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Chemical characterization, antimicrobial and cytotoxic activity of essential oil and post-distillation waste of juniper (Juniperus communis L.) and their combinations with conventional antibiotics and cytostatics
No full textU ovom radu ispitivana su etarska ulja i postdestilacioni ostatak (PDO) Juniperus communis L. GC-MS analiza ulja je pokazala dominantno učešće α-pinena, a LC-MS/MS je identifikovala hinsku kiselinu i rutin kao najzastupljenije u PDO. Bolja antioksidativna aktivnost PDO utvrđena je u TBA i naročito DPPH eseju (IC50 4,4 μg/mL), što potvrđuju i rezultati FRAP testa. Citotoksičnost je bila veća u slučaju ulja: IC50 vrednosti na ćelijskim linijama HT-29, HCT 116, A549 i MRC-5 bile su u opsegu 69-120 μg/mL. Efekat kombinacija oba derivata sa doksorubicinom na kancerskim A549 i normalnim MRC-5 ćelijama bio je sinergistički, naročito u slučaju A549. Citometrijska analiza i alkalni komet test su pokazali da PDO indukuje apoptozu, zaustavlja ćelijski ciklus i ima genotoksični efekat. MIK testom je utvrđeno da su Listeria monocytogenes i Aspergillus versicolor najosetljivije, posebno prema PDO (MIK 0,39 i 0,12 mg/mL). Sinergizam je uočen u slučaju tretmana listerije kombinacijama ulja/PDO i streptomicina/ampicilina/azitromicina, kao i Candida albicans kombinacijom PDO i bifonazola. Adhezija patogena na ćelije kolona u kulturi bila je inhibirana dejstvom PDO ili ulja, u slučaju listerije ili kandide. Etarska ulja kleke i čubra, primenjena u mikrodilucionim testovima odvojeno ili u kombinacijama, pokazala su dobar potencijal i postojanje sinergizma u slučaju primoizolata L. monocytogenes. Time kill test je potvrdio sinergizam i ukazao na veću osetljivost listerije u aerobnim nego hipoksičnim uslovima. Ulja primenjena na mariniranom mesu inhibirala su rast ukupnih aerobnih heterotrofa, enterobakterija, bakterija mlečne kiseline i inokulisane listerije. Uočeni citotoksični i antimikrobni efekat preporučuju derivate kleke za adjuvante koji pojačavaju aktivnost konvencionalnih lekova.In this work essential oils and post-distillation waste (PDW) of Juniperus communis L. were screened. GC-MS analysis pointed at α-pinene as dominant constituent of oils; LC-MS/MS identified quinic acid and rutin as the most prevalent in PDW. Higher antioxidativity of PDW was shown in TBA and especially DPPH assay (IC50 4.4 μg/mL), which is additionally confirmed by FRAP test. Oil induced higher cytotoxicity against HT-29, HCT 116, A549 and MRC-5 cells, with IC50 values ranging 69-120 μg/mL. Combinations of each derivative with doxorubicin, applied on cancer A549 and normal MRC-5 cells induced synergism especially pronounced in A549. Cytometric analysis and alkaline comet test showed that PDW induced apoptosis and cell cycle arrest, and was genotoxic. Microdilution assay pointed at Listeria monocytogenes and Aspergillus versicolor as the most sensitive, especially to PDW (MIC 0.39 and 0.12 mg/mL). Synergism was observed in the case of listeria treated with combinations of oil/PDW and streptomycin/ampicillin/azithromycin, as well as Candida albicans with a combination of PDW and bifonazole. In vitro adhesion of L. monocytogenes/C. albicans to colon cells was inhibited by PDW and oil, respectively. Additionaly, juniper and savory essential oils, applied in microdilution tests separately or in combinations, showed notable potential and synergism in the case of primoisolates of listeria. Time kill assay confirmed synergism and indicated a higher bacterial sensitivity under aerobic than hypoxic conditions. Oils applied to marinated meat inhibited the growth of total aerobic heterotrophs, enterobacteria, lactic acid bacteria and inoculated listeria. Observed cytotoxic and antimicrobial effect recommends juniper derivatives as adjuvants that enhance conventional drugs activity
Characterisation of indigenous apple accessions with respect to polymorphism of ACS1 and ACO1 genes
No full textEthylene is the simplest signalling molecule with a hormone-like function that plays a major role in many developmental processes, including ripening of climacteric apple fruit. The allelic polymorphisms of ACS1 and ACO1 genes, encoding for ACC synthase and ACC oxidase, which catalyse the last two steps in the ethylene biochemical pathway, were analysed in nineteen indigenous apple accessions grown in individual growers' orchards in the regions of central and southwestern Serbia. A polymorphism was detected using the polymerase chain reaction (PCR) for the ACS1 gene and the additional enzymatic digestion of the PCR product with BamH1 and RsaI for the ACO1 gene. The ACS1-1 and ACS1-2 alleles of the ACS1 gene, as well as the a and c alleles of the ACO1 gene were identified. The polymorphisms observed upon PCRs and digestion with restriction enzymes were generated in two genotypes for both genes, i.e. the ACS1 gene - ACS1-1/1 and ACS1-1/2, and the ACO1 gene - aa and ac. Out of nineteen apple accessions, sixteen were homozygous for the allele ACS1-1 and three were heterozygous (ACS1-1/2); regarding the ACO1 genotype, fourteen were homozygous for the allele a and five were heterozygous (ac allelic constitution). The molecular survey in the current study provides an increase in the number of apple accessions with potential to be used as parents in breeding programmes, aiming to obtain high quality cultivars that retain fruit texture during long storage. Therefore, the heterozygous accessions for the ACS1 gene - 'J-LuN/1', 'Kraljica' and 'Šumatovka', may have an important position in future breeding programmes.Etilen je najjednostavniji signalni molekul sa hormonskom funkcijom koji utiče na brojne procese tokom rastenja i razvića, uključujući i dozrevanje klimakteričnog ploda jabuke. Alelni polimorfizam ACS1 i ACO1 gena koji kodiraju 1-aminociklopropan-1-karboksilat (ACC) sintazu i ACC oksidazu, ključne enzime u biosintezi etilena, analiziran je kod devetnaest autohtonih genotipova jabuke gajenih u zasadima individualnih proizvođača na području pojedinih teritorijalnih jedinica centralne i jugozapadne Srbije. Polimorfizam ACS1 gena je detektovan primenom lančane reakcije polimeraze (PCR metoda), dok je digestija PCR proizvoda sa BamH1 i RsaI restrikcionim enzimima korišćena za detektovanje alela ACO1 gena. U ispitivanom materijalu identifikovana su po dva alela ACS1 (ACS1-1 i ACS1-2) i ACO1 (a i c) gena. Uočeni polimorfizam nakon PCR amplifikacije i restrikcione analize omogućio je identifikovanje po dve alelne konstitucije ACS1 gena - ACS1-1/1 i ACS1-1/2, kao i ACO1 gena - aa i ac. Od devetnaest analiziranih autohtonih genotipova jabuke, šesnaest su homozigoti za alel ACS1-1 (ACS1-1/1), dok su tri genotipa heterozigoti (ACS1-1/2); u pogledu ACO1 alelne konstitucije, četrnaest genotipova su homozigoti za alel a (aa alelna konstitucija) i pet genotipova su heterozigoti (ac alelna konstitucija). Molekularno istraživanje u okviru ovog rada pruža novi kvalitet u oblasti očuvanja i održivog korišćenja genetičkih resursa, rezultirajući povećanjem broja genotipova koji mogu biti interesantni za budući oplemenjivački rad na stvaranju novih sorti jabuke dobrih skladišnih sposobnosti. U tom pogledu, genotipovi heterozigotni za ACS1 gen - ‘J-LuN/1’, ‘Kraljica’ i ‘Šumatovka’, mogu predstavljati važan početni materijal za savremene programe oplemenjivanja, kao i programe uvođenja u proizvodnju autohtonih genotipova jabuke željenih agronomskih osobina
Morphological and pathogenic characteristics of Alternaria alternata isolates from plum (Prunus domestica L.)
No full textDuring June and July 2020, numerous 'Stanley' plants were observed with small, brown spots on leaves in the plum orchard in locality Bačka Topola. Later on, leaf lesions became dark brown and irregular, resulting in necrosis that was followed by defoliation. Collected samples were analyzed using standard mycological procedures. Ten fungal isolates from leaves and branches were obtained from infected plums. Two isolates originating from branches (ŠG) and leaves (ŠL), were selected for further studies. The pathogenicity test was performed on five plum cultivars: 'Stanley', 'Čačanska Lepotica', 'Čačanska Rodna', 'Čačanska Najbolja' and 'Požegača'. Morphological characteristics of two selected isolates were studied on three nutrient media: cornmeal agar (CMA), carnation leaf agar (CLA) and potato dextrose agar (PDA). Both tested isolates formed airy, dark olive-green colonies on PDA medium. The colonies were light olive-green in all tested isolates on CMA. On CLA, all studied isolates formed olive-green substrate mycelium. Conidia size for tested isolate ŠG on PDA medium ranged from 19.20 to 46.24×7.95-12.88 μm (mean 25.85×10.42 μm), and for isolate ŠL from 18.17 to 36.16×8.09-12.59 μm, (mean 24.98×11.10 μm). Both studied isolates on all tested plant organs (leaf, branch and fruit) in all plum cultivars caused symptoms typical for fungi of the genus Alternaria, the appearance of dark brown necrotic lesions on leaves, branches and fruits. Based on preliminary research of morphological characteristics and pathogenic tests, analyzed isolates were identified as belonging to the species Alternaria alternata
Early tree performances, precocity and fruit quality attributes of newly introduced apricot cultivars grown under western Serbian conditions
No full textIn this work, 19 newly introduced and some traditional apricot cultivars were evaluated by 20 phenological and agronomical traits and fruit quality attributes. 'the results showed a wide variation in phenological data, tree vigour (TCSA), productivity [yield per tree, cumulative yield (CY) and yield efficiency (YE)], and fruit quality attributes such as fruit and stone weight, flesh/stone ratio, fruit dimensions, size, shape index, soluble solids content (SSC), titratable acidity (TA) and ripening index (RI). The average onset of blossoming varied from 16 March to 20 March, whereas harvest was between 1 June and 12 September. The most vigorous trees were 'Ketch Pshar'. The best productivity was observed in 'Fardao' and the poorest in 'Farbaly'. More apricots were relatively small to medium in fruit size, whereas 'Candela' had very large fruits. Most cultivars tended towards a round shape, whereas some had round/ flat and/or ovoid-shaped fruits. The highest values for SSC were observed in 'Ketch Pshar', 'Candela' and 'Fardao', TA in 'Candela' and RI in 'Hungarian Best' There was a medium to high correlation between yield properties, fruit and stone size and flesh/seed ratio, also between SSC versus acidity and RI. As observed by PCA, the first three components represented 74.3% of total variance (38.3%, 22.1% and 19.8% for PC1, PC2 and PC3, respectively)
Polyphenols content of berry leaves and calluses
No full textSecondary plant metabolites are synthetized as stress-responding compounds and provide selective advantages to plants. This is a huge group of compounds with heterogeneous structures. One of the biggest classes of these compounds are polyphenols. Most of them also act as antioxidants, providing benefits for human health. Consequently, food scientists explore possibilities for the isolation and application of these bioactive compounds in food industry. In order to obtain polyphenols, which are present in the plant in low quantities, conventional plant cultivation is not efficient enough. In vitro plant tissue culture techniques, in highly controlled conditions and using appropriate medium, allow, in shorter period of time, production of the whole plant or the plant tissue with ability to synthesize desired compounds, overcoming this problem.
In order to determine whether in vitro tissues can be a good source of polyphenols, in this paper, in vitro shoot and callus cultures were established. It was done with two berry fruit genotypes – blackberry (Rubus subg. Rubus Watson ‘Čačanska Bestrna’) and blueberry (Vaccinium corumbosum L. ‘Toro’). Extraction and quantification of total phenolics (TPC) and total flavonoid content (TFC) were done both from callus and leaves tissues obtained from in vitro plants, as well as from field-grown plants. TPC and TFC were the highest in leaves of blueberry (13.47–14.06 mg GAE/g DW, for in vitro and field-grown plants, respectively) whereas low TPC and TFC values (0,78–2,39 mgGAE/g DW) for both callus cultures were obtained.
Results show that in vitro tissues could be a good source of polyphenols. Considering the cultivation time reduction and the process simplification, tissues are rentable sources of polyphenols
Effect of soil management systems on the generative potential and fruit quality of black chokeberry
No full textThis experiment evaluated the effect of soil management systems (bare soil and black foil mulch)
on the generative potential (number of clusters per inflorescence, number of flowers per inflorescence, number of berries per cluster, berry set percentage, cluster weight, berry weight, yield per bush, yield per unit area) and fruit quality (soluble solids content, total sugars, total acids, pH value and protein content) in black chokeberry [Aronia melanocarpa (Michx.) Elliott] of cultivar 'Nero'. Significant differences in generative potential were detected among soil management systems. The experimental results showed that black foil mulch, due to its ability to increase temperature and moisture in the soil directly led to an increase in the number of clusters per inflorescence, number of flowers per inflorescence and number of berries per cluster. In contrast, bare soil, with lower soil water content and temperature stimulated an increase in the cluster weight and berry weight, as well as yields per bush and per unit area. Also, soil management systems had a significant effect on the chemical composition of the fruit. Black foil mulch promoted the synthesis of soluble solids, sugars and protein, while bare soil favoured the accumulation of acids in chokeberries. Generally, the results suggested that black chokeberry grow well under varied soil management systems, and indicated that soil management systems had a significant effect on the generative potential and fruit quality
Cytotoxicity and genotoxicity of Juniperus communis essential oil and post-distillation waste
No full textCytotoxicity of essential oil (EO) and post-distillation waste (PDW) of Juniperus communis was investigated in MTT assay on human lung carcinoma (A549), colorectal adenocarcinoma (HT-29) and normal lung fibroblast (MRC-5) cells. EO was remarkably more cytotoxic comparing to PDW, with determined IC50 values in the ranges 69-120 and 1270-2860 µg mL-1, respectively. In addition, both test substances were more cytotoxic to cancer than to normal cells. However, calculated selectivity indexes which ranged from 1.4 to 2.2 revealed that selectivity towards cancer cells was not remarkable. Genotoxicity was tested in comet assay on MRC-5 cells. Obtained results showed that only PDW, applied in concentrations 500 and 1000 µg mL-1, was genotoxic
ʽLedaʼ a new sour cherry cultivar
No full textSour cherry is one of the most economically important temperate fruit species worldwide, which is widely used both fresh and processed. Due to the consumer demand for antioxidant-rich products, the development of new tart cherry selections with desirable quality characteristics is critical for the sustainability of cherry producers and processors. ‘Leda’ is a new sour cherry cultivar released from the sour cherry breeding program at the Institute PKB Agroekonomik. It was selected in mixed sour cherry orchard, from spontaneus seedlings population, and was recognized as cultivar in 2018 by the Serbian Ministry of Agriculture, Forestry and Water Management. The evaluation of this cultivar was done in comparison with the control cultivar ‘Oblačinska’ during a two-year period (2015 and 2017). Averagely, it bloomed a day or two after the control cultivar, while the ripening time was a day earlier than ‘Oblačinska’. The productivity is higher than the standard cultivar. The attractive bright red coloured fruits are larger (3.8 g), compared to ‘Oblačinska’ (3.3 g), with shorter fruit stalk length (2.9 and 3.4 cm, respectively). It has rich, mild flavour, sweet-acidic and harmonic taste, having slightly higher value of soluble solids/total acidity ratio (10.4) compared to the standard cultivar (9.5). ‘Leda’ is highly resistant to economically important diseases and pests (Monilinia laxa, Monilinia fructigena, Blumeriella jaapii, Wilsonomyces carpophilus and Rhagolethis cerasi). This cultivar could be used both for processing and fresh consumption
Freezing as a pre-treatment in air drying of plums
No full textAccording to its composition, primarily for its high phenolic content and high antioxidant capacity, prune is among highly valuable nutritional foods. Because of using whole fresh fruit, drying of plums is considered a slow and time-consuming process. Thus, various pre-treatments are applied in order to reduce drying time with no effect on declining nutritional value. The most simple and commonly used pretreatment is dipping - immersing fruit into boiling water before drying. Since the plum drying season is timely restricted, we had to come up with the idea to extend it by previously freezing the fruit intended for drying and then, after a few months of storage at -20°C, use them for drying. Considering the physical changes of fruit during freezing that could affect drying speed, freezing can be regarded as pre-treatment in the production of prune. Drying of fruit was carried out in an experimental dryer at an air temperature of 90°C until reaching 75% of the total dry matter. Plum cultivars created at the Fruit Research Institute recommended for drying were used ('Nada', 'Čačanska Rodna' and 'Valjevka') and 'Stanley', which is the most frequently used for drying in Serbia. The aim of this work was to study the effect of pre-treatment (dipping and freezing) on quality of prune in comparison with non-treated dried fruit. The following quality parameters of prune: contents of sugars, acids, sugar/acid ratio, total phenols and antioxidant capacity as well as organoleptic properties (appearance, taste, aroma, consistence), were tested. Contents of total phenols, antioxidant capacity as well as total organoleptic evaluations in dried fruit obtained by pre-treatments (dipping and freezing) do not differ compared to non-treated fruit (control) in all tested cultivars
Impact of rootstock on fruit physical properties of 'Pink Lady' apple
No full textTokom 2018. i 2019. godine ocenjivali smo glavna fizička svojstva ploda [masa (FW), dužina (L), prečnik (D), srednji geometrijski prečnik (Dg), odnos dužina/prečnik (L/D odnos), sferičnost ( ph), površina (S), zapremina (V) i specifična masa (r)] sorte 'Pink Lady' kalemljene na kržljavoj M.9 T337, polukržljavoj M.26 i srednjebujnoj MM.106 podlozi. Rezultati su pokazali da su podloge značajno menjale ispitivane osobine sa izuzetkom L/D odnosa, ph i r. Među podlogama, M.9 T337 je uslovila najbolje vrednosti, dok su manje i statistički slične vrednosti promovisale podloge M.26 i MM.106. Sve podloge su izazvale prečnik ploda veći od 66 mm, dok je samo M.9 T337 indukovala prečnik ploda ≥70 mm. Podloge nisu imale sposobnost da promene oblik ploda klub sorte 'Pink Lady'.During 2018 and 2019, we evaluated the main fruit physical properties [weight (FW), length (L), diameter (D), geometric mean diameter (Dg), length/diameter ratio (L/D ratio), sphericity (ph), surface area (S), fruit volume (V) and solid density (r)] of 'Pink Lady' apple grafted on dwarf M.9 T337, semi-dwarf M.26 and semi-vigour MM.106 rootstocks. Results showed that rootstocks significantly changed the properties evaluated with the exception of L/D ratio, ph and r. Among rootstocks, M.9 T33 7 produced the best properties, whereas lower and statistically similar values were obtained from M.26 and MM.106 rootstocks. All rootstocks promoted D higher than 66 mm, whereas only M.9 T337 induced D ≥70 mm. The rootstocks did not have the capacity to change the fruit shape of the 'Pink Lady' club cultivar