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    Volume 44
    Issue 2

Diversity and the role of endophytic bacteria: a review
Sofia S. KHAN, Vijeshwar VERMA and Shafaq RASOOL
Pages 103-120
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    ABSTRACT:  Endophytes belong to a widespread group of microorganisms that colonise intracellular and intercellular spaces in all known plant parts but do not cause diseases or major morphological changes to the host. Endophytic bacteria ubiquitously colonise plant internal tissues, where they can form a variety of interactions, including commensalistic, symbiotic, trophobiotic and mutualistic. Endophytic bacteria produce pharmaceutically important compounds such as antimicrobials, antioxidants, industrial enzymes, antidiabetics and anti-cancer agents. In addition, endophytes can also support their host by producing a variety of natural products for potential use in medicine, agriculture or industry. This group of bacteria can have a tremendous impact on plant communities, raising their fitness by endowing tolerance to biotic and abiotic stress. There are great prospects for searching, selecting and studying new endophytic bacteria species in order to create new microbial preparations for adaptive crop production, while reducing the environmental impacts of agriculture. The present review summarises studies to date about endophytic bacteria, including topics such as isolation methods, the diversity of these bacteria and their biological roles.

    KEY WORDS:  endophytes, biological roles, histological localisation, biocontrol, antimicrobial activity

CRISPR/Cas-based genome editing to improve abiotic stress tolerance in plants
Tae Kyung HYUN
Pages 121-127
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    ABSTRACT:  Climate change is affecting agriculture in a number of ways, such as changing water distribution, daily temperatures and salinity patterns. In this regard, plant breeding innovations and genetic engineering approaches to improve abiotic stress tolerance are necessary to avoid a decline in crop yields caused by climate change during the 21st century. In the last few years, genome editing using the CRISPR/Cas system has attracted attention as a powerful tool that can generate hereditary mutations. So far, only a few studies using the CRISPR/Cas system have been reported to improve abiotic stress tolerance, but they have clearly suggested its effective role for future applications in molecular breeding to improve abiotic stress tolerance. Accordingly, the CRISPR/Cas system application is introduced in this mini-review as a way to improve abiotic stress tolerance. Although editing efficiency and target discovery for plant CRISPR/Cas systems require further improvement, CRISPR/Cas systems will be the key approach to maintaining global food security during climate change.

    KEY WORDS:  CRISPR/Cas system, climate change, drought, salt, high temperature

Intraspecific variability of some functional traits of Trigonocaryum involucratum (Steven) Medw., a Caucasus endemic plant
Pages 129-136
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    ABSTRACT:  Plant traits have been used to predict species and community responses to environmental gradients. We studied variation of leaf area (LA), specific leaf area (SLA), flowering shoot number and seed mass along an elevation gradient in the case of Trigonocaryum involucratum, a scree-growing plant endemic to the Caucasus ecoregion. The study had two major aims: (1) to compare the role of intra-population and inter-population variation of functional trait values; and (2) to ascertain dependence between elevation and trait values and their variation. We collected trait data in several populations in Dagestan (Russian Federation), where the species has about a 1000-m amplitude of elevational distribution. The intraspecific variability of trait values was assessed via standard statistical tools (one-way ANOVA and linear regression analysis). The trait values mostly have high inter-population variation (more than 90% for each of the trait values compared to intra-population variation of each trait), indicating adaptation of populations to site conditions. Much higher intra- vs. inter-population variation in SLA at subnival elevations indicates local micro-site diversity and may serve as a buffer against future stress related to climate change. All the trait values negatively but significantly correlate (weakly or moderately) with elevation. Negative correlation presumably shows a certain increase in the limiting effect of the elevation gradient associated with changes in temperature, soil nutrient availability and soil water content, as well as with high solar radiation. More evidence from a broader study of the species throughout its distribution range is needed for firmer conclusions about the intraspecific variability of T. involucratum.

    KEY WORDS:  leaf functional traits, specific leaf area (SLA), Myosotis involucrata, Dagestan, the North Caucasus

Morphological variability of leaf and shoot traits of four barberry taxa (Berberis L.) from the Balkan Peninsula and Sicily
Pages 137-148
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    ABSTRACT: Leaf and shoot characteristics of the following four European barberry taxa from the Balkan Peninsula and Sicily were investigated in the present study: Berberis croatica, B. vulgaris, B. aetnensis and B. cretica. Analyses were based on 10 populations of B. croatica, five of B. vulgaris and two populations of both B. aetnensis and B. cretica. Populations were randomly selected within the natural distribution area of these species. Eight leaf traits, three shoot traits and the blade length/width ratio were analysed. Multivariate analysis (principal component analysis, canonical discriminant analysis and cluster analysis) distinguished B. cretica and B. aetnensis populations and, to a lesser extent, the populations of B. croatica and B. vulgaris. ANOVA showed that the analysed populations of both B. aetnensis and B. cretica were homogeneous within the species. All populations of both B. croatica and B. vulgaris showed different degrees of intraspecies variability. Lack of complete separation, the observed grouping of populations and high intraspecies variability in B. vulgaris and B. croatica may reflect the fact that the sampled B. vulgaris and B.croatica populations were located at environmentally variable sites (unlike B. aetnensis and B. cretica), resulting in high phenotypic plasticity in those populations. Even though the observed patterns of morphological variation support the idea of four barberry taxa on the Balkan Peninsula and in Sicily, because of suspected adaptive phenotypic plasticity of the analysed Berberis taxa, the true taxonomic status of these taxa needs to be additionally confirmed by molecular methods.

    KEY WORDS:  Berberidacae, Berberis aetnensis, Berberis cretica, Berberis croatica, Berberis vulgaris, morphology, morphometry, multivariate analysis, taxonomy

Stress-induced carbon starvation in Rhizophora mucronata Lam. seedlings under conditions of prolonged submergence and water deficiency: survive or succumb
Pages 149-162
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    ABSTRACT:  The behaviour of carbohydrate metabolism in a plant, particularly its total starch content, total soluble sugar (TSS) content and their utilisation, is of great importance in coping with abiotic stress conditions. With this in mind, we studied total starch and TSS contents, survival, growth, biomass accumulation and stomatal conductance in Rhizophora mucronata under conditions of prolonged submergence and water stress for a period of 11 months. The experiment was designed in such a way as to include three replicates per each treatment level, about 1600 young mangrove plants being subjected to study in the process. Under conditions of prolonged submergence and high levels of water stress, a small number of mangrove plants survived and they were promptly exhausted due to higher starch utilisation rates (0.75-1.05% dry mass/month). Although TSS content was increased under these intense stress conditions, it was not matched by increased seedling growth or biomass production; instead, a significant reduction in growth (i.e., ~78%) and dry matter content was observed in stressed seedlings as compared to young plants in the respective controls. It follows that the intense increase of TSS content might be due to the direct conversion of starch to soluble sugars in order to produce metabolic energy for tolerance mechanisms like osmoregulation and root anatomical adaptations under stress conditions. This indicates that more energy is allocated for plant maintenance than for growth and biomass production under stress conditions, which might be a good acclimatory strategy to rescue young mangrove plants at the early phase. However, stomatal closure under stress conditions may have caused restricted photosynthesis. Therefore, stress–induced starch degradation may upsurge, which in turn might lead in the long-run to carbon starvation, a condition lethal to mangrove seedlings.
    KEY WORDS:  acclimation, mangrove, plant maintenance, growth, starch, total soluble sugars

Effect of plant growth regulators and explants on callus induction and study of antioxidant potentials and phenolic metabolites in Salvia tebesana Bunge
Pages 163-173
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    ABSTRACT: This study was undertaken to investigate the effect of different plant growth regulators (PGRs) on callus induction in Salvia tebesana explants grown in vitro and to evaluate the content of secondary phenolic compounds and their antioxidant potential. The explants (shoot apical meristem, leaf and petiole) were dissected from an 8-week-old plant of S. tebesana growing in vitro and cultured on MS media containing different concentrations of 2,4-D (0, 0.5, 1, 1.5 and 2 mg L-1), NAA (0, 0.5 and 1 mg L-1) and BAP (0, 0.5 and 1 mg L-1), either alone or in a blend with each other. Morphological characteristics of the callus (consistency and colour), biomass increase based on fresh and dry weight and the percentage of induction were recorded after 56 days. Levels of total phenols, ortho-diphenols, phenolic acids, flavonoids, proanthocyanidins and flavonols of callus, as well as antioxidant activities, were evaluated in vitro. The maximum callus formation (100%) was obtained from shoot apical meristem on MS medium supplemented with 0.5 and 1.5 mg L-1 2,4-D + 1 mg L BAP and with 1 and 1.5 mg L-1 2,4-D + 0.5 mg L-1 BAP, whereas the highest fresh (15.06 ± 0.88 g) and dry (0.33 ± 0.02 g) weights of call were observed in a medium containing1.5 mg L-1 2,4-D + 0.5 mg L-1 NAA. It was noted that MS media augmented with combined PGRs had the highest accumulation of polyphenols, phenolic acids and flavonoid compounds, with levels of content varying in the following order: 2,4-D + BAP > NAA + BAP > 2,4-D + NAA. Strong linear correlations were established between total phenolic content of callus extracts and results of the DPPH and FRAP assays (r2 = 0.896 and r2 = 0.946, p < 0.01, respectively). The obtained results suggest that the described method could be utilised as a tool for large-scale production of medicinal metabolites of S. tebesana by tissue culture.

    KEY WORDS:  antioxidant activity, callus induction, reducing power, polyphenols, Salvia tebesana, secondary metabolites

Concentration- and time-dependent effects of strontium on Lens culinaris Medik
Hakan SEPET and Murat ÇANLI
Pages 175-182
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    ABSTRACT: This study investigates how strontium (Sr) ions act on meristematic root tip cells of lentil (Lens culinaris) with changing parameters (time and concentration). Plant seeds were exposed to both a standard solution of Sr for different lengths of time (1/4, 1/2, 1, 2, 4, 8, 12, 16, 20 and 24 hours) at a fixed concentration of 1.0 molL-1 (M) and Sr ions at various concentrations (0.05, 0.1, 0.25, 0.5 and 1.0 M) for a certain length of time (12 hours). The seeds treated with Sr were made to sprout and microscopic examination focused on the root tips. The aim of microscopic examination was to clarify chromosomal abnormalities of cell division. Microscopic examination showed that various abnormalities occurred in cells of the seedlings, abnormalities such as chromosome adherence, chromosome breakings, bridge chromosomes, chromosome dispersion, chromosome shrinking, fish bones and ring chromosomes. Those abnormalities were detected several times for each treatment depending on the different periods and concentrations. Adsorption and absorption of Sr inside lentil seeds were detected by the spectroscopic method. Removed and excess amounts of Sr ions were found by spectroscopic determination. Statistical evaluation of the results was used in order to reveal the differences and similarities. The results showed that while there is a positive correlation with the concentration parameter, there is a negative correlation with the time parameter. Over 90% of Sr was removed from the solution during 12-hour exposure. Lentil seeds can be accepted as good bioaccumulators of Sr ions only for an exposure period shorter than 12 hours at an Sr concentration of 1 M.
    KEY WORDS:  chromosome abnormalities, bioaccumulator, lentil, cytogenetic effects

Physiological response of Moringa oleifera exposed to bisphenol A
Gulcan CINAR and Muhittin DOGAN
Pages 183-189
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    ABSTRACT: The physiological and morphological effects of different concentrations of bisphenol A (BPA) on Moringa oleifera seedlings were determined in this study. Significant chlorosis and abscission were observed in leaves exposed to 50 mg/L of BPA. Photosynthetic pigment levels were affected differently by varying doses of BPA. Although the total carbohydrate content of seedling parts was increased by BPA, protein content was lowered by it, except in the case of roots at 1.5 mg/L of BPA. However, it was determined that the content of nonprotein sulphhydryl groups of seedling parts did not change significantly. The total phenolic content of root tissues showed an insignificant change; however, it was found that phenolic content increased in the stems and leaves following application of BPA. The content of hydrogen peroxide (H2O2) in seedling tissues increased with increasing concentrations of BPA. Statistical analysis indicated that H2O2 content was significantly correlated with malondialdehyde content. These results clearly show that the application of BPA causes oxidative stress in seedling tissues.
    KEY WORDS:  bisphenol A, Moringa oleifera, growth, physiological effect

Effect of N-acetyl-L-cysteine (NAC) on soluble sugar and polyamine content in wheat seedlings exposed to heavy metal stress (Cd, Hg and Pb)
Pages 191-201
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    ABSTRACT: Heavy metal stress adversely affects plant growth and productivity worldwide. Alleviating the stress effect through the exogenous use of various chemical substances has become an interesting area of study in the field of plant stress tolerance. As a thiol compound, the cysteine derivative N-acetylcysteine (N-acetyl- L-cysteine, NAC) is the precursor of glutathione synthesis and a potent ROS scavenger with powerful antioxidant and free radical scavenging capabilities. This study investigated the effects of heavy metals (Cd, Hg and Pb, 100 μM) on accumulation of soluble sugars and polyamine content in roots and shoots of wheat seedlings, the water potential and proline content in shoots and the role of NAC in protection against heavy metal toxicity. The addition of 1 mM NAC significantly increased root content of glucose, fructose and sucrose in varying degrees (avg. 1.34-, 1.20- and 1.51-fold, respectively) in comparison with heavy metals alone. The treatments led to a significant reduction of sugar content in shoots. Water potential values were highly correlated with proline and sugar content in wheat seedling shoots. Heavy metal stress significantly reduced polyamine content in both plant parts. The addition of NAC increased polyamine content in seedlings in comparison with heavy metals alone in both roots and shoots. These results suggest that NAC may protect plants from oxidative stress damage in heavy metal stress, and this enhancement of stress tolerance seems to involve soluble sugar and polyamine biosynthesis.
    KEY WORDS:  leaf water potential, N-acetyl-L-cysteine, Triticum aestivum, osmolytes, stress tolerance

Susceptibility of Serbian plum cultivars to indigenous bacterial and Monilinia laxa isolates
Pages 203-210
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    ABSTRACT: The susceptibility of Serbian plum cultivars to pathogens originating from their phyllosphere was evaluated by inoculating detached young leaves and mature fruits. The virulence of indigenous isolates of the bacteria Pseudomonas syringae, Pseudomonas congelans, Erwinia persicina, Clavibacter michiganensis and Rhizobium nepotum was tested on detached leaves of four Serbian plum cultivars (Ranka, Požegača, Čačanska lepotica and Čačanska rodna). The Pseudomonas syringae isolates formed intense symptoms within 48 hours on all tested cultivars with severity index values in the range of 41 – 47%. The other isolates had significantly lower severity values or no symptoms were developed. This study demonstrates for the first time pathogenicity of Pseudomonas congelans on plum, with symptom intensity not significantly different from P. syringae after 96 h of incubation. Virulence of Monilinia laxa isolates was tested on mature fruit of the Čačanska rodna and Požegača cultivars and was detected in both of them. Higher susceptibility to M. laxa was recorded for the Požegača cultivar, with a fruit infection rate between 43 and 66%. In the case of the Rodna cultivar, no statistically significant difference in the fruit infection rate was detected between the four tested M. laxa isolates. These data indicate significant susceptibility of Serbian plum cultivars to indigenous P. syringae and M. laxa isolates. KEY WORDS:  plum, leaf, fruit, bacterial phytopathogens, Monilinia laxa

Variations in the water potential of stem xylem in Russian olive (Elaeagnus angustifolia) seedlings treated with mycorrhizal fungi under drought conditions
Pages 211-217
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    ABSTRACT: Measures must be taken to reduce the stress caused by water scarcity, which is the greatest obstacle to increasing the success of afforestation in arid areas. Precautions such as site preparation and species change do not ensure sufficient benefits. For this, it is necessary to try alternative methods such as using mycorrhization of seedlings for afforestation. The aim of the present study was to obtain Russian olive (Elaeagnus angustifolia) seedlings with high resistance to water stress and ascertain the effects of mycorrhizae on the water potential of water- stressed seedlings. Accordingly, we determined the water potentials of seedlings inoculated with arbuscular mycorrhizal fungi. Reduction in soil water content caused a reduction in the water potential of seedlings in all treatment variants. Mycorrhization reduced stress by increasing the water potential of seedlings in drought conditions, thereby enhancing their resistance to water
    KEY WORDS:  mycorrhizae, water potential, water stress, Elaeagnus angustifolia

Jumping the barrier: does a glacier tongue affect species distribution along the elevation gradient in the subnival and nival belts? A case study on Mt. Kazbegi, Georgia, Central Great Caucasus Mountains
Pages 219-229
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    ABSTRACT: Glaciers are a prominent feature in high mountains and can affect plant distribution along the gradients. However, the possible effect of glaciers on plant community structure at landscape scale has been little studied. We asked: if a glacier tongue crosses a slope laterally and potentially blocks dispersal and migrations, how can this affect vegetation structure and species composition below and above this barrier? A suitable study system is offered by slopes on Mt. Kazbegi, where we established a transect through the subnival and nival belts. We sampled vegetation below and above the glacier tongue and conducted direct gradient analyses to reveal possible effects of the glacier on patterns of species distribution and vegetation structure such as the ratio of solitary plants in vegetation patches. The obtained results indicate that the glacier tongue in our study does not cause a “vegetation switch” in the usual sense of this phrase. However, it might contribute to an abrupt change in the share of solitary plants, as well as to a very rapid decline of plant abundance and species numbers above the glacier.
    KEY WORDS:  elevation gradient, available plant nutrients, plant diversity, subnival-nival vegetation patch

Contribution to study of the pyrophilous fungi of Ukraine
Pages 231-241
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    ABSTRACT: The paper reports data on pyrophilous ascomycetes collected in 2017-2019 during a mycological survey in the Holosiivskyi National Nature Park (Kyiv, Ukraine). Sixteen species belonging to the Pezizomycetes were found. Among them, Tricharina praecox and Octospora similis were collected in Ukraine for the first time. Substrates, localities, data on general distribution and some notes about these species are presented. Ecological peculiarities of all species are briefly discussed.
    KEY WORDS:  biodiversity, distribution, Holosiivskyi National Nature Park, new records, Pezizales, post-fire fungi

Records of lichen species new for Ukraine from steppe habitats of the country
Alexander Ye. KHODOSOVTSEV and Valery V. DARMOSTUK
Pages 243-250
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    ABSTRACT: The lichens Arthonia cretacea, Bacidia viridescens, Candelariella blastidiata, C. xanthostigmoides, Enchylium bachmanianum, Haematomma nemetzii, Lecania triseptata, Lecidea sarcogynoides, Ramalina europaea, Sarcogyne praetermissa and Xanthocarpia diffusa are reported for the first time from Ukraine. Xanthocarpia interfulgens is confirmed from an arid Ukrainian landscape. Locations, herbarium deposits and substrates are given, together with notes on worldwide distribution of the reported taxa and morphological differences from similar species.
    KEY WORDS:  Arthonia, Bacidia, Candelariella, Enchylium, Haematomma, Lecania, Lecidea, Ramalina, Sarcogyne, Xanthocarpia, loess, granite, limestone, bark

New records and noteworthy data of plants, algae and fungi in SE Europe and adjacent regions, 2
Pages 251-259
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    ABSTRACT: This paper presents new records and noteworthy data on the following taxa in SE Europe and adjacent regions: the diatom alga Eunotia boreoalpina; the saprotrophic fungus Clitocybe truncicola; the liverwort Haplomitrium hookeri; the moss Leptodon smithii: the monocots Epipactis purpurata, Stipa tirsa, Typha laxmannii and T. shuttleworthii; and the dicots Krascheninnikovia ceratoides, Polygonum albanicum and Sorbus latifolia.
    KEY WORDS:  new record, Clitocybe truncicola, Eunotia boreoalpina, Epipactis purpurata, Haplomitrium hookeri, Krascheninnikovia ceratoides, Leptodon smithii, Polygonum albanicum, Sorbus latifolia, Stipa tirsa, Typha laxmannii, Typha shuttleworthii