sclerotinia sclerotiorum, Macrophomina phaseolina, Fusarium oxysporum f. sp. lycopersici, and F. culmorum. Moreover, they were tested on two soil beneficial bacteria—Bacillus mycoides and Bradyrhizobium japonicum—as well as two entomopathogenic nematodes (EPNs)—Heterorhabditis bacteriophora and Steinernema feltiae. The molecular docking was performed on the three enzymes responsible for fungal growth, the three plant cell wall-degrading enzymes, and acetylcholinesterase (AChE). The most active compounds against fungi S. sclerotiorum were 2-chlorophenyl derivative (H9) (43.07% of inhibition) and 2,5-dimethoxyphenyl derivative (H7) (42.23% of inhibition), as well as H9 against F. culmorum (46.75% of inhibition). Compounds were shown to be safe for beneficial soil bacteria and nematodes, except for compound H9 on EPN H. bacteriophora (18.75% mortality), which also showed the strongest inhibition against AChE (79.50% of inhibition). The molecular docking study revealed that antifungal activity is possible through the inhibition of proteinase K, and nematicidal activity is possible through the inhibition of AChE. The fluorinated pyrazole aldehydes are promising components of future plant protection products that could be environmentally and toxicologically acceptable.

Sclerotinia sclerotiorum (Lib.) De Bary is the most significant pathogen which cause significant damage to sunflower, soybean and rapeseed worldwide. Great economics losses can occur in areas with humid and colder climate. Characteristic symptoms are white coatings of mycelium on infected parts of plants and the appearance of sclerotia. Since there are no effective protection measures, it is necessary to implement agro-technical control measures.

Vrsta Sclerotinia sclerotiorum (Lib.) De Bary najznačajniji je patogen koji uzrokuje značajne štete na uljaricama (suncokret, soja i uljana repica) širom svijeta. U područjima s vlažnijom i hladnijom klimom može doći do velikih ekonomskih gubitaka. Karakteristični simptomi su bijele prevlake micelija na zaraženim dijelovima biljaka i pojava sklerocija. Budući da efikasnih mjera zaštite nema, nužno je provoditi agrotehničke mjere suzbijanja.

During cultivation, wheat is exposed to several abiotic and/or biotic stress conditions that may adversely impact the wheat yield and quality. The impact of abiotic stress caused by nitrogen deficiency and biotic stress caused by phytopathogenic fungus Fusarium culmorum on biomarkers of oxidative stress in the flag leaf of nine winter wheat varieties (Ficko, U-1, Galloper, BC Mandica, BC Opsesija, Ingenio, Isengrain, Felix, and Bezostaya-1) was analyzed in this study. Hydrogen peroxide concentration and lipid peroxidation level were measured as indicators of oxidative stress, while the antioxidant response was determined by measuring the concentration of phenolic compounds and activities of antioxidant enzymes. Wheat variety and nitrogen treatment had a significant effect on all examined biomarkers of oxidative stress in the flag leaf, while the impact of Fusarium treatment was less pronounced. The most significant impact on the measured stress biomarkers had a low nitrogen level, which mainly increased hydrogen peroxide concentration and lipid peroxidation level and decreased activities of antioxidant enzymes in most varieties. The obtained results were discussed and compared with the previous study in which biochemical analyzes were performed on the wheat spike. There was no significant strong correlation between flag leaf and spike response in the measured parameters, which, in addition to the variety-specific response, also indicates a tissue-specific antioxidant response.

Fuzarijska palež klasova (FHB) pšenice ekonomski je značajna bolest koja može imati nepovoljne posljedice na prinos i kvalitetu pšenice. Do danas ne postoji mjera zaštite koja osigurava potpunu zaštitu usjeva pšenice, ali kombinacija više mjera može dati dobre rezultate. Učinkovitost svake pojedine mjere pod utjecajem je okolišnih uvjeta koji prevladavaju tijekom vegetacijske sezone, a učinkovitost će ovisiti i o osjetljivosti pojedine sorte na FHB. Sve veći broj istraživanja potvrđuje kako je integrirana zaštita bilja (IZB) ključ za što uspješniju borbu protiv FHB-a. Kako bi primijenjene mjere zaštite bile što uspješnije, od iznimne je važnosti dobro poznavati uzročnika bolesti, njegov životni ciklus i ekologiju. Budući da je F. graminearum dominantni uzročnik FHB-a na području Republike Hrvatske, osobiti naglasak je stavljen na sve dostupne mjere zaštite u njegovu suzbijanju.

U istraživanju je ispitivano djelovanje eteričnih ulja domicilnoga bilja Hrvatske na supresiju rasta micelija Botrytis cinerea izoliranoga s vinove loze. U pokusu su korištena ulja sljedećih biljaka: metvice paprene (Mentha x piperita), kadulje (Salvia officinalis), ružmarina (Rosmarinus officinalis), lavandina (Lavandula hybrida), mravinaca (Origanum compactum), timijana (Thymus vulgaris), gospine trave (Hiperici oleum), stolisnika (Achillea millefolium) i smilja (Helichrysum italicum). Rezultati in vitro volatilnogq testa i makrodilucijskoga testa pokazali su da ulja timijana i paprene metvice mogu imati pozitivan utjecaj na inhibiciju rasta gljive Botrytis cinerea pri svim ispitivanim koncentracijama te da fungistatski učinak traje i preko 96 h. Primjena ulja stolisnika i gospine trave u makrodilucijskoj metodi dala je bolje rezultate u odnosu na volatilnu metodu. Eterično ulje smilja stimuliralo je porast micelija B. cinerea u obje metode. S obzirom na smanjenje porasta micelija B. cinerea u uvjetima in vitro, eterična ulja, kao biofungicidi, predstavljaju moguću zamjenu za sintetske pripravke u suzbijanju razvoja sive plijesni na vinovoj lozi, no potrebna su dodatna istraživanja kako u uvjetima in vitro, tako i u uvjetima in vivo.

In an olive orchard on the western part of Istria, Croatia, twig and branch dieback was observed on several olive trees. In total, seven samples from symptomatic trees were collected. Samples were analyzed, and four fungal isolates showed morphological similarities to the species Phaeoacremonium. One isolate, chosen as a representative, was taken for molecular identification and pathogenicity tests. Based on the DNA sequence data of the ITS, TUB, and EF1α gene regions, the isolate was identified as P. iranianum. Pathogenicity tests were conducted on detached olive branches and olive trees in the greenhouse. To the best of our knowledge, this is the first report of twig and branch dieback on olive caused by Phaeoacremonium iranianum.

The effect of different conservation soil tillage (CST) treatments and different level of nitrogen fertilization on Fusarium wheat grain infection on two locations are presented in this paper. The research was conducted on winter wheat with different soil tillage treatments: conventional tillage to 30 cm depth, subsoiling to 35-40 cm depth, chiselling up to 25 cm, disk-harrowing to 10-15 cm and no-tillage, without any tillage treatments and three different nitrogen rates (N1 - amount reduced by 30% of recommended; N2 - according to standard recommendation and N3 - amount increased by 30% of recommended). The occurrence of Fusarium species was determined in all the tested variants of tillage and fertilization treatments. The effect of reduced fertilization (N1) on Fusarium sp. grain infection in all tillage treatments was statistically lower in comparison with other (N2 and N3) nitrogen treatments. The lowest percentage of wheat grains infected with Fusarium sp. was recorded in conventional tillage on location Magadenovac and no tillage treatments on location Cacinci.

Fusarium head blight and inadequate nitrogen fertilization can cause numerous biochemical changes in wheat. The main aim of this study was to determine the effects of Fusarium inoculation and a broader range of different nitrogen fertilization on the defense response in the spikes of four wheat varieties, varying in FHB susceptibility. Total soluble phenolics content, activities of enzymes involved in phenol metabolism (PAL and PPO), and total antioxidant capacity were determined as indicators of defense response. In both growing seasons, Fusarium inoculation altered PHE content in wheat spikes, indicating involvement of PHE in the defense response to Fusarium attack. Increased PHE content in the partially resistant varieties (Apache and Graindor) indicates involvement of PHE in the defense response and better disease tolerance in the more resistant varieties. Breeding wheat varieties with enhanced PHE synthesis could be a promising strategy to control FHB. To the best of our knowledge, this is the first study that emphasizes the effects of Fusarium infection and a broader range of different nitrogen fertilization on PHE and enzymes involved in PHE metabolism. In addition, this is the first study using the FRAP method to determine the antioxidant capacity of wheat tissues under the influence of Fusarium infection and different nitrogen fertilization.

Coumarin derivatives have been reported as strong antifungal agents against various phytopathogenic fungi. In this study, inhibitory effects of nine coumarinyl Schiff bases were evaluated against the plant pathogenic fungi (Fusarium oxysporum f. sp. lycopersici, Fusarium culmorum, Macrophomina phaseolina and Sclerotinia sclerotiourum). The compounds were demonstrated to be efficient antifungal agents against Macrophomina phaseolina. The results of molecular docking on the six enzymes related to the antifungal activity suggested that the tested compounds act against plant pathogenic fungi, inhibiting plant cell-wall-degrading enzymes such as endoglucanase I and pectinase. Neither compound exhibited inhibitory effects against two beneficial bacteria (Bacillus mycoides and Bradyrhizobium japonicum) and two entomopathogenic nematodes. However, compound 9 was lethal (46.25%) for nematode Heterorhabditis bacteriophora and showed an inhibitory effect against acetylcholinesterase (AChE) (31.45%), confirming the relationship between these two activities. Calculated toxicity and the pesticide-likeness study showed that compound 9 was the least lipophilic compound with the highest aquatic toxicity. A molecular docking study showed that compounds 9 and 8 bind directly to the active site of AChE. Coumarinyl Schiff bases are promising active components of plant protection products, safe for the environment, human health, and nontarget organisms.

The effect of five essential oils (anise, peppermint, basil, rosemary and true lavander) and their two most common components on the mycelial growth in Colletotrichum coccodes, economically important phytopathogic fungi and compared with fungicides have been investigated in the study. Tests were conducted in vitro conditions in eight volumes on a PDA substrate in four replicates. The increase in mycelium was measured on the eighth and fifteenth day after the mycelium inoculation. It was found out that some essential oils and components applied at a given volume have a significant antifungal effect, in some examples comparable to fungicides. Anise and peppermint essential oils as well as the anethole component had the best activity and the lowest IC50. Fifteen days after the inoculation of the mycelium, the essential oils had a significantly better antifungal activity compared to their second most represented component.

The functioning of soil ecosystems greatly depends on the interactions occurring between soil biota communities. It is well known that earthworms are an important soil component that substantially affects its function, including their meaningful impact on the development of different phytopathogenic soil fungi. Phytopathogenic fungi are responsible for crop disease and cause great economic damage. It has previously been established that earthworms’ coelomic fluid can suppress the growth of phytopathogenic fungi, but the exact molecular mechanism is unknown. The present study aimed at broadening the proof of this observed phenomenon by investigating the effects of the coelomic fluid extract of three different earthworm species (Eisenia andrei, Dendrobaena veneta and Allolobophora chlorotica) on the growth of six different phytopathogenic fungi species (Berkeleyomyces basicola, Fusarium culmorum, Globisporangium irregulare, Rhizoctonia solani, Macrophomina phaseolina, and Sclerotinia sclerotiorum). Coelomic fluid extract was obtained by electrostimulation or usage of extraction buffer (only in case of A. chlorotica) and prepared in three different concentrations by diluting the obtained coelomic fluid with physiological saline. The coelomic fluid extract of the three investigated earthworm species had an inhibitory effect on the growth of all six phytopathogenic fungi species. The greatest inhibitory effect was achieved with the E. andrei coelomic fluid extract reducing the growth of R. solani fungi. The findings of this research confirm the antifungal activity of coelomic fluid obtained from earthworm species belonging to different ecological categories and may be of potential use in crop protection against phytopathogenic fungi.

The aim was to study the inhibitory effects of coumarin derivatives on the plant pathogenic fungi, as well as beneficial bacteria and nematodes. The antifungal assay was performed on four cultures of phytopathogenic fungi by measuring the radial growth of the fungal colonies. Antibacterial activity was determined by the broth microdilution method performed on two beneficial soil organisms. Nematicidal activity was tested on two entomopathogenic nematodes. The quantitative structure-activity relationship (QSAR) model was generated by genetic algorithm, and toxicity was estimated by T.E.S.T. software. The mode of inhibition of enzymes related to the antifungal activity is elucidated by molecular docking. Coumarin derivatives were most effective against Macrophomina phaseolina and Sclerotinia sclerotiorum, but were not harmful against beneficial nematodes and bacteria. A predictive QSAR model was obtained for the activity against M. phaseolina (R2tr = 0.78; R2ext = 0.67; Q2loo = 0.67). A QSAR study showed that multiple electron-withdrawal groups, especially at position C-3, enhanced activities against M. phaseolina, while the hydrophobic benzoyl group at the pyrone ring, and –Br, –OH, –OCH3, at the benzene ring, may increase inhibition of S. sclerotiourum. Tested compounds possibly act inhibitory against plant wall-degrading enzymes, proteinase K. Coumarin derivatives are the potentially active ingredient of environmentally friendly plant-protection products.

Zbog brojnih nutritivnih vrijednosti i potencijalnoga pozitivnoga učinka na ljudsko zdravlje, šparoga se pokazala kao interesantna kultura za konzumaciju te predmet brojnih znanstvenih ispitivanja. Samonikla šparoga (Asparagus acutifolius L.) raste duž Jadranske obale i otoka, dok se kultivirana šparoga (Asparagus officinalis L.) uglavnom uzgaja u istočnome dijelu Hrvatske. Ovim istraživanjem određivan je ukupni sadržaj polifenola (spektofotometrijski) i antioksidativna aktivnost (DPPH) kod samoniklih i kultiviranih šparoga. Rezultati su pokazali da nema statistički značajne razlike u sadržaju ukupnih polifenola između samoniklih i kultiviranih šparoga. Najviša vrijednost ukupnih polifenola samoniklih šparoga iznosila je 6,20 [mg GAE/g svježe tvari], odnosno 49,60 [mg GAE/g suhe tvari], dok je kod kultiviranih šparoga bila 4,52 [mg GAE/g svježe tvari], odnosno 50,93 [mg GAE/g suhe tvari]. Antioksidativna aktivnost mjerena DPPH (2,2-difenil-1-pikrilhidrazil) metodom, pokazala je statistički značajne razlike. Veća je antioksidativna aktivnost izmjerena kod kultiviranih, nego kod samoniklih šparoga. Također je utvrđeno da porastom polifenola raste antioksidativna aktivnost, ali više kod kultiviranih šparoga. Temeljem rezultata, zaključujemo da je kultivirana šparoga odlična alternativa samonikloj, koja bi trebala biti zaštićena od pretjeranog ubiranja, kako bi se zaštitila prirodna raznolikost obale i otoka.

Asparagus is an interesting vegetable as a part of a healthy diet and has been widely studied due to the high nutritional value and potential positive effects on human health. Wild asparagus (Asparagus acutifolius L.) grows along the Adriatic coast and islands, while the cultivated asparagus (A. officinalis L.) is mostly grown in the eastern region of Croatia. The aim of this study was to determine total polyphenol content (measured spectrophotometrically) and antioxidant activity (DPPH) in wild and cultivated asparagus. No significant differences in total polyphenol content were found between wild and cultivated asparagus. The highest content of polyphenols in wild asparagus was 6.20 [mg GAE/g fresh weight] and 49.60 [mg GAE/g dry weight], and in cultivated asparagus it was 4.52 [mg GAE/g fresh weight] and 50.93 [mg GAE/g dry weight]. Antioxidant activity measured using DPPH (2,2-diphenyl-1-picrylhydrazyl) method showed statistically significant difference. Higher antioxidant activity was measured in cultivated than in wild asparagus. The results also indicate that increased polyphenol content leads to higher antioxidant activity, but more so in the cultivated asparagus. Based on the results, the cultivated asparagus is an excellent alternative to wild asparagus that should be protected from over-harvesting, thus preventing devastation of plant diversity of islands and coast.

Abiotic and biotic stresses, such as mineral nutrition deficiency (especially nitrogen) and Fusarium attack, pose a global threat with devastating impact on wheat yield and quality losses worldwide. This preliminary study aimed to determine the effect of Fusarium inoculation and two different nitrogen levels on oxidative status and antioxidative response in nine wheat varieties. Level of lipid peroxidation, activities of antioxidant enzymes (catalase, ascorbate peroxidase, glutathione reductase), phenolics, and chloroplast pigments content were measured. In general, wheat variety, nitrogen, and Fusarium treatment had an impact on all tested parameters. The most significant effect had a low nitrogen level itself, which mostly decreased activities of all antioxidant enzymes and reduced the chloroplast pigment content. At low nitrogen level, Fusarium treatment increased activities of some antioxidative enzymes, while in a condition of high nitrogen levels, antioxidative enzyme activities were mostly decreased due to Fusarium treatment. The obtained results provided a better understanding on wheat defense mechanisms against F. culmorum, under different nitrogen treatments and can serve as an additional tool in assessing wheat tolerance to various environmental stress conditions.

The aim of this study was to investigate the influence of nutrition media (PDA, carrot agar) and temperature (15, 22 and 30ºC) on growth and mycelium development As well as sclerotium formation of Sclerotinia sclerotiorum. The experiment was set up under laboratory conditions in year 2013. Statistical analysis showed that nutrition media and temperature had significant influence on mycelim development and formation on sclerotia. Mycelial growth was significantly higher on PDA and at 15 and 20ºC. Sclerotia are formed only on PDA and the two lower temperatures. Sclerotia formed at 15ºC were bigger than sclerotia formed at 20ºC.

survey was conducted in 2012 and 2013 to detect the presence and distribution of Alfalfa mosaic virus (AMV) in lavandin crops growing in continental parts of Croatia. A total of 73 lavandin samples from six crops in different localities were collected and analyzed for the presence of AMV and Cucumber mosaic virus (CMV) using commercial double-antibody sandwich (DAS)-ELISA kits. AMV was detected serologically in 62 samples collected at three different localities, and none of the samples tested positive for CMV. For further analyses, six selected samples of naturally infected lavandin plants originating from different localities were mechanically transmitted to test plants: Chenopodium quinoa, C. amaranticolor, Nicotiana benthamiana and Ocimum basilicum, confirming the infectious nature of the disease. Molecular detection was performed by amplification of a 751 bp fragment in all tested samples, using the specific primers CP AMV1/CP AMV2 that amplify the part of the coat protein (CP) gene and 3’-UTR. The RT-PCR products derived from the isolates 371-13 and 373-13 were sequenced (KJ504107 and KJ504108, respectively) and compared with the AMV sequences available in GenBank. CP sequence analysis, conducted using the MEGA5 software, revealed that the isolate 371-13 had the highest nucleotide identity of 99.5% (100% amino acid identity) with an isolate from Argentina originating from Medicago sativa (KC881010), while the sequence of isolate 373-13 had the highest identity with an Italian AMV isolate from Lavandula stoechas (FN667967) of 98.6% (99% amino acid identity). Phylogenetic analysis revealed the clustering of selected isolates into four molecular groups and the lavandin AMV isolates from Croatia grouped into two distinct groups, implying a significant variability within the AMV lavandin population.

The quaternization reactions of nicotinamide, with different electrophiles: methyl iodide and substituted 2-bromoacetophenones (4-Cl, 4-Br, 4-H, 4-CH3, 4-F, 4-OCH3, 4-Ph, 2-OCH3, 4-NO2) are reported. The preparations were carried out by conventional synthesis and under microwave irradiation in absolute ethanol and acetone. The synthesis performed by microwave dielectric heating significantly improved yield, up to 8 times, and shortened down the reaction time from ca. one day in conventional, to 10–20 min. The structures of the synthesized compounds were confirmed by IR, 1H- and 13C-NMR spectroscopy, mass spectrometry and elemental analysis. The compounds have been screened for antifungal activities against Fusarium oxysporum, Fusarium culmorum, Macrophomina phaseolina and Sclerotinia sclerotiorum at concentrations of 10 µg/mL and 100 µg/mL. Six compounds showed the strong inhibition of mycelium growth at a concentration of 10 µg/mL. All tested compounds revealed the great inhibitory activities against S. sclerotiorum at a concentration of 100 µg/mL.

Different formae speciales of Fusarium oxysporum are the main causal agent of Fusarium wilts. In 2008 and 2009 we collected F. oxysporum samples from symptomless Abutilon theophrasti, Xanthium strumarium, Chenopodium album, Matricaria perforata, Ambrosia artemisiifolia, Polygonum lapathifolium, Sonchus arvensis, Amaranthus blitoides, Amaranthus retroflexus, Datura stramonium, Sorghum halepense and Hibiscus trionum. Only 16 out of 41 isolates of F. oxysporum yielded nit mutants. The frequency of nit3 mutants was higher (43%) than the frequency of nit1 (35%) and NitM (22%) mutants. Two vegetative compatibility groups (VCGs) of F. oxysporum were determined in the complementation tests. These results stress out the problem with isolation of nit mutants and show a high genetic diversity of F. oxysporum isolated from weeds.

A research was conducted during 2011 and 2012 on five locations of Eastern Croatia in order to investigate the occurrence of Septoria leaf bloch of wheat caused by Mycosphaerella graminicola. The first symptoms of the infection during 2011 were determined at the beginning of April, and during 2012 in early March. After statistical analysis has been conducted results showed that there were statistically significant differences in the number of spots on the leaf and number of pycnidia on the spots between locations in both years of the investigation.

Pathogenicity of thirty isolates representing 14 Fusarium species isolated from weeds and plant debris in eastern Croatia was investigated in the laboratory. Pathogenicity tests were performed on wheat and maize seedlings. The most pathogenic Fusarium spp. was F. graminearum isolated from Amaranthus retroflexus, Abutilon theophrasti and Chenopodium album. There was a noticeable inter- and intraspecies variability in pathogenicity towards wheat and maize. Isolates of F. solani from Sonchus arvensis and F. verticillioides from C. album were highly pathogenic to wheat seedlings and apathogenic to maize seedlings. Isolates of F. venenatum were very pathogenic to wheat and maize being the first report about pathogenicity of this species. This experiment proves that weeds and plant debris can serve as alternate hosts and source of inoculum of plant pathogens.

There has not been a systematic research of medicinal plants mycoflora in Croatia. This paper aims to present the results of preliminary research of mycopopulation of 14 species of medicinal plants. Total of 393 plant parts has been examined and 10 genera of fungi were isolated: Penicillium, Aspergillus, Sordaria, Phoma, Cladosporium, Rhizopus, Stemphillium, Fusarium, Phomopsis and one unidentified genus. Penicillium sp. (from 11 of 14 plant species) was isolated from the majority of samples. The plants fungi were isolated from did not show any macroscopically visible symptoms of infection, except plant parts of Lavandula x intermedia and Foeniculum vulgare, from which Phomopsis sp. and Fusarium sp. were isolated

The obligate fungi inside the family Albuginaceae are widespread world wide and cause white rust or white blister disease. Mycopopulation of weeds has been researched within the project „The role of weeds in epidemiology of row-crop diseases“. The aim of this research was to identify white blister species occurring on weeds in Eastern Croatia. Weed plants with disease symptoms characteristic for white blister species have been collected since 2001 on location Slavonia and Baranja country. Determination of white blister species was based on morphological characters of pathogen and the host. Wilsoniana bliti was determined on Amaranthus retroflexus and Amaranthus hybridus leaves. Capsella bursa pastoris is a host for Albugo candida. Ambrosia artemisiifolia is a host for Pustula sp. and Cirsium arvense was found to be host for Pustula spinulosa. Wilsoniana portulaceae was determined on Portulaca oleracea.

Growth and sporulation of selected isolates of F. graminearum, F. verticillioides and F. subglutinans were examined over 15 days on seven different media of diverse pH levels and under two light conditions. All isolates formed the most densd mycelia and had the fastest growth on PDA media and under dark conditions. F. graminearum had the weakest sporulation on all media. F. verticillioides and F. subglutinans sporulated well on the all investigated media. Statisticaly significant slower mycelial growth was determined on pH 4.5 for all species.

Eleven essential oils (clove, rosemary, cinnamon leaf, sage, scots pine, neroli, peppermint, aniseed, caraway, lavander, common thyme) were tested for in vitro antifungal activity on twelve plant pathogenic fungi (Fusarium graminearum, F. verticillioides, F. subglutinans, F. oxysporum, F. avenaceum, Diaporthe helianthi, Diaporthe phaseolorum var. caulivora, Phomopsis longicolla, P. viticola, Helminthosporium sativum, Colletotrichum coccodes, Thanatephorus cucumeris). The results indicated that all oils except scots pine and neroli had antifungal activity against some or all tested fungi. The best antifungal activity had common thyme, cinnamon leaf, clove and aniseed oils. When compared to control, scots pine, neroli and sage oils stimulated mycelium growth of some investigated fungi.

During a three year period (2004 to 2006) on locations of Eastern Croatia (Kneževi Vinogradi, Osijek, Brođanci and Krndija) we collected plants of Achillea millefolium L. from which we isolated Phomopsis sp. Many of the collected plants exhibited no visible symptoms of infection but in spring pycnidia were noticed on overwintered plants. Isolation was done on PDA from naturally infected plants and morphological and molecular characteristics were studied. Biometrical values of reproductive structures developed on naturally infected plants and grown on PDA were measured. Internal transcribed spacers ITS rDNA of the examined isolates were sequenced. Although molecular analysis showed that isolates were grouped with Diaporthe arctii, we decided to classify isolates from A. millefolium as Phomopsis sp., the details of which were discussed in the paper.

Diaporthe helianthi isolates were grown on various media (water agar with various plant tissues, potatoes dextrose agar, malt agar and V-8 juice agar) with the aim to examine media influence on the development of reproductive structures. Isolates were distinguished by whether they form teleomorphic stage or not, and by the number of formed pycnidia and perithecia. Water agar with various plant parts was proved as a suitable media for perithecia production.

Disease appearance on soybean can influence quality and quantity of yield. Different spieces of saprophyte and parasite fungi can be isolated from stems, pods and grain of soybean. The aim of the research was to evaluate the incidence of important disease on natural soybean grain over the period of 4 years (2004-2007) of experiment held on the location Sopot-Vinkovci and included 9 cultivars of soybean. The following plant pathogenic fungi were identified: Peronospora, Sclerotinia, Cercospora, Fusarium and Diaporthe/Phomopsis. The most frequent fungi on soybean grains were: Cladosporium, Alternaria, Penicillium, Aspergillus and Epicoccum. The health condition of the natural soybean grains over the four years period on all cultivars was good.

Diaporthe/Phomopsis sp. was isolated from Xanthium italicum (Italian cockleburr) for the first time in Eastern Croatia during 2004 year. As Diaporthe/Phomopsis species are known as pathogens of soybean, sunflower and other arable crops, it is important to study alternative hosts as a possible source of inoculum from an epidemiological point of view. This paper describes symptoms of infection on X. italicum, pathogen morphological and cultural characters on potato-dextrose agar (PDA), biometrical values of reproductive structures (pycnidia, beta conidia, perithecia, asci and ascospores) of naturally infected plants and of cultures grown on PDA. Results obtained from undertaken studies indicate that our Phomopsis sp. isolates from X. italicum belong to the group of P. helianthi.

Botrytis gray mold caused by Botrytis cinerea is an important pathogen of grapevines all over the world. In order to determine the intensity of B. cinerea attack on mature grapes and efficiency of botryticides (cv. Traminac - iprodion, pyrimethanil, fludioxinil + cyprodinil; cv. Graševina - fenhexamid, fludioxinil+cyprodinil, tebuconazol+diclofluanid) a two-year experiment (2001-2002) was set up on locations Mladice (Traminac) and Škomić (Graševina). By evaluation of the intensity of B. cinerea infection on the Graševina and Traminac grapes the strongest infection determined on the control, and the weakest on the variant with botryticides Switch. Comparison of infection intensity on the Traminac grapes with the Graševina grapes led to no differences concerning the sensitivity of varieties. The disease intensity primarily depended on climate conditions. Both experimental years were charactized by the quantity of precipitation in the vine-yards which were in favour of the high humidity allowing high disease intensity.

During a three–year period (2001-2003), occurrence of diseases on sunflower stems was monitored in fields of PIK Vinkovci (on the Sopot location). The experiment involved 24 genotypes planted in 2001, 30 genotypes in 2002, and 20 genotypes in 2003, each of them repeated in two trials: treated and non-treated. The Konker fungicide preparation, dosed 1.5 l/ha, was used for sunflower protection. The treatment was applied at the R1-R2 development stage of sunflower (the size of immature bud was up to 2 cm). Based on the conducted survey, it was found out that Sclerotinia sclerotiorum was the most dominant disease agent to sunflower stems in 2001. Percentage of infection for the non-treated and treated trials was between 0 and 32.5%, and 0 and 17.5%, respectively. In 2002 and 2003, Phomopsis helianthi was the most dominant disease agent. Average value rate in 2002 fluctuated between 0.65 and 4.00 in non-treated trial and between 0.05 and 2.60 in treated trial. In 2003, occurrence of P. helianthi was of weak intensity, mostly because of unfavorable weather conditions for parasite development. Average ratings were 0.35 to 3.30 in the non-treated trial, and 0.20 to 1.85 in the treated trial. Application of fungicide during the whole experiment period had positive effects on the decrease of infection intensity and the increase of grain and oil yields.

Fusarium species from section Discolor are widespread and well-known and play an important role in disease etiology of wheat, barley and maize. F. graminearum and F. culmorum were isolated during a four-year period at several locations in Eastern Croatia and from different hosts. The mycelium development of 236isolates of F. graminearum and 2 isolates of F. culmorum was cultered during an eight day period on water agar, PDA, Bilai, Czapek's and CLA agar at temperatures 5°, 15°, 20°, 25° and 30°C and a 12 hour dark/light regime. The results show that agar medium does not influence colony diameter significantly. The agar medium influences the richness and density of the aerial mycelium significantly, although the shape and compactness of the mycelium is not only the result of the medium on which the fungus is developed, but also of the characteristics of the species itself. The sporulation of F. culmorum was abundant on all investigated medium, whereas the sporulation of F. graminearum was very weak on PDA and Bilai agar and it was medium on CLA.

A large number of weeds are alternative hosts to numerous pathogenic agents of fungus diseases to arable crops and they represent inoculum source to cultivated plants. The aim of our investigation was to determine weed mycopopulation, to establish pathogenicity of some fungi to cultivated plants as well as to choose potential parasites for biological control of weeds. During a two year investigation of weed mycopopulation obtained from root crops at five localities in East Slavonia and Baranya 32 fungus species were established at 25 weeds that were characterized by disease symptoms. Seven fungi species were determined on roots of 18 weeds, although there were no obvious disease symptoms. Obligated parasites along with 21 determined fungi are of Oomycetes, Plectomycetes and Hemibasidiomycetes genus. Facultative parasites from 18 determined fungus species are of Discomycetes, Pyrenomycetes, Coelomycetes and Hyphomycetes genus. Isolates of Sclerotinia sclerotiorum were tested for their pathogenicity to soybean. The results showed that there were no significant differences in pathogenicity of isolates in artificial conditions in laboratory. In natural conditions isolates from soybean were more pathogenic to soybean than the isolates from weeds. Experiments done with sunflower showed that the isolates from weeds were more pathogenic than isolates from sunflower. The isolates of Phomopsis/Diaporthe complex affected the length of germ, the length of necrosis and seed disease differently. Results showed that the isolates from weeds of Phomopsis species are pathogenic to soybean representing an important source of inoculum to soybean. Isolates of Fusarium species isolated from weeds were pathogenic for popcorn seedlings. Artificial infection of Abutilon theophrasti by Colletotrichum coccodes showed that foliar mass wilted earlier and whole plants died. For the first time in Croatia the presence of 14 fungus species was determined on 27 new hosts.

Fusarium species from section Liseola play an important role in disease etiology of wheat, barley and maize. F. moniliforme, F. subglutinans and F. proliferatum were isolated during seven year period at several locations in Eastern Croatia and from different hosts. The mycelium development of 6 isolates of F. moniliforme, 3 isolates of F. subglutinans and 3 isolates of F. proliferatum was cultered during an eight day period on water agar, PDA, Bilai, Czapek's and CLA agar at temperatures 5°, 15°, 20°, 25° and 30°C and a 12 hour dark/light regime. The results show that agar medium only insignificantly affected diameter of colonies, and that all isolates developed well on all media. After an 8-day period, at the temperature of 25ºC, all examined isolates filled out the Petri dish. The most abundant mycelium was developed on PDA, a weaker one on Bilai medium, and the weakest one on CLA medium and water agar. Sporulation of examined species was abundant on all media.

The academic textbook Diseases of Flowers and Decorative Plants introduces the most important casual agents of flower and decorative plant diseases to students at all study levels, as well as to floriculturists. Floriculture as a branch of plant production is a very old discipline that was developed along with human mankind. Although it may seem otherwise, diseases are extremely important in plant production cycle. Diseases elaborated in the present handbook are classified according to their casual agents (fungi, bacteria and viruses). Parasites are organized according to their systematic grouping within separate classes, thus avoiding repetitions, as one parasite can attack different plant sorts. Disease symptoms appearing on attacked plants, development stages and protection measures are presented for each casual agent. The textbook is also well illustrated with photographs. The list of professional terminology and the list of scientific and common names for flowers and decorative plants are provided at the end of the handbook. Besides results of the authors’ own researches, this academic textbook comprises up-to-date findings of well known scientists and professionals.

Wilted hemp (Cannabis sativa L.) plants were observed in August 2019 in commercial fields around Osijek, Croatia. Plants and roots with disease symptoms were collected. The single-spored isolates produced septate cottony white to light pink aerial mycelium and purple undersurface on potato dextrose agar (PDA). Smooth and hyaline hyphae were branched and septate. Macroconidia were fusiform to sickle-shaped with foot-shaped basal cells, elongated apical cells and three to five septa. Sequencing of the internal transcribed spacer and the partial elongation factor 1-α gene identified the species as Fusarium oxysporum. Artificial infection fulfills Koch’s postulates, producing plants which show stunted growth and wilt symptoms similar to those observed in the commercial fields. Control seedlings remained symptomless and healthy. To the best of our knowledge, this is the first report of hemp Fusarium wilt causing F. oxysporum in Croatia. Considering that F. oxysporum has been reported in main field crops in Croatia, the presence of this pathogen could cause economically significant hemp production decreases, especially in humid and cold springs and susceptible varieties.