Diferencijacija Pseudomonas syringae patogenih varijeteta poreklom iz koštičavih voćaka

Due to an overlapping host range, similar symptomatology and many common characteristics, Pseudomonas syringae pathovars originating from stone fruits can easily be misidentified. In order to select tests for rapid and efficient differentiation of P. s. pvs. syringae, morsprunorum and persicae, we studied the suitability and differentiating potential of some standard bacteriological and molecular methods. Differentiation of the strains was performed using LOPAT, GATTa and ice nucleation tests, nutrient sucrose broth growth and utilization of various carbon sources. PCR method enabled the detection of toxin-producing genes: syrB and syrD in P. s. pv. syringae, and cfl gene in P. s. pv. morsprunorum race 1. Syringomycin production by pv. syringae was confirmed in bioassay using Geotrichum candidum, Saccharomyces cerevisiae and Rhodotorula pilimanae as indicator organisms. Pathogenicity test on lemon and immature nectarine fruits, as well as on string bean pods, showed different intensity of reaction of the inoculated material which could separate pv. syringae from the other two pathovars. PCR-based repetitive sequences, Rep-PCR with REP, ERIC and BOX primers revealed different genetic profiles within P. syringae pathovars.Patogeni varijeteti Pseudomonas syringae poreklom sa koštičavih voćaka poseduju brojne zajedničke karakteristike u pogledu kruga domaćina, simptomatologije i biohemijskofizioloških osobina, što otežava njihovu identifikaciju. U cilju odabira testova pogodnih za brzu i pouzdanu identifikaciju P. s. pv. syringae, morsprunorum i persicae, primenjeni su standardni bakteriološki i molekularni testovi. Diferencijacija sojeva izvršena je LOPAT i GATTa testovima, posmatranjem razvoja u hranljivom rastvoru sa saharozom, sposobnošću sojeva da formiraju čestice leda, kao i mogućnošću korišćenja različitih ugljenikovih jedinjenja. PCR metod korišćen je u detekciji gena odgovornih za proizvodnju toksina siringomicina kod soja P. s. pv. syringae (syrB i syrD geni) i koronatina kod soja P. s. pv. morsprunorum rase 1 (cfl gen). Proizvodnja siringomicina potvrđena je i biotestom, korišćenjem gljiva Geotrichum candidum, Saccharomyces cerevisiae i Rhodotorula pilimanae kao indikatora. Proverom patogenosti sojeva na plodovima limuna, nesazrelim plodovima nektarine i mahunama boranije, došlo je do ispoljavanja simptoma različitog intenziteta, na osnovu kojih se može izdvojiti pv. syringae od ostala dva patovara. Primenom Rep-PCR metode, uz korišćenje REP, ERIC i BOX prajmera, ustanovljene su razlike u genetskim profilima proučavanih P. syringae patogenih varijeteta

Bakteriozna plamenjača vinove loze - Xylophilus ampelinus

Bacterial blight of grapevine, caused by phytopathogenic bacterium Xylophilus ampelinus, is economically important disease that can significantly affect grapevine production, leading to the overall yield reduction and reduced vitality of infected grapevines. In regions where disease was recorded, losses in production, especially in susceptible varieties, can reach up to 80% of total yield. The bacterium infects only grapevine. In infected plants, the pathogen is located in the vascular tissues (xylem) from where it spreads further, causing a systemic infection of the host plant. During vegetation it is transmitted from plant to plant to short distance and the main source of inoculum for long distance dissemination are infected cuttings used either as rootstocks or grafting material. There are no completely resistant varieties or effective methods for controlling the disease. In Serbia X. ampelinus has a quarantine status. Therefore, it is of utmost importance to prevent introduction of the pathogen by inspecting the health of reproductive plant material, especially of those originating from countries where the pathogen is already present.Bakteriozna plamenjača vinove loze, koju prouzrokuje fitopatogena bakterija Xylophilus ampelinus, je ekonomski značajno oboljenje koje može ugroziti proizvodnju vinove loze umanjujući ukupan prinos i smanjujući dugovečnost zaraženih biljaka. U regionima gde je zabeleženo prisustvo bolesti gubici u proizvodnji, naročito kod osetljivih sorti, mogu dostići i do 80% ukupnog prinosa. Krug domaćina ove bakterije ograničen je samo na vinovu lozu. U zaraženim biljkama patogen se nalazi u sudovnom sistemu (ksilemu) odakle se dalje širi prouzrokujući sistemičnu infekciju biljke domaćina. Tokom vegetacije prenosi se u neposrednoj okolini izvora zaraze sa biljke na biljku, a na veću udaljenost zaraženim sadnim materijalom koji predstavlja primarni izvor inokuluma. Ne postoje potpuno otporne sorte ni dovoljno efikasne metode zaštite od ove bolesti. U Srbiji X. ampelinus ima karantinski status. Stoga je od najvećeg značaja sprečavanje unošenja patogena kontrolom zdravstvene ispravnosti biljnog materijala za reprodukciju, posebno iz zemalja gde je utvrđeno prisustvo patogena

Bakterioze tikava

Production of cucurbits has a long tradition worldwide. Cucumber, watermelon and melon production is very important for many producers in some parts of Serbia. These crops are especially susceptible to phytopathogenic bacteria which can cause substantial losses in years with favorable weather conditions. In this paper major bacterial diseases of cucurbits are described, such as bacterial fruit blotch caused by Acidovorax citrulli, angular leaf spot caused by Pseudomonas syringae pv. lachrymans, bacterial spot caused by Xanthomonas campestris, bacterial wilt caused by Erwinia tracheiphila, cucurbit yellow wine disease caused by Serratia marcescens, bacterial leaf spot, caused by Pseudomonas syringae pv. syringae, inducing bacterial soft rot caused by Pectobacterium species.Biljke familije Cucurbitaceae imaju dugu tradiciju proizvodnje širom sveta. U pojedinim krajevima Srbije, naročito severnim, proizvodnja krastavca, lubenice i dinje predstavlja primarno zanimanje mnogih poljoprivrednih gazdinstava. Pojava bakterioza u godinama sa uslovima koji pogoduju nastanku infekcije, može značajno ugroziti proizvodnju ovih biljaka. U ovom radu predstavljena su najznačajnija bakteriozna oboljenja biljaka familije Cucurbitaceae, među kojima su: bakteriozna mrljavost plodova lubenice, čiji je prouzrokovač Acidovorax citrulli, uglasta pegavost lišća krastavca, prouzrokovana bakterijom Pseudomonas syringae pv. lachrymans; bakteriozna uvelost krastavca prouzrokovana vrstom Erwinia tracheiphila; bakteriozna pegavost koju prouzrokuje Xanthomonas cucurbitae; potom žutilo vreža, čiji je prouzrokovač Serratia marcescens; zatim bakteriozna lisna pegavost prouzrokovana Pseudomonas syringae pv. syringae; kao i bakteriozna vlažna trulež prouzrokovana vrstama roda Pectobacterium

Specificity and sensitivity of three pcr-based methods for detection of erwinia amylovora in pure culture and plant material

Three PCR methods, referred in this study as,conventional" "nested" and,chromosomal" PCR and suggested for routine detection of Erwinia amylovora in pure culture and plant material, were evaluated according to their specificity and sensitivity. Specificity of PCR methods was analyzed by using 42 strains of E. amylovora, originating from different locations and plant species, with diverse PFGE profiles, representing distant populations of the pathogen. Sensitivity of PCR protocols in pure culture was studied by using nine different concentrations of E. amylovora in sterile ultrapure water as a template in PCR reactions. In order to study inhibitory effect of plant DNA and other inhibitors on sensitivity of the three PCR methods bacterial dilutions were mixed with plant macerate of pear, apple and quince prior to the PCR reaction. In specificity assays, tested PCR protocols were able to detect all E. amylovora strains regardless of the host of the strain, its origin or PFGE group, indicating primer specificity. On the other hand, sensitivity among tested methods varied, depending on bacterial concentration and selected plant material used in the PCR. When working with pure cultures nested PCR showed the greatest sensitivity by detecting 1.9 bacterial cells per PCR reaction, followed by detection limit of 9.5 cells per PCR reaction with conventional PCR and 1.9.105 cells/PCR reaction with chromosomal PCR. In spiked samples plant inhibitors either did not affect or they decreased the sensitivity of the PCR reaction, depending on the protocol and/or type of plant macerate. In our experiments, inhibitors from pear and quince macerates did not affect sensitivity of nested PCR, while apple macerate reduced its sensitivity by a factor of 10. Conventional PCR protocol was able to detect 95 cells/PCR reaction in pear and apple macerate, but only 9.5.103 cells/PCR in quince macerate. Greatest decrease in sensitivity of the PCR method was observed in spiked samples with chromosomal PCR since bacterial DNA was not detected in each of the spiked samples. Our research shows that all three PCR protocols are specific for detection of E. amylovora, but nested PCR proved to be most sensitive when working with pure cultures and plant material

Draft Genome Sequences of Agrobacterium nepotum Strain 39/7(T) and Agrobacterium sp. Strain KFB 330

Tumorigenic strains of Agrobacterium spp. are responsible for crown gall disease of numerous plant species. We present here draft genome sequences of nonpathogenic Agrobacterium nepotum strain 39/7(T) (CFBP 7436(T), LMG 26435(T)), isolated from crown gall tumor on Prunus cerasifera, and tumorigenic Agrobacterium sp. strain KFB 330 (CFBP 8308, LMG 28674), isolated from galls on raspberry

'Candidatus Liberibacter solanacearum' - prouzrokovač 'zebrastog čipsa' krompira

Candidatus Liberibacter solanacearum (CaLsol) is a phloem-limited pathogen and it was found associated with the 'zebra chip' potato disease. This pathogen causes signifi cant economic losses by reducing the yield and quality of potato crops. It is widespread in commercial potato fi elds in USA, Mexico, Central America ,New Zealand. CaLsol can also affect other solanaceous crops, including tomato, pepper, eggplant, tamarillo. Recently, it was detected in several Northern European countries in carrot and celery crops. Chips made from zebra chip-infected tubers show dark stripes that become more visible upon frying, and are therefore commercially unacceptable. Bearing in mind the importance of potato production in our country, it is necessary to raise the awareness about the disease occurrence and distribution.Među najznačajnijim štetnim organizmima koji ugrožavaju proizvodnju krompira od nedavno se ubraja fastidiozna bakterija 'Candidatus Liberibacter solanacearum' (CaLsol). Ovaj patogen odgovoran je za pojavu novog oboljenja krompira pod nazivom 'zebrasti čips' (ZČ), rasprostranjenog u proizvodnim područjima Amerike i Novog Zelanda. Iako je glavni domaćin patogena krompir, ozbiljne štete zabeležene su i na biljkama paprike, paradajza i drugim gajenim vrstama iz familije Solanaceae. U Evropi isti patogen nedavno je otkriven na obolelim biljkama mrkve i celera. Na krompiru, kao posledica infekcije dolazi do smanjenja prinosa zaraženih biljaka, a najznačajnije štete nastaju usled pojave simptoma na krtolama i gubitka njihove tržišne vrednosti. S obzirom na ekonomski značaj proizvodnje krompira u našoj zemlji, kao i na mogućnost unošenja bakterije putem vegetativnog sadnog materijala, postoji opravdana zabrinutost od širenja ovog patogena na teritoriji naše zemlje

Novo destruktivno oboljenje krompira - zebrasti čips

Potato production provides one of the main sources for food production, processed either at home or by the industry. Plant diseases are often limiting factor in cultivation of this crop, leading to reduction of yield and tuber quality. Zebra chip disease, caused by a floem-limited bacterium, belonging to the genus Candidatus Liberibacter, is a new, economically important disease of potato and other solanaceous plants. It is widespread in commercial potato fields in USA, Mexico, Central America, New Zealand, and was recently reported in Europe. The disease reduces yield, but most significant damage occurs due to the symptom development on tubers and loss of their marketability. Given the importance of potato production in our country, the goal of this paper is to raise the awareness about the disease occurrence and distribution, discuss the main characteristics of this relatively unknown pathogen, detection and identification methods necessary for the successful and timely diagnosis, epidemiology, symptomatology and disease management.Krompir je biljna vrsta čija proizvodnja ima veoma značajnu ulogu u obezbeđivanju hrane, a takođe i sirovine za industrijsku preradu. Pojava štetnih organizama neretko predstavlja ograničavajući faktor gajenja ove biljke, jer dovodi do smanjenja prinosa i pogoršanja kvaliteta krtola. Oboljenje nazvano zebrasti čips, koje prouzrokuje fastidiozna bakterija iz roda Candidatus Liberibacter, je novo, ekonomski značajno oboljenje biljaka iz familije Solanaceae, čiji je glavni domaćin krompir. Patogen, takođe, zaražava i srodne vrste kao što su paradajz i paprika. Ovo destruktivno oboljenje rasprostranjeno je u proizvodnim područjima Amerike, Novog Zelanda, a nedavno je otkriveno i na evropskom kontinentu. Kao posledica infekcije, dolazi do smanjenja prinosa zaraženih biljaka, dok najznačajnije štete nastaju usled pojave simptoma na krtolama i gubitka njihove tržišne vrednosti. S obzirom na značaj proizvodnje krompira, paradajza i paprike u našoj zemlji, cilj ovog rada je da se ukaže na pojavu i rasprostranjenost oboljenja, glavne karakteristike ovog malo poznatog patogena, metode detekcije i identifikacije neophodne za pouzdanu i pravovremenu dijagnozu oboljenja, epidemiologiju, simptomatologiju i mere zaštite

Bakteriozna oboljenja biljaka familije Cucurbitaceae

Cucurbits production has a long tradition in our country. In some parts of the country, cucumber, watermelon and melon are the most important agricultural crops. High yield and good profit are often compromized by various biotic and abiotic factors. In years with favorable weather conditions cucurbit bacterial diseases can cause serious damage. This paper describes major bacterial diseases of cucurbit plants, such as angular leaf spot caused by Pseudomonas syringae pv. lachrymans, bacterial fruit blotch caused by Acidovorax citrulli, bacterial spot caused by Xanthomonas campestris, bacterial wilt caused by Erwinia tracheiphila and cucurbit yellow wine disease caused by Serratia marcescens, relatively new and invasive disease across the United States. In addition, bacterial leaf spot, caused by Pseudomonas syringae pv. syringae and Pectobacterium carotovorum subsp. carotovorum, inducing bacterial soft rot, are becoming more important in cucurbit production worldwide.Biljke familije Cucurbitaceae se dugi niz godina gaje širom naše zemlje. U pojedinim krajevima, proizvodnja krastavca, lubenice i dinje predstavlja osnovni izvor prihoda za mnoga poljoprivredna domaćinstva. Visok prinos i ekonomsku dobit često ugrožavaju različiti biotski i abiotski faktori. U godinama sa uslovima povoljnim za nastanak infekcije, bakteriozna oboljenja mogu prouzrokovati velike gubitke. U ovom radu predstavljene su najznačajnije bakterioze biljaka familije Cucurbitaceae kao što su uglasta pegavost lišća krastavca, prouzrokovana bakterijom Pseudomonas syringae pv. lachrymans; bakteriozna mrljavost plodova lubenice, čiji je prouzrokovač Acidovorax citrulli; zatim bakteriozna pegavost koju prouzrokuje Xanthomonas cucurbitae; bakteriozna uvelost krastavca prouzrokovana vrstom Erwinia tracheiphila i žutilo vreža, čiji je prouzrokovač Serratia marcescens. Gubicima u proizvodnji doprinose i prouzrokovači bakteriozne pegavosti, Pseudomonas syringae pv. syringae, kao i bakteriozne vlažne truleži, Pectobacterium carotovorum subsp. carotovorum

Rasprostranjenost, biologija i štetnost mrkvine lisne buve Bactericera trigonica (Hodkinson, 1981) (Hemiptera, Triozidae) u Srbiji

Carrot psyllid Bactericera trigonica belongs to the family Triozidae, superfamily Psylloidea, order Hemiptera. Together with onion psyllid (B. tremblayi) and potato psyllid (B. nigricornis), it is a member of the complex Bactericera nigricornis Foerster with morphologically and bioecologically very similar species. It was originally described by Hodkinson (1981) as Trioza trigonica based on the analysis of adults collected from the carrots in Portugal, Italy, Cyprus, Turkey, Egypt and Iran. Later it has been transferred to the genus Bactericera together with the two other species from this complex. Adults and larvae B. trigonica, feed on carrot leaf and other Apiaceae. If they appears in large number, they can lead to chlorosis and leaf yellowing, which causes a yield reduction. Besides direct damage, its vector role of bacteria “Candidatus Liberibacter solanacearum” has been determined, which leads to root proliferation and weakening of plants from Apiaceae family due to chlorosis. Ca. Liberibacter solanacearum has been known in North Africa and New Zealand for a long time on plants from Solanacae family, potato in the first place, while in Europe it has been determined on carrot and cellery for the first time, with B. trigonica as its vector. The presence, biology and harmfulness of B. trigonica in the region of Serbia were studyed in the period from 2014 to 2017. It was found in 2014 for the first time and since then, its presence has been determined in all researched localities, during the whole vegetative period of carrot. Besides carrot, B. trigonica has been registered on celery, parsley and wild carrot as well. It has a several generations per year, overwintering as adults.Mrkvina lisna buva Bactericera trigonica pripada familiji Triozidae, nadfamiliji Psylloidea, redu Hemiptera. Zajedno sa lukovom lisnom buvom (B. tremblayi) i krompirovom lisnom buvom (B. nigricornis) pripada kompleksu Bactericera nigricornis Foerster, u morfološkom i bioekološkom smislu veoma sličnih vrsta. Opisana je 1981. godine kao Trioza trigonica na osnovu analize imaga sakupljanih sa mrkve iz Portugalije, Italije, Kipra, Turske, Egipta i Irana. Kasnije je prebačena u rod Bactericera zajedno sa druge dve vrste ovog kompleksa. Imaga i larve se hrane na listu mrkve i drugih štitonoša. Ukoliko se jave u povećanoj brojnosti mogu dovesti do hloroze i žućenja lista, što ima za posledicu smanjenje prinosa. Pored direktnih šteta, utvrđena je njena vektorska uloga fastidiozne bakterije «Candidatus Liberibacter solanacearum», koja na biljkama iz familije Apiaceae dovodi do proliferacije korena i hlorotičnog slabljenja. Ca. L. solanacearum je odavno poznata u Severnoj Americi i Novom Zelandu na biljkama iz familije Solanacae, prevashodno na krompiru, a u Evropi je prvi put utvrđena na mrkvi i celeru, a B. trigonica kao njen vektor. U periodu od 2014. do 2017. godine proučavano je prisustvo, biologija i štetnost B. trigonica na području Srbije. Prvi put je registrovana 2014. godine i od tada je utvrđeno njeno prisustvo u svim lokalitetima istraživanja tokom celog vegetacionog perioda mrkve. Pored mrkve, B. trigonica je registrovana na celeru, peršunu i divljoj mrkvi. U toku godine ima veći broj generacija, prezimljava u stadijumu imaga na skrovitim mestima

Bakteriozna mrljavost plodova lubenice u Srbiji

During the summer of 2014, watermelon producers in the north of Serbia (Srem district), observed the occurrence of irregularly shaped, water-soaked lesions on mature watermelon fruits indicating possible bacterial nature of the disease. Bacteria were isolated from symptomatic samples and identified as Acidovorax citrulli, previously described as a causal agent of bacterial fruit blotch. In the past few years, this pathogen caused considerable damage to the watermelon industry of the United States where, in some commercial production sectors, the loss of marketable fruit reached 90%. In Serbia, it is considered a quarantine pathogen and was not recorded until 2014.U leto 2014. godine, proizvođači lubenice iz sremskog okruga uočili su pojavu simptoma u vidu pega vodenastog izgleda i nepravilnog oblika na zrelim plodovima lubenice, koji su ukazivali na oboljenje bakteriozne prirode. Izolacijom i identifikacijom patogena, utvrđeno je prisustvo bakterije Acidovorax citrulli, prouzrokovača bakteriozne mrljavosti plodova lubenice. Ova vrsta je u skorije vreme dosta dobila na značaju, posebno zbog gubitaka u proizvodnji koji su u pojedinim oblastima Sjedinjenih Američkih Država (SAD) dostizali i 90% od ukupnog prinosa. U Srbiji A. citrulli ima karantinski status i nalazi se na A1 listi Evropske i mediteranske organizacije za zaštitu bilja (EPPO)