Efikasnost fungicida različitog mehanizma delovanja u suzbijanju kestenjaste pegavosti izdanaka maline (Didymella applanata)

Efficacy trials of four multi-site fungicides (copper hydroxide, mancozeb, chlorothalonil and dithianon), as well as six fungicides with specific modes of action (fluopyram, boscalid, fluazinam, tebuconazole, azoxystrobin and pyraclostrobin) in raspberry spur blight (Didymella applanata) control were carried out in the seasons 2014 and 2016. The experiments were conducted as a randomized block design with four replicates in a commercial raspberry orchard in the locality Trešnjevica (Arilje) in western Serbia. All fungicides were applied preventively, four times until the beginning of harvest and once after harvest. The effects of the products tested were assessed three weeks after the last fungicide application according to the intensity of cane infection. Disease severity in control (untreated) plots were 53.7 (2014) and 76.3% (2016). In both years, the highest efficacy was achieved by tebuconazole (96.3 and 99.6%), followed by fluopyram (95.7 and 99.3%) and boscalid (94.7 and 95.9%). The broad-spectrum multi-site fungicides mancozeb, chlorothalonil, copper hydroxide and dithianon were effective against D. applanata and they reduced disease severity significantly, in comparison with the untreated plots. The efficacy of these compounds was between 64.4 and 81.7%. Conversely, azoxystrobin, pyraclostrobin, and fluazinam showed very low efficacy (13.7-37.8%) in control of raspberry spur blight.U toku 2014. i 2016. godine, ispitivana je efikasnost četiri fungicida sa nespecifičnim (bakar-hidroksid, makozeb, hlorotalonil i ditianon) i šest sa specifičnim (fluopiram, boskalid, fluazinam, tebukonazol, azoksistrobin i piraklostrobin) mehanizmom delovanja, u suzbijanju prouzrokovača kestenjaste pegavosti izdanaka maline (Didymella applanta). Ogledi su izvedeni u komercijalnom zasadu maline na lokalitetu Trešnjevica (Arilje), u zapadnoj Srbiji, po tipu potpunog slučajnog blok sistema sa četiri ponavljanja. Svi fungicidi su primenjeni preventivno, a obavljeno je ukupno pet tretiranja, četiri pre berbe i jedno nakon završene berbe. Efikasnost testiranih preparata ocenjena je tri nedelje nakon poslednje primene fungicida. Intenzitet oboljenja u kontrolnim (netretiranim) parcelama iznosio je 53,7% (2014) i 76,3% (2016). Tokom obe godine ispitivanja, najviša efikasnost postignuta je primenom tebukonazola (96,3% i 99,6%), fluopirama (95,7% i 99.3%) i boskalida (94,7% i 95,9%). Fungicidi nespecifičnog mehanizma delovanja mankozeb, hlorotalonil, bakar-hiroksid i ditianon bili su efikasni u suzbijanju D. applanata i značajno su smanjili intenzitet oboljenja u poređenju sa netretiranim parcelama. Efikasnost ovih preparata bila je između 64,4% i 81,7%, dok su azoksistrobin, piraklostrobin i fluazinam pokazali veoma nisku efikasnost (13,7-37,8%) u subijanju prouzrokovača kestenjaste pegavosti izdanaka maline

Validation Method for Pesticide Residue after Gamma Irradiation

Pesticide residues include the remaining substances or mixtures, metabolites and impurities in food which are considered to be toxicological significance. The goal of this work was validation method for the determination of residue by UPLC PDA and GC MS techniques. Special scope has been chlorpyrifos from apple after oil and emulsion formulation Radar EW and Radar EC treatment. In order to fully respond to this task, it has been induced the degradation products of active component -chlorpyrifos from these formulations. The degradation products chlorpyrifosoxon, 3,5,6-trichloro-2-pyridinol (TCP) and o,o -diethyl thiophosphate (DTPA) received after gamma radiation samples in a permanent for 66 hours with 2,5 KGray from Co-60 source. Gamma radiation is the closest degradation to natural conditions, followed degradation path, based on the chemical and microbiological processes. Four different extraction procedures have been used: liquid-liquid extraction (Dutch procedure), solid phase extraction with HLB cartridge, multiwall carbon nano tubes and Quechers extraction. The recovery for these clean up methods has been in range 75-94%. The validation parameters for UPLC PDA has been: LOD 0,06 ppm, LOQ 0,18 ppm. The linearity was confirmed by correlation coefficient, R=0.998 for UPLC. The parameter for GC MS has been on ppm level also. The correlation coefficient for GC MS is 0.999. The result on apple samples showed that for pesticide residue quantifications is more adequate GC MS technique that is characterized by highest recovery, the lowest standard deviation and better LOQ.Fourth International Conferenceon Radiation and Applications in Various Fields of Research, RAD 2016, May 23-27, 2016, Niš, Serbi

Rod Botrytis i vrsta Botrytis cinerea: Patogene, morfološke i epidemiološke karakteristike

Species of the genus Botrytis occur wherever their hosts are grown, ranging from cold areas of Alaska to warm and dry areas in Israel. They have a necrotrophic life style which is often associated with phenology of the host plant. The genus comprises 22 species, most of which have a narrow host range. Polifagous species Botritys cinerea, a causal agent of grey mould disease, is the most important and the most extensively studied representative of this genus. More than 350 papers related to all aspects of the research of this necrotrophic pathogen are published each year. In this paper up-to-date knowledge about pathogenic, morphological and epidemic characteristics of the genus Botrytis and, particularly, species B. cinerea are summarized. Symptoms caused by B. cinerea on various plant species and various plant parts are shown. Morphological and genetic variability of the species is described. The possible mechanisms of variability, as well as the attempts to divide the species into Group I (B. 'pseudocinerea') and Group II (B. cinerea 'sensu-stricto') are pointed out.Vrste roda Botrytis rasprostranjene su svuda gde se gaje ili su prisutne njihove biljke-domaćini - od hladnih zona Aljaske do toplih i suvih područja u Izraelu. Imaju nekrotrofni životni ciklus koji je najčešće tesno povezan sa fenologijom domaćina. Rod obuhvata 22 vrste od kojih većina ima uzak krug domaćina - parazitira biljke jednog roda ili jedne familije. Polifagna vrsta Botrytis cinerea, prouzrokovač sive truleži, najznačajniji je i najproučavaniji predstavnik ovog roda. Godišnje se objavi više od 350 radova koji se odnose na sve aspekte istraživanja ovog nekrotrofnog patogena. U ovom radu je dat pregled dosadašnjih saznanja o patogenim, morfološkim i epidemiološkim karakteristikama vrsta roda Botrytis, a posebno vrste B. cinerea. Detaljno su obrađeni simptomi oboljenja koje prouzrokuje B. cinerea na različitim biljnim vrstama i različitim delovima biljaka, morfološka i genetička varijabilnost vrste. Takođe, ukazano je i na moguće izvore varijabilnosti, kao i na pokušaje podele vrste na dve grupe - Grupu I (B. 'pseudocinerea') i Grupu II (B. cinerea 'sensu-stricto')

Fungicide sensitivity of selected Verticillium fungicola isolates from Agaricus bisporus farms

Five isolates of Verticillium fungicola, isolated from diseased fruiting bodies of Agaricus bisporus collected from mushroom farms in Serbia during 2002-2003, were studied. By observing their colony morphology under different growth conditions and their pathogenic characteristics, the isolates were identified as V. fungicola var. fungicola. The peat/lime casing was the primary source of infection. Testing of sensitivity to selected fungicides showed that all isolates were highly resistant to benomyl (EC50 values were higher than 200.00 mg/l), moderately sensitive to iprodione (EC50 values were between 11.93 and 22.80 mg/l), and highly sensitive to prochloraz-Mn (EC50 values were less than 3.00 mg/l)

Botrytis squamosa - prouzrokovač lisne pegavosti luka u Bosni i Herzegovini

Over the past several decades, necrotic spots, lesions and blight symptoms have been observed on onion leaves in several locations in Bosnia and Herzegovina, where the crop is grown intensively. The type of symptoms indicated a possible infection with Botrytis squamosa, a widespread pathogen of onion. As symptoms of leaf spots and necrotic lesions can also be caused by some other biotic and abiotic factors, our research focused on identifying the causal agent of the observed symptoms. The pathogen was isolated from diseased tissue using standard phytopathological procedure and identified based on pathogenic and morphological features. Identification was confirmed by amplification and sequencing of the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA). The influence of temperature and growth medium on mycelial growth rate of the isolates was also studied.U poslednjih nekoliko godina, na nekoliko lokaliteta u području intenzivne proizvodnje crnog luka u Bosni i Hercegovini uočena je pojava nekrotičnih pega i sušenja lišća. Simptomi oboljenja ukazali su na moguće prisustvo široko rasprostranjenog patogena luka, vrste Botrytis squamosa. S obzirom da pegavost i nekrotične lezije na listu luka mogu biti prouzrokovane različitim biotičkim i abiotičkim faktorima, cilj ovog istraživanja bio je da se precizno identifikuje prouzrokovač oboljenja. Patogen je izolovan primenom standardnih fitopatoloških metoda i identifikovan na osnovu proučenih patogenih i morfoloških karakteristika dobijenih izolata. Identifikacija je potvrđena amplifikacijom i sekvenciranjem ITS rDNA genomnog regiona. Takođe, proučen je uticaj temperature i hranljive podloge na porast izolata

Integralni pristup u suzbijanju najvažnijih prouzrokovača bolesti cveća u zaštićenom prostoru

Disease causal agents are serious treat for the production of ornamentals under both open fi led and protected cultivation. Success in the disease prevention depends mainly on the level of knowledge on the specific requirements of the host plants, biology of the pathogen, disease epidemiology and the relationship between the parasite and the host plant. The selection of effective control measures is based on the comprehensive knowledge on all the factors contributing directly or indirectly to the disease development. The most important disease causal agents of ornamental plants under protected cultivation, required environmental conditions for disease development and cost-effective and environmentally safe control measures are discussed in this paper.Prouzrokovači bolesti predstavljaju veliki problem u proizvodnji cveća, kako na otvorenom polju, tako i u zaštićenom prostoru. Uspeh u sprečavanju pojave i razvoja oboljenja u velikoj meri zavisi od stepena poznavanja specifi čnih zahteva biljke domaćina, biologije patogena, epidemiologije bolesti i odnosa parazit i biljka- domaćin. Samo na osnovu dobrog poznavanja svih činilaca koji direktno ili indirektno utiču na razvoj oboljenja moguće je primeniti kompleks mera kojima se sprečavaju štete. U radu je dat pregled najvažnijih prouzrokovača bolesti cveća u zaštićenom prostoru, uslovi u kojima nanose štete, mera koje je neophodno primeniti sa ciljem njihovog efikasnog, ekološki i ekonomski prihvatljivog suzbijanja, kao i mera koje umanjuju rizik razvoja rezistentnosti na fungicide

Konvencionalni i real-time PCR protokoli za detekciju i identifikaciju Rhizoctonia solani AG-2-2 prouzrokovača truleži korena šećerne repe u Srbiji

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

Molekularna identifikacija i karakterizacija dvojedarnih Rhizoctonia spp. prouzrokovača crne truleži korena jagode u Srbiji

Strawberry production is a popular, fast-growing agricultural business in Serbia. Its cultivar selection has been changing fast, following market demands. One of the limiting factors of strawberry production worldwide is black root rot, primarily caused by binucleate Rhizoctonia. Recently, outbreaks of black root rot of strawberry have occurred in Serbia and the estimated disease incidence was up to 30%. Isolates of binucleate Rhizoctonia AG-A were recovered from symptomatic strawberry plants, and characterized on the bases of morphological, molecular and pathogenic features. Despite their uniform morphological characteristics, the isolates demonstrated genetic variability within ITS rDNA, grouping into three different phylogenetic sub-clusters which comprise AG-A isolates originating from Italy, Israel, Japan and the USA. The binucleate Rhizoctonia AG-A from Serbia exhibited uniform virulence on strawberry after inoculation of daughter plants and detached leaf petioles, as well as on seedlings of bean, carrot and sunflower, while they were non-pathogenic to wheat, maize, tomato, pepper, tobacco, cucumber, lettuce, peas, cabbage, rapeseed and sugar beet.Proizvodnja jagode je popularna grana poljoprivrede u Srbiji koja se brzo razvija. Sastav sorti uključenih u proizvodnju podleže stalnim promenama u skladu sa zahtevima tržišta. Jedan od ograničavajućih faktora u proizvodnji jagode širom sveta je crna trulež korena, bolest koju najčešće izazivaju dvojedarne vrste roda Rhizoctonia. U Srbiji je poslednjih godina nekoliko puta zabeležena pojava crne truleži jagode uz učestalost koja je iznosila do 30%. Iz biljaka jagode sa simptomima izdvojeni su izolati dvojedarne Rhizoctonia AG-A i okarakterisani na osnovu morfološih i molekularnih osobina i patogenosti. Uprkos uniformnim morfološkim osobinama, izolati su ispoljili genetičku varijabilnost na osnovu sekvence ITS rDNK, grupišući se u tri različita filogentska sub-klastera koji su obuhvatali isolate iz Italije, Izraela, Japana i SAD. Izolati dvojedarnih Rhizoctonia AG-A poreklom iz Srbije ispoljili su uniformnu virulentnost na jagodi nakon inokulacija živića i fragmenata lisnih peteljki, kao i na sejancima pasulja, mrkve i suncokreta, dok nisu bili patogeni za pšenicu, kukuruz, paradajz, papriku, duvan, krastavac, salatu, grašak, kupus i šećernu repu

Molekularna detekcija Monilinia fructigena prouzrokovača truleži ploda dunje

Species of the genus Monilinia are important causal agents of fruit rot on pome and stone fruits in Serbia. The pathogen is very harmful, especially on small properties and cottage plantations where intensive control measures are not applied. Quince is important host for the pathogens of this genus. During spring 2010, intensive occurrence of mummified fruits overwintering on branches of the quince was observed. The pathogen was isolated using standard phytopathological methods. Pathogenicity of eight obtained isolates was tested by artificial inoculation of injured apple fruits. Identification was performed according to pathogenic, morphological and ecological properties, and was confirmed by Multiplex polimeraze chain reaction, PCR. All the isolates studied caused brown rot on inoculated apple fruits. The isolates form light yellow colonies with lobate margins, with single-celled, transparent, elliptical or oval conidia in chains, regardless temperature or light presence. Sclerotia are observed in 14 days old cultures. The highest growth rate of most of the isolates is at 27°C and in dark. Based on studied pathogenic, morphological and ecological characteristics, it was found that the Monilinia fructigena is causal agent of brown rot of quince. Using specific primers (MO368-5, MO368-8R, MO368-10R, Laxa-R2) for detection of Monilinia species in Multiplex PCR reaction, the expected fragment 402 bp in size was amplified, which confirmed that the studied isolates belonged to the species M. fructigena.Vrste roda Monilinia su značajni prouzrokovači truleži ploda jabučastog i koštičavog voća u našoj zemlji. Parazit nanosi velike štete, posebno na malim posedima i u vikend zasadima u kojima se ne primenjuje redovna zaštita. Dunja je značajan domaćin patogena ovog roda. Tokom proleća 2010. godine ustanovljena je intenzivnija pojava mumificiranih plodova prezimelih na granama. Cilj ovog rada bio je da se identifikuje prouzrokovač mumifikacije plodova dunje, prikupljenih sa različitih lokaliteta. Patogen je izolovan primenom standardnih fitopatoloških metoda. Patogenost osam dobijenih izolata proverena je veštačkom inokulacijom povređenih plodova jabuke. Identifikacija je obavljena na osnovu patogenih, morfoloških i ekoloških osobina, a potvrđena je primenom Multiplex PCR. Svi proučavani izolati su na inokulisanim plodovima jabuke prouzrokovali trulež smeđe boje. Proučavani izolati na KDA podlozi formiraju svetlo-žute kolonije režnjevitog oboda. U kulturi gljiva, nezavisno od uticaja temperature i prisustva ili odsustva svetlosti, formira jednoćelijske, providne, eliptične ili ovalne konidije u nizovima. Sklerocije se uočavaju u kulturama starosti 14 dana. Većina izolata ostvaruje najbolji porast na temperaturi od 27°C i u odsustvu svetlosti. Na osnovu proučenih patogenih, morfoloških i odgajivačkih odlika, utvrđeno je da izolati pripadaju vrsti Monilinia fructigena. Korišćenjem specifičnih prajmera (MO368-5, MO368-8R, MO368-10R, Laxa-R2) za detekciju vrsta roda Monilinia u Multiplex PCR reakciji, amplifikovan je očekivani fragment veličine oko 402 bp čime je potvrđeno da proučavani izolati pripadaju vrsti M. fructigena

First Report of Brown Rot Caused by Monilinia fructicola on Nectarine in Serbia

In August 2011, nectarine (Prunus persica (L.) Batsch var. nucipersica (Suckow) C. K. Schneid) fruit originated from Oplenac region with symptoms of fruit rot was collected at a green market in Belgrade. Fruit had large, brown, sunken lesions covered with grayish brown tufts. Symptoms resembled those caused by species of Monilinia including M. laxa, M. fructigena, or M. fructicola (2). In order to isolate the causal organism, small superficial fragments of pericarp were superficially disinfected with commercial bleach and placed on potato dextrose agar (PDA). The majority (32 out of 33) isolates formed rosetted non-sporulating colonies with lobed margins resembling those of M. laxa. However, one isolate (Npgm) produced an abundant, grayish-white colony with even margins and concentric rings of sporogenous mycelium, resembling those described for M. fructicola (2). Conidia were one-celled, hyaline, ellipsoid to lemon shaped, 7.38 to 14.76 × 4.92 to 9.84 μm, and borne in branched monilioid chains. The average daily growth on PDA at 24°C was 10.9 mm. A single-spore isolate of Npgm was identified as M. fructicola based on the morphology of colony and conidia, temperature requirements, and growth rate (2). Morphological identification was confirmed by an amplified product of 535 bp using genomic DNA extracted from the mycelium of pure culture and species-specific PCR for the detection of M. fructicola (2). The ribosomal internal transcribed spacer (ITS) region of rDNA of Npgm was amplified and sequenced using primers ITS1/ITS4. Sequence analysis of ITS region revealed 100% nucleotide identity between the isolate Npgm (GenBank Accession No. JX127303) and 17 isolates of M. fructicola from different parts of the world, including four from Europe (FJ411109, FJ411110, GU967379, JN176564). Pathogenicity of the isolate Npgm was confirmed by inoculating five surface-disinfected mature nectarine and five apple fruits by placing a mycelial plug under the wounded skin of the fruit. Nectarine and apple fruits inoculated with sterile PDA plugs served as a negative controls. After a 3-day incubation at 22°C, inoculated sites developed brown lesions and the pathogen was succesfully reisolated. There were no symptoms on the control nectarine or apple fruits. M. fructicola is commonly present in Asia, North and South America, New Zealand, and Australia, while in the EPPO Region the pathogen is listed as an A2 quarantine organism (3). In Europe, the first discovery of M. fructicola was reported in France and since then, it has been found in Hungary, Switzerland, the Czech Republic, Spain, Slovenia, Italy, Austria, Poland, Romania, Germany, and Slovakia (1). Most recently, M. fructicola was found on stored apple fruits in Serbia (4). To our knowledge, this is the first report of M. fructicola decaying peach fruit in Serbia. These findings suggest that the pathogen is spreading on its principal host plants and causing substantial economic losses in the Serbian fruit production. References: (1) R. Baker et al. European Food Safety Authority. Online publication. www.efsa.europa.eu/efsajournal . EFSA J. 9:2119, 2011. (2) M. J. Côté. Plant Dis. 88:1219, 2004. (3) OEPP/EPPO. EPPO A2 list of pests recommended for regulation as quarantine pests. Version 2009-09. http://www.eppo.org/QUARANTINE/listA2.htm . (4). M. Vasic et al. Plant Dis. 96:456, 2012. </jats:p