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Morphological and molecular characterization of Ascochyta species

dc.contributor.advisorStojšin, Vera
dc.contributor.otherMaširević, Stevan
dc.contributor.otherJevtić, Radivoje
dc.contributor.otherKaragić, Đura
dc.contributor.otherNagl, Nevena
dc.creatorŽivanov, Dalibor
dc.date.accessioned2020-02-26T16:23:30Z
dc.date.available2020-02-26T16:23:30Z
dc.date.available2020-07-03T13:33:49Z
dc.date.issued2018-11-09
dc.identifier.urihttps://nardus.mpn.gov.rs/handle/123456789/12055
dc.identifier.urihttps://www.cris.uns.ac.rs/DownloadFileServlet/Disertacija153147992459180.pdf?controlNumber=(BISIS)107594&fileName=153147992459180.pdf&id=11706&source=NaRDuS&language=srsr
dc.identifier.urihttps://www.cris.uns.ac.rs/record.jsf?recordId=107594&source=NaRDuS&language=srsr
dc.identifier.urihttps://www.cris.uns.ac.rs/DownloadFileServlet/IzvestajKomisije153147992999751.pdf?controlNumber=(BISIS)107594&fileName=153147992999751.pdf&id=11707&source=NaRDuS&language=srsr
dc.description.abstractPoslednjih godina u različitim delovima Srbije zapažena je manja ili veća pojava patogena iz Ascochyta spp. kompleksa, kako na stočnom, tako i na konzumnom grašku. Navedeni kompleks čine tri fitopatogene gljive: Didymella pinodes, Didymella pinodella i Ascochyta pisi. Kompleksno oboljenje prouzrokovano gljivama iz Ascochyta spp. kompleksa se naziva Crna pegavost graška. Cilj istraživanja je bio da se prouče morfološke, molekularne i odgajivačke odlike gljiva D. pinodes, D. pinodella i A. pisi, kao i njihova patogenost i rasprostranjenost na teritoriji Republike Srbije, kao i da se ispita tolerantnost domaćeg sortimenta stočnog graška prema navedenim patogenima. U periodu od 2013. do 2015. godine, utvrđeno je prisustvo Ascochyta spp. na 27 različitih lokaliteta u Srbiji, kao i na po jednom lokalitetu u Bosni i Hercegovini, Turskoj i Francuskoj. Morfološko - molekularnim analizama sakupljenih izolata, prvi put je potvrđeno prisustvo D. pinodes i D. pinodella na teritoriji Srbije, kao i prisustvo A. pisi na teritoriji BiH. Od ukupnog broja izolata poreklom iz Srbije, 48 izolata (62%) pripada vrsti D. pinodes, 26 izolata (34%) vrsti A. pisi i 3 izolata (4%) vrsti D. pinodella. Molekularna identifikacija svih 89 izolata sprovedena je na osnovu ITS i TEF1-α sekvenci, kao i na osnovu dužine amplikona IGS1 prajmera. IGS1 prajmeri su omogućili pouzdanu i nedvosmislenu identifikaciju pojedinačnih vrsta D. pinodes, D. pinodella i A. pisi. Navedeni prajmerski par je po prvi put uspešno primenjen na vrsti A. pisi. Sagledavanjem interakcija između različitih uslova gajenja i testiranih izolata Ascochyta spp., uočene su statistički značajne intraspecijske varijacije u pogledu porasta kolonija. Kod ispitivanih izolata D. pinodes, D. pinodella i A. pisi, u zavisnosti od svetlosno-temperaturnog režima i hranljive podloge uočena je velika varijabilnost u boji supstratne i vazdušne micelije. Kod pojedinih izolata vrste D. pinodella registrovano je formiranje belih kristala lepezastog, pahuljastog i paperjastog oblika na PDA hranljivoj podlozi pri temperaturi od 20ºC. Peritecije gljive D. pinodes su se formirale samo na Conn’s hranljivoj podlozi na 15ºC u tami, kao i na 20ºC pri oba svetlosna režima. Izolati D. pinodes, D. pinodella i A. pisi ispoljili su veliku varijabilnost u dužini i širini obrazovanih askospora i piknospora kao i hlamidospora u različitim uslovima gajenja. U ovom radu je po prvi put zabeleženo formiranje sklerocija kod izolata vrsta D. pinodes i D. pinodella u ispitivanim laboratorijskim uslovima. Veštačkom inokulacijom uočena je i velika varijabilnost u patogenosti između izolata D. pinodes, D. pinodella i A. pisi, prema domaćim sortama stočnog graška. Sorta Dukat je bila najosetljivija na ispitivane patogene Ascochyta spp. kompleksa, dok je sorta Kosmaj bila najtolerantnija.sr
dc.description.abstractThe presence of pathogens from the complex Ascochyta spp., either in low or high levels, has been confirmed on both forage pea and field pea in different regions of Serbia in the previous several years. This complex consists of three phytopathogenic fungi: Didymella pinodes, Didymella pinodella and Ascochyta pisi. Ascochyta blight (blackspot) of field pea is a composite disease caused by the fungi belonging to Ascochyta spp. complex. The aim of this dissertation was to study the morphological, molecular and selection characteristics of the fungi D. pinodes, D. pinodella and A. pisi, and their pathogenicity and distribution on the territory of the Republic of Serbia, as well as to test the tolerance of domestic forage pea cultivars to the given pathogens. In the period 2013-2015, the presence of Ascochyta spp. was confirmed on 27 different sites in Serbia, as well as on one site in Bosnia and Herzegovina, one in Turkey, and one in France. Morphological and molecular analyses of the collected isolates confirmed the first appearance of D. pinodes and D. pinodella in Serbia, and A. pisi in Bosnia and Herzegovina. Out of the total number of isolates that originated from Serbia, 48 isolates (62%) belong to the species D. pinodes, 26 isolates (34%) belong to A. pisi and 3 isolates (4%) belong to D. pinodella. Molecular identification of all 89 isolates was based on ITS and TEF1-α sequences and amplicon length of IGS1 primer. IGS1 primers enabled reliable and unambiguous identification of individual species of D. pinodes, D. pinodella and A. pisi. The mentioned primer pair was successfully applied to A. pisi for the first time. The interactions between different growing conditions and the tested isolates of Ascochyta spp. were studied, and statistically significant intraspecies variations in colony growth were found. Depending on the light-temperature regime and growth medium, the tested isolates of D. pinodes, D. pinodella and A. pisi showed high variability in the colour of substrate and aerial mycelia. Certain isolates of D. pinodella formed white fan-shaped, dendritic and plumose crystals on PDA growth medium at 20ºC. Perithecia of D. pinodes formed only on Conn’s medium at 15ºC in the dark, and at 20ºC in both lights regimes. Isolates of D. pinodes, D. pinodella and A. pisi showed high variability in length and width of the formed ascospores and picnospores, as well as chlamydospores under different growing conditions. This study is the first report on the formation of sclerotia in isolates of D. pinodes and D. pinodella under the tested laboratory conditions. Artificial inoculation also showed a high variability in pathogenicity among the isolates of D. pinodes, D. pinodella and A. pisi to domestic cultivars of forage pea. The cultivar Dukat was the most susceptible to the tested pathogens of Ascochyta spp. complex, and the cultivar Kosmaj was the most tolerant.en
dc.languagesr (latin script)
dc.publisherУниверзитет у Новом Саду, Пољопривредни факултетsr
dc.relationinfo:eu-repo/grantAgreement/EC/FP7/613551/EU//
dc.relationinfo:eu-repo/grantAgreement/MESTD/Technological Development (TD or TR)/31024/RS//
dc.rightsopenAccessen
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/
dc.sourceУниверзитет у Новом Садуsr
dc.subjectDidymella pinodessr
dc.subjectDidymella pinodesen
dc.subjectDidymella pinodellasr
dc.subjectAscochyta pisisr
dc.subjectITSsr
dc.subjectTEF1-αsr
dc.subjectIGS1sr
dc.subjectmorfologijasr
dc.subjectpatogenostsr
dc.subjectDidymella pinodellaen
dc.subjectAscochyta pisien
dc.subjectITSen
dc.subjectTEF1-αen
dc.subjectIGS1en
dc.subjectmorphologyen
dc.subjectpathogenicityen
dc.titleMorfološko - molekularna karakterizacija patogena iz Ascochyta spp. kompleksasr
dc.title.alternativeMorphological and molecular characterization of Ascochyta speciesen
dc.typedoctoralThesisen
dc.rights.licenseBY-NC-ND
dc.identifier.fulltexthttps://nardus.mpn.gov.rs/bitstream/id/36254/IzvestajKomisije.pdf
dc.identifier.fulltexthttps://nardus.mpn.gov.rs/bitstream/id/36253/Disertacija.pdf
dc.identifier.fulltexthttp://nardus.mpn.gov.rs/bitstream/id/36253/Disertacija.pdf
dc.identifier.fulltexthttp://nardus.mpn.gov.rs/bitstream/id/36254/IzvestajKomisije.pdf
dc.identifier.rcubhttps://hdl.handle.net/21.15107/rcub_nardus_12055


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