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Secondary Somatic Embryogenesis in Plants: From Cellular Mechanisms to Biotechnological Potential
No full textSecondary somatic embryogenesis (SSE) is a powerful tool in plant biotechnology, enabling the continuous production of embryos from primary somatic embryos (PSEs) and offering broad applications across agriculture, forestry, horticulture, and pharmaceutical industries. Depending on culture conditions, SSE may proceed directly from the surface of PSEs or indirectly via callus formation, with the outcome strongly influenced by exogenous plant growth regulators (PGRs). A key advantage of SSE is its cyclic nature, which offers a valuable strategy to maintain embryogenic potential over extended culture periods, generating true-to-type embryos without reliance on the original explant, while significantly increasing the multiplication rate, often making SSE more productive than PSE in many species. This review explores in detail the cellular origin and developmental pathways of secondary embryos, the maintenance of embryogenic competence through cyclic embryogenesis, as well as genetic and epigenetic aspects and the biotechnological applications of this process. Moreover, it addresses challenges regarding strong genotype dependence, variability in embryo quality and morphology, limitations in maturation and conversion potential, and the gradual decline of embryogenic competence with successive cycles, all of which need to be overcome to ensure the stability and reproducibility of SSE and maximize its impact
Relationship between photosynthetic efficiency and total carotenoids and the content of Zn and Sr in Aesculus hippocastanum L.
No full textAesculus hippocastanum L., known as the European horse chestnut, is a deciduous tree naturally occurring in the mountainous regions of the Balkans and southeastern Europe, at altitudes between 700 and 1200 meters. Although it adapts well to different growing conditions, its resistance to pollution is limited, resulting in visible damage in urban environments and a decline in natural populations due to disease and environmental stress. Given its widespread occurrence, frequent planting, and the ease of monitoring physiological parameters, A. hippocastanum is often used as a bioindicator of pollution, particularly in urban environments. In this study, the influence of Zn, a cofactor of numerous enzymes involved in carotenoid and antioxidant synthesis, and Sr²⁺, a chemical analog of Ca²⁺ that can interfere with thylakoid membrane stability and PSII enzyme function, was investigated for their effects on photosynthetic efficiency and total carotenoid content at a polluted - Pionirski Park, Belgrade and an unpolluted site – Kosmaj. The leaf samples were analysed using ICP-OES for Mn and Zn, chlorophyll fluorometry for photosynthetic efficiency (Fv/Fm), and spectrophotometry for total carotenoid content. The results showed that Sr concentrations at both sites exceeded levels considered toxic for plant tissues (>30 mg kg⁻¹; Shacklette et al., 1978), measuring 37.17 mg kg⁻¹ at Belgrade and significantly higher at Kosmaj (62.41 mg kg⁻¹). In contrast,
Zn concentrations were below the threshold required for normal physiological functioning of plants (<20 mg kg⁻¹; Kabata-Pendias and Pendias, 2001), with values of only 11.81 mg kg⁻¹ in A.hippocastanum leaves at both Belgrade and Kosmaj. Consequently, photosynthetic efficiency values were lower than optimal at both sites—0.671 in Pionirski Park and 0.692 at Kosmaj, as well as the total carotenoid content (1.59 mg g⁻¹ and 1.87 mg g⁻¹, respectively), probably due to the combined effects of toxic Sr concentrations, which can disrupt thylakoid membrane integrity and PSII activity, and Zn deficiency, which impairs antioxidant defence and enhances oxidative pigment degradation.
The results confirm that A. hippocastanum responds to Zn deficiency and toxic Sr accumulation through reduced photosynthetic efficiency and lower carotenoid content, indicating sensitivity to both micronutrient imbalance and excess metal exposure. This highlights its potential as a reliable bioindicator for assessing the impact of trace element availability in different environments.Pavlović M, Pantović Pavlović M, editors. Proceedings : Meeting point of the science and practice in the fields of corrosion, materials and environmental protection : XXVI YuCorr International Conference; 2025 Nov 3-5; Palić, Serbia. Belgrade: Serbian Society of Corrosion and Materials Protection, UISKOZAM; 2025. p. 62
The effects of novel diclofenac-based carborane-substituted prodrug on colon cancer cell lines and mouse-derived organoids
No full textBackground: Drug repurposing is a promising strategy in drug development in oncology. A notable example of this approach is diclofenac, a nonsteroidal anti-inflammatory drug (NSAID) traditionally used for pain relief, which has shown anticancer potential via COX-dependent and COX-independent mechanisms. In this study, a novel anticancer compound, a unique carborane derivative of diclofenac with a zwitterionic amidine structure and a nido-carborane cluster, was developed using fragment-based drug discovery. Designed as an engineered prodrug, it aims to overcome traditional NSAID limitations and enhance biological effects.
Material and Methods: The anticancer properties of the compound were tested against three cancer cell lines: murine colon adenocarcinoma (MC38), human colorectal carcinoma (HCT116), and human colorectal adenocarcinoma (HT29), as well as peritoneal exudate cells (PECs) isolated from healthy mice and on the human fetal lung fibroblast cell line MRC-5. Furthermore, the effects of the experimental compound were tested on colon cancer mouse-derived organoids. Cell viability was measured using MTT and CV assays on cell lines and the CellTiter-Glo® assay on organoids. The mechanism of action of the substance on the MC38 cell culture was evaluated by flow cytometry and fluorescence microscopy.
Results and Conclusions: The tested compound showed strong cytotoxicity on all cancer lines after 72 h, with IC50 values for PECs and MRC-5 being 2–3 times higher, indicating moderate selectivity towards malignant cells. Treatment also decreased the viability of organoids. Flow cytometry analysis revealed that the cytotoxicity of the substance is based on caspase-independent apoptosis preceded by cell division blockage. Fluorescence microscopy after PI staining confirmed apoptosis, cells with possibly undivided genetic material, and senescence cells. Acridine orange staining showed increased presence of autophagosomes, but co-treatment with autophagy inhibitors further reduced viability, suggesting a pro-survival role of autophagy. The production of reactive oxygen and nitrogen species (ROS/RNS) in the presence of the tested compound was strongly reduced, and it was in concordance with the efficient inhibition of COX-2. Our findings suggest that carborane-based prodrugs offer a promising route for developing new anticancer therapies, especially as adjuvant or combination therapies. The presented results open valuable possibilities for further research.Čavić M, editor. Proceedings book: The 1st Regional SDIR-HDIR-MOKAD Congress; 2025 Oct 8-10; Belgrade, Serbia. Belgrade : Serbian Association for Cancer Research; 2025. p. 144-5. (Oncology Insights; No. 3)
Milestone for the ENLIGHT project: Dataset on physiological stress response in juveniles
No full textThis data set is a milestone of the WP2 work package of the Development under artificial light at night. Biological impact of light pollution on amphibians (ENLIGHT) project number #11120 financed by the Science Fund of the Republic of Serbia. Data set contains measurements of physiological response of juveniles of Triturus ivanbureschi exposed to cold and warm LED artificial light at night
Deficits in spatial and recognition memory in rat models of Parkinson’s disease – cellular and synaptic changes in the hippocampus
No full textWe investigated spatial (SM) and recognition memory (RM) and the underlying
cellular and synaptic changes in the hippocampus in rat models of Parkinson’s
disease (PD). Adult male Wistar rats were divided into four experimental groups:
Control (n=17), PD cholinopathy (n=12), hemiparkinsonism (n=11) and
hemiparkinsonism with PD cholinopathy (n=13). The memory deficit was examined
at day 14 and 42 after the lesion-induced PD model with a spatial habituation test
and a novel object recognition test. Cellular and synaptic changes in the
hippocampus were analyzed 14, 42 and 100 days after the lesions using PV, MAP,
PSD-95 and Cx36 immunohistochemistry. The onset of PD cholinopathy was
characterized by MAP suppression in the hippocampus, followed by SM (p=0.83)
and RM (p=0.91) impairments from day 42. Although hippocampal cellular and
synaptic function remained unchanged in hemiparkinsonian rats, we detected a
long lasting deficit in SM (p≥0.22) but not in RM (p≤0.03). The onset of
hemiparkinsonism with PD cholinopathy was characterized by overexpression of
PSD-95 in the hippocampus, followed by long-lasting impairment in SM (p≥0.22)
and delayed impairment in RM (p=0.82), which was correlated with hippocampal
PV suppression (r=0.52, p=0.04) but not with Cx36 suppression (r=0.08, p=0.80).
Our results suggest an important role of hippocampal PV interneurons in the
progression of hemiparkinsonism with PD choliopathy; differential synaptic
changes in the hippocampus at the onset and during the progression of distinct
PD neuropathology underlying specific memory deficits; and an important role of
dopaminergic neurodegeneration in SM deficit and cholinergic
neurodegeneration in RM deficit in PD models.Dragić M, editor. Book of abstracts: 9th Congress of Serbian Neuroscience Society; 2025 Oct 6-8; Belgrade, Serbia. Belgrade: Serbian Neuroscience Society; 2025
Assessing the role of mitonuclear and genotype-by-environment interactions for the Trojan Female Technique in pest biocontrol
No full textThe seed beetle (Acanthoscelides obtectus) is a globally distributed pest affecting common beans and various legume species. It feeds on seeds and causes considerable damage to stored products, resulting in substantial economic losses. Currently, insecticides are mainly used to control this species. However, the detrimental effects of these chemicals on the environment and non-target species, as well as the evolution of resistance, necessitate the development of safer and more specific biocontrol methods.
One such method is the recently proposed Trojan Female Technique (TFT), which leverages the evolutionary phenomenon known as the Mother’s Curse. This phenomenon predicts the accumulation of male-only harming mutations in the mitochondrial genome due to its matrilineal inheritance. The release of Trojan Females - females carrying naturally occurring mitochondrial genotypes (mitotypes) that negatively impact male reproductive potential - facilitates self-sustaining transgenerational population suppression. A candidate TFT mitotype that induces male subfertility has recently been identified in seed beetles.
However, a crucial criterion for the successful application of TFT is that the TFT mitotype must render males subfertile across a range of nuclear and environmental backgrounds, such as different temperatures. To investigate this, we tested the effects of the candidate TFT mitotype (MG3b) and two control mitotypes (MG1a and MG1d) on male and female fertility in three different nuclear backgrounds and under two temperature regimes.
Our results show that the candidate TFT mitotype at standard temperature reduces male fertility across all nuclear environments compared to control mitotypes. Conversely, MG3b males have similar reproductive performance to MG1a and MG1d males at elevated temperatures. On the other hand, female beetles carrying MG3b remain fully fertile independently of nuclear background or temperature. The results in A. obtectus suggest that the sex-specific effect of mitochondrial haplotype on fertility depends on complex interactions between the mitochondrial DNA, the nuclear DNA and the thermal environment.Abstract Book: Congress of the European Sociey for Evolutionary Biology; 2025 Aug 17-22; Barcelona, Spain. European Society for Evolutionary Biology; 2025. p. 589
Unveiling the Balkans’ advances: In vitro biotechnology of woody plants in the early 21st century
No full textThe Balkan Peninsula is a European biodiversity hotspot, home to 6,500 native vascular plant species, many of which are endemic. The region has diverse range of climates and complex topography, creating conditions that suit many woody ornamental, fruit, and forest species. Nevertheless, climate change, habitat destruction, invasive species, plant diseases, and agricultural practices threaten natural ecosystems and cultivated species. Many Balkan countries have addressed these challenges using advanced biotechnological approaches, including micropropagation, in vitro conservation, and in vitro selection for stress-tolerant genotypes. This paper provides a comprehensive overview of in vitro plant biotechnology progress in the Balkan countries from the beginning of the 21st century to the present, with a focus on woody horticultural and forest species. The primary objectives of scientific research include optimizing media composition for all components and micropropagation stages, as well as effective initial explant selection. In addition, temporary immersion bioreactors, synthetic seed technology, and cryopreservation techniques have been explored to enhance plant production and conservation. Scientific research for woody ornamentals and fruit tree species has progressed in Albania, Bulgaria, Croatia, Greece, and Serbia while remaining limited in other Balkan countries. Forest tree production research is currently conducted in Bulgaria, Greece, and Serbia, with labs focusing on micropropagation and ex situ conservation. In addition to advances in scientific research, several commercial companies operate in the Balkans, with establishments in Serbia, Greece, Bulgaria, and North Macedonia, that produce in vitro-derived planting material for fruit trees and woody ornamental plants. Despite this progress, research in the Balkan countries remains fragmented, emphasizing the need to strengthen regional collaboration and knowledge exchange to promote agricultural development processes and biotechnological applications in this region. This review represents the first exploration of this topic in the Balkans and successfully unites researchers from several countries. It highlights key scientific advances in in vitro biotechnology for woody plants, identifies challenges, and proposes solutions. The value of interdisciplinary collaboration is emphasized through the optimization of in vitro methodologies, the promotion of germplasm conservation, and the sustainable use of plant genetic resources in the Balkans
Diversity and distribution of black flies (Diptera: Simuliidae) in the Balkan Peninsula
No full textThese files contains revised data sets and code (R) used for preparation of the manuscript "Global drivers of the diversity and distribution of black flies (Diptera: Simuliidae) in the Balkan Peninsula". Data set 1 is organised as a single sheet table in .csv file format. Rows represents data for a single sampling location, while recorded species and variables for each locality are organised as columns. Data set 2 is organised as a single sheet table in .csv file format. Rows represents data for a single sampling location, while recorded species and variables for each locality are organised as columns with aditional variables for land use and bioclimatic variables. File Code R is a script for R software used for analyses in this study
Refugial habitats of paleoendemic Picea omorika: soil and plant attributes
No full textRefugial habitats occupied by palaeoendemic species provide valuable insights into the adaptive strategies of populations shaped by long-term climatic and environmental change. They also help to assess the conservation potential of refugia under future climate scenarios. In this study, two refugia of Picea omorika on Mount Tara, Serbia, Trenice (S) on serpentine substrate and Vranjak (NS) on non-serpentine substrate, were investigated in terms of soil properties (both under natural and climate-simulated conditions) and plant traits (leaf stoichiometry and morphophysiological traits). The soils at the two sites differed significantly in terms of nutrient content, total organic carbon, moisture content and trace metal concentrations, while the pH values were similar and both had a high Ca:Mg ratio. Serpentine (S) and non-serpentine (NS) populations differed significantly in trace metal concentrations in needles, soil-to-leaf stoichiometric patterns, morphophysiological traits and water status. The serpentine population had higher values in traits associated with a conservative strategy, such as specific leaf mass, relative water content and leaf thickness, suggesting a potential for better adaptation to future climatic conditions. Soil incubation in the climate chamber showed that trace metal concentrations remained stable, while other physico-chemical properties changed in different ways. Our findings highlight the importance of considering site-specific conditions and integrating more ecosystem indices (geological substrate, edaphic factors, functional plant traits) into climate-driven species distribution models
Food restriction counteracts the decrease in the number of parvalbumin-expressing neurons in the 5xFAD mouse model of Alzheimer's disease
No full textAlzheimer’s disease (AD), characterized by amyloid-β (Aβ) accumulation, synaptic dysfunction, and progressive neuronal loss, disrupts inhibitory networks critical for cognitive function. Cortical parvalbumin-expressing (PV) interneurons, essential for maintaining inhibitory-excitatory balance, are highly vulnerable to Aβ toxicity, contributing to network hyperexcitability and cognitive decline. Dietary interventions like every-other-day (EOD) feeding show promise in mitigating age-related neurodegeneration, but their impact on PV interneuron survival in AD remains unclear. This study investigates EOD feeding effects on PV interneurons and neurotrophic signaling in female 5xFAD mice, a model of amyloid pathology. Mice were assigned to ad libitum (AL) or EOD feeding regimen from 2 months of age. Immunohistochemical analysis in 6-month-old mice revealed significant PV interneuron loss in retrosplenial, parietal, and somatosensory cortices of AL-fed 5xFAD mice, paralleling reduced brain-derived neurotrophic factor (BDNF). Strikingly, EOD feeding restored PV neuron density to non-transgenic levels, despite further BDNF reduction in Tg-EOD mice. Western blot analysis also showed increased TrkB receptor levels in Tg-EOD mice, suggesting compensatory neurotrophic signaling, while phosphorylated TrkB (pTrkB) decreased only in Tg-AL mice. These findings demonstrate that EOD feeding preserves PV interneurons in Aβ-rich environments, potentially stabilizing inhibitory networks. However, persistent deficits in downstream plasticity markers (e.g., pCaMKII, CBP) highlight unresolved synaptic dysfunction. The study underscores EOD feeding as a non-pharmacological strategy to counteract early AD related PV interneuron loss, though synergistic approaches may be needed to address broader molecular deficits. These insights advance understanding of dietary interventions in AD and their potential to modulate GABAergic vulnerability.Society for Neuroscience meeting: Neuroscience 2025; 2025 Nov 15-19; San Diego, SAD. Society for Neuroscience; 2025. PSTR111.11/D32