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Detection and identification of causal agents of cucurbit bacterial diseases by classical and molecular methods

dc.contributor.advisorObradović, Aleksa
dc.contributor.otherIvanović, Milan
dc.contributor.otherGašić, Katarina
dc.contributor.otherMoravčević, Đorđe
dc.contributor.otherGrahovac, Mila
dc.creatorZlatković, Nevena Z.
dc.date.accessioned2018-09-06T08:39:40Z
dc.date.available2018-09-06T08:39:40Z
dc.date.available2020-07-03T09:03:09Z
dc.date.issued2018-06-21
dc.identifier.urihttp://eteze.bg.ac.rs/application/showtheses?thesesId=5928
dc.identifier.urihttp://nardus.mpn.gov.rs/handle/123456789/9779
dc.identifier.urihttps://fedorabg.bg.ac.rs/fedora/get/o:18059/bdef:Content/download
dc.identifier.urihttp://vbs.rs/scripts/cobiss?command=DISPLAY&base=70036&RID=512159965
dc.description.abstractFamilija Cucurbitaceae broji preko 800 biljnih vrsta, od kojih neke imaju široku primenu u ishrani ljudi, prerađivačkoj i farmaceutskoj industriji. S obzirom da su biljke familije Cucurbitaceae posebno osetljive prema nekoliko vrsta fitopatogenih bakterija, u godinama sa uslovima pogodnim za nastanak infekcije, gubici mogu dosegnuti velike razmere. U periodu od 2013-2016. godine, sa različitih lokaliteta širom Srbije, prikupljeni su simptomatični uzorci biljaka familije Cucurbitaceae. Iz obolelog tkiva, izolovano je ukupno 63 soja fitopatogenih bakterija. Sojevi su proučeni primenom standardnih i savremenih metoda. Svi proučavani sojevi bili su Gram-negativni i katalaza pozitivni. Od 63 testirana soja, 61 soj je prouzrokovao pojavu hipersenzitivne reakcije biljaka duvana. Izuzetak su predstavljala 2 soja izolovana iz ploda lubenice sa simptomima bakteriozne mrljavosti koje prouzrokuje vrsta Acidovorax citrulli. Na osnovu patogenih, morfoloških, biohemijsko-fizioloških i molekularnih odlika, izolovani sojevi svrstani su u 3 roda: Acidovorax, Pseudomonas i Pectobacterium. Nepektolitički, oksidaza pozitivni sojevi, koji se razvijaju pri 41˚C, pripadaju rodu Acidovorax. Sojevi na YDC podlozi obrazuju okrugle, ispupčene kolonije, krem boje, sa tendencijom porasta i promene boje kolonija u tamniju. Primenom prajmera BX-L1/BX-S-R2 specifičnih za vrstu A. citrulli, došlo je do umnožavanja fragmenta veličine 279 bp, čime je potvrđena pripadnost izolovanih sojeva ovoj vrsti. Proučavanjem sekvenci 16S rRNK gena, i njihovim poređenjem sa sekvencama iz NCBI baze, takođe je utvrđen visok stepen sličnosti (100%) sa drugim sojevima A. citrulli. U cilju proučavanja genetičkog diverziteta sojeva, primenjen je rep-PCR test. Iako je ispoljena potpuna homogenost sojeva, u BOX-PCR profilu jednog soja, umnožen je fragment koji nije uočen kod ostalih sojeva. Stoga se zaključuje da izolovani sojevi A. citrulli pripadaju populaciji zajedničkog porekla, koja je kod nas dospela zaraženim semenom. Sojevi su ispoljili su visok stepen virulentnosti u testovima patogenosti na različitim domaćinima familije Cucurbitaceae, kao i na različitim sortama lubenice. Bakteriozna mrljavost plodova lubenice do 2014. godine nije zabeležena u nas...sr
dc.description.abstractThe Cucurbitaceae family consists of over 800 plant species, some of which play an important role in human nutrition, processing and pharmaceutical industries. Bearing in mind these plants are especially susceptible to several types of phytopathogenic bacteria, losses can be substantial in years with favorable conditions for the emergence of infection. In the period 2013‒2016, symptomatic cucurbit plant samples were collected from different locations throughout Serbia. A total of 63 strains of phytopathogenic bacteria were isolated from diseased tissues. The strains were studied using the standard and modern methods. All studied strains were Gram-negative and catalase positive. Out of the 63 strains tested, 61 strains caused the hypersensitivity reaction of tobacco plants. The exceptions were two strains isolated from a watermelon fruit showing the symptoms of bacterial fruit blotch, caused by Acidovorax citrulli. Based on pathogenic, morphological, biochemical-physiological and molecular characteristics, isolated strains were classified into 3 genera: Acidovorax, Pseudomonas and Pectobacterium. Isolated strains, which did not produce potato soft rot, but were oxidase positive and grew at 41˚C, were classified into the genus Acidovorax. They produced beige to tancoloured, round, nonmucoid, convex colonies on yeast extract-dextrose-CaCO3 (YDC) agar. After three days, spreding of the colony margins was noticed, and the colour of the colony became darker. The identity of the strains was also confirmed by using the A. citrulli-specific primer set BX-L1/BX-S-R2. A 279-bp DNA fragment was amplified. The 16S rRNA gene sequence from two strains showed 100% identity to A. citrulli strains, previously deposited in the NCBI GenBank database. Genetic relatedness among strains was investigated by the rep- PCR method. All tested strains, except one, showed the same BOX-PCR profiles. The obtained results show that A. citrulli strains isolated in Serbia belonged to a relatively homogeneous population...en
dc.formatapplication/pdf
dc.languagesr
dc.publisherУниверзитет у Београду, Пољопривредни факултетsr
dc.relationinfo:eu-repo/grantAgreement/EC/FP7/316004/EU//
dc.relationinfo:eu-repo/grantAgreement/EC/H2020/635646/EU//
dc.relationinfo:eu-repo/grantAgreement/MESTD/Integrated and Interdisciplinary Research (IIR or III)/46008/RS//
dc.rightsopenAccessen
dc.sourceУниверзитет у Београдуsr
dc.subjectbiljke familije Cucurbitaceaesr
dc.subjectplant family Cucurbitaceaeen
dc.subjectbacterial diseasesen
dc.subjectAcidovorax citrullien
dc.subjectPseudomonas syringae sensu strictoen
dc.subjectPseudomonas viridiflavaen
dc.subjectPectobacterium carotovorum subsp. brasilienseen
dc.subjectidentificationen
dc.subjectbacteriological characteristicsen
dc.subjectPCRen
dc.subjectMLSAen
dc.subjectbakteriozesr
dc.subjectAcidovorax citrullisr
dc.subjectPseudomonas syringaesr
dc.subjectPseudomonas viridiflavasr
dc.subjectPectobacterium carotovorum subsp. brasiliensesr
dc.subjectidentifikacijasr
dc.subjectbakteriološke odlikesr
dc.subjectPCRsr
dc.subjectMLSAsr
dc.titleDetekcija i identifikacija bakterija parazita biljaka familije Cucurbitaceae klasičnim i molekularnim metodamasr
dc.title.alternativeDetection and identification of causal agents of cucurbit bacterial diseases by classical and molecular methodsen
dc.typedoctoralThesisen
dc.rights.licenseBY-NC-ND
dcterms.abstractОбрадовић, Aлекса; Граховац, Мила; Моравчевић, Ђорђе; Гашић, Катарина; Ивановић, Милан; Златковић, Невена З.; Детекција и идентификација бактерија паразита биљака фамилије Цуцурбитацеае класичним и молекуларним методама; Детекција и идентификација бактерија паразита биљака фамилије Цуцурбитацеае класичним и молекуларним методама;
dc.identifier.fulltexthttp://nardus.mpn.gov.rs/bitstream/id/13711/Disertacija.pdf
dc.identifier.fulltexthttp://nardus.mpn.gov.rs/bitstream/id/13712/IzvestajKomisije17458.pdf


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