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Molekularna karakterizacija 'Candidatus Phytoplasma solani' na celeru: studija u Futogu
No full text‘Candidatus Phytoplasma solani’, known by its trivial name stolbur phytoplasma, is a plant pathogen
infecting numerous crops in Serbia. Celery plants with prominent leaf yellowing and chlorosis, sporadically with tissue
necrosis, were observed during august 2020 in Futog, situated in Novi Sad suburbia in Vojvodina. Total of 12 sampled
celery plants, 8 symptomatic and 4 asymptomatic ones, were analysed for ‘Ca. P. solani’ presence. All symptomatic
celery plants were infected with stolbur phytoplasma according to the stol11 gene detection and therefore were further
subjected to multigene molecular characterization on three genes: tuf, stamp and vmp1. Combining molecular tools
PCR/RFLP and sequencing reviled two ‘Ca. P. solani’ multilocus genotypes in celery: tuf-b/Rqg31/V14 and tuf-b/
Rpm35/V14, present in 4 samples each. Obtained results of the strain genotyping are in concordance with previous
data on the ‘Ca. P. solani’ diversity on celery, but supplemented with genotyping of the vmp1 gene. Outbreak of stolbur
phytoplasma in the assessed locality in Futog is linked to tuf-b epidemiological cycle correlated in Serbia mainly with
weed Convolvulus arvensis which was present in the subjected celery plot, and could have been the phytoplasma
inoculum source. Visual evaluation of the symptom occurrence suggests on 10-15% of ‘Ca. P. solani’ affected celery
plants scattered throughout the plot corresponding to the pathogen dispersal in crop by cixiid planthoper Hyalesthes
obsoletus (Hemiptera: Auchenorrhyncha) associated with C. arvensis, main vector of stolbur phytoplasma in Serbia.‘Candidatus Phytoplasma solani’, poznata po trivijalnom nazivu stolbur fitoplazma, biljni je patogen koji
inficira brojne kulture u Srbiji. Biljke celera sa izraženim žutilom i hlorozom listova, sporadično i sa nekrozom tkiva,
uočene su u avgustu 2020. u Futogu, predgrađu Novog Sada u Vojvodini. Ukupno 12 uzoraka celera, 8
simptomatičnih i 4 asimptomatične biljke, analizirane su na prisustvo ‘Ca. P. solani’. Svi simptomatični uzorci su se
pokazali inficiranim stolbur fitoplazmom na osnovu analize stol11 gena i dalje su molekularno okarakterisani na 3
gena: tuf, stamp i vmp1. Kombinacijom molekularnih tehnika PCR/RFLP i sekvenciranja utvrđeno je prisustvo dva
‘Ca. P. solani’ multigenska genotipa, tuf-b/Rqg31/V14 i tuf-b/Rpm35/V14, prisutnih u po četiri uzorka. Dobijeni
rezultati genotipizacije uzoraka stolbur fitoplazme odgovaraju prethodnim podacima o diverzitetu ovog patogena na
celeru u Srbiji i dopunjeni su sa genotipizacijom vmp1 gena. Pojava stolbur fitoplazme u Futogu je vezana za tuf-b
epidemiologiju, koja uglavnom izvire iz korovske vrste Convolvulus arvensis (poponac), koja je bila prisutna u
istraživanom zasadu celera i mogla je biti izvor inokuluma. Vizuelna procena pojave simptoma pokazala je 10-15%
prisustva simptomatičnih biljaka celera, nepravilno raspoređenih unutar zasada što ukazuje da je odgovorni vektor
najverovatnije cikada Hyalesthes obsoletus (Hemiptera: Auchenorrhyncha) asocirana sa poponcem, najznačajniji vektor stolbur fitoplazme u Srbiji
The effectiveness of low temperature (5 degrees C) on Sitophilus oryzae (L.), Sitophilus zeamais (Motch.) and Sitophilus granarius (L.) in wheat grain: The impact of pre-acclimation
No full textLaboratory trials were conducted in wheat grain to assess the effectiveness of low temperature (5 degrees C), applied at intervals of 5, 7, 9, 11, 20, 25, 30 and 35 days (d), as a control measure for stored product insect pests of the genus Sitophilus, and compared the results with the impacts of 7 d pre-acclimation at 15 degrees C and non-acclimation. Mortality was determined after 1, 2 and 7 d of recovery in the laboratory at 25 +/- 1 degrees C and 60 +/- 5% r. h., while progeny were assessed 7 weeks later. Acclimation had the strongest impact on S. oryzae after 5 d exposure to 5 degrees C and 7 d recovery, while the same effect on S. zeamais was shown after 20 and 25 d of exposure and 1 d recovery, and the same impact on S. granarius occurred after 30 and 20 d of exposure and 1 and 7 d recovery, respectively. Low temperature was found to be the most effective against S. oryzae, and the least effective against S. granarius. Mortality was high ( gt = 96%) in both S. oryzae test groups after 11 d of exposure and 7 d recovery, S. zeamais mortality was high ( gt 97%) after 35 and 30 d of exposure and 2 and 7 d of recovery, respectively, and S. granarius mortality was lt = 87% in all test variants. Total reduction (100%) of non-acclimated S. oryzae, S. zeamais and S. granarius progeny required 8, 25 and 30 d of exposure to 5 degrees C, respectively, while acclimated weevils required 11, 30 and 30 d, respectively. (C) 2020 Elsevier Ltd. All rights reserved
Resistance of Monilinia fructicola to thiophanate-methyl in Croatia
No full textCroatian isolates ofMonilinia fructicola,M.laxaandM.fructigenahave been collected from peach, nectarine, sweet cherry, plum and apricot fruits and assessed for their sensitivity to thiophanate-methyl. Out of 66 isolates collected, 34 were identified asM.fructicola(52%), 22 asM. fructigena(33%) and 10 asM. laxa(15%). Mycelial radial growth on water agar amended with thiophanate-methyl was measured to determine half maximal effective concentration values (EC50). All isolates ofM.laxaandM.fructigenashowed as sensitive, with EC(50)values lower than 1 mu g ml(- 1)in 31 from 32 isolates. InM. fructicola, 19 isolates (56%) were sensitive, nine (26%) were resistant (EC(50)2-30 mu g ml(- 1)), and six (18%) were highly resistant (EC50 gt 30 mu g ml(- 1)). Positive allele-specific PCR with primer pair HRR/HRF, detecting resistant point mutations in beta-tubulin gene, was recorded in 15 out of 16 highly resistant, resistant and less sensitiveM.fructicolaisolates. Among all species, PCR assay coincided with agar assay in 95% of cases. Highly resistantM.fructicolaisolates AL 24/19 and VR 8/18 caused typical brown rot on nectarine fruits inoculated with conidia suspended in 5 or 10 mu g ml(- 1)of thiophanate-methyl. Four sensitive isolates of all three species did not develop on inoculated fruits. Besides implications for brown rot management, the finding ofM. fructicolastrains resistant to thiophanate-methyl may indicate that this species could be adapting to fungicides more rapidly thanM. laxaorM. fructigena. As resistantM. fructicolaisolates were also found in orchards where benzimidazoles have never been used, there is a possibility they originate from already benzimidazole-resistant strains which entered and established in Croatia
Genotypic differentiation of Monilinia spp. populations in Serbia using a high-resolution melting (HRM) analysis
No full textMonilinia laxa, Monilinia fructicola and Monilinia fructigena are the three main causal agents of brown rot, which is one of the most important diseases of stone fruits in pre- and postharvest conditions. Nowadays, the need for the precise genotyping of these Monilinia species in terms of the genetic diversity of their populations or differences in their pathogenicity and host range is a prerequisite for any efficient disease management. In our study, the genetic structure of Monilinia populations in Serbia from three geographically distinct regions was investigated employing a high-resolution melting (HRM) analysis which is a sensitive and rapid molecular approach in fungal genotyping and diagnostics. Using species-specific primer pairs genotype-specific HRM melting curve profiles were generated allowing to efficiently decipher the genetic diversity of the Monilinia populations. The Monilinia genotypes could be easily distinguished according to their melting curves. The isolates from the northern region were assigned to distinct genotypes and grouped rather independently compared to the isolates of the other two regions among all three tested Monilinia spp. M. fructicola and M. fructigena showed a higher genetic diversity among their populations (44%) compared with the genetic diversity among the M. laxa populations (7%). In contrast, the genetic variance within the pathogen populations was higher in the case of M. laxa (93%). Our data revealed an absence of host specificity in the Monilinia spp. populations
Vilina kosica: stari problem traži nova rešenja
No full textVilina kosica je najrasprostranjenija parazitna cvetnica u Srbiji. Za sada se može
tvrditi da su ekonomski najvažnije vrste, prisutne kod nas, Cuscuta campestris Yunck.
i Cuscuta epythimum L. Ove dve vrste se po načinu vezivanja za biljku domaćina
razlikuju i to utiče na izbor mera njihovog suzbijanja, pa je pre svega, potrebno
determinisati koja vrsta je prisutna i tome prilagoditi način suzbijanja. Imajući u vidu
značaj viline kosice i štete koje može da nanese, u mnogim zemljama, takođe i kod
nas, ona je svrstana u kategoriju karantinskih biljnih parazita (A2 grupa), čije je
sprečavanje širenja i suzbijanje obavezno u celoj zemlji. Uprkos tome, zakonska
regulativa, kojom se sprečava širenje viline kosice se nedovoljno poštuje, što ovoj
parazitnoj cvetnici omogućava veću ekspanziju. Najveću pažnju treba obratiti na
useve u polju. Suzbijanje viline kosice treba početi čim se uoče početna žarišta
zaraze i završiti ga pre nego što vilina kosica cveta i obrazuje seme. Na jednoj biljci
može se obrazovati i do 15.000 semena koje može da sačuva vitalnost u zemljištu i
preko 10 godina, što predstavlja glavni mehanizam održavanja ove parazitne cvetnice
u prirodi. Prisustvo viline kosice na antropogenim staništima (različiti tipovi useva i
ruderalna staništa urbanih i ruralnih područja) pre više od jedne decenije u Srbiji je
zabeleženo na 25% ocenjenih površina, a danas su te površine značajno uvećane.
Najveće štete kod nas vilina kosica pravi kada se u velikim infestacijama javi na tek
zasnovanim višegodišnjim leguminozama (lucerištima, deteliništima), koji ujedno
spadaju u najčešće parazitirane useve od strane ove parazitne cvetnice. Vilina kosica
je jedan od glavnih uzroka proređivanja lucerišta i može izazvati smanjenje prinosa i
do 80%. Takođe, poslednjih godina vilina kosica je u ekspanziji u usevu šećerne
repe, gde pravi sve veće probleme. Štete koje vilina kosica pravi u usevu šećerne
repe se prvenstveno odnose na redukciju prinosa (i do 40%) i sadržaja šećera (i do
5%). Pored ovih, problemi sa vilinom kosicom se javljaju i pri proizvodnji rasada
povrtarskih biljaka (paradajz, paprika, kupus), kao i plasteničkoj proizvodnji, krompiru,
šargarepi i lekovitom bilju. Primena samo hemijskog suzbijanja često ne daje
zadovoljavajuće rezultate, pa se pod najefikasnijim suzbijanjem viline kosice
podrazumeva sistematski pristup integralnim merama zaštite. Treba početi od
monitoringa viline kosice kako u usevima, tako i na ruderalnim površinama, a zatim
adekvatne rotacije useva koja podrazumeva gajenje biljaka koje nisu podobni
domaćini viline kosice, kao i primene svih preventivnih mera (čist semenski materijal,
čišćenje međa, okolnih parcela, mašina za kombajniranje i dr.). Direktne mere
suzbijanja su nehemijske mere uklanjanja (košenje i sagorevanje propanom) i na
kraju primena hemijskih mera (herbicida), kada se ovaj problem ne može rešiti drugim
putem. U poslednje vreme, usled nedostatka rešenja za suzbijanje ove parazitne
cvetnice, razvija se mogućnost primene bioloških agenasa (pojedine rizosferne
bakterije i aktinomicete, kao i eterična ulja
Crna trulež korena rasada Viola x wittrockiana
No full textU rasadnicima JKP „Zelenilo Beograd“ proizvodi se oko 500 000 komada biljaka i isto
toliko jesenjeg cvetnog rasada čime se zadovoljavaju potrebe za sadnju cvetnih površina,
žardinjera, saksija na stubovima rasvete i zelenih zidova na svim opštinama grada Beograda.
Proizvodnja se obavlja u lejama i delom na njivi na ukupnim površinama od oko 6 000 m2 u
dva proizvodna ciklusa: letnjem (od februara do maja) i jesenjem (od jula do septembara).
Gajenje biljaka na istom zemljištu u uskom plodoredu i ograničenom broju dozvoljenih
sredstava za hemijsku sterilizaciju zemljišta, opterećeno je brojnim problemima u ishrani i
zaštiti biljaka od štetnih organizama. S obzirom da u proizvodnji ukrasnog bilja, svaki
simptom na listu ili cvetu značajno umanjuje njegovu dekorativnu vrednost, primenjuje se
intezivna hemijska zaštita koja obuhvata sprečavanje pojave najčešćih problema u vidu
poleganja rasada, pegavosti lista, sive truleži biljaka i bele leptiraste vaši. U 2020. godini, setvi
cvetnog rasada Viola x wittrockiana prethodila je obrada zemljišta i hemijska sterilizacija
dazometom. Nakon setve vršena je preventivna hemijska zaštita protiv prouzrokovača
poleganja rasada u više navrata propamokarb-hidrohloridom i fosetil-aluminijumom. Cvetni
rasad bio je odličnog zdravstvenog statusa do faze razvoja prvog para stalnih listova. Nakon
toga, u kratkom vremenskom intervalu došlo je do žućenja donjih listova, pojave istanjenih
korenovih vratova i korena tamno-smeđe do crne boje, što je ubrzo uzrokovalo masovno
“košenja rasada”. Na osnovu simtoma, bolest se nije mogla pripisati ni jednom prouzrokovaču
oboljenja. Na obolelim korenovima, mikroskopskim pregledom uočeno je prisustvo
hlamidospora Thielaviopsis basicola, poznatog prouzrokovača crne truleži korena. Međutim,
izolacija patogena iz obolelih biljnih delova nije bila uspešna, tako da su dalja labaratoriska
ispitivanja u toku.
S obzirom da je proizvodnji pretila velika šteta, a da u Srbiji nema registrovanih
preparata za suzbijanje fitopatogenih gljiva prouzrokovača bolesti korena u toku vegetacije,
na osnovu literalnih podataka, primenjeni su preparati na bazi tiofanat-metila, fludioksinila, i
triadimenola, u preporučenim dozama, u više navrata alternativno, poštujući strategiju o sprečavanju razvoja rezistentnosti, što je zaustavilo dalje širenje oboljenja. Primećeno je da su
nakon tretmana pojedine biljke V. x wittrockiana koje su se nalazile u zahvaćenim lejama i
ispoljavale simptome žućenja donjih listova i slabljenja čitavog nadzemnog dela, iznad mesta
tanjenja korena i korenovog vrata razvile adventivne korenove. Te biljke su u početku slabije
napredovale i bile su blago hlorotične, ali kasnije su se potpuno oporavile tako da se razlika u
odnosu na druge biljke nije videla
Savremeni pristup u suzbijanju biljnih patogena u zemljištu
No full textFitopatogene glјive i pseudoglјive koje se odrţavaju u zemlјištu,
predstavlјaju znaĉajan problem u proizvodnji ukrasnih bilјaka.
Dezinfekcija supstrata hemijskim fumigantima i primena fungicida
ĉinili su osnovu tradicionalnog naĉina suzbijanja ovih patogena u
našoj zemlјi dugi niz godina. MeĊutim, pokazalo se da hemijski
fungicidi mogu samo privremeno da pomognu u spreĉavanju razvoja
nekih obolјenja i nastajanja šteta u proizvodnji. Nјihova ĉesta i
neselektivna primena moţe da dovede do razvoja rezistentnosti
patogena, a povećana zabrinutost zbog mogućeg negativnog uticaja
na zdravlјe lјudi i ţivotnu sredinu dodatno ograniĉava njihovu
primenu. Noviji, sistematiĉni pristup u zaštiti bilјa od patogena iz
zemlјišta podrazumeva pre svega primenu mera kojima se
unapreĊuje mikrobiološka zajednica u zemlјištu i podstiĉu prirodni
mehanizmi regulacije populacija patogena, a koje obuhvataju:
primenu plodoreda, korišćenje pokrovnih useva i zelenišnog Ċubriva,
kao i primenu organskih dodataka za zemlјište. Osim ovih, u radu će
biti prikazane i neke ekološki prihvatlјive korektivne mere za direktno
suzbijanja patogena kao što su: sterilizacija zemlјišta vodenom parom i solarizacija, biofumigacija, anaerobna dezinfestacija
zemlјišta i primena preparata na bazi antagonistiĉkih
mikroorganizama.Mesto i datum održavanja: Univerzitet u Beogradu - Šumarski fakultet Beograd, 18-19. februar 2021. godin
Osetljivost populacija Brassicogethes aeneus (f.) (Coleoptera, Nitidulidae) na insekticide: rezultati testiranja 2021. godine
No full textRepičin sjajnik, Brassicogethes aeneus (F.) je najznačajniji štetni insekt uljane repice
koji se često suzbija insekticidima. Radi praćenja stanja i pravovremenog utvrđivanja
promene osetljivosti i rezistentnosti populacija na insekticide u Srbiji, tokom aprila 2021. godine su testirane populacije prikupljene sa ozime uljane repice na području Bajše, Bačke
Topole (Malog Beograda), Novog Sada (Rimskih šančeva) i Dobanovaca.
Prikupljena imaga B. aeneus su u skladu sa metodom IRAC (No. 027) do testiranja
čuvane u frižideru na 5°C. U biotestu, realizovanom u laboratorijskim uslovima, su korišćeni
insekticidi na bazi lambda-cihalotrina, cipermetrina, alfa-cipermetrina, deltametrina i
hlorpirifosa + cipermetrina, rastvoreni u smeši voda + aceton (5%+95%). Primenjeni su u
dozama od 100% i 25% od preporučene, tako što je na zidove staklenih posuda, dimenzija 5,5
cm x 2,5 cm i 48 cm2 unutrašnje površine, nanošeno po 0,5 ml rastvora svakog insekticida, uz
ručno rolovanje do sušenja depozita. U kontroli je korišćena samo smeša vode i acetona. Po
deset imaga je ubacivano u svaku posudu, a testiranje svake doze i insekticida je realizovano u
tri ponavljanja na temperaturi 22±2°C, relativnoj vlažnosti vazduha 60±5% i fotoperiodu
16:8. Beležen je broj uginulih insekata posle 24 časa od početka izlaganja svakom depozitu.
Prema klasifikaciji datoj u metodi IRAC (No. 011), dobijeni rezultati testiranja pokazuju
da populacije iz Novog Sada, Dobanovaca i Bajše spadaju u prvu grupu, visoko osetljive, na
insekticid hlorpirifos + cipermetrin, jer je smrtnost jedinki bila 100% kod obe koncentracije
(100% i 25% od preporučene). U drugu grupu, osetljive, spadaju sve četiri ispitivane
populacije sjajnika na alfa-cipermetrin, zatim populacija iz Bačke Topole na lambda-cihalotrin
i cipermetrin, i populacije iz Dobanovaca i Bajše na cipermetrin. U grupu tri, umereno
rezistentne, spada populacija iz Novog Sada na lambda-cihalotrin i cipermetrin, i populacije iz
Dobanovaca i Bajše na lambda-cihalotrin. Međutim, u grupu četiri, rezistentne, spadaju sve
populacije repičinog sjajnika na insekticid deltametrin, jer je smrtnost jedinki posle kontakta
sa koncentracijom 100% od preporučene bila oko 70%, a posle kontakta sa koncentracijom
25% od preporučene, u rasponu od 17 do 50%. Ove sezone ni za jednu od ispitivanih
populacija repičinog sjajnika na ozimoj uljanoj repici nije utvrđeno da pripada petoj grupi,
odnosno da je visoko rezistentna na insekticide
The Embryotoxic Potential and Photocatalytic Degradation of Thiophanate-Methyl
No full textPesticides are substances designed to protect plants
from various types of diseases and pests [1]under UV irradiation
is studied using synthesized Zinc oxide (ZnO.
Synthetic organic pesticides, in addition to the high
efficiency, have led to frequent adverse environmental
impact, as a consequence of their high accumulation
and toxicity. Due to the increased pollution of water
with mixture pesticides, it is necessary to use different
processes for their removal and degradation. Therefore,
oxidative processes have been developed, commonly
named as Advanced Oxidation Processes (AOPs).
Among them, special attention was attributed to photocatalysis,
as a process that enables the degradation
of difficult-to-decompose organic molecules under the
action of UV radiation in the presence of catalysts [2].
In this study the photocatalytic degradation of thiophanate-
methyl (TM) in the presence of the TiO2 Degussa
P-25 catalyst was investigated. Different experimental
conditions were varied, such as the concentration of
the pesticide solution, the mass of the catalyst and the
influence of the anions (chloride, sulphate, nitrate, etc.).
The pesticide concentration in the reaction system was
monitored based on the decrease in system absorbance
using a Shimadzu 1800 UV spectrophotometer. For optimized
conditions of complete photodegradation, the
environmental acceptability of the defined degradation
process was examined. The toxic effect of the TM solution
before and after degradation was examined using
the embryotoxicity test with Danio rerio, in order to
prove the reduction of toxicity and the success of the
degradation process [3]which raises the issue of potential
influence of different formulation types on herbicide
toxicity. The present study evaluated the toxicity and
teratogenic effects of the active ingredient clomazone
and its two formulations (Rampa® EC and GAT Cenit
36 CS, both containing 360 g a.i./l of clomazone.
Comparing the obtained results of the influence of
ions on the processes of photocatalysis, it was noticed
that all ions have catalytic effects on the kinetics of the
degradation process of TM. The presence of sulphates
and carbonates had the greatest catalytic effect, while
hydrogen phosphates and bicarbonates showed the lowest
catalytic capacity.
The optimal experimental conditions were obtained
using 0,2 g/L of TiO2 and 5 mg/L of TM solution. In
addition, the embryotoxicity test followed the analytical
examination. Comparison of results obtained in embryotoxicity
assay testing of the initial solution, partly
and completely degraded samples confirmed suitability
of applied degradation method. Increase in toxicity,
compared to the initial solution, was registered in partly
degraded sample. This observation can be attributed to
increase in concentration of carbendazim (TM metabolite)
more toxic than parent substance. Finally, completely
degraded sample caused no mortality or adverse
effects in D. rerio embryos after 120 h exposure. Toxicity
of samples, in decreasing order is half degraded >
initial >completely degraded sample.
Based on the obtained results it can be concluded
that used photocatalytic degradation process can be successfully
applied in pesticide contaminated water management
Acaricide resistance in Panonychus citri and P. ulmi (Acari: Tetranychidae): Molecular mechanisms and management implications
No full textThe European red mite, Panonychus ulmi (Koch), and the citrus red mite, P. citri (McGregor) (Acari: Tetranychidae), are destructive mite pests in pome/stone fruit orchards and citrus groves, respectively, the management of these two spider mites species has relied largely on the use of synthetic acaricides. However, frequent, long-term use of acaricides has caused rapid development of resistance in P. ulmi and P. citri populations worldwide. Levels of resistance in P. ulmi and P. citri are exceeded only by that in the two-spotted spider mite, Tetranychus urticae Koch. Recent research on T. urticae has provided new insights into the molecular mechanisms of acaricide resistance. The molecular basis of resistance in P. ulmi and P. citri populations has been less thoroughly studied. In this brief review, the current understanding of the molecular mechanisms of toxicokinetic and toxicodynamic resistance of P. ulmi and P. citri to acaricides, as well as implications of the findings for the resistance management, are discussed