Prvi nalaz tumorogenih sojeva Agrobacterium radiobacter na malini u Srbiji

During the spring of 2003, gall symptoms on the roots and crowns of young raspberry plants cv. Vilamette were observed near Valjevo. Phytopathogenic bacteria were isolated from diseased plant samples. Based on the pathogenic, morphological, differential biochemical and physiological characteristics, the isolated strains were identified as tumorigenic Agrobacterium radiobacter (biovar 1 Agrobacterium). In order to confirm the identity of isolated strains by polymerase chain reaction (PCR) primers complementary to tms2 gene located on the Ti plasmid were used. In the first PCR protocol using a tms2F1 + tms2R2 primer pair, 617 bp products specific for tumorigenic Agrobacterium strains were amplified. The second PCR protocol, using a tms2F1 + tms2B primer pair, amplified the expected 458 bp products. On the basis of multiplex PCR with primers complementary to chromosomal gene coding for 23S rRNA, the isolated strains were classified as biovar 1 Agrobacterium (A. radiobacter). This is the first report of tumorigenic A. radiobacter on raspberry in Serbia.Tokom proleća 2003. godine, zapaženi su tumori na korenu i korenovom vratu mladih biljaka maline sorte Vilamette u okolini Valjeva. Iz obolelih uzoraka izolovane su fitopatogene bakterije. Proučavanjem patogenih, morfoloških i diferencijalnih biohemijsko-fizioloških karakteristika izolovanih sojeva, utvrđeno je da proučavani sojevi pripadaju bakteriji Agrobacterium radiobacter (biovar 1). U cilju potvrde identiteta izolovanih sojeva lančanom reakcijom polimeraze (PCR), korišćeni su prajmeri komplementarni tms2 genu lociranom na Ti plazmidu. U prvoj PCR reakciji izvedenoj korišćenjem tms2F1 + tms2R2 para prajmera, detektovani su fragmenti nukleinske kiseline veličine 617 baznih parova (bp), specifični za tumorogene sojeve A. tumefaciens. U drugoj PCR reakciji korišćenjem para prajmera tms2F1 i tms2B su amplifikovani PCR produkti očekivane veličine 458 bp, karakteristični za ovu bakteriju. Na osnovu multiplex PCR reakcije korišćenjem prajmera komplementarnih na hromozomalni gen koji kodira 23S rRNA sojevi izolovani iz tumora na korenu maline svrstani su u biovar 1 Agrobacterium (A. radiobacter). Ovo je prvi nalaz ove bakterije na malini u Srbiji

Pseudomonas syringae - prouzrokovač nekroze plodova trešnje

Severe symptoms of necrosis sweet cherry fruits was recorded during 2014. in region of Šabac. About 30 % of fruits (cv. Summit) were affected and necrosis cover 1/3 of fruit surface in average. Gram negative, fluorescent, oxidative bacterial strains were isolated from diseased tissues. Symptoms similar to those, observed in natural infection were recorded on artificial inoculated cherry fruits (cv. Summit and cv. Summbarst). Isolated strains were HR positive, oxidase, pectinase, arginin dehidrolase negative and levan positive (LOPAT +---+). Same characteristics also own check strains Pseudomonas syringae (CFBP 11). According obtained results it was concluded that necrosis of sweet cherry fruits is caused by Pseudomonas syringae. Further characteristic in order proper detection of pathogens, including molecular methods are underway.U radu su prikazani rezultati proučavanja sojeva bakterija izolovanih iz nekrotičnih plodova trešnje sorte Samit (područje Šapca). Nekroza zahvata u proseku oko 1/3 ploda koji potpuno gubi tržišnu vrednost. Bolešću biva zahvaćeno oko 30% plodova. Primenom standardnih bakterioloških metoda (izolacijom na hranljivim podlogama), proverom patogenosti i proučavanjem bakterioloških karakteristika, zaključeno je da pomenute simptome prouzrokuje fitopatogena bakterija Pseudomonas syringae, široko rasprostranjen patogen naročito koštičavih voćaka

Characteristics of Phomopsis sp. isolates of plum trees origin

Twelve isolates of Phomopsis sp. were obtained from the branches and the trunk of plums (Prunus domestica L) with decay symptoms in Valjevo, Ljig Koceljeva and Ub vicinity during 2004-2006. Morphological, pathogenic and growing characteristics were studied. Pathogen caused tissue necrosis of branches around the inoculate seats, and wrinkling and watering of plum fruits. All media were suitable for pathogen development, except prune agar. The best growth of isolates was at medium pH 5,5. The optimal temperature for growth and germination of pycnidiospores was 25°C

Provera antagonističkog delovanja mikroorganizama na Colletotrichum acutatum i Colletotrichum gloeosporioides

The antagonistic activities of five biocontrol agents: Trichoderma harzianum, Gliocladium roseum, Bacillus subtilis, Streptomyces noursei and Streptomyces natalensis, were tested in vitro against Colletotrichum acutatum and Colletotrichum gloeosporioides, the causal agents of anthracnose disease in fruit crops. The microbial antagonists inhibited mycelial growth in the dual culture assay and conidial germination of Colletotrichum isolates. The two Streptomyces species exhibited the strongest antagonism against isolates of C. acutatum and C. gloeosporioides. Microscopic examination showed that the most common mode of action was antibiosis. The results of this study identify T. harzianum, G. roseum, B. subtilis, S. natalensis and S. noursei as promising biological control agents for further testing against anthracnose disease in fruits.Antagonističko delovanje pet agenasa biološke zaštite: Trichoderma harzianum, Gliocladium roseum, Bacillus subtilis, Streptomyces noursei i Streptomyces natalensis, testirano je in vitro na Colletotrichum acutatum i Colletotrichum gloeosporioides, prouzrokovače antraknoze plodova. Svi mikrobni antagonisti inhibiraju porast micelije u testovima dvojne kultivacije i klijavost konidija Colletotrichum izolata. Dve vrste roda Streptomyces ispoljavaju najjači stepen antagonizma prema izolatima C. acutatum i C. gloeosporioides. Mikroskopsko ispitivanje pokazuje da je antibioza najčešći način delovanja antagonista. T. harzianum, G. roseum, B. subtilis, S. natalensis i S. noursei su rezultatima ovih istraživanja identifikovani kao biološki agensi koji se mogu uspešno uključiti u buduća testiranja u cilju suzbijanja antraknoze plodova

Proučavanje raznovrsnosti bakterije Pseudomonas syringae poreklom sa različitih voćaka u Srbiji

Pseudomonas syringae is a widespread and economically important plant pathogen, one found on a number of hosts, including fruit trees, field crops, vegetables, and ornamental plants. This bacterium has been experimentally identified as a parasite of pear, apple, apricot, peach, cherry, sour cherry, plum, and raspberry. The present study was designed to establish differences between strains isolated from fruit trees in Serbia. The pathogenic and biochemical characteristics of isolates were studied. The BOX-PCR method was used to generate genomic fingerprints of Pseudomonas syringae isolates and to identify strains that were previously not distinguishable by other classification methods. Different Bacillus sp. strains were tested for in vitro inhibitory activity against Pseudononas syringae isolates. Bacillus sp. strains show inhibitory activity only against P. syringae isolates that originated from peach. The obtained results demonstrate that the population of the bacterium Pseudomonas syringae from the fruit trees in Serbia is very diverse.Pseudomonas syringae je široko rasprostranjena i ekonomski značajna fitopatogena bakterija, sa širokim krugom domaćina koji uključuje voćke, ratarske, povrtarske i ukrasne biljke. Pseudomonas syringae u Srbiji je eksperimentalno potvrđen kao parazit kruške, jabuke, kajsije, breskve, trešnje, višnje, šljive i maline. Cilj ove studije bio je da utvrdi postojanje eventualnih razlika između sojeva izolovanih sa različitih vrsta voćaka u Srbiji. Proučavane su patogene i biohemijske osobine sojeva. BOX-PCR je korišćen za dobijanje profila izolata Pseudomonas syringae u cilju identifikacije sojeva koji se ne mogu utvrditi drugim metodama. Različiti sojevi roda Bacillus su testirani u cilju utvrđivanja njihove in vitro inhibitorne aktivnosti. Sojevi roda Bacillus su pokazali inhibitornu aktivnost samo na P. syringae izolovanih sa breskve. Dobijeni rezultati pokazali suda je populacija bakterije Pseudomonas syringae poreklom sa voća u Srbiji vrlo raznovrsna

Vegetativna inkompatibilnost izolata Phomopsis spp. izolovanih sa stabla šljive

Isolates of Phomopsis spp. obtained from plum trees (Prunus domestica L.) with symptoms of die-back. There were significant morpho-physiological and pathogenic differences between the isolates. The genetic variability (12 isolates of Phomopsis spp. and isolate of Photopia pernicious, which was used as control ) was proved by study of vegetative compatibility. These isolates were inoculated in Petri dishes in pairs in all possible cross combinations, on potato dextrose agar. After 5 and 10 days the interaction between colonies were scored according to appearance of contact lines of paired cultures. On the contact line on two different isolates distinct barrage zones of inhibition - vegetative incompatibility reaction. The barrage zones generally became black and sometimes showed formation of pycnidia. Necrotic parts of hyphen that formed line of reaction were observed. A compatible reaction was observed only when cultures of the same isolate were paired.Izolati Phomopsis spp. dobijeni sa grana i stabla šljive (Prunus domestica L.) sa simptomima sušenja i izumiranja ispoljili su značajne razlike u morfolo-fiziološkim i patogenim karakteristikama. Primenom metode vegetativne kompatibilnosti potvrđena je genetska varijabilnost 12 izolata Phomopsis spp. i kontrolnog izolata Phomopsis perniciosa. Isečci ispitivanih kultura zasejavani su u Petri kutije po parovima i u svim mogućim unakrsnim kombinacijama, na podlozi KGA. Očitavanje rezultata je vršeno petog i desetog dana i podrazumevalo je praćenje izgleda linije međusobnog kontakta uparivanih izolata. Nakon perioda inkubacije između ispitivanih kultura dolazi do formiranja crne reakcione linije - barijere, a u pojedinim slučajevima i reproduktivnih organa - piknida (reakcija vegetativne inkompatibilnosti). Delovi hifa koje sačinjavaju crnu reakcionu liniju bili su nekrotirani. Vegetativna kompatibilnost konstatovana je samo u slučajevima kada su izolati uparivani sami sa sobom

Sorbus domestica i S.torminalis novi domaćini Erwinia amylovora u Srbiji

U radu su prikazani rezultati proučavnja izolovanih sojeva bakterije iz plamenjačom zahvaćenih mladara ukrasnih biljka Sorbus domestica i Sorbus torminalis. Proučavani sojevi su Gram negativni, ne stvaraju fluorescentni pigment na King-ovoj podlozi B, a glukozu metabolišu i oksidativno i fermentativno. Prouzrokuju HR duvana i nekrozu nesazrelih, inokulisanih plodova kruške (viljamova) praćenu obilnom produkcijom bakterijskog eksudata.. Prvi simptomi na inokulisanim mladarima biljaka domaćina u očavaju se posle 2-3 dana, ispoljavaju se u vidu vlažnih pega koje se vremenom šire zhavatjući ga u celosti Na osnovu patogenih, biohemijskih i seroloških odlika (ELISA) proučavanih izolata zaključeno je da patološke promene na ukrasnim biljkama Sorbus domestica i Sorbus torminalis prouzrokuje Erwinia amylovora. To je potvrđeno i primenom molekularne metode (PCR) korišćenjem specifičnog prajmera za bakteriju E. amylovora. Ovo je prvi nalaz E. amylovora na ukrsnim biljkama roda Sorbus u Srbij

Etiološka proučavanja bakteriozne vlažne truleži uskladištenih glavica komorača

Soft rot bacterial strains were isolated from diseased fennel bulbs. They were characterised on the basis of its pathogenicity morphological, cultural and biochemical properties. All investigated strains caused the soft rot of inoculated fenel bulbs, pepper fruits as well as clices of potato and carrot. Results of bacteriological properties show that soft rot of fennel bulbs were caused by Gram negative, nonfluorescent, facultative anaerobic, levan and oxidase negative bacteria belonging to the genus Pectobacterium. The investigared strains utilised lactose, grew at 5% NaCl and weak growth was recorded on 37oC; they are resistent to erytromicin. Negative results were recorded in indol, phospathase, lecithinase and reducing compounds pf sucrose tests. These results as well as the characteristics growth on Logan differential medium indicated that sof rot of fenel bulbs was caused by Pectobacterium carotovorum subsp. carotovorum. This is the first report of this bacterium affecting fennel bulbs in Serbia.U radu su proučene patogene i biohemijske odlike fitopatogenih bakterija izolovanih iz glavica komorača sa simptomima truleži. Proučavani sojevi prouzrokuju trulež glavica komorača, veštački inokulisanih, plodova paprike, kao i kriški mrkve i krompira i pri tome ispoljavaju izraženu pektolitičku aktivnost. Proučavanjem bakterioloških odlika utvrđeno je da su izolovani sojevi Gramnegativni, ne fluoresciraju na Kingovoj podlozi B, glukozu metabolišu i u aerobnim i anaerobnim uslovima, ne stvaraju levan i oksidazu. Na osnovu navedenih karakteristika zaključeno je da ispitivani sojevi pripadaju rodu Pectobacterium. Proučavani sojevi ne stvaraju fosfatazu, lecitinazu, indol, redukujuće supstance iz saharoze, a na Loganovoj podlozi formiraju karakteristične kolonije ružičaste boje sa tamnim centrom. Na osnovu ovih, ali i rezultata ostalih diferencijalnih testova za vrste roda Pectobacterium (metabolizam ugljenih hidrata, razvoj pri 5% NaCl, osetljivost prema eritromicinu), zaključeno je da vlažnu trulež komorača prouzrokuje bakterija Pectobacterium carotovorum subsp. carotovorum. Ovo je prvo saopštenje o nalazu ove bakterije kao patogena komorača u Srbiji

Proučavanje proteinskih profila bakterije Pseudomonas syringae izolovane sa različitih vrsta voćaka

Bacterium Pseudomonas syringae has numerous varieties with differentiated properties and is abundant in genetic diversity. Pseudomonas syringe has been experimentally identified in Serbia as a parasite of pear, apple, cherry, sour cherry, plum and raspberry. This study was designed to establish differences between the strains isolated from the fruit trees in Serbia with whole cell protein profiles analysis. The paghogenic and bacteriologycal characteristics of the isolates, cultivating, morphological and biochemical characteristics were studied. The obtained results demonstrate that the population of the bacterium Pseudomonas syringae from the fruit trees in Serbia is diverse, but do not show any difference in their whole cell protein profiles.Bakterija Pseudomonas syringae ima veliki broj varijeteta sa različitim osobinama i sa velikom genetičkom raznovrsnošću. Pseudomonas syringae u Srbiji je eksperimentalno potvrđen kao parazit kruške, jabuke, breskve, trešnje, višnje, šljive i maline. Cilj ove studije bio je da utvrdi postojanje eventualnih razlika između sojeva izolovanih sa različitih vrsta voćaka u Srbiji proučavanjem elektroforetskih profila ukupnih ćelijskih proteina. Proučavane su takođe patogene i biohemijske odlike sojeva poreklom sa raznih vrsta voćaka. Dobijeni rezultati pokazali su da su populacije bakterije Pseudomonas syringae poreklom sa voća u Srbiji raznovrsne, ali da ne pokazuju razlike u elektroforetskim profilima ukupnih ćelijskih proteina

Pectobacterium carotovorum subsp. Carotovorum - prouzrokovač vlažne truleži biljaka kale u Srbiji i Crnoj Gori

Bacterial strains were isolated from above- and underground parts of diseased calla plants originating from different localities in Serbia and one locality in Montenegro. They were characterized by studying their pathogenic, cultural, biochemical and physiological characteristics. All investigated strains caused soft rot of calla leaf stalks, potato slices and aloe leaves, and induced hypersensitive reaction on tobacco. Bacteriological properties of the strains indicated that symptoms on calla plants were caused by Gram-negative, nonfluorescent, oxidase negative, catalase positive and facultatively anaerobic bacterium belonging to the genus Pectobacterium. The investigated strains grew at 37ºC and in 5% NaCl, utilised lactose and trechalose, and produced neither indol nor lecitinase. These results, as well as the characteristic growth on Logan's differential medium indicated that soft rot of tuber and stem base of calla plants was caused by Pectobacterium carotovorum subsp. carotovorum. This is the first report of this pathogen affecting calla plants in Serbia.Proučene su patogene, odgajivačke i biohemijsko-fiziološke odlike sojeva izolovanih iz nadzemnih i podzemnih delova obolelih biljaka kale gajene u različitim lokalitetima u Srbiji i jednom lokalitetu u Crnoj Gori. Svi proučavani sojevi ispoljili su izraženu pektolitičku aktivnost prouzrokujući vlažnu trulež lisnih drški kale, kriški krompira, liski aloje ili sanseverije, a prouzrokovali su i hipersenzitivnu reakciju duvana. Proučavanjem bakterioloških karakteristika utvrđeno je da promene na biljkama kale prouzrokuju Gram-negativni, nefluorescentni, oksidaza-negativni, katalaza-pozitivni i fakultativno-anaerobni sojevi bakterije, koja prema navedenim karakteristikama pripada rodu Pectobacterium. Proučavani sojevi se razvijaju pri 37ºC i u prisustvu 5% NaCl, razlažu laktozu i trehalozu, ne stvaraju indol i lecitinazu. Navedeni rezultati, kao i karakterističan razvoj na Loganovoj diferencijalnoj podlozi ukazuju da je vlažnu trulež korena i prizemnog dela biljaka kale prouzrokovala bakterija Pectobacterium carotovorum subsp. carotovorum. Ovo je prvo saopštenje o pojavi P. c. ssp. carotovorum kao patogena kale u Srbiji