A search of Brassica SI-involved orthologs in buckwheat leads to novel buckwheat sequence identification: MLPK possibly involved in SI response

Kod biljaka cvetnica postoje genetički određeni sistemi self-inkompatibilnosti (SI), koji sprečavaju samooprašivanje i ukrštanje u srodstvu održavajući genetičku raznovrsnost vrsta. SI se javlja u dva oblika, kao gametofitna i sporofitna SI, koje se razlikuju u načinu određivanja SI fenotipa polena - kod GSI je SI fenotip polena određen polenovim sopstvenim haploidnim genomom, dok je kod SSI određen dipolidnim genotipom majke biljke. SSI se javlja kao homomorfna (jedan tip cveta u biljaka jedne vrste) i heteromorfna (dva ili tri tipa cveta u biljaka jedne vrste). Heteromorfna SSI je u poređenju sa homomorfnom SSI i GSI izuzetno malo proučena i za sada je upoznavanje na molekularnom nivou tek započelo. Kod heljde je prisutna distilna heteromorfna SSI, o kojoj je sakupljeno dosta podataka na fiziološkom nivou, ali o kojoj za sada nema molekularnih podataka. Na osnovu fiziološke sličnosti SI odgovora biljaka rodova Brassica i Prunus sa tram i pin morfom heljde, respektivno, zatim na osnovu toga što postoje dokazi da slični biohemijski mehanizmi leže u osnovi različitih SI odgovora i na osnovu toga što i evolutivno udaljene SI vrste mogu posedovati iste ili slične predačke SI gene, mi smo odlučili da ispitamo prisustvo ortologih gena uključenih u SI odgovore Brassica i Prunus u genomu heljde. Upotrebom izrođenih prajmera dizajniranih na osnovu evolutivno očuvanih regiona SRK, SLG, SP11 i MLPK sekvenci Brassica rapa, kao i S-RNaza i SFB gena roda Prunus, dostupnih u NCBI bazi podataka, ispitano je prisustvo ortologa ovih gena u genomu heljde. Takođe je prisustvo S-RNaza ispitano u proteinskim izolatima neoprašenih i kompatibilno i inkompatibilno oprašenih tučkova heljde oba morfa. Rezultati su pokazali da nema ortologa SRK, SLG, SP11, kao ni S-RNaza i SFB u genomu heljde, ali da postoji MLPK ortolog kod heljde. Izvedena aminokiselinska sekvenca pokazala je 80 % sličnosti sa MLPKf2 sekvencom Brassica rapa i APK1A Arabidopsis thaliana, potvrđujući da su u pitanju ortolozi koji bi mogli da imaju i sličnu ulogu. Naš sledeći korak je dobijanje cele nukleotidne sekvence MLPK heljde uz is- pitivanje postojanja alternativnih mesta iskrajanja i određivanje nivoa ekspresije po tkivima, kao i ispitivanje moguće uloge u SI odgovoru heljde. Ovi odgovori omogućiće bolje upoznavanje heteromorfnih SSI sistema koji su još uvek u svojoj najranijoj fazi istraživanja i obezbediće podatke nužne za uvid u evoluciju SI sistema biljaka cvetnica. Najzad, rasvetljavanjem SSI sistema heljde, koja se koristi u ishrani, biće moguće genetički kontrolisati ukrštanje heljde i dobijanje linija sa željenim hranljivim i/ili fiziološkim osobinama.Self-incompatibility (SI) systems, gamethophytic (GSI) and sporophytic (SSI), prevent self-pollination in angiosperms. Buckwheat displays heteromorphic SSI, with pollination allowed only between different flower morphs - thrum and pin. The physiology of thrum and pin morph SI responses are entirely different, resembling homomorphic Brassica SSI and Prunus GSI responses, respectively. Considering angiosperm species may share ancestral SI genes, we examined the presence of Brassica and Prunus SI-involved gene orthologs in the buckwheat genome. We did not find evidence of SRK, SLG and SP11 Brassica or S-RNase and SFB Prunus orthologs in the buckwheat genome, but we found a Brassica MLPK ortholog. We report the partial nucleotide sequence of the buckwheat MLPK and discuss the possible implications of this finding

Uticaj ekstrakta rizoma i korena kukureka (Helleborus odorus W. et K.) na parametre bele krvne slike i stepen fagocitoze kod wistar pacova

The objective of this research was to study the effects that the extract of rhizome and root of Helleborus odorus W. et K. (Ranunculaceae) can have on modifications in the parameter values of white blood cells count and degree of phagocytosis by peritoneal macrophages and neutrophil granulocytes in Wistar rats. The trial was conducted on 28 rats divided into 4 groups with 7 animals in each group. To the control group of rats sterile physiological solution in the quantity of 0.25 mL/100 g BW was applied intramuscularly. For the purpose of monitoring the effect of the extract of rhizome and root of hellebore (HE) during a time period, the HE was applied intramuscularly to rats in a dose of 10 mg/100 g BW, while the blood samples for analysis were taken after 24h, 48h and 72h. The consequence of intramuscular application of HE was an increased count of total leukocytes in all trial groups, the most expressed leukocytosis being registered 24h after application of HE. Statistically significant higher value in the count and percent of neutrophil granulocytes in the blood was recorded 24h after treatment in relation to the control and two other trial groups (p lt 0.001), among which a statistical significance was not established. The extract of hellebore rhizome and root has led to lymphopenia, resulting in the increase of the neutrophil/limphocyte index in the trial groups 24h and 48h after treatment. The application of HE had no significant effect on the count of monocytes in treated animals. The applied extract has caused a significant increase in the degree of phagocytosis by residing peritoneal macrophages and neutrophil granulocytes in blood.Cilj ovog rada je bio da se ispita uticaj ekstrakta rizoma i korena H. odorus W. et K. na promenu vrednosti parametara bele krvne slike i stepen fagocitoze od strane peritonealnih makrofaga i neutrofilnih granulocita kod pacova soja Wistar. Ogled je izveden na 28 pacova podeljenih u 4 grupe po 7 jedinki. Kontrolnoj grupi pacova je intramuskularno aplikovan sterilan fiziološki rastvor u količini od 0,25 ml/100 g TM. U cilju praćenja efekta ekstrakta rizoma i korena kukureka (EK) u toku vremena, pacovima je intramuskularno aplikovan EK u dozi od 10mg/100g TM, a krv za analizu je uzimana posle 24h, 48h i 72h. Intramuskularna aplikacija EK imala je za posledicu povećanje broja ukupnih leukocita u svim oglednim grupama, pri čemu je najizraženija leukocitoza registrovana 24h nakon aplikovanja EK. Statistički značajno veća vrednost broja i procenta neutrofilnih granulocita u krvi zabeležena je 24h posle tretmana u odnosu na kontrolnu i ostale dve ogledne grupe (p lt 0,001), između kojih nije utvrđena statistička značajnost. Ekstrakt rizoma i korena kukureka doveo je do nastanka limfopenije, što je imalo za posledicu povećanje neutrofilno/limfocitnog indeksa u oglednim grupama 24h i 48h nakon tretmana. Aplikacija EK nije značajno uticala na broj monocita kod tretiranih životinja. Upotrebljeni ekstrakt doveo je do značajnog povećanja stepena fagocitoze od strane rezidentnih peritonealnih makrofaga i neutrofilnih granulocita krvi

Transient expression in tobacco Bright Yellow 2 cells and pollen grains: A fast, efficient and reliable system for functional promoter analysis of plant genes

Ekspresija gena je regulisana posredstvom promotora-DNK sekvenci uzvodno od kodirajućeg regiona. Promotori svojom sekvencom određuju mesta vezivanja transkripcionih faktora, regulatora i RNK polimeraze. Strukturna i funkcionalna analiza promotora neophodna je za razumevanje mehanizama genske ekspresije. Cilj ovog rada je razvijanje brzog, efikasnog i pouzdanog sistema za testiranje bazalne promotorske aktivnosti kao i otkrivanje sekvenci polen-specifičnih elemenata promotora. U ovom radu je testirana funkcionalnost promotora za metalotionein heljde, posredstvom histohemijskog GUS-eseja u dva tranzijentna sistema za ekspresiju: BY2 ćelijama i polenovim zrnima. U oba sistema je uočena izražena aktivnost ispitivanog promotora.Gene expression is mediated by DNA sequences directly upstream from the coding sequences, recruited transcription factors and RNA polymerase in a spatially-defined manner. Understanding promoter strength and regulation would enhance our understanding of gene expression. The goal of this study was to develop a fast, efficient and reliable method for testing basal promoter activity and identifying core sequences within its pollen specific elements. In this paper we examined the functionality of buckwheat metallothionein promoter by a histochemical GUS assay in two transient expression systems: BY2 cells and pollen grains. Strong promoter activity was observed in both systems

Electrodeposition of cobalt powders with novel three-dimensional structure

Novel three-dimensional cobalt powder structures were successfully prepared by electrodeposition. Electrodeposited cobalt powder was characterized by scanning electron microscopy (SEM) and light microscopy. It was possible to control the morphology and structure of cobalt particles by adjusting process parameters of electrodeposition such as current density and type of working electrode. The morphology and structure of cobalt powders were investigated and the formation mechanism of agglomerate was also discussed

Influence of Ni2+/Co2+ ratio in electrolyte on morphology, structure and magnetic properties of electrolytically produced Ni−Co alloy powders

Nickel−cobalt (Ni−Co) alloy powders were produced galvanostatically by using sulphate electrolytes with various ratios of Ni2+/Co2+ (mole ratios). The morphology, phase structure, chemical composition and magnetic properties were examined by scanning electron microscope (SEM), X-ray diffractometer (XRD), atomic emission spectrometer (AES), and SQUID-based magnetometer, respectively. Morphology of the particles changed from cauliflower-like and dendritic to coral-like and spongy-like ones with increasing Ni2+/Co2+ ratio from 0.25 to 4.0. XRD analysis of the Ni−Co powders revealed that the decrease of Ni2+/Co2+ ratios (the increase of Co content) caused a change of structure from face centered cubic (FCC) obtained for the ratios of 4.0, 1.5 and 0.67 to a mixture of FCC and hexagonal closed-packed (HCP) phases for the ratio of 0.25. The increasing content of nickel led to change of mechanism of electrolysis from irregular (up to ~40 wt.% Ni in the electrolytes) to close to equilibrium (between ~40 and 60 wt.% Ni in the electrolytes) and anomalous co-deposition (over 60 wt.% Ni in the electrolytes) type. All of the obtained Ni−Co alloy samples behaved as soft magnetic materials while their magnetic parameters showed immediate composition dependence since both coercivity and saturation magnetization almost linearly increased with increase of the Co content

Uticaj parametra i režima elektrohemijskog taloženja na morfologiju i strukturu dendrita kalaja

Tin dendrites were synthesized via electrochemical route from the alkaline electrolyte applying both potentiostatic and galvanostatic regimes of the electrodeposition. Various both cathodic potentials and amounts of the passed electricity were used for Sn electrodeposition in the potentiostatic regime. Morphology and structure of synthesized tin dendritic particles were characterized by scanning electron microscopy (SEM) and X-ray diffraction (XRD) techniques, respectively. Depending on the applied cathodic potential, morphology of tin dendrites changed from the needle-like and the spear-like to very ramified dendrites of various shape. The branchy Sn dendrites were of two dimensional (2D) shape constructed from stalk and branches developed from it (primary (P) branches), classifying them into 2D(P) type according to Wranglen`s definition of a dendrite. The XRD analysis of produced tin particles showed that the needle-like and the spear-like dendrites represented monocrystals (200) orientation, while Sn crystallites in the potenciostatically obtained 2D(P) dendrites were predominately oriented in (440) plane. Morphology of tin particles was also correlated with polarization characteristics for this system, confirming belonging tin to the group of the normal metals, characterized by the high values of the exchange current density and overpotential for hydrogen evolution reaction, and by the low melting point.Dendriti kalaja su bili sintetizovani elektrohemijskim putem iz alkalnog elektrolita primenjujući i potenciostatski i galvanostatski režim elektrohemijskog taloženja. Elektrohemijsko taloženje kalaja u potenciostatskom režimu je vršeno na različitim katodnim potencijalima i sa različitim količinama naelektrisanja. Morfologija i struktura sintetizovanih dendritičnih čestica kalaja su okarakterisane tehnikom skenirajuće elektronske mikroskopije (SEM) i rentgensko difrakcionom analizom. U zavisnosti od primenjenog katodnog potencijala, morfologija dendrita kalaja se menjala od igličastih i dendrita nalik koplju do veoma razgranatih dendrita. Veoma razgranati kalajni dendriti su bili dvodimenzionalnog (2D) oblika izgrađeni od stabla i grana razvijenih iz stabla (primarne (P) grane), klasifikujući ih u 2D(P) tip prema Vranglenovoj definiciji dendrita. Rentgensko-difrakciona analiza je pokazala da igličasti i dendriti nalik koplju su predstavljali monokristale (200) orijentacije, dok su kristaliti kalaja u potenciostatski dobijenim 2D(P) dendritima bili predominantno orijentisani u (440) ravni. Morfologija čestica kalaja je bila takođe korelisana sa polarizacionim arakteristikama za ovaj sistem, potvrđujući pripadnost kalaja grupi normalnih metala okarakterisanih visokim vrednostima gustine struje izmene i prenapetosti za reakciju izdvajanja vodonika, i niskom tačkom topljenja

Correlation of Morphology and Crystal Structure of Metal Powders Produced by Electrolysis Processes

In this review paper, morphologies of metal powders produced by the constant (potentiostatic and galvanostatic) regimes of electrolysis from aqueous electrolytes are correlated with their crystal structure at the semiquantitative level. The main parameters affecting the shape of powder particles are the exchange current density (rate of electrochemical process) and overpotential for hydrogen evolution reaction. Depending on them, various shapes of dendrites (the needles, the two-dimensional (2D) fern-like, and the three-dimensional (3D) pine-like dendrites), and the particles formed under vigorous hydrogen evolution (cauliflower-like and spongy-like particles) are produced by these regimes of electrolysis. By decreasing the exchange current density value, the crystal structure of the powder particles is changed from the strong (111) preferred orientation obtained for the needle-like (silver) and the 2D (lead) dendrites to the randomly orientated crystallites in particles with the spherical morphology (the 3D dendrites and the cauliflower-like and the spongylike particles). The formation of metal powders by molten salt electrolysis and by electrolysis in deep eutectic solvents (DESs) and the crystallographic aspects of dendritic growth are also mentioned in this review

Morphological and crystallographic characteristics of electrodeposited lead from a concentrated electrolyte

Electrodeposition of lead from a concentrated nitrate solution was examined by scanning electron microscopy (SEM) and by X-ray diffraction (XRD) analysis of the obtained powder particles. Single crystals of the (111) preferred orientation were formed at a low overpotential by ohmic controlled electrodeposition. Irregular crystals, needle-like and fern-like dendrites, predominantly of the (111) preferred orientation, were formed at high overpotentials (the diffusion control of the electrodeposition). The ratio of Pb crystallites oriented in the (200), (220), (311) and (331) planes increased with increasing electrodeposition overpotential. The correlation between the morphologies and crystallographic structures of the lead deposits was discussed by the consideration of general characteristics of growth layers in electrodeposition processes

Detection and genotyping of classical swine fever virus isolates in Serbia

Classical swine fever (CSF) is a highly contagious disease of pigs leading to significant economic losses worldwide. Classical swine fever virus can be classified into three genogroups, each consisting of three or four subgroups. However, there is a lack of knowledge on the genotypes of CSFV isolates in Republic of Serbia. This study, based on the sequences analysis of partial E2 gene and 5' non coding region (NCR) of 15 CSFV isolated during 2006-2008 from domestic pigs, revealed that all were clustered into genetic group 2.3. Additionally, we showed that the two most often used real time RT-PCR assays were able to detect all local CSF viruses circulated in Serbia in the last years during intensive vaccination campaign against CSF. [Projekat Ministarstva nauke Republike Srbije, br. TR 31075 and TR 31088

Morphology and Structure of Electrolytically Synthesized Tin Dendritic Nanostructures

The formation of tin dendritic nanostructures by electrolysis from the alkaline electrolytehas been investigated. Morphology and structure of Sn dendrites produced applying both potentiostatic and galvanostatic regimes of the electrolysis are characterized by SEM and XRD, respectively. Depending on the applied cathodic potentials, three types of Sn dendrites were obtained: (a) needle like and spear-like, (b) fern-like, and (c) stem-like dendrites. The very branchy dendrites with branches of the prismatic shape obtained by the galvanostatic regime of electrolysis represented anovel type of Sn dendrites, not previously reported in the literature. To explain the formation ofvarious dendritic forms, correlation with the polarization characteristics for this electrodepositionsystem is considered. The needle-like and the spear-like dendrites represented monocrystals of(200),(400) preferred orientation, the fern-like dendrites exhibited the predominant (220),(440) preferredorientation, while in the stem-like particles Sn crystallites were oriented to a greater extent inthe (440) crystal plane than in other planes. The galvanostatically synthesized Sn particles possessed the strong (200),(400) preferred orientation. The strong influence of parameters and regimes ofelectrodeposition on structural characteristics of Sn dendrites is explained by the fundamental lawsof electrocrystallization taking into consideration the concept of slow-growing and fast-growingcrystal planes