The Influence of Pesticides and Biopesticides on Morphological Changes in Raspberry (Rubus ideaus L. „POLKA“) Infected with the Bacterium Agrobaterium tumefaciens

Proizvodnja maline (Rubus idaeus L.), kao jedna od najperspektivnijih grana voćarstva u Bosni i Hercegovini, mogla bi biti trajno ugrožena zbog sve češćega pojavljivanja bolesti bakterioznoga tumora korijena, čiji je uzročnik Agrobacterium tumefaciens. Cilj ovoga rada bio je ispitati morfološke promjene na sadnicama mali ne izazvane bakterijom A. tumefaciens, kao i veličinu i brojnost tumora u zavisnosti od primijenjenoga biološkog ili kemijskog tretmana. U radu je korišten referentni soj Agrobacterium tumefaciens MTCC 431 (ATCC 15955). Morfometrijski parametri na sadnicama maline izmjereni su uz pomoć programa ImageJ 1.48v. Za mjerenje dijametra guka korišteno je binokularno povećalo pri uvećanju od 20x. Statistička analiza rezultata podrazumijevala je analizu glavnih komponenata na osnovi korela cijske matrice u računalnome programu R v. 3.4.0. Sadnice maline koje su inokuli rane bakterijom A. tumefaciens imale su niži potencijal rasta u odnosu na kontrolnu skupinu. Soj K1026 Rhizobium rhizogenes kao aktivna tvar biopesticida pozitivno je utjecao na rast zaraženih biljaka maline suzbijajući infekciju. Primijenjeno kemijsko sredstvo na bazi bakrova (I) oksida pokazalo je nešto manji učinak na intenzitet pro mjena uzrokovanih A. tumefaciens u odnosu na biološko sredstvo. U ovome je radu zabilježen i niži intenzitet promjena kod istodobne primjene biopesticida i fungicida u odnosu na pojedinačne tretmane.The production of raspberry (Rubus idaeus L.) as one of the most promising branches of fruit growing in Bosnia and Herzegovina could be permanently endangered due to the increasing incidence of crown gall disease caused by Agrobacterium tumefaciens. The aim of this paper was to examine the morphological changes on raspberry seedlings caused by the bacterium A. tumefaciens, as well as the size and number of tumors depending on the applied biological or chemical treatment. The strain Agrobacterium tumefaciens MTCC 431 (ATCC 15955) was used. Morphometric parameters on raspberry seedlings were measured using the ImageJ 1.48v program. A binocular magnifier at 20x magnification was used to measure the the gook diameter. The computer program R c. 3.4.0 was used for a statistical analysis of results and an analysis of main components. It was observed that the raspberries inoculated with A. tumefaciens had a lower growth potential when compared to the control line. Rhizobium rhizogenes strain K1026, as an active substance of biopesticide, had a positive effect on the growth of infected raspberry plants, reducing bacterial infection. The applied chemical agent based on copper (I) oxide demonstrated lesser impact on the intensity of A. tumefaciens infection when compared to the biological agent. In this paper, a lower intensity of changes in raspberry plants was observed when the biopesticides and fungicides were applied simultaneously, in comparison with the individual treatments

Antimicrobial and antioxidant activity of the extracts from Origanum vulgare L. growing wild in Bosnia and Herzegovina

Antifungal, antimicrobial, insecticidal and antioxidant activities of Origanum vulgare L. provide the basis for suggesting that oregano plant extracts may be useful for prevention and treatment of many infection. The main goal of this study was to determine antimicrobial and antioxidant activities of methanolic and aqueous extracts from the leaves and flowers of Origanum vulgare. Antimicrobial testing of plant extracts was done using well diffusion method. Activity of extracts were tested against Gram positive bacteria: Staphylococcus aureus ATCC 25923, methicillin-resistant Staphylococcus aureus (MRSA) ATCC 33591, Bacillus subtilis ATCC 6633, Enterococcus faecalis ATCC 29212 and five Gram-negative bacteria: Salmonella abony ATCC 6017, Salmonella enterica serovar Enteritidis ATCC 31194, Pseudomonas aeruginosa ATCC 9027, Escherichia coli ATCC 25922, extended-spectrum β-lactamase (ESBL) producing Escherichia coli ATCC 35218 and fungi Candida albicans ATCC 1023. Antibiotics ampicillin, streptomycin and antimycotic nystatin were used as positive controle. The antioxidant activity was determined by using the DPPH (1,1-diphenyl-2-picrylhydrazyl) method. The highest values for inhibition zone for methanolic and aqueous extracts were recorded for Gram positive Bacillus subtilis, Staphylococcus aureus and MRSA. Methanolic extracts exhibited antibacterial activity against tested Gram negative bacteria in variable degree while the growth of these bacteria was not inhibited by aqueous extracts. Tested fungi Candida albicans was not susceptible to investigated oregano extracts. All the extracts showed moderate to potent antioxidant activity, among which the methanolic flower extract demonstrated the strongest antioxidant activity with the IC50 value of 0.205 mg/mL. Therefore it can be concluded that flower and leaf oregano extracts have great antibacterial and antioxidant potential

Phenolics of Aerial Parts of Gentiana lutea L. and Their Biological Activity

Gentiana lutea L. is a medicinal plant the roots of which are primarily used in treatments of various human digestive disorders, but also the production of alcoholic liquors. The roots (radix) of G. lutea are described in the European Pharmacopeia, but knowledge about the chemical composition and biological activities of its aerial parts is still limited. Thus, until today aerial parts of this species have not been used in medical treatments or consumed. Therefore, flowers, leaves, and stems extracts of G. lutea obtained by using four different extraction solvents (petrol ether, chloroform, ethanol, and water) were examined for their chemical composition and biological activities. High concentrations of salicylic acid, apigenin, and naringenin were recorded for ethanol stem extracts, while significant amounts of kaempferol were detected in leaves and flowers in chloroform and water extracts, respectively. The highest antioxidant potential was recorded for flower and stem petrol ether extracts with the lowest IC50 values, ranging from 94.46 ± 9.45 to 105.38 ± 10.54 μg/mL. Ethanol extracts of flowers and stems showed moderate antioxidant activity (IC50 143.15 ± 14.32 and 146.90 ± 14.69 μg/mL) as well as strong antimicrobial activity against Candida albicans (21.00 ± 1.00 and 27.50 ± 1.78 mm inhibition zones, respectively). In addition, ethanol extracts had higher antimycotic activity compared to naturally occurring phenolic compounds that are used as positive controls. Moreover, statistical analysis of the activities of plant extracts and single compounds showed that levels of chlorogenic and caffeic acids strongly correlate with the biological activities of the extracts, i.e., they are the main carriers of these biological activities. The presented results indicate the possible use of aerial parts of G. lutea as a natural preservative, as well as a antimicrobial agent, which significantly amplifies the benefits of this medicinal crop and greatly affects the sustainability of cultivated Gentiana plantation

Influence of Extraction Solvent on the Phenolic Profile and Bioactivity of Two Achillea Species

The phenolic composition, as well as the antioxidant and antimicrobial activities of two poorly investigated Achillea species, Achillea lingulata Waldst. and the endemic Achillea abrotanoides Vis., were studied. To obtain a more detailed phytochemical profile, four solvents with different polarities were used for the preparation of the plant extracts whose phenolic composition was analyzed using UHPLC-MS/MS (ultra-high performance liquid chromatography-tandem mass spectrometry). The results indicate that both of the investigated Achillea species are very rich in both phenolic acids and flavonoids, but that their profiles differ significantly. Chloroform extracts from both species had the highest yields and were the most chemically versatile. The majority of the examined extracts showed antimicrobial activity, while ethanolic extracts from both species were potent against all tested microorganisms. Furthermore, the antioxidant activity of the extracts was evaluated. It was found that the ethanolic extracts possessed the strongest antioxidant activities, although these extracts did not contain the highest amounts of detected phenolic compounds. In addition, several representatives of phenolic compounds were also assayed for these biological activities. Results suggest that ethanol is a sufficient solvent for the isolation of biologically active compounds from both Achillea species. Moreover, it was shown that the flavonoids naringenin and morin are mainly responsible for these antimicrobial activities, while caffeic, salicylic, chlorogenic, p-coumaric, p-hydroxybenzoic, and rosmarinic acid are responsible for the antioxidant activities of the Achillea extracts

Exploring New Sources of Bioactive Phenolic Compounds from Western Balkan Mountains

This study presents the first report on phenolic composition and bioactivity of ethanolic extracts of three plant species that grow in the western Balkan mountains and are used in traditional folk medicine: Valeriana montana, Salix retusa, and Campanula hercegovina. Phenolics were extracted from different aerial plant parts using 80% ethanol to assess the possibility of sustainable use of these plants as a source of bioactive compounds without disruption to the roots (for V. montana) or destruction of whole habitats (for S. retusa and C. hercegovina). The ethanolic extract of V. montana flower contained noticeable levels of apigenin and quercetin. The branches and bark of S. retusa were significantly rich in catechin, while rutin was the major phenolic found in the leaf extract of C. hercegovina. Furthermore, the flower extract of V. montana revealed the best antioxidant activity, which was comparable to 4-hydroxybenzoic acid and quercetin. Considering antimicrobial activity, the leaf extracts of V. montana and C. hercegovina demonstrated potent activity against all microbes tested, while the extracts of S. retusa were moderately effective. The presented results emphasize the potential of these plants as novel sources of bioactive compounds