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    Volume 45
    Issue 1

The role of epigenetic modifications in plant responses to stress
Xuefeng LU and Tae Kyung HYUN
Pages 3-12
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    ABSTRACT:  Epigenetics is the study of hereditary changes in gene expression under the premise that the nucleotide sequence is not changed. Such hereditary changes mainly involve DNA methylation, histone modification, and chromatin remodeling. These covalent modifications play indispensable roles in regulating gene expression; DNA replication, recombination, and repair; and cell differentiation. Epigenetic modifications can be partially inherited by daughter cells during mitosis and meiosis and influenced by external factors, such as environmental stresses and supply deficits. In this review, we summarize the current knowledge regarding epigenetic factors, such as DNA methylation, histone acetylation, and regulation by non-coding RNAs, in the development and stress response of plants.

    KEY WORDS:  DNA methylation, environmental stress, epigenetics, histone acetylation, non-coding RNAs

A new species of Centaurea L. subgen. Cyanus Mill. (Asteraceae) from Turkey
Emrah ŞIRIN, Hasan YILDIRIM, Tuna UYSAL and Kuddisi ERTUĞRUL
Pages 13‒22
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    ABSTRACT:  Here we describe Centaurea hekimhanensis, a new species found in Hekimhan (Turkey, Malatya Province). This Centaurea grows on the stony-gravel mountain slopes of Yamadağ and is morphologically similar to C. reuteriana, C. bourgaei, and C. pichleri in terms of their flowering lateral stem at the base of the rosette, but differs regarding the shape of the stem and rosette leaves, and the size of the rosette leaves and median appendages; the achene ornamentation is irregularly sulcate. The chromosome number of the new species is 2n = 20.

    KEY WORDS:  achene, Compositae, Centaureinae, Cyanus, karyology

Anatomical traits of Artemisia umbelliformis subsp. eriantha (Asteraceae) alpine glacial relict from Mt. Durmitor (Montenegro)
Pages 23‒30
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    ABSTRACT:  In this study, the vegetative anatomy of Artemisia umbelliformis subsp. eriantha, a Central European alpine glacial relict, wild-growing on Mt. Durmitor (Montenegro) was examined for the first time. The aim was to investigate the general anatomy and particular anatomical traits which might have possible taxonomic value. Microscopic slides were prepared according to the standard histological procedures. The adventitious young root showed a primary structure, while the older root showed a secondary structure with a well-developed periderm on its surface. The rhizome showed a primary structure with elements of a secondary structure (periderm). The stem cross section is characterized by a round shape with a well-developed periderm at certain stages, and collateral vascular bundles arranged in a circle. The petiole is concave in shape with a single-layered epidermis and parenchyma tissue with one large and two small vascular bundles. Druses and rhombohedral crystals are observed inside some petiole parenchyma cells. The leaf lobe cross section has an oblong-linear shape and is isolateral and amphistomatous in structure. The anticlinal walls of the leaf epidermal cells are sinuate. Secretory canals are present in the root cortex parenchyma (endodermal secretory canals) and the rhizome cortical parenchyma. The stem, petiole and leaf lack secretory canals. The stem and leaf are covered with T-shaped non-glandular and glandular trichomes. The taxonomic value of the analyzed characteristics is briefly discussed.

    KEY WORDS:  anatomy, crystals, secretory canals

The toxic metal stress in two mosses of different growth forms under axenic and controlled conditions
Jelena D. STANKOVIĆ, Sladjana JANKOVIĆ, Ingeborg LANG, Milorad M. VUJIČIĆ, Marko S. SABOVLJEVIĆ and Aneta D. SABOVLJEVIĆ
Pages 31-47
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    ABSTRACT: Bryophytes are widely used in monitoring environmental changes and various types of pollution. Features such as longevity and direct intake by the whole body make them very good models for bioindication. However, there is a considerable lack of knowledge on the coping, resistance and tolerance mechanisms to toxic elements within representatives of this second largest group of terrestrial plants. We tested two different growth form mosses, acrocarpous Atrichum undulatum and pleurocarpous Hypnum cupressiforme in axenic in vitro fully controlled laboratory conditions, thus eliminating additional side, antagonistic or synergistic effects. These mosses were exposed to three concentrations (including control) of zinc-, copper- or cadmium-acetate for a short and long period, and their growth, development and biochemical parameters were recorded. The results showed that reactions to stress are both species and metal-specific. Also, the short and long-term exposure times and the level of toxic elements played a role in species response and showed the differences between the two moss growth forms and species. Among the investigated parameters considered in the two mosses tested, the multiplication index proved to be the most useful and sensitive in detecting the metal pollution effects. On the other hand, the concentration of chl a seemed to vary over a rather small range between the stressed moss species. Overall, copper was shown to be quite toxic compared to the other two metals at lower applied concentration. However, the studied mosses demonstrated decreased viability according to all of the parameters tested in the media enriched with high concentrations of cadmium when compared with copper and zinc. These results suggest that species studies related to different metals/pollutants in small scale controlled conditions are urgently needed before application in broad areas monitored by mosses.

    KEY WORDS:  Hypnum cupressiforme, Atrichum undulatum, zinc, copper, cadmium, in vitro

The phytochemical composition and biological activities of different types of extracts of Achillea ageratifolia subsp. serbica
Ivana D. RADOJEVIĆ, Mirjana Ž. GRUJOVIĆ, Ljiljana R. ČOMIĆ, Milan S. DEKIĆ, Gorica T. DJELIĆ and Nevena R. MIHAILOVIĆ
Pages 49-59
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    ABSTRACT:  Achillea ageratifolia subsp. serbica is a poorly investigated endemic of the Balkan area and there are almost no data about the potential application of this plant. The purpose of this study was the investigation of the phytochemical composition, antioxidant, antimicrobial, and antibiofilm activity of its water, acetone, and ethyl acetate extracts. The phenolic composition and flavonoids were established using high-performance liquid chromatography with photodiode array detection (HPLC-PDA) analysis, while the chemical composition of the ethyl acetate extracts of the aerial parts of the plant was investigated by gas chromatography–mass spectrometry (GC and GC-MS). Antioxidant and antimicrobial activity, as well as the inhibition ability of biofilm formation on Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 27853 biofilms, were also investigated. The total phenolic content ranged from 18.61 mgGAE/g to 43.78 mgGAE/g of extract. The total flavonoid content ranged from 10.79 mgRUE/g to 34.02 mgRUE/g. The acetone and ethyl acetate extracts of the aerial parts contained chlorogenic acid, myricetin, apigenin, and luteolin. Analysis of the volatile specialized metabolites in the ethyl acetate extract of the aerial parts allowed for the identification of thirty-two constituents. The highest antioxidant activity was detected in the water extract of the aerial parts of the plant (IC50 = 641.06 μg/ml) and in the ethyl acetate extract of the root (IC50 = 675.33 μg/ml). Bacillus subtilis and S. aureus showed higher sensitivity to the tested extracts, while the antifungal activity of the marked extracts was significant. All extracts showed a moderate inhibitory effect on the ability of biofilm formation of P. aeruginosa ATCC 27853, while only the ethyl acetate inflorescence extract showed an effect on the biofilm formation of P. aeruginosa ATCC 27853. The tested extracts showed the potential for further investigation and possible application as biofungicides.
    KEY WORDS:  antimicrobial activity, antibiofilm activity, antioxidant activity, phenols, flavonoids, Asteraceae

Differences in proline accumulation between wheat varieties in response to heat stress
Nevena H. DJUKIĆ, Stefan M. MARKOVIĆ, Jasna S. MASTILOVIĆ and Predrag SIMOVIĆ
Pages 61-69
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    ABSTRACT: Proline is one of the organic osmolytes that accumulates in plants in response to stressful environmental conditions. The aim of this study was to determine the concentration of proline in the grain filling phase in ten winter wheat cultivars under conditions of prolonged periods of air temperatures above 30°C and to assess the variability of wheat variety responses. The correlation between proline accumulation and photosynthetic pigments, accumulated starch and yield was also determined. In the experimental field, flag leaves were sampled for each variety in the grain filling phase. Statistical data analysis and parameter correlation were performed using the SPSS program. The results showed that there is a statistically significant difference between proline values in moderate and high air temperature conditions when proline accumulation occurs. The significant correlation of accumulated starch content and yield with proline concentration showed that in addition to higher proline values under heat stress, wheat varieties Apač, Talas and Futura also have higher yield and starch content. A correlation was also found between proline concentration and photosynthetic pigment contents, where the Apač wheat variety showed the best adaptive response to the investigated traits and was characterized by a smaller reduction in photosynthetic pigment content under heat stress. The interrelation of proline with the photosynthetic pigment content and wheat quality parameters may be important in breeding technologies aimed at improving wheat stress tolerance.

    KEY WORDS:  Triticum aestivum, amino acid, photosynthetic pigments, adaptation, starch

A biochemical and proteomic approach to the analysis of tomato mutant fruit growth
Pages 71-85
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    ABSTRACT: To assess the effects of ABA deficiency on tomato fruit growth, the ABA mutant flacca was grown in an optimal soil water regime and various analyzes were performed, including morphological (fruit number, diameter and fruit biomass), physiological (duration of growth and fruit growth rate), biochemical (ABA accumulation, enzyme cell wall peroxidase activity) as well as proteomics. The fruit growth analysis showed that the slower fruit growth rate and development resulted in smaller flacca fruits in comparison to the wild-type fruits. The comparison of the temporal dynamics of cell wall peroxidase activity and ABA content in our experiment indicated an opposite relationship during fruit development. Proteomic analysis and the down-regulation of most proteins from carbon and amino acid metabolism, the translation and processing of proteins, energy metabolism and cell wall-related metabolism in the flacca fruits compared to the wild type, indicated reduced metabolic flux which reflected a slower fruit growth and development and reduced fruit size in the ABA mutant. These findings also indicated that ABA limited carbon sources, which could be responsible for the reduced fruit growth and size of ABA-deficient tomato fruits. The up-regulation of sulfur and oxygen-evolving enhancer proteins in the flacca fruits implicated the maintenance of photosynthesis in the late expansion phase, which slows down transition to the ripening stage. The majority of antioxidative and stress defence proteins were down-regulated in the flacca fruits, which could be related to the role of ABA in the activity of different antioxidative enzymes as well as in regulating cell wall expansion and the cessation of fruit growth.
    KEY WORDS:  ABA, cell wall peroxidase, flacca mutant

Alterations in the root proteomes of Brassica napus cultivars under salt stress
Hakan TERZI and Mustafa YILDIZ
Pages 87-96
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    ABSTRACT: Soil salinization is an important environmental problem affecting agricultural production worldwide. Seed germination is a critical process, and seedling establishment under saline conditions can be achieved by successful germination. In the present study, comparative proteomics combined with physiological analyses were used to investigate the protein alterations in germinating Brassica napus cultivars (Caravel and Sary) under NaCl stress. Seed germination declined with the increasing NaCl concentration. However, Caravel exhibited better performance in terms of seed germination and seedling growth under salinity stress. Therefore, Caravel was found to be more tolerant to salinity than Sary. The root proteins were extracted from B. napus cultivars germinating on a plant growth medium with or without 100 mM NaCl for seven days. After the root proteins had been separated by two-dimensional (2-D) gel electrophoresis, the differentially accumulated proteins were identified using MALDI-TOF/TOF MS. The comparative proteomics analysis revealed 12 and 27 statistically significant proteins accumulated in the NaCl-treated roots of Caravel and Sary, respectively. The identified proteins were mostly involved in protein metabolism, stress defense, cytoskeleton and cell wall metabolism, and energy metabolism. The salt-sensitive cultivar Sary displayed an elevated accumulation of proteins involved in antioxidant defense and the protein catabolic process such as superoxide dismutase [Fe], L-ascorbate peroxidase 1, and different components of the proteasome system. On the other hand, the levels of molecular chaperones including 20 kDa chaperonin, chaperonin CPN60, heat shock cognate protein HSC70, and heat shock 70 kDa protein 1 were higher in Caravel than Sary under salt stress. These findings will provide the possible mechanisms which contribute to salt tolerance and may serve as the basis for improving salinity tolerance in rapeseed.
    KEY WORDS:  Brassica napus, germination, proteomics, salt tolerance

Estimation of arsenic-induced genotoxicity in melon (Cucumis melo) by using RAPD-PCR and comet assays
Pages 97-106
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    ABSTRACT: In this study, arsenic (As)-induced genotoxicity in the roots and shoots of Cucumis melo (melon) seedlings were investigated by using the random amplified polymorphic DNA - polymerase chain reaction (RAPD-PCR) and comet assays. For this purpose, melon seedlings were exposed to arsenate [As(V)] at 0, 100, 200, 300, and 400 μM concentrations in the hydroponic system for 14 days to examine the level of As accumulation, alterations in growth parameters, and DNA damage. A reduction in growth with increasing As(V) concentration was observed in the melon seedlings. Total As accumulations in the shoot and root tissue increased in a dose-dependent manner; however, the level was higher in the roots than the shoots. In RAPD-PCR analysis, 26 primers gave reproducible and scorable results and produced a total of 128 bands in the control seedlings. Alterations in RAPD profiles, including the loss or appearance of new bands, were determined in the As-treated seedlings when compared to the control. The values of genomic template stability (GTS) were decreased by increasing the concentration of the As(V) in both tissue types. DNA strand breaks were observed in all the tested As(V) concentrations in the alkaline comet assay; furthermore, the loss of DNA integrity was higher with 300 and 400 μM As(V) treatments. The results clearly indicate that the combination of DNA-based molecular and cytogenetic techniques (e.g. the comet assay) may be proposed as a reliable evaluation of genotoxicity in plants after exposure to heavy metal pollution.
    KEY WORDS:  DNA damage, genomic stability, genotoxicity tests, heavy metal stress

The wild raspberry in Serbia: an ethnobotanical study
Pages 107-117
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    ABSTRACT: An ethnobotanical survey of the traditional use of the wild raspberry (Rubus ideaus) was conducted on nine sites in the regions of southwest (SWS) and southeast (SES) Serbia. The regions studied belong to mountainous rural areas in Serbia, mainly populated by Serbian people of the Orthodox faith. In total, 93 respondents were interviewed about the medicinal, food and cosmetic uses of the wild raspberry. In terms of plant parts in traditional use, the leaf and fruit are known to be used frequently. The dry leaf is more commonly used in the SES (78%) than in the SWS (25%) region. In the SWS region, the most frequent use of the wild raspberry leaf is linked to gastrointestinal disorders and prostate inflammation. In the SES region, the leaves are mostly used for the prevention and healing of gynecological disorders during both pregnancy and lactation. The local population of the SWS region uses the fruits mainly for the preparation of syrup juice (92%), fruit preserves (42%) and in desserts (62%), while in the SES area the fruits are most frequently used for the preparation of nonalcoholic (97.4%) and alcoholic beverages (35.9%). Our results provide some novel information on the use of the wild raspberry in Serbia and in the Balkans, such as for the treatment of prostate inflammation, use during breastfeeding and to alleviate inflammation of the eyes. KEY WORDS:  Rubus idaeus, ethnobotany, leaf and fruit, folk medicine, traditional products

New records and noteworthy data of plants, algae and fungi in SE Europe and adjacent regions, 3
Marko S. SABOVLJEVIĆ, Gordana TOMOVIĆ, Petya BOYCHEVA, Dobri IVANOV, Teodor T. DENCHEV, Cvetomir M. DENCHEV, Ivana STEVANOSKI, Aleksandra MARKOVIĆ, Sanja Z. DJUROVIĆ, Uroš BUZUROVIĆ, Galina YANEVA, Sorin ȘTEFĂNUȚ, Miruna-Maria ȘTEFĂNUȚ, Aleksandar KNEŽEVIĆ, Predrag PETROVIĆ, Boris ASSYOV, Jovana PANTOVIĆ, Marjan NIKETIĆ, Snežana VUKOJIČIĆ, Roxana ION and Gabriela TAMAS
Pages 119-127
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    ABSTRACT: This paper presents new records and noteworthy data on the following taxa in SE Europe and adjacent regions: parasitic fungus Antherospora hortensis, saprotrophic fungi Loweomyces fractipes and Pholiota henningsii, stonewort Chara canescens, mosses Grimmia caespiticia and Rhodobryum ontariense, fern Woodsia alpina, monocots Aegilops triuncialis, Epipactis purpurata, Galanthus elwesii and Typha shuttleworthii and dicot Umbilicus luteus.
    KEY WORDS:  new report, Aegilops triuncialis, Antherospora hortensis, Chara canescens, Epipactis purpurata, Galanthus elwesii, Grimmia caespiticia, Loweomyces fractipes, Pholiota henningsii, Rhodobryum ontariense, Typha shuttleworthii, Umbilicus luteus, Woodsia alpina

New records and noteworthy data of plants, algae and fungi in SE Europe and adjacent regions, 4
Pages 129‒136
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    ABSTRACT: This paper presents new records and noteworthy data on the following taxa in SE Europe and adjacent regions: diatom alga Stauroneis neofossilis, parasitic fungus Anthracoidea arenariae, horsetail Equisetum hyemale, liverwort Harpanthus flotovianus, mosses Fissidens exilis and Rhizomnium punctatum, monocots Epipactis helleborine subsp. orbicularis, Himantoglossum calcaratum subsp. rumelicum and Schoenus nigricans and dicots Calluna vulgaris, Mahonia aquifolium and Willemetia stipitata subsp. albanica.
    KEY WORDS:  new report, Anthracoidea arenariae, Calluna vulgaris, Epipactis helleborine subsp. orbicularis, Equisetum hyemale, Fissidens exilis, Harpanthus flotovianus, Himantoglossum calcaratum subsp. rumelicum, Mahonia aquifolium, Rhizomnium punctatum, Schoenus nigricans, Stauroneis neofossilis, Willemetia stipitata subsp. albanica, SE Europe

The chemical composition and anti-inflammatory effect of the essential oil obtained from Abeliophyllum distichum flowers
Hyun Ju JU, Heung Bin LIM and Tae Kyung HYUN
Pages 137‒142
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    ABSTRACT: In the present study, the chemical composition of the essential oil (EO) obtained from the flowers of Abeliophyllum distichum, commonly known as white forsythia, was examined by gas chromatography–mass spectrometry (GC/MS). Thirty-five components including epoxy linalool, methyl salicylate, linalool oxide (pyranoid), and L-linalool were identified in the EO of A. distichum flowers (AfEO). In addition, the AfEO exhibited a remarkable anti-inflammatory effect in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. We found that this anti-inflammatory effect was mediated by inhibiting the expression of proinflammatory mediators, including IL-1β, IL-6, and IL-18. Taken together, these results confirm the potential use of the AfEO as an anti-inflammatory agent for topical application.
    KEY WORDS:  Abeliophyllum distichum, anti-inflammation, essential oil, pro-inflammatory mediator

A contribution to Amanita alseides, a recently described European species in the section Vaginatae
Boris ASSYOV, Fuat BOZOK, Mahmut YARAR, Monica SLAVOVA and Hatıra TAŞKIN
Pages 143‒148
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    ABSTRACT: The authors present a contribution to Amanita alseides, so far known from its type specimens from France and Italy. A detailed description is included, based on molecularly supported collections from Bulgaria and Greece, expanding the knowledge on the morphological variability and the distribution of the species.
    KEY WORDS:  Amanitaceae, Amanitopsis, Balkan mycota, biogeography, taxonomy

In memoriam: Prof. Dr. Slobodan Jovanović (1955-2021)
Pages 149-159
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List of reviewers for Botanica Serbica in 2020 — Acknowledgements
Pages 161-162
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