Effect of salycilic acid on biochemical tolerance mechanisams of Impatiens walleriana (Balsaminaceae) to water stress

U prvom delu doktorske disertacije ispitivan je odgovor biljaka Impatiens walleriana na vodni stres indukovan polietilen glikolom (PEG) i uticaj salicilne kiseline (SA) u in vitro uslovima. Kulture izdanaka I. walleriana su gajene na hranljivoj podlozi koja je sadržala različite kombinacije i koncentracije PEG (1-3%) i SA (1-3 mM). Nakon dugotrajnog delovanja vodnog stresa na biljke I. walleriana utvrđen je uticaj PEG i SA na parametre hidratacije, indikatore oksidativnog stresa, kao i na parametre rastenja izdanaka. PEG indukovani vodni stres je uzrokovao redukciju rastenja, smanjenje mase svežih izdanaka, broja listova i izdanaka, relativnog sadržaja vode (RWC) i sadržaja hlorofila, kao i akumulaciju prolina, H2O2, i malondialdehida (MDA). Aktivnosti antioksidativnih enzima, katalaza (CAT), superoksid dismutaza (SOD) i peroksidaza (POX), su povećane kao odgovor na PEG tretman i to zavisno od primenjene koncentracije PEG. U drugom delu doktorske disertacije ispitivan je uticaj folijarne primene 2 mM SA na I. walleriana gajene ex vitro u uslovima suše. Biljke su podeljene u 4 grupe: zalivane biljke, zalivane biljke tretirane sa SA, nezalivane biljke i nezalivane biljke tretirane sa SA. Utvrđen je broj cvetnih pupoljaka i cvetova, RWC, sadržaj MDA, prolina, kao i aktivnosti SOD, CAT i POX u različitim vremenskim tačkama tokom trajanja eksperimenta. Suša je dovela do venjenja, odbacivanja cvetova, smanjenja RWC i porasta MDA. Kao odgovor na sušu kod I. walleraina je pojačana aktivnost hloroplastne Cu/ZnSOD i dve izoforme gvajakol peroksidaze. Identifikovane su tri de novo sekvence dehidrina korišćenjem transkriptoma iz listova I. walleriana: IwDhn1, IwDhn2.1 i IwDhn2.2. Suša je indukovala ekspresiju IwDhn2.1 i IwDhn2.2. Primena SA je pozitivno uticala na biljke koje su bile izložene suši, sprečavajući venjenje, održavajući RWC i aktivnost antioksidativnih enzima, ali nije imala uticaj na sprečavanje odbacivanja cvetova.In the first part of dissertation the responses of Impatiens walleriana to polyethylene glycol (PEG) induced water stress and the potential of exogenous salicylic acid (SA) as stress-ameliorating agent was described. Impatiens shoot culture was established on medium containing different concentrations and combinations of PEG (1–3% ) and SA (1–3 mM). After prolonged drought, water relation parameters, oxidative stress indicators, and growth responses of the shoots to PEG and/or SA were recorded. PEG reduced growth, fresh weight of shoots, the number of developed leaves and shoots, relative water content (RWC), and chlorophyll content. PEG caused accumulation of proline, H2O2, and malondialdehyde (MDA). The activities of catalase (CAT), superoxide dismutase (SOD), and peroxidase (POX) were increased in response to PEG in a dose-dependent manner. In the second part of dissertation it was investigated whether the foliar application of 2 mM SA could protect ex vitro grown I. walleriana plants from drought stress. The plants were divided into 4 groups: watered plants, drought-stressed plants, watered plants treated with SA and drought-stressed plants treated with SA. The number of flower buds and flowers, RWC, contents of MDA, proline and the activities of superoxide dismutases, catalases and peroxidases were recorded at different time points. Three dehydrin sequences were identified in de novo assembled leaf transcriptome: IwDhn1, IwDhn2.1 and IwDhn2.2. Drought stress caused wilting, floral abortion, reduction of RWC and increased MDA. In response to drought, I. walleriana induced chloroplastic Cu/ZnSOD and two guaiacol peroxidase isoforms. The remarkable drought response was induction of IwDhn2.1 and IwDhn2.2. SA had ameliorating effects on plants exposed to drought, including prevention of wilting, preservation of RWC, modulation of antioxidative activities, but had no effects on flowers preservation

Krioprezervacija vrhova izdanaka Impatiens hawkeri W. Bull

Impatiens hawkeri (Balsaminaceae) je ukrasna biljna vrsta poreklom sa Nove Gvineje. Krioprezervacija predstavlja metod za dugotrajno čuvanje biljnog materijala na -196C u tečnom azotu koji se koristi i za eliminaciju patogena kod biljaka.1 Vrste roda Impatiens su izrazito osetljive na tretmane dehidratacije koji prethode zamrzavanju.2 Takođe, koncentracija saharoze u hranljivim podlogama utiče na morfogenetski potencijal tokom gajenja u kulturi in vitro.3 U ovom radu smo ispitivali uticaj prekulture vrhova izdanaka na povišenoj koncentraciji saharoze (6%), kao i vitamina C na preživljavanje i regeneraciju biljaka posle hemijske dehidratacije vrhova izdanaka sa rastvorima za vitrifikaciju (PVS2 i PVS3). Vrhovi izdanaka I. hawkeri su izrazito osetljivi na tretman sa 100% PVS2 rastvorom. Najveći morfogenetski potencijal posle izlaganja PVS2 rastvoru su pokazali vrhovi tretirani sa 50% PVS2 rastvorom kada je dolazilo samo do kalusiranja vrhova izdanaka. Potpuna regeneracija biljaka posle PVS2 tretmana postignuta je uz dodatak vitamina C u hranljivoj podlozi u koncentraciji od 100 mg/l. Najbolji rezultati preživljavanja i regeneracije biljaka posle odmrzavanja postignuti su korišćenjem metode vitrifikacije sa 100% PVS3 rastvorom u trajanju od 45 min. Izdanci gajeni na hranljivoj podlozi sa 6% saharoze su pokazali veće preživljavanje (49,4%) posle izlaganja ultra niskim temperaturama u odnosu na izdanke gajene na podlogama sa 3% saharoze (26,5 %)

Cryopreservation of Viola cornuta shoot tips using vitrification procedure

Cryopreservation represents a suitable method for long term storage of different plant genetic resources. The aim of this study was to develop protocol for cryopreservation of Viola cornuta shoot tips using one step freezing method with chemical dehydration of tissue with modified Plant Vitrification Solutions (PVS2 or PVS3). Shoot tips (1-2 mm) of two-week cold acclimated shoots were cultured on ½MS medium with 0.3 M sucrose for one day before treatment with loading solution (2 M glycerol, 0.4 M sucrose) for 30 min. Osmotic dehydration with PVS2 solution (30% glycerol, 15% ethylene glycol and 15% DMSO in liquid ½MS medium with 0.4 M sucrose) were tested at 0 °C or 24 °C. Osmotic dehydration with PVS3 (50% sucrose, 50% glycerol in liquid ½MS medium) were tested at 24 °C for 45 min. After the treatment the explants were directly immersed in liquid nitrogen (LN) for at least one day. Re-warming was performed at 42 °C in water bath for 2 min. After re-warming, the PVS solutions were replaced with unloading solution containing 1.2 M sucrose for 20 min. Re-warmed shoot tips were cultured on ½MS medium with 0.1 mg L-1 BAP. We observed that PVS2 solution is cytotoxic for V. cornuta shoot tips and cannot be used for cryopreservation. However, cryopreservation with PVS3 solution was successful, where 71.9-100% shoot tips survived treatment before immersion to LN and 31-40% survived after re-warming from LN. Regrowth of cryopreserved shoot tips with new well-formed leaves was obtained after four weeks of culture

Pedijatrijski nefrotski sindrom: međusobna interakcija oksidativnog stresa i inflamacije

Background: The pathophysiological mechanisms crucial in the development of nephrotic syndrome (NS) in the pediatric population are still not fully understood. This study aimed to investigate the relationship between hypertension, oxidative stress, and inflammation in pediatric patients during the acute phase of the disease. Methods: The study included 33 children, aged 2 to 9 years, with nephrotic syndrome. Blood samples were collected during the acute phase and remission. Parameters of oxidative status were determined, including total oxidative status (TOS), advanced oxidation protein products (AOPP), prooxidant-antioxidant balance (PAB), sulfhydryl groups (- SH), paraoxonase 1 (PON1), and total antioxidant status (TAS) in serum, measured spectrophotometrically. Inflam- matory parameters such as pentraxin 3 (PTX3), leptin, programmed cell death ligand 1 (PD-L1), and E-cadherin were determined using enzyme-linked immunosorbent assay (ELISA). Results: Patients with nephrotic syndrome and hypertension had significantly higher levels of advanced oxidation protein products and total antioxidant status (p=0.029 and p=0.003, respectively). During the acute phase of the dis- ease, lower activity of sulfhydryl groups and paraoxonase 1 was observed compared to remission (p<0.001, for both). Pentraxin 3 levels were higher, while leptin levels were lower during the acute phase (p<0.001, for both). Pentraxin 3 correlated with advanced oxidation protein products and total antioxidant status during the acute phase but not in remission (r s =0.42, p=0.027 and r s =0.43, p=0.025, respectively). A negative correlation between Advanced oxidation protein products and leptin was observed during the acute phase, which disappeared in remission (rs=-0.42, p=0.028). Conclusions: Results of this study show that hypertension influences oxidative stress markers, and decreased antioxi- dant capacity may contribute to nephrotic syndrome devel- opment. Pentraxin 3 appears as a potential disease activity marker, indicating a dynamic connection between inflam- mation and oxidative stress. Leptin may also play a role in oxidative stress in nephrotic syndrome.Uvod: Patofiziološki mehanizmi ključni u razvoju nefrotskog sindroma (NS) u pedijatrijskoj populaciji još uvek nisu u potpunosti razjašnjeni. Ova studija ima za cilj proučavanje sinergističkog delovanja oksidativnog stresa i inflamacije u patogenezi NS. Takođe, jedan od ciljeva ove studije je i ispitivanje veze hipertenzije sa stepenom oksidativnog stresa i inflama - cije kod pacijenata u akutnoj fazi bolesti. Metode: U studiju je uključeno 33 dece sa NS uzrasta od 2 do 9 godina. Uzorci krvi su prikupljeni tokom akutne faze i remisije. Od parametara oksidativnog statusa određivani su: totalni oksidativni status (TOS), uznapredovali proizvodi oksidacije proteina (AOPP), balans prooksidans-antioksidans (PAB), sulfhidrilne grupe (-SH), paraoksonaza 1 (PON1) i ukupan antioksidativni status (TAS) u serumu su mereni spektrofometrijski, a od parametara inflamacije su pentraksin 3 (PTX3), leptin, ligand programirane smrti ćelije 1 (PD-L1) i E-kadherin određivani metodom enzimskog imunosorbentnog testa (ELISA). Rezultati: Pacijenti sa NS i hipertenzijom imali su značajno više nivoe AOPP i TOS (p=0.029 i p=0.003, respektivno). U akutnoj fazi bolesti su uočene nižu aktivnost -SH i PON1 u poređenju sa remisijom (p<0.001, za oba). Nivoi PTX 3 su bili viši, dok su nivoi leptina bili niži tokom akutne faze (p<0.001, za oba). PTX 3 je korelirao sa AOPP i TAS u akutnoj fazi, ali ne i u remisiji (rs=0.42, p=0.027 i rs=0.43, p=0.025,respektivno). U akutnooj fazi utvrđena je negativna korelacija između AOPP i leptina, koja je nestala u remisiji (rs=-0.42, p=0.028). Zaključak: Rezultati ove studije ukazuju da hipertenzija utiče na markere oksidativnog stresa, a smanjeni antioksidativni kapacitet može doprineti razvoju NS. PTX3 se pojavljuje kao potencijalni marker aktivnosti bolesti, što ukazuje na dinamičku vezu između inflamacije i oksidativnog stresa. Leptin može igrati ulogu u oksidativnom stresu u NS

Effect of salycilic acid on biochemical tolerance mechanisams of Impatiens walleriana (Balsaminaceae) to water stress

U prvom delu doktorske disertacije ispitivan je odgovor biljaka Impatiens walleriana na vodni stres indukovan polietilen glikolom (PEG) i uticaj salicilne kiseline (SA) u in vitro uslovima. Kulture izdanaka I. walleriana su gajene na hranljivoj podlozi koja je sadržala različite kombinacije i koncentracije PEG (1-3%) i SA (1-3 mM). Nakon dugotrajnog delovanja vodnog stresa na biljke I. walleriana utvrđen je uticaj PEG i SA na parametre hidratacije, indikatore oksidativnog stresa, kao i na parametre rastenja izdanaka. PEG indukovani vodni stres je uzrokovao redukciju rastenja, smanjenje mase svežih izdanaka, broja listova i izdanaka, relativnog sadržaja vode (RWC) i sadržaja hlorofila, kao i akumulaciju prolina, H2O2, i malondialdehida (MDA). Aktivnosti antioksidativnih enzima, katalaza (CAT), superoksid dismutaza (SOD) i peroksidaza (POX), su povećane kao odgovor na PEG tretman i to zavisno od primenjene koncentracije PEG. U drugom delu doktorske disertacije ispitivan je uticaj folijarne primene 2 mM SA na I. walleriana gajene ex vitro u uslovima suše. Biljke su podeljene u 4 grupe: zalivane biljke, zalivane biljke tretirane sa SA, nezalivane biljke i nezalivane biljke tretirane sa SA. Utvrđen je broj cvetnih pupoljaka i cvetova, RWC, sadržaj MDA, prolina, kao i aktivnosti SOD, CAT i POX u različitim vremenskim tačkama tokom trajanja eksperimenta. Suša je dovela do venjenja, odbacivanja cvetova, smanjenja RWC i porasta MDA. Kao odgovor na sušu kod I. walleraina je pojačana aktivnost hloroplastne Cu/ZnSOD i dve izoforme gvajakol peroksidaze. Identifikovane su tri de novo sekvence dehidrina korišćenjem transkriptoma iz listova I. walleriana: IwDhn1, IwDhn2.1 i IwDhn2.2. Suša je indukovala ekspresiju IwDhn2.1 i IwDhn2.2. Primena SA je pozitivno uticala na biljke koje su bile izložene suši, sprečavajući venjenje, održavajući RWC i aktivnost antioksidativnih enzima, ali nije imala uticaj na sprečavanje odbacivanja cvetova.In the first part of dissertation the responses of Impatiens walleriana to polyethylene glycol (PEG) induced water stress and the potential of exogenous salicylic acid (SA) as stress-ameliorating agent was described. Impatiens shoot culture was established on medium containing different concentrations and combinations of PEG (1–3% ) and SA (1–3 mM). After prolonged drought, water relation parameters, oxidative stress indicators, and growth responses of the shoots to PEG and/or SA were recorded. PEG reduced growth, fresh weight of shoots, the number of developed leaves and shoots, relative water content (RWC), and chlorophyll content. PEG caused accumulation of proline, H2O2, and malondialdehyde (MDA). The activities of catalase (CAT), superoxide dismutase (SOD), and peroxidase (POX) were increased in response to PEG in a dose-dependent manner. In the second part of dissertation it was investigated whether the foliar application of 2 mM SA could protect ex vitro grown I. walleriana plants from drought stress. The plants were divided into 4 groups: watered plants, drought-stressed plants, watered plants treated with SA and drought-stressed plants treated with SA. The number of flower buds and flowers, RWC, contents of MDA, proline and the activities of superoxide dismutases, catalases and peroxidases were recorded at different time points. Three dehydrin sequences were identified in de novo assembled leaf transcriptome: IwDhn1, IwDhn2.1 and IwDhn2.2. Drought stress caused wilting, floral abortion, reduction of RWC and increased MDA. In response to drought, I. walleriana induced chloroplastic Cu/ZnSOD and two guaiacol peroxidase isoforms. The remarkable drought response was induction of IwDhn2.1 and IwDhn2.2. SA had ameliorating effects on plants exposed to drought, including prevention of wilting, preservation of RWC, modulation of antioxidative activities, but had no effects on flowers preservation

Implementing the concept of Albergo Diffuso through urban planning: the case of “venac” historic core in Sombor, Serbia

‘Albergo Diffuso’ is an innovative concept in urban planning and design, where a hotel is dispersed through urban fabric, usually occupying several historic buildings under one ownership in a smaller community – village, town or small city. In this way, Albergo Diffuso enables their guests to easily participate in local urban life. The roots of the concept are were related to the urban decline of small historic settlements in Italy in the late 20th century. Hence, this concept belongs to international efforts to regenerate small shrinking cities as a new normality in urbanisation. However, two concepts – Albergo Diffuso and shrinking cities – have not been adequately interlinked; usually the physical aspect (‘design’) of Albergo Diffuso has been in spotlight. The other important issues, such as the adaptations of local urban planning and regulative documents and their impact on local property market have not been properly considered. This is the presentation of one such attempt in the City of Sombor in northwestern Serbia. Sombor ‘old town’, known as “Venac/Coronet”, is under state protection as the best-preserved core of any medium sized city at national level. Nevertheless, the relative isolation of Sombor from major communication corridors has caused its significant economical and demographic shrinkage last decades. The recent development of cultural tourism along the nearby Danube has slowed this process, but it has also imposed new challenges for the urban heritage of Sombor, such as a collision between developing property market and the aforementioned protective measures for local heritage. One of the efforts of local authorities to mediate this is to create a new urban plan for “Venac”, where local urban experts propose innovative measures, including regulations for Albergo Diffuso projects. The aim is to explain work on this plan and expected effects on urban development and local property market. The Sombor case is even more important, as it can be a national pilot to address sustainable tourism development in shrinking cities.[https://plpr2024.bole.ed.tum.de

Changes of antioxidative enzymes in Impatiens walleriana L. shoots in response to genetic transformation

Impatiens walleriana L. shoots were inoculated with Agrobacterium rhizogenes A4M70GUS and the effects of genetic transformation on the catalase (CAT), superoxide dismutase (SOD) and peroxidase (POX) activities in wounded region of stems and unwounded leaves were evaluated 10, 24, 240 and 720 hours after inoculation. Following Agrobacterum infection activities of plant antioxidative enzymes changed in a time-dependent manner indicating that dynamic processes occurred during plant-Agrobacterium interaction, plant cell transformation and formation of hairy roots. Appearance of hairy roots on wound sites of shoots was observed ten days after inoculation with A. rhizogenes and the root induction frequency was 100%. Among selected hairy root lines significant differences in growth rate and biomass production were observed and an average 3-fold increase in biomass production was observed for the best growing hairy root line compared with the untransformed roots. PCR analysis showed presence of uidA, rolB, rolC and rolD genes in all analyzed I. walleriana L. hairy root lines, while amplification fragment of rolA gene was detected in 83.3% transformed lines. Efficient transformation protocol for I. walleriana L described in this work offer possibilities to generate hairy root cultures for in vitro propagation of plant viruses. [Projekat Ministarstva nauke Republike Srbije, br. TR-31019

ODREDIVANJE INTENZITETA BUKE NA TERITORIJI GRADA BANJA LUKA

Buka u zivotnoj sredini, ili kako se veoma cesto zove komunalna buka, definite se kao buka koju stvaraju svi izvori buke koji se javljaju u covjekovom okruzenju. Glavni izvori komunalne buke su izvore buke na otvorenom pro- storu i izvori buke u zatvorenom prostoru. Kontrola buke, odnosno zvuka se mjeri raznim velicinama i jedinicama, ali najcesci je intezitet koji se definite kao protok energije u jedinici i vremenu i kroz jedinicu povrsine. Cilj rada je da se na osnovu izvrsenih mjerenja nivoa buke na podrucju grada Banja Luka za petomjesecni period analizira odnos ekvivalentnog i vrsnog nivoa buke u zivotnoj sredini i izvrsiporedenje intenziteta bukeprema zakonskoj regulati- vi

Plant regeneration from in vitro-derived leaf and petiole explants of Viola cornuta L. 'Lutea Splendens'

In vitro-derived petiole and leaf explants of Viola cornuta L. 'Lutea Splendens' were cultured on half-strength Murashige and Skoog (MS) medium with several concentrations of 6-benzylaminopurin (BAP), 6-furfurylaminopurine (KIN), N-phenyl-N'(1,2,3-thidiazol-5-yl) urea (TDZ) or N-(2-chloro-4-pyridyl)-N'phenylurea (CPPU) alone or supplemented with indole-3-acetic acid (IAA) or a-naphthaleneacetic acid (NAA). The most efficient direct adventitious shoot induction (21%) without callus formation was obtained using petiole explants cultured on half-strength MS medium containing 2.02 μM CPPU. Significantly higher shoot multiplication was achieved on medium supplemented with TDZ (7.6 shoots) or CPPU (7.1 shoots) than on medium supplemented with NAA and BAP (2.9 shoots). Shoots were rooted on half-strength MS medium without plant growth regulators

Plant regeneration from in vitro-derived leaf and petiole explants of Viola cornuta L. 'Lutea Splendens'

In vitro-derived petiole and leaf explants of Viola cornuta L. 'Lutea Splendens' were cultured on half-strength Murashige and Skoog (MS) medium with several concentrations of 6-benzylaminopurin (BAP), 6-furfurylaminopurine (KIN), N-phenyl-N'(1,2,3-thidiazol-5-yl) urea (TDZ) or N-(2-chloro-4-pyridyl)-N'phenylurea (CPPU) alone or supplemented with indole-3-acetic acid (IAA) or a-naphthaleneacetic acid (NAA). The most efficient direct adventitious shoot induction (21%) without callus formation was obtained using petiole explants cultured on half-strength MS medium containing 2.02 μM CPPU. Significantly higher shoot multiplication was achieved on medium supplemented with TDZ (7.6 shoots) or CPPU (7.1 shoots) than on medium supplemented with NAA and BAP (2.9 shoots). Shoots were rooted on half-strength MS medium without plant growth regulators