Razlikovanje Rhizoctonia spp. Na osnovu antigenih osobina

Antigenic properties and serological relationship was investigated in binucleate and multinucleate Rhizoctonia spp. isolates from strawberries soybean, alfalfa and potato plants from Serbia, from Spain, anastomosis group testers and in strawberry roots inoculated with binucleate Rhizoctonia AG A and AG I. Two polyclonal antisera, unabsorbed and cross absorbed, were used in dot-immunobinding assay for these investigations. Antisera were produced against mycelial antigens of two isolates, which belong to different anastomosis groups (AG) of binucleate Rhizoctonia - AG A and AG I. Both unabsorbed antisera reacted positively with all tested Rhizoctonia spp. isolates, and the reaction was absent with control isolates (Pythium sp. Agaricus sp. and Fusarium sp). The results prove a close serological relationship among Rhizoctonia spp. isolates, and diversity between Rhizoctonia spp. and isolates from different taxonomic groups. Also, both unabsorbed antisera reacted with higher intensity with closely related antigens (belonging to the same AG) than with ones from another AG of binucleate Rhizoctonia or R. solani (multinucleate Rhizoctonia). After cross absorption specificity of the antisera was enhanced, especially with the antiserum raised against mycelial proteins of binucleate Rhizoctonia AG I. This antiserum reacted positively only with antigens from the same AG, after cross absorption with antigens from AG A of binucleate Rhizoctonia and from R. solani AG 2-2. It proved to be specific to AG I of binucleate Rhizoctonia, and able to differentiate isolates of this AG from others. In this way the serological homology among isolates of one AG was proven, and also the diversity among isolates which belong to different AGs of binucleate Rhizoctonia as well as isolates of R. solani.Antigene osobine i serološki medjuodnosi ispitivani su kod dvojedarnih i višejedarnih Rhizoctonia spp., izolovanih iz jagode, soje, lucerke i krompira prikupljenih na teritoriji Srbije, izolata iz Španije, standard izolata anastomoznih grupa i izolata prisutnih u zaraženom korenu jagode. Ispitivanja su obavljena dot-blot metodom primenom dva poliklonalna antiseruma, pre i posle unakrsne apsorpcije. Antiserumi su proizvedeni na miceliju dva izolata, pripadnika različitih anastomoznih grupa (AG) dvojedarnih Rhizoctonia spp. - AG A i AG I. Neapsorbovani antiserumi pozitivno su reagovali sa svim ispitivanim izolatima Rhizoctonia spp.,dok su reakcije izostale sa kontrolnim izolatima (Pythium sp., Agaricus sp. i Fusarium sp), što dokazuje serološku srodnost izmedju izolata Rhizoctonia spp., kao i serološku različitost Rhizoctonia spp. od izolata drugih taksonomskih grupa. Osim toga, reakcija oba neapsorbovana antiseruma bila je intenzivnija sa srodnim izolatima (pripadnicima iste AG), nego sa izolatima druge AG dvojedarnih Rhizoctonia ili R. solani (višejedarna Rhizoctonia) što govori o većoj serološkoj srodnosti u okviru jedne AG. Unakrsnom apsorpcijom antiseruma srodnost izolata iz iste grupe je i potvrdjena. Specifičnost antiseruma na ovaj način se povećava, što je naročito izraženo kod antiseruma proizvedenog na proteine micelije dvojedarne Rhizoctonia AG I. Ovaj antiserum je, nakon apsorpcije antigenom iz AG A i R. solani AG-2-2 pozitivno reagovao samo sa homologim antigenima (izolati iz AG I), odnosno ovaj antiserum može razlikovati izolate AG I dvojedarnih Rhizoctonia od ostalih Rhizoctonia spp. Na ovaj način potvrdjena je serološka srodnost izmedju izolata iste AG, i serološke razlike u odnosu na pripadnike drugih AG kako dvojedarnih Rhizoctonia spp. tako i R. solani

Antigene osobine kao taksonomski kriterijum za razlikovanje Alternaria spp. patogenih za mrkvu i peršun

Identification of Alternaria genus species is a very complicated process which demands broadly designed investigations and studying of great number of properties which together can be considered as satisfying taxonomic criteria. The main objective of these investigations was examining the possibilities of applying the antigenic characteristics of Alternaria spp. phytopathogenic fungi as a taxonomic criterion, as well as introducing the serological methods for their identification. Conducting the examination of Alternaria spp., pathogenic for Apiaceae plants in Serbia, several isolates were obtained and identified as Alternaria radicina, A. petroselini, A. dauci and A. alternata, based on the conventional mycological methods and host range, as well as on molecular detection and partial characterization. The investigation included 12 isolates from plant leaves, seeds and soil which were pathogenic mainly to carrot and parsley and were identified as A. radicina, A. petroselini, A. dauci and A. alternate. Investigated isolates were compared with each other, as well as with standard isolates for the mentioned species (a total of 5 isolates, originating from USA and EU). During the investigation of serological characteristics of Alternaria spp. firstly a polyclonal antiserum was prepared against one isolate from Serbia identified as A. dauci. This antiserum was specific to Alternaria genus while there was no reaction with antigens from other phytopathogenic fungi genera (Fusarium, Rhizoctonia and Agaricus). Antiserum titer, determined by slide agglutination test, was 1/32. Antigenic characteristics of Alternaria genus fungi were examined by Electro-Blot-Immunoassay serological method (EBIA, Western blot), i.e. their protein profiles were compared. Investigated Alternaria spp. isolates showed different protein band profiles in gel and on nitrocellulose paper, and the observed differences were in complete correlation with the results of the previous identification. All investigated isolates, both domestic and the standards, were similar to each other, and they could be correctly identified to the species level using EBIA. Besides grouping to the species level, antigenic characteristics indicated similarities and differences among the isolates within the same and different species, showing their complex relationships which properly reflect their diversity in nature. In all the previous investigations of Alternaria genus fungi up to now, there have been no data about their serological characteristics as possible taxonomic criteria. Introduction of this group of characteristics represents an important contribution both to the taxonomy and implementation of fast and accurate methods of phytopathogenic fungi identification.Identifikacija vrsta u okviru roda Alternaria je veoma složen proces koji zahteva široko postavljena ispitivanja i proučavanje većeg broja osobina koje zajedno predstavljaju zadovoljavajuće taksonomske kriterijume. Osnovni cilj ovog rada je bio da se ispita mogućnost primene antigenih osobina fitopatogenih gljiva iz roda Alternaria kao taksonomskog kriterijuma, kao i uvođenje seroloških metoda za njihovu identifikaciju. Proučavajući gljive iz roda Alternaria koje su patogene za gajene biljke iz fam. Apiaceae u našoj zemlji, dobijeno je više izolata koji su, na osnovu proučavanja konvencionalnim metodama i kruga domaćina, kao i molekularne detekcije i delimične karakterizacije, razvrstani u četiri vrste: Alternaria radicina A. petroselini, A. dauci i A. alternata. U ispitivanja je bilo uključeno 12 izolata poreklom sa lista, semena ili iz zemlje, koji su ispoljavali patogenost prvenstveno prema mrkvi i peršunu i identifikovani da pripadaju vrstama A. radicina, A. petroselini, A. dauci i A. alternata. Ispitivani izolati su međusobno upoređivani kao i sa standardima za navedene vrste (ukupno 5 izolata, poreklom iz SAD i EU). Prilikom ispitivanja seroloških osobina Alternaria spp., prvo je pripremljen poliklonalni antiserum na izolat iz Srbije, koji je identifikovan kao A. dauci. Antiserum je ispoljio specifičnost za rod Alternaria, dok nije reagovao sa antigenima iz drugih rodova fitopatogenih gljiva (Fusarium, Rhizoctonia, Agaricus). Titar antiseruma, ispitivan metodom aglutinacije na pločici, bio je 1/32. Antigene osobine gljiva iz roda Alternaria proučavane su primenom elektroforetsko-adsorpciono-imunoenzimske serološke metode (EBIA, Western blot), odnosno upoređivani su njihovi proteinski profili. Ispitivani izolati Alternaria spp. ispoljili su različite profile proteinskih traka u gelu i na nitroceluloznom papiru, a uočene razlike su, u potpunosti, odgovarale rezultatima prethodne identifikacije. Navedeni izolati, domaći kao i standardi, ispoljili su međusobne sličnosti i mogli su biti pravilno identifikovane do nivoa vrste, primenom EBIA. Pored grupisanja do nivoa vrsta, antigene osobine su ukazale na postojanje sličnosti i razlika između izolata unutar istih, odnosno različitih vrsta, ukazujući na njihove veoma složene međuodnose koji verno oslikavaju diverzitet postojanja ovih gljiva u prirodi. U dosadašnjim proučavanjima gljiva iz roda Alternaria, ne postoje podaci o njihovim serološkim osobinama kao mogućim taksonomskim kriterijumima. Uvođenje ove grupe osobina, predstavlja značajan doprinos kako u taksonomiji, tako i u primeni brzih i tačnih metoda identifikacije fitopatogenih gljiva

Prisustvo Alternaria spp. na semenu biljaka iz familije Apiaceae i njihov uticaj na nicanje

Considerable damping-off of the seedlings of several commercial Apiaceae plant species was observed in Serbia. The infection of a total of 48 seed samples of nine vegetable and spice plants with phytopathogenic Alternaria spp. was established using the deep-freeze-blotter method. Identification of Alternaria species was performed using both conventional methods and PCR. Four different plant-pathogenic Alternaria species were detected in Serbia: A. dauci, A. radicina, A. petroselini, and A. alternata, all of which caused reduction of carrot, parsley, parsnip, and celery seed emergence. Alternaria dauci, A. radicina, and A. petroselini were relatively more aggressive compared to A. alternata. Substantial seed infection levels and strong influence of Alternaria spp. on seed emergence indicated that production of Apiaceae seed needs to be improved in order to obtain pathogen-free seed.U proizvodnji nekoliko useva biljaka izfamilije Apiaceae, uočeno je intenzivno propadanje sejanaca u Srbiji. Primenom metode zamrzavanja na filter papiru, ustanovljena je zaraza ukupno 48 uzoraka semena 9 gajenih biljaka povrća i začinskih biljaka fitopatogenim vrstama izroda Alternaria. Identifikacija Alternaria spp. obavljena je primenom konvencionalnih metoda i PCR. U Srbiji je detektovano prisustvo ukupno četiri fitopatogene vrste izroda Alternaria: A. dauci, A. radicina, A. petroselini i A. alternata koje su izazvale smanjeno nicanje mrkve, peršuna, paštrnaka i celera. A. dauci, A. radicina i A. petroselini bile su relativno agresivnije u poređenju sa A. alternata. Značajan nivo zaraze semena, kao i ispoljeni uticaj Alternaria spp. na nicanje, ukazali su da bi proizvodnju semena vrsta familije Apiaceae trebalo poboljšati u cilju dobijanja zdravog semena

Viroze kukuruza

Over 40 plant viruses has been known to cause diseases of maize, but economically the most important yield looses, which in certain years can be total, are caused by viruses from Potyvirus genera, known to be aphid-transmitted in a non-persistant maner. The most important viruses, pathogens of maize, sugar cane and sorghum are considered to be Maize dwarf mosaic virus (MDMV), Sorghum mosaic virus (SrMV), Sugarcane mosaic virus (SCMV), and Johnsongrass mosaic virus (JGMV). In Serbia, the presence of three viruses as maize pathogens have been established, MDMV, SCMV and Barley yellow dwarf virus (BYDV), with MDMV proved to be the most widely distributed and economically important. MDMV has been detected for the first time in 1962 and the causal agent has been attributed the name Maize dwarf mosaic virus. For longer than two decades, MDMV has been considered to be a strain of previously described SCMV until the contemporary research has clarified that a majority of previously described strains of this virus in fact represents clearly defined separate plant virus species. Thus, four viruses within the Potyvirus genera capable of infecting maize, sugar cane and sorghum has been defined and widely accepted. Economic damage of MDMV is reflected as direct yield reduction followed by lower yield quality. MDMV control must include simultaneous practicing of several control measures. The most important is control of Johnson grass population since it represents the main source of infection, coupled with growing the resistant or tolerant hybrids, as well as selecting appropriate sowing period in order to avoid aphid migration flights from weeds to maize.Do sada je opisano više od 40 vrsta virusa infektivnih za kukuruz, ali ekonomski najznačajnije štete, koje pojedinih godina mogu potpuno da unište proizvodnju ove kulture, prouzrokuju virusi roda Potyvirus koji se na neperzistentan način prenose vašima. Najznačajniji Potyvirus-i infektivni za kukuruz, šećernu trsku i sirak su: virus mozaične kržljavosti kukuruza (Maize dwarf mosaic virus, MDMV), virus mozaika šećerne trske (Sugarcane mosaic virus, SCMV), virus mozaika sirka (Sorghum mosaic virus, SrMV) i virus mozaika divljeg sirka (Johnsongrass mosaic virus, JGMV). U Srbiji na kukuruzu je ustanovljeno prisustvo tri virusa: MDMV, SCMV i virus žute patuljavosti ječma (Barley yellow dwarf virus, BYDV). Najrasprostranjeniji i najčešće zastupljen virus u našoj zemlji je MDMV. Mozaična kržljavost kukuruza prvi put je opisana 1962. godine, a prouzrokovač tog oboljenja imenovan je kao virus mozaične kržljavosti kukuruza. Više od dve decenije, ovaj virus smatran je sojem ranije opisanog virusa mozaika šećerne trske, ali savremena istraživanja su pokazala da su mnogi prethodno opisani sojevi virusa mozaika šećerne trske zapravo jasno defi nisane posebne vrste. Izdvojena su četiri virusa u okviru roda Potyvirus koji su infektivni za kukuruz, sirak i šećernu trsku. Ekonomska štetnost MDMV ogleda se u smanjenju prinosa i lošijem kvalitetu zrna. Kontrola oboljenja koje MDMV izaziva podrazumeva istovremenu primenu više mera. Osnovne mere su uništavanje populacije divljeg sirka koji predstavlja osnovni izvor zaraze, zatim primena otpornih ili toleratnih hibrida, kao i odabir vremena setve u cilju izbegavanja preleta vaši sa korova na kukuruz

Viroze vrežastih kultura

There are over 200 known cucurbit diseases of diverse etiologies. Viruses are the most common causes of diseases affecting cucurbits, causing a signifi cant yield reduction. A large number of viruses have been reported to infect cucurbits. In Serbia, three economically signifi cant viruses are present on cucurbits: Zucchini yellow mosaic (ZYMV), Cucumber mosaic (CMV) and Watermelon mosaic (WMV). They cause very severe and destructive symptoms on the whole plants, leaves and fruit, leading to signifi cant yield losses. The main symptoms are: plant stunting, leaf deformation and severe young fruit deformation. These viruses are transmitted by several aphid species in a nonpersistent manner with variation in the virus prevalence. They occurred in single or mixed infections. Mixed infection are common in cucurbits causing a notable symptom enhancement due to synergistic effects. In addition to these viruses, Tomato spotted wilt tospovirus (TSWV) and Cucurbit aphid-borne yellows virus (CABYV) have been detected in some pumpkin growing sites in Serbia, whose frequency has varied over the years. Over the last two decade, intensive spread of virus infection of oilseed pumpkin and other cucurbits has resulted in significant losses in their crop production in Serbia.U usevu tikava širom sveta zabeleženo je preko 200 oboljenja različite etiologije. Virusi koji mogu da zaraze tikve su brojni i smatraju se najčešćim prouzrokovačima oboljenja tikava izazivajući značajno smanjenje prinosa. U našoj zemlji na tikvama prisutna su tri ekonomski značajna virusa: virus žutog mozaika cukinija (Zucchini yellow mosaic virus, ZYMV), virus mozaika krastavca (Cucumber mosaic virus, CMV) i virus mozaika lubenice (Watermelon mosaic virus, WMV). Najznačajniji virusi tikava izazivaju izražene i destruktivne simptome u vidu kržljavosti biljaka, deformacije lišća i izražene malformacije tek formiranih plodova, zbog čega su štete ogromne. Ovi virusi se prenose vašima na neperzistentan način, smenjuju se u pogledu prevalentnosti ili se često javljaju zajedno u kompleksnim zarazama. Kompleksne zaraze su česte u usevu tikava i imaju za posledicu značajno izraženije simtome usled sinergističkog delovanja. Pored ovih virusa, u Srbiji su na pojedinim lokalitetima gajenja tikava otkriveni i Tomato spotted wilt tospovirus (TSWV) i Cucurbit aphid-borne yellows virus (CABYV), čija se učestalost po godinama menjala. Tokom poslednje dve decenije intenzivno širenje virusa tikava u našoj zemlji dovelo je do značajnih gubitaka u proizvodnji uljane tikve i drugih tikava u Srbiji

Višejedarna Rhizoctonia sp. - patogen šećerne repe i osetljivost sorti u polju

Sugar beet root rot has severely occurred in our country recently, especially in localities of Pazova, Pećinci, Ruma, Sremska Mitrovica and Šid. From diseased roots as well as from soil collected from the localities where decay occurred, fungal isolates were obtained by bait plant method. Based on their characteristics, they were identified as multinucleate Rhizoctonia sp. During the year of 2004 in Mitrosrem trial field T-11, where the presence of multinucleate Rhizoctonia sp. was confirmed, an experiment under the coordination of Committee for Acknowledgement and Registration of New Cultivars in our country was conducted in order to determine cultivars' tolerance, i.e. their susceptibility and possibility for growing on infested fields. Six cultivars of sugar beet, Laetitia (as standard) and five new ones were included in the investigation. The trial was conducted in accordance with the established and accepted method (Ministry of Agriculture, Forestry and Water Resources, Republic of Serbia). Susceptibility of investigated cultivars was evaluated according to significant production characteristics root yield, sugar content, corrected sugar content, thick juice Q, molasses sugar, content of K, Na and amino-N, polarized sugar yield and white sugar yield, as it was recommended by the method. Conducted investigations have revealed that tested sugar beet cultivars showed different reactions to natural infection with multinucleate Rhizoctonia sp. Concerning root yield as the most important agricultural characteristic, statistically significantly higher yield was obtained with the cultivar under code mark 5 (61.120 kg/ha) whereas the cultivar marked under code 6 had significantly lower yield comparing to the standard (38.100 kg/ha).Poslednjih godina uočeno je masovno propadanje šećerne repe u našoj zemlji i to u lokalitetima Pazove, Pećinaca, Rume, Sremske Mitrovice i Šida. Iz obolelih korenova i iz zemljišta prikupljenog sa terena gde je propadanje uočeno, metodom mamaka izolovana je gljiva koja je po svojim osobinama identifikovana kao višejedarna Rhizoctonia sp. U toku 2004. godine na parceli Mitrosrema T-11 gde je izolacijom dokazano prisustvo višejedarne Rhizoctonia sp., postavljen je ogled u okviru sortne komisije za priznavanje i registraciju novih sorti u našoj zemlji sa ciljem utvrđivanja tolerantnosti, odnosno osetljivosti pojedinih sorti i time njihove pogodnosti za gajenje na infestiranom zemljištu. U ispitivanja je uključeno 6 sorti šećerne repe: Laetitia (kao standard) i još pet novih sorti. Ogled je posejan po utvrđenoj i prihvaćenoj metodi sortne komisije (Ministarstvo poljoprivrede, šumarstva i vodoprivrede Republike Srbije). Osetljivost ispitivanih sorti ocenjivana je na osnovu značajnih proizvodnih osobina: prinos korena, polarizacija, korigovani sadržaj šećera Q gustog soka, sadržaja sećera u melasi, sadržaja K, Na i amino N, prinos polarizacionog šećera i prinos kristalnog šećera, kako to metoda i zahteva. Na osnovu obavljenih istraživanja ustanovljeno je da se ispitivane sorte šećerne repe različito ponašaju u uslovima prirodne zaraze višejedarnom Rhizoctonia sp. U pogledu prinosa, kao najvažnije proizvodne karakteristike, statistički značajno viši prinos od standarda ispoljila je sorta koja se vodi pod šifrom 5 (61.120 kg/ha), dok je sorta koja se vodi pod šifrom 6 imala statistički značajno niži prinos u poređenju sa standardom (38.100 kg/ha)

ANALIZA STVARNIH I PLANIRANIH TROŠKOVA ODRŽAVANJA I UPORABE GRAĐEVINA

Analiza održavanja i uporabe građevina opisuje se kao skup trajnih aktivnosti koje sadrže tehnička i pridružena im administrativna djelovanja, kako bi se tijekom cijeloga vijeka uporabe građevini u cjelini i svim njezinim dijelovima osigurala razina služnosti u dostupnim (zadovoljavajućim) granicama. Uporabom metode proračuna troškova održavanja i uporabe u ranoj fazi projektiranja moguće je racionalizirati troškove održavanja i uporabe, planirati iste buduće troškove te spriječiti ili umanjiti utjecaj prekida poslovanja zbog potreba održavanja i funkcionalne uporabivosti. Rezultati proračuna troškova održavanja i uporabe građevina znatno ovise o duljini vremenskog razdoblja za koje se radi proračun i primijenjenoj diskontnoj stopi pri svođenju troškova na neto sadašnju vrijednost. U ovom radu predstavljena je studija slučaja kojom se uspoređuju stvarni i planirani troškovi desetogodišnjeg održavanja i uporabe građevine namijenjene fakultetskoj funkciji. Definiraju se aktivnosti planiranog preventivnog održavanja, reaktivnog održavanja i zamjene istrošenih materijala, te troškovi uporabe na osnovi kojih se uspoređuju isti troškovi za razdoblja od 10 i 50 godina

Synthesis, characterization and biological study of new dinuclear zinc(ii) and nickel(ii) octaaza macrocyclic complexes

Two new nitrato complexes of zinc and nickel with 1,4,8,11-tetrakis(2-pyridylmethyl)-1,4,8,11-tetraazacyclotetradecane (tpmc), have been synthesized and characterized. The IR spectral peaks showed that the coordinated and ionic nitrate ions are in agreement with the formula proposed by elemental analysis. Conductometric titrations predicted methanol to be a convenient solvent for synthesis and revealed the stoichiometry of the complexes, while molar electrical conductivities indicated a 1 : 3 complex electrolyte type for the zinc complex, and a 1 : 2 complex electrolyte type for the nickel complex. The optimized complex structure was obtained by molecular modeling and density functional theory calculations. The biological activity of the novel complexes was examined by screening eight different bacterial strains and two cancer cell lines. The zinc complex showed better antimicrobial activity against the bacterial strains, while the complexes did not show significance antiproliferative activity toward cancer cells MCF-7 and MDA-MB-231

Filamentous fungi isolated from grape marc as antagonists of botrytis cinerea

In this paper we report on the isolation and identification of three filamentous fungi from grape marc, and antifungal effect of their cell-free culture filtrates on the growth of Botrytis cinerea, causal agent of gray mold. Grape marc is a waste material that has been used as soil amendment in sustainable agriculture. Isolates originating from grape marc were identified on the basis of morphological features and internal transcribed spacer rDNA or beta-tubulin gene sequencing. The presence of three different species, Penicillium paneum, Penicillium chrysogenum and Aspergillus fumigatus has been detected expressing different effect on the growth of B. cinerea. The effect of crude culture filtrates of selected fungi on B. cinerea growth was tested. Heat sensitivity of the established inhibition effect was examined by autoclaving the crude culture filtrate prior to testing. Additional aim was to determine whether antifungal effect was influenced by previous exposure to B. cinerea in dual liquid cultures. Crude culture filtrate of A. fumigatus K16/2 showed the lowest suppression of B. cinerea growth. A maximal percentage inhibition achieved within the study was 38.2%, 39.8% and 23.8 for crude filtrates of P. paneum K7/1, P. chrysogenum K11/1 and A. fumigatus K16/2, respectively. Presence of B. cinerea in dual liquid culture induced significant increase in antifungal capacity of the culture filtrates in comparison to pure culture filtrates of the chosen isolates. The antifungal activity of all of the isolates' culture filtrates retained after heat treatment suggesting the presence of some thermostable antifungal metabolites. The results indicate the complexity and specificity of the interaction between filamentous fungi and B. cinerea. Grape marc is a good source for isolation od B. cinerea fungal antagonists and their antifungal metabolites. Specificity of fungal-fungal interactions suggests that further research on the antagonistic mechanisms and factors affecting them should be studied separately for each pair of antagonists