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Waitea circinata var. zeae, a pathogen with a relatively narrow host range, has recently been detected in cabbage and oilseed rape in Europe and worldwide. In this study, we developed specific conventional and real-time PCR protocols for direct detection of W. circinata var. zeae from mycelium and diseased plant tissue. The newly developed primer pair zeaefor1/zeaerew1, used in PCR protocols, specifically amplified only target isolates of W. circinata var. zeae when tested against isolates of 11 different binucleate and multinucleate anastomosis groups of Rhizoctonia spp. including AG-A, AG-G, AG-F, AG-U, AG-2-1, AG-2-2, AG-3, AG-4 HGI, AG-4 HGII, AG-4 HGIII, and AG-6 and common soil-borne pathogens. Total of nine previously published primer pairs designed for the detection of various Rhizoctonia spp. were also tested and did not amplify target isolates of W. circinata var. zeae. The detection limit of conventional and real-time PCR protocols was 10–2 and 10–5 (with starting concentration 9.5 ng/µl), respectively, and both methods are the first available tools for direct detection and identification of W. circinata var. zeae from mycelium and diseased oilseed rape seedlings. Both conventional and SYBR-Green-based real-time PCR protocols are cost-effective and provide a solid basis for further investigations of W. circinata var. zeae, particularly in relation to distribution, host range, and epidemiology.

Waitea circinata var. zeae (anamorf Rhizoctonia zeae) je značajan patogen uglavnom onokotiledonih biljaka, koji je prisutan u mnogim delovima sveta, a održava se u zemljištu duži niz godina. Poslednjih godina krug domaćina W. circinata var. zeae širi se i na neke dikotiledone biljke. U okviru nedavnih istraživanja u Srbiji W. circinata var. zeae je utvrđen kao prouzrokovač truleži korena i korenovog vrata, i propadanja biljaka kupusa i uljane repice na lokalitetima Futog i Rimski Šančevi. U radu su sistematizovani publikovani podaci o rasprostranjenosti, krugu domaćina, načinu održavanja i širenja, kao i suzbijanju W. circinata var. zeae, kao i rezultati istraživanja koja su obavljena u Srbiji.

Proučavanja fitopatogenih gljiva i pseudogljiva u našoj zemlji imaju dugu tradiciju, a početak se uglavnom povezuje sa osnivanjem Poljoprivredno-šumarskog fakulteta davne 1919. godine. Fitopatogene gljive su danas u svetu široko poznati patogeni koji pričinjavaju oko 70% šteta od svih biljnih bolesti. U Srbiji, fitopatogene gljive i pseudogljive takođe redovno nanose velike štete u biljnoj proizvodnji pa su među proizvođačima prepoznate kao važni uzročnici gubitaka koji se mogu sprečiti ili ublažiti samo uz podršku nauke. Po svemu, istraživanja fitopatogenih gljiva i pseudogljiva danas predstavljaju jednu dobro profilisanu i značajnu oblast primenjenih istaživanja.

Grey mould, which is caused by Botrytis cinerea Pers., is among the most damaging diseases of cultivated plants worldwide. The use of fungicides dominates in the protection of tomatoes against grey mould. Due to the emergence of resistance of pathogens to the active ingredients of fungicides, the possibility of using bacteria of the genus Bacillus was studied. A total of 666 isolates were isolated from different agricultural soils, and a preliminary screening dual culture test with Alternaria dauci was done. A total of 77 isolates were dual culture tested against Fusarium tricinctum and F. proliferatum. Based on preliminary dual culture testing, eight Bacillus isolates were selected for testing their biological potential against two isolates of B. cinerea. The effect of Bacillus spp. isolates was studied for seed germination and plant growth promotion (PGP) on the seedlings of tomato (Solanum lycopersicum L.). Using molecular identification (16S rDNA), five bacterial isolates were identified as Bacillus subtilis, two isolates as B. amyloliquefaciens, and one isolate as B. pumilus. Using the Internal transcribed spacer (ITS) region, two fungal isolates from an infected leaf and a tomato fruit were identified as B. cinerea. The results of the dual culture test showed that eight bacterial isolates could significantly inhibit the growth of B. cinerea mycelia from 50% to 80%. Two isolates B. amyloliquefaciens 28.3 and B. amyloliquefaciens 21/IV increased seed germination by 85.66% and 86.16%, respectively, compared to the control (82.66%). All bacterial isolates positively affected the morphological parameters of the seedlings. In the greenhouse experiment, three isolates B. subtillis 20.10, B. amyloliquefaciens 28.3, and B. amyloliquefaciens 21/IV and their mixture increased all measured PGP parameters. Isolates Bac 20.10 and Bac 28.3 reduced the severity of grey mould disease by 27.47% and 30.36%, respectively. These results show that Bacillus isolates have the potenti

Paradajz (Solanum lycopersicum L.) je značajna povrtarska vrsta čiju proizvodnju može ugroziti veliki broj bolesti umanjujući prinos i kvalitet plodova. Siva trulež paradajza, koju prouzrokuje gljiva Botrytis cinerea Pers., izaziva značajne štete na paradajzu, ali i na više od 1400 drugih domaćina širom sveta. Upotreba fungicida i dalje predstavlja najvažniju meru kontrole ovog patogena. Međutim, zabrinutost javnosti zbog ostataka fungicida u plodovima i pojava rezistentnosti patogena dovele su do stroži regulative u pogledu upotrebe fungicida. Stoga se pojavila potreba za izučavanjem alternativnih mera suzbijanja patogena, a jedna od njih je primena antagonističkih bakterija roda Bacillus kao bioloških agensa za kontrolu B. cinerea. Ciljevi istraživanja bili su ispitivanje antagonističkog potencijala tri autohtona soja Bacillus spp., poreklom iz Srbije, kao i njihove smeše prema izolatu B. cinerea 463-19 na listovima i plodovima paradajza u in vitro uslovima. Listovi i plodovi paradajza dezinfikovani su površinski 70% etil alkoholom u trajanju od 40 sekundi, a zatim tretirani prskanjem suspenzije pojedinačnog soja ili smešom sojeva Bacillus spp. približne koncentracije 107 ćel/ml. Suspenzije su pripremljene od bakterijske kulture stare 24h gajene u LB (engl. Lysogeny broth) podlozi. Nakon sušenja micelijarni diskovi B. cinerea, veličine 9 mm, dobijeni od kulture stare 14 dana gajene na PDA (engl. Potato Dextrose Agar-krompir dekstrozni agar) podlozi, postavljeni su u blizini lisnih nerava, ili na povređenu pokožicu ploda paradajza. Intezitet oboljenja (engl. disease severity-DS %) na listovima izračunat je prema formuli: DS(%)=((∑(broj zaraženih listova x intenzitet oboljenja))/(4xbroj ocenjenih listova))x100. Na plodovima paradajza ocenjivan je procenat smanjenja razvoja truleži. Listovi i plodovi inkubirani su pri 25 °C (12 h dan/12 hnoć), a ogled je ocenjen nakon 3 dana. Za svaki tretman u ogledu korišćeno je po 10 listova ili 6 plodova, u tri ponavljanja. U kon

Fungal diseases of cucurbits are caused by various fungi and fungus-like organisms which are frequent limiting factor in the production of cucurbit crops all over the world. Mycoses of cucurbits are usually categorized into three overlapping groups, disease of subterranean parts, diseases of aerial parts and fruit rots. Although each group comprises numerous, nonrelated fungal species with different taxonomic position, they share several epidemiological features demanding similar control strategies which can be successfully applied. Diseases of subterranean organs of cucurbits are caused by several soil-borne fungi and fungus-like organisms. The most frequent diseases of this group are seed rot and seedling root rot (caused by Pythium spp.), seedling damping-off and root and crown rot (Phytophthora spp., Rhizoctonia spp., Sclerotinia sclerotiorum), fusarium wilt (Fusarium spp.), verticilium wilt (Verticillium albo-atrum i V. dahliae) and other. The most common diseases of aerial parts, also known as diseases of cucurbits leaves are alternaria leaf spot and leaf blight (caused by Alternaria cucumerina, A. alternata), gummy stem blight (Didymella bryoniae), anthracnose (Colletotrichum orbiculare), downy mildew (Pseudoperonospora cubensis), powdery mildew (Podosphaera xanthii and Golovinomyces cichoracearum), cercospora leaf spot (Cercospora citrullina) and others. Diseases of cucurbit fruits are economically very important and are caused either as fi nal stage of several diseases of leaves or by fruit infection and colonization by soil-borne fungi. The most common causing agents worldwide are Phytophthora capsici, several Fusarium spp., Alternaria alternata, Rhizoctonia solani, Didymella bryoniae, Sclerotium rolfsii and others. Managing strategies for successful control of cucurbit diseases are based to target disease prevention, on one hand, and to slow down disease spreading. Starting the production with pathogenfree seed is crucial fi rst step in disease prevention, Mikoze vrežastih kultura mogu biti izazvane raznorodnim vrstama gljiva i pseudogljiva i čest su ograničavajući faktor u proizvodnji u mnogim delovima sveta. Uobičajeno je da se svrstavaju u tri osnovne grupe, bolesti podzemnih organa, bolesti nadzemnih organa i bolesti ploda pre i posle berbe. U svakoj od ovih grupa gljive i pseudogljive prouzrokovači su brojni i taksonomski veoma udaljeni, ali među njima postoje izvesne sličnosti u epidemiologiji zbog čega se za suzbijanje mogu primeniti neke relativno slične strategije. Više vrsta gljiva i pseudogljiva koje se održavaju u zemljištu (eng. soilborne fungi) obično su prouzrokovači bolesti podzemnih organa vrežastih kultura. Uobičajene bolesti podzemnih organa su: rano propadanje tokom klijanja i nicanja (prouzrokovači vrste roda Pythium spp.), propadanje ili topljenje sejanaca (Phytophthora spp., Rhizoctonia spp., Sclerotinia sclerotiorum), fuzariozno uvenuće i sušenje biljaka (Fusarium spp.), verticiliozna uvelost (Verticillium alboatrum i V. dahliae) i druge. Bolesti lista vrežastih kultura javljaju se ili isključivo na listovima ili tu započinju razvoj, a potom napadaju i vrežu i ostale biljne organe. Među značajnije bolesti lista vrežastih kultura ubrajaju se alternarijska pegavost lista (prouzrokovači Alternaria cucumerina, A. alternata), gumozno sušenje stabla (Didymella bryoniae), antraknoza (Colletotrichum orbiculare), plamenjača (Pseudoperonospora cubensis), pepelnica (Podosphaera xanthii i Golovinomyces cichoracearum), pegavost lista (Cercospora citrullina) i druge. Bolesti ploda vrežastih kultura su ekonomski veoma značajne i nastaju usled zaraze listova i stabla ili gljiva i pseudogljiva poreklom iz zemljišta sa kojima plod može biti u direktnom kontaktu. Najčešći prouzrokovači su Phytophthora capsici, vekoliko vrsta Fusarium spp., Alternaria alternata, Rhizoctonia solani, Didymella bryoniae, Sclerotium rolfsii i druge. Efi kasna kontrola bolesti vrežastih kultura zasniva se pre svega na preventivnim i ag

U periodu 2013-2020. godine, pregledom 22 useva na 15 lokaliteta i šest partija krompira u Srbiji, ustanovljena je pojava nekroze i truleži korena različitih biljaka. Sakupljeno je 330 uzoraka i dobijeno 177 izolata Rhizoctonia spp., koji su morfološki i molekularno razdvojeni u 11 anastomoznih grupa (AG). Rezultati su potvrđeni sekvenciranjem ITS regiona rDNA, kao i LSU, rpb2, tef-1α, atp6 i β-tubulin genskih markera. Detaljno su okarakterisana 64 izolata i generisano je 77 sekvenci koje su omogućile prvi uvid u diverzitet i strukturu populacije Rhizoctonia spp. u Srbiji. Kao prouzrokovači bolesti u Srbiji javljaju se dvojedarne Rhizoctonia spp. AG-A na jagodi, AG-G na paradajzu, trešnji i pasulju, AG-F na paprici, AG-U na jabuci i Poa sp., a od višejedarnih R. solani potvrđeno je prisustvo AG-2-1 na paprici, AG-2-2 na šećernoj repi, AG-3 na krompiru, AG-4 HGI na mrkvi, paprici, uljanoj repici i kupusu, AG-4 HGII na jabuci, AG-6 na malini, kao i Waitea circinata var. zeae na kupusu i uljanoj repici. Od toga, osam AG dokazano je u Srbiji prvi put ili prvi put na novom domaćinu u Srbiji, a osam AG je prvi put opisano na novim domaćinima u svetu. Filogenetske analize ukazale su na njihove međuodnose, na moguće puteve introdukcije i postojanje varijabilnosti u okviru roda Rhizoctonia u Srbiji. Razvijen je protokol za direktnu detekciju AG primenom specifičnih prajmera, kao i qPCR protokol za specifičnu detekciju R. solani AG-2-2. Ustanovljena je različita virulentnost i krug domaćina pojedinih AG inokulacijama 14 biljaka iz osam familija. Identifikovan je izolat Trichoderma harzianum sa agresivnošću prema različitim AG u in vitro i in vivo eksperimentima. Dobijeni rezultati pokazuju da su vrste roda Rhizoctonia veoma značajna i raznovrsna grupa patogena u Srbiji.

Kupus i uljana repica su dva vrlo značajna useva u našoj zemlji i njihova proizvodnja ugrožena je brojnim patogenim organizmima. Višejedarne anamorfne Rhizoctonia spp. sa teleomorfom opisanim kao Waitea circinata su biljni patogeni koji se dugo održavaju u zemljištu i rasprostranjeni su i vrlo štetni širom sveta. Od pet opisanih varijeteta W. circinata var. zeae je najrasprostranjeniji. Za sada poznat krug domaćina W. circinata var. zeae obuhvata monokotiledone i mali broj dikotiledonih biljaka, među kojima soju, pasulj, grašak, šećernu repu i mrkvu.

Cabbage, a widely used and popular vegetable, and oilseed rape, the second most valuable oilseed crop in the world, are two important species from the Brassicaceae family. Two geographically separated outbreaks of cabbage and oilseed rape root rot with estimated incidence of 15 and 20%, respectively, were recorded during 2017 in the Vojvodina region, Serbia. Twelve hyphal-tip isolates were obtained from symptomatic cabbage and oilseed rape plants and identified as Waitea circinata var. zeae based on morphological and molecular features. This indicates that W. circinata var. zeae has expanded its host range to the Brassicaceae family. Sequence analyses of internal transcribed spacer (ITS) and large subunit of the ribosomal DNA, RPB2, and B-tubulin genes revealed the highest similarity with multiple W. circinata var. zeae. Neighbor-joining analyses of ITS sequences resulted in a phylogenetic tree with one welldefined branch of W. circinata var. zeae, with two separate groups. All Serbian isolates and the majority of isolates originating from natural infection of dicotyledonous plants grouped together in group I. Following artificial inoculation, W. circinata var. zeae isolates caused mild to medium root necrosis of seedlings of 2 monocotyledonous and 12 dicotyledonous plant species, implying a wider host range than was known for W. circinata var. zeae. Additionally, this is the first occurrence of W. circinata var. zeae on dicotyledonous host plants in Europe. Because cabbage and oilseed rape are important crops grown worldwide, the occurrence of this new soilborne pathogen with a broad host range imposes the necessity for changes in routine disease control practices, particularly crop rotation.

Species of the genus Alternaria are among the most important plant pathogens that occur worldwide on many host plants, as well as plants of the Apiaceae family, including important vegetables such as carrots, parsley, celery, parsnips and fennel, and popular herbs such as anise, cumin, dill, coriander and others. These plants are attacked by different species of phytopathogenic fungi of the genus Alternaria, and this paper offers current review of nine different species. However, there is still insufficient information on both these species and the nu-merous interactions between them and their host plants and environmental factors. This information are necessary as the basis for adequate disease control measures and thus enabling successful and profitable cultivation of these high valued crops. Therefore, we present pathogen profiles of A. dauci, A. radicina, A. carotiincultae, A. petroselini, A. selini, A. smyrnii, A. alternata, A. longipes and A. burnsii as pathogens of Apiaceae plants, as well as their full current names and synonyms, taxonomic position, distribution and symptoms these pathogens cause to their host plants, as well as their biology and mode of transmission and spread, with special emphasis on control measures., Vrste roda Alternaria spadaju među najvažnije biljne patogene koji se pojavljuju širom svijeta na mnogim biljkama domaćinima, uključujući i biljke iz familije Apiaceae. Među njima posebno su značajne povrtrarske biljke kao što su mrkva, peršun, celer, paštrnak i komorač, kao i brojne i često upotrebljavane začinske biljke, kao što su anis, kim, mirođija, korijander i druge. Ove biljke napadaju različite vrste fitopatogenih gljiva iz roda Alternaria, pri čemu je u ovom radu opisano devet vrsta. Međutim, još uvijek je nedovoljno informacija kako o ovim vrstama, tako i o brojnim interakcijama između njih i njihovih biljaka domaćina i faktora spoljne sredine. Ove informacije su neophodne kao osnova za preduzimanje adekvatnih mjera za njihovo suzbijanje i omogućavanje uspješnog i profitabilnog uzgoja ovih na tržištu izuzetno traženih i cijenjenih biljnih vrsta. Zbog toga su u ovom radu prikazane vrste roda Alternaria prourokovači bolesti povrtarskih i začinskih biljaka iz familije Apiaceae, pre svega A. dauci, A. radicina, A. carotiincultae, A. petroselini, A. selini, A. smyrnii, A. alternata, A. longipes i A. burnsii, te njihovi puni aktuelni nazivi i sinonimi, taksonomska pozicija, rasprostranjenost i simptomi koje prouzrokuju na biljkama domaćinima, kao i njihova biologija i način prenošenja i širenja, uz poseban naglasak na mjere za njihovo suzbijanje.

After its first detection in 2005, Alternaria petroselini, the causal agent of black leaf spot, has become one of the most important pathogens of parsley (Petroselinum crispum (Mill.) Nym.) in Serbia. During almost a decade, isolates of A. petroselini originating from parsley leaves, seed and infested soil were collected and identified on the bases of morphological, pathogenic and molecular features. All morphological features were consistent with those described previously, including dark brown colonies on potato dextrose agar (PDA) and subsurface microsclerotia production, and the presence of pigmented, broadly ellipsoidal, singly produced feodyctiosporic conidia. Pathogenicity and host range studies using spray inoculation of spore suspensions, revealed that isolates were pathogenic for parsley, parsnip, celery and coriander, weakly pathogenic for carrot, fennel and anise, and not pathogenic for onion, cabbage, pepper, tomato, and cucumber. Molecular detection utilizing polymerase chain reaction (PCR) and A. radicina specific primers failed to demonstrate a difference between A. petroselini and A. radicina, amplifying isolates belonging to both species. For further confirmation of Serbian A. petroselini isolates, sequencing of selected genomic fragments including ITS and mitochondrial small subunit (mtSSU) rDNA and major allergen Alt a1, b-tubulin, and translation elongation factor-1 alpha (EF1a) protein coding genes was performed. All obtained sequences were deposited in GenBank and BLAST search for the closest matching revealed several A. petroselini isolates from GenBank to share 99-100% nucleotide identity. Parsley is traditionally grown herb and vegetable plant in Serbia and A. petroselini causes considerable damage every year and occasionally very severe yield losses, resulting in significant economic impact, especially in parsley leaf production. Further investigation of the epidemiology and population structure of A. petroselini is needed in order to esta

Blackberry cane diseases with the symptoms of necrosis, canker, and wilting are caused by several fungi worldwide. Surveys conducted from 2013 to 2016 in Serbia revealed the occurrence of Gnomoniopsis idaeicola, the causal agent of cane canker and wilting, which was found to be distributed in almost half of the surveyed orchards, in three blackberry cultivars, and with disease incidence of up to 80%. Wide distribution and high disease incidence suggest that G. idaeicola has been present in Serbia for some time. Out of 427 samples, a total of 65 G. idaeicola isolates were obtained (isolation rate of 34.19%). Reference isolates, originating from different localities, were conventionally and molecularly identified and characterized. G. idaeicola was detected in single and mixed infections with fungi from genera Paraconiothyrium, Colletotrichum, Diaporthe, Botryosphaeria, Botrytis, Septoria, Neofusicoccum, and Discostroma, and no diagnostically specific symptoms could be related directly to the G. idaeicola infection. In orchards solely infected with G. idaeicola, blackberry plant mortality was up to 40%, and yield loses were estimated at 50%. G. idaeicola isolates included in this study demonstrated intraspecies diversity in morphological, biological, pathogenic, and molecular features, which indicates that population in Serbia may be of different origin. This is the first record of a massive outbreak of G. idaeicola infection, illustrating its capability of harmful influence on blackberry production. This study represents the initial step in studying G. idaeicola as a new blackberry pathogen in Serbia, aiming at developing efficient control measures.

Soil-borne fungi belonging to the genus Rhizoctonia are considered to be among the most destructive sugar beet pathogens. Although multinucleate R. solani AG-2-2 is frequently detected as the main causal agent of root rot of sugar beet worldwide, several binucleate (AG-A, AG-E and AG-K) and multinucleate Rhizoctonia (R. solani AG-4, AG-5 and AG-8) have also been included in the disease complex. Due to their soil-borne nature and wide host range, the management of Rhizoctonia root rot of sugar beet is highly demanding. Identification of Rhizoctonia AG associated with root rot of sugar beet is the essential first step in determining a successful disease management strategy. In this paper we report a highly specific and sensitive real-time PCR protocol for detection of R. solani AG-2-2 which showed a high level of specificity after testing against 10 different anastomosis groups and subgroups, including AG-2-1 as the most closely related. Moreover, a similar conventional PCR assay showed the same specificity but proved to be at least a 100 times less sensitive. Future research will include further testing and adaptation of this protocol for direct detection and quantification of R. solani AG-2-2 in different substrates, including plant tissue and soil samples., Smatra se da su gljive iz roda Rhizoctonia koje se održavaju u zemljištu, najdestruktivniji patogeni šećerne repe. Mada je širom sveta višejedarna R. solani AG-2-2 najčešće označena kao najvažniji prouzrokovač truleži korena šećerne repe, nekoliko dvojedarnih (AG-A, AG-E i AG-K) kao i višejedarnih Rhizoctonia (R. solani AG-4, AG-5 i AG-8) takođe mogu da učestvuju u kompleksu bolesti. Kako se održavaju u zemljištu i imaju širok krug domaćina, suzbijanje vrsta Rhizoctonia koje izazivaju trulež korena šećerne repe veoma je zahtevno. Identifikacija anastomoznih grupa prouzrokovača oboljenja predstavlja neophodan prvi korak u uspostavljanju uspešnog suzbijanja. U ovom radu razvijen je visokospecifičan i osetljiv real-time PCR protokol za detekciju i identifikaciju R. solani AG-2-2 koji se pokazao kao visokospecifičan nakon testiranja koja su obuhvatila 10 različitih AG grupa i podgrupa, uključujući i AG-2-1 kao najsrodniju. Sličan konvencionalni PCR protokol ispoljio je istu specifičnost, ali i najmanje 100 × manju osetljivost. Buduća istraživanja uključiće dalje testiranje i adaptaciju ovog protokola za direktnu detekciju i kvantifikaciju R. solani AG-2-2 u različitim uzorcima, uključujući biljno tkivo i zemljište.

U peroiodu od 2004. do 2012. godine, na teritoriji Srbije, uočeni su simptomi odumiranja čokota vinove loze. Simptomi se najpre ispoljavaju u vidu sitnih, hlorotičnih i nekrotičnih pega po obodu listova, deformacijom listova, pojavom skraćenih lastara, često sa tzv. cik-cak porastom internodija. Vremenom dolazi do delimičnog ili potpunog sušenja čokota vinove loze. Primenom standardnih fitopatoloških metoda iz simptomatičnih biljaka vinove loze dobijeno je 47 izolata Eutypa sp. iz 14 lokaliteta. Izolati potiču iz sledećih sorti vinove loze: Kaberne sovinjon, Rkaciteli, Prokupac, Rizling rani, Burgundac crni, Italijanski rizling, Sovinjon, Šardone, Frankovka i Kardinal. Patogenost izolata proverena je veštačkom inokulacijom zdravih reznica vinove loze sorte Kaberne sovinjon. Svi proučavani izolati prouzrokuju nekrozu tkiva oko mesta inokulacije, hlorozu i deformaciju listova kao i pojavu sitnih, nekrotičnih pega po obodu liski koji vremenom opadaju. Inokulisane reznice zaostaju u porastu dobijajući žbunast izgled sa cik-cak porastom internodija. Identifikacija proučavanih izolata Eutypa sp. izvršena je na osnovu morfoloških osobina i prisustvo piknida u kulturi. Piknidi su loptastog ili nepravilnog oblika, tamnosmeđe do crne boje, veličine 0,5-1 mm. Iz piknida se izlučuje konidijalna masa krem do bledo narandžaste boje. Konidije su jednoćelijske, neseptirane, hijalinske, izdužene i umereno krive sa spljoštenom osnovom, dimenzija 36,10 μm × 1,71 μm. Različite hranljive podloge i tipovi svetlosti imaju veliki uticaj na sporulaciju proučavanih izolata Eutypa sp. Najintenzivnija sporulacija svih proučavanih i kontrolnih izolata odvijala se na PDA podlozi, zatim na MA, GWA i WA, dok na podlogama YA i TA ni nakon 3 meseca ne dolazi do sporulacije. Izlaganje kultura režimu od 24 h UV svetlosti se pokazalo kao najpogodnije za razvoj konidija, potom 12 h UV i 12 h tame, dok je najnepovoljniji režim svetlosti za razvoj konidija je 12 h svetla i 12 h tame. Pročavane kulture ne, During the period from 2004 to 2012, symptoms of grapevine dieback were observed in Serbia. Symptoms initially appear as small, chlorotic and necrotic spots along the rim of the leaves, deformation of leafs and the appearance of shortened shoots, often with the so-called zig-zag internodes. Over time, partial or complete dying of the vines develops. Standard phytopathological methods were used and 47 isolates were obtained from symptomatic grapevine plants from 14 localities. The isolates originate from the following cultivars: Cabernet Sauvignon, Rkaciteli, Prokupac, Riesling early, Pinot noir, Italian Riesling, Sauvignon, Chardonnay, Franconien noir and Cardinal. Pathogenicity of isolates was tested by artificial inoculation of healthy grapevine cuttings cv. Cabernet Sauvignon. All studied isolates cause tissue necrosis around the site of inoculation, chlorosis and deformation of the leaves, along with the appearance of tiny, necrotic spots on the periphery of the leaf that falls of over time. Inoculated cuttings are stunted, dwarf-like with zig-zag internodes. Identification of the studied isolates was performed on the basis of morphological properties and the presence of pycnidia in the culture. Pycnidia are spherical or irregular shape, ranging from dark brown to black, measuring 0,5-1 mm. Pycnidia exude conidial mass pale brown to pale orange color. Conidia are single celled, non-septate, hyaline, elongated and curved with flat basis, measuring 36,10 μm × 1,71 μm. Different artificial mediums and types of light have major influence on sporulation of tested isolates Eutypa sp. Most intense sporulation of all studied and control isolates was on PDA medium, followed by MA, GWA and WA, while on YA and TA mediums no sporulation occurred after 3 months. Exposure of cultures to 24h UV light proved to be the most suitable for the development of conidia, followed by 12h UV and 12h darkness, while the most unfavorable regime of light for the development of conidia was 1

Tokom ispitivanja sprovedenog u periodu od 2013. do 2016. godine, pregledom pet komercijalno značajnijih sorti kupine na 24 lokaliteta u 13 okruga u Srbiji, ustanovljena je široka rasprostranjenost bolesti stabla kupine (incidenca 5 - 80%, prosečno 37,7%), a nekroza i pucanje kore bili su najčešći simptomi. U pojedinim lokalitetima zabeleženo je sušenje 40% biljaka uz direktno smanjenje prinosa od oko 50%. Rasvetljavanje etiologije bolesti stabla kupine u Srbiji obavljeno je kroz morfološku i molekularnu identifikaciju i karakterizaciju 468 izolata dobijenih iz 427 uzoraka. Nakon ispunjenja Kohovih postulata, na osnovu makroskopskih i mikroskopskih osobina dobijeni izolati izdvojeni su u 10 morfoloških grupa što je potvrđeno sekvenciranjem ITS regiona rDNA. Identifikovano je 10 vrsta gljiva prouzrokovača bolesti stabla kupine u Srbiji: Paraconiothyrium fuckelii, Gnomoniopsis idaeicola, Septoria spp., Botryosphaeria dothidea, Diplodia seriata, Botrytis cinerea, Colletotrichum kahawae, Diaporthe eres, Diaporthe betulae i Neofusicoccum parvum. Dodatna karakterizacija na osnovu filogenetskih analiza urađena je za vrste G. idaeicola, B. dothidea, D. seriata, C. kahawae, D. eres, D. betulae i N. parvum čime je određena njihova srodnost i međuodnos sa ostalim vrstama rodova Gnomoniopsis, Botryosphaeria, Diplodia, Colletotrichum, Diaporthe i Neofusicoccum. Izolate Septoria spp. moguće je bilo identifikovani do nivoa roda na osnovu morfoloških, patogenih i molekularnih odlika i biće predmet daljih istraživanja uz sekvenciranje dodatnih molekularnih markera. Tokom ovih istraživanja vrsta G. idaeicola prvi put je opisana na kupini u Srbiji, a kako se pokazalo da je rasprostranjena u skoro polovini ispitivanih zasada, to predstavlja i prvu masovnu pojavu na kupini u svetu kao i prvi uvid u moguću epidemiologiju nekroze stabla i sušenja kupine izazvane sa G. idaeicola. Izolati G. idaeicola iz Srbije ispoljili su morfološku, molekularnu i biološku varijabilnost što je ustanovljen, A Survey was conducted from 2013 to 2016 which included a total of 24 orchards planted with five of the most commonly grown blackberry cultivars in 13 administrative districts in Serbia. A wide distribution of blackberry cane diseases (incidence of 5 - 80%, with an average of 37,7%) was observed with the most frequent symptoms of bark necrosis and canker. In some of the localities, the wilting of up to 40% of plants was recorded with yield reduction of about 50%. The etiology of blackberry cane diseases was investigated through morphological and molecular identification and characterization of 468 isolates obtained from 427 samples. After fulfilling the Koch’s postulates, the isolates were grouped into 10 morphological groups based on macroscopic and microscopic features which were confirmed by the sequencing of the ITS region rDNA. Ten species of fungi causing blackberry cane diseases were identified including Paraconiothyrium fuckelii, Gnomoniopsis idaeicola, Septoria spp., Botryosphaeria dothidea, Diplodia seriata, Botrytis cinerea, Colletotrichum kahawae, Diaporthe eres, Diaporthe betulae and Neofusicoccum parvum. Additional characterization based on phylogenetic analysis was performed for G. idaeicola, B. dothidea, D. seriata, C. kahawae, D. eres, D. betulae and N. parvum and revealed their relationship to the additional species within the genera of Gnomoniopsis, Botryosphaeria, Diplodia, Colletotrichum, Diaporthe and Neofusicoccum. Isolates of Septoria spp. could be identified to the genus level based on morphological, pathogenic and molecular features and will be subjected to further research through sequencing of additional molecular markers. During this research G. idaeicola was detected on blackberries in Serbia for the first time. As it was found that G. idaeicola was distributed in almost half of the orchards, this is the first record of a massive outbreak of G. idaeicola on blackberries in the world as well as the first insight into the possible epidemi