Dulić-Marković, Ivana

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Methods for detection of Phytophthora fragariae var. rubi on raspberry

Koprivica, Mirjana; Dulić-Marković, Ivana; Jevtić, Radivoje; Cooke, Dave E.L.

(Institut za pesticide i zaštitu životne sredine, Beograd i Društvo za zaštitu bilja Srbije, Beograd, 2009)

TY  - JOUR
AU  - Koprivica, Mirjana
AU  - Dulić-Marković, Ivana
AU  - Jevtić, Radivoje
AU  - Cooke, Dave E.L.
PY  - 2009
UR  - http://fiver.ifvcns.rs/handle/123456789/696
AB  - Phytophthora fragariae var. rubi (Wilcox & Duncan), a causal agent of raspberry root rot, is a serious soil-borne pathogen listed by EPPO as an A2 quarantine pest. Root samples were collected from badly diseased raspberry plants showing a variety of characteristic and often dramatic symptoms during surveys carried out in western Serbia in 2002. Identification of the causal agent was performed in collaboration work with the Scottish Crop Research Institute (S.C.R.I.), Dundee, UK. Necrotic roots were plated on selective French bean agar (incorporating ampicilin, ryfamicin, bavistin and hymexasol). Detection of isolates was based on cultural and morphological features compared with referent cultures. DNA was extracted directly from the sampled roots using extraction buffer (200 mM Tris-HCl pH 8.5, 250 mM NaCl, 25 mM EDTA, 0.5% SDS), purified by multi spin separation columns [Thistle Scientific (Axygen)] or in 24:1 mixture of chlorophorm-isoamyl alcohol and amplified by nested PCR (ITS 4 and DC 6 for first round, DC 1 and DC 5 for second round). Diluted DNA extracts were also amplified by conventional PCR with modified 'universal' Phytophthora primers (ITS 6, ITS 7 and ITS 8, Cooke et al., 2000) and digested with Msp1. Digestion patterns of the universal primers PCR product from infected roots matched those of Scottish strains. P. fragariae var. rubi occurred on 8 out of 14 sites. Our results indicate that nested PCR (ITS 4 and DC 6 for first round, DC 1 and DC 5 for second round) or digestion of the 'universal' Phytophthora primers PCR product for detection of P. fragariae var. rubi are more sensitive and less time-consuming and therefore recommended for use.
AB  - Phytophthora fragariae var. rubi (Wilcox & Duncan), prouzrokovač truleži korena maline, je veoma značajan zemljišni patogen A2 karantinskog statusa u EPPO. Uzorci korena su sakupljeni sa jako zaraženih biljaka maline koje su ispoljavale čitav niz karakterističnih i često dramatičnih simptoma tokom pregleda koji su vršeni u zapadnoj Srbiji 2002. godine. Identifikacija prouzrokovača je izvršena u saradnji sa Scottish Crop Research Institute (S.C.R.I.), Dundee, UK. Nekrotični korenčići su unošeni u selektivnu French bean podlogu, koja je sadržala ampicilin, rifamicin, bavistin i himeksazol. Detekcija dobijenih izolata je vršena posmatranjem kulture i morfoloških karakteristika i poređenjem sa autentičnim kulturama. DNK je ekstrahovana direktno iz uzorkovanih korenova pomoću ekstrakcionog pufera (200 mM Tris-HCl pH 8,5, 250 mM NaCl, 25 mM EDTA, 0,5% SDS), prečišćena Multi spin separacionim kolonama (Thistle Scientific (Axygen) ili 24:1 mešavinom hloroforma i izoamil alkohola i amplifikovana nested PCR (ITS 4 i DC 6 za prvi krug, DC 1 i DC 5 za drugi krug). Rastvoreni DNK ekstrakti su takođe amplifikovani putem konvencionalnog PCR sa modifikovanim 'univerzalnim' Phytophthora prajmerima (ITS 6, ITS 7 i ITS 8, Cooke i sar., 2000) i di gestirani sa Msp1. Digestioni paterni proizvoda univerzalnih prajmera iz zaraženih korenova su se poklapali sa škotskim izolatima. P. fragariae var. rubi se javila na 8 od 14 lokaliteta. Naši rezultati ukazuju da su nested PCR (ITS 4 i DC 6 za prvi krug, DC 1 i DC 5 za drugi krug) i digestija produkta univerzalnih Phytophthora prajmera u cilju detekcije P. fragariae var. rubi senzitivnije i zahtevaju manje vremena, pa ih stoga preporučujemo za korišćenje.
PB  - Institut za pesticide i zaštitu životne sredine, Beograd i Društvo za zaštitu bilja Srbije, Beograd
T2  - Pesticidi i fitomedicina
T1  - Methods for detection of Phytophthora fragariae var. rubi on raspberry
T1  - Metode za detekciju Phytophthora fragariae var. rubi na malini
EP  - 184
IS  - 3
SP  - 177
VL  - 24
DO  - 10.2298/PIF0903177K
ER  - 
@article{
author = "Koprivica, Mirjana and Dulić-Marković, Ivana and Jevtić, Radivoje and Cooke, Dave E.L.",
year = "2009",
abstract = "Phytophthora fragariae var. rubi (Wilcox & Duncan), a causal agent of raspberry root rot, is a serious soil-borne pathogen listed by EPPO as an A2 quarantine pest. Root samples were collected from badly diseased raspberry plants showing a variety of characteristic and often dramatic symptoms during surveys carried out in western Serbia in 2002. Identification of the causal agent was performed in collaboration work with the Scottish Crop Research Institute (S.C.R.I.), Dundee, UK. Necrotic roots were plated on selective French bean agar (incorporating ampicilin, ryfamicin, bavistin and hymexasol). Detection of isolates was based on cultural and morphological features compared with referent cultures. DNA was extracted directly from the sampled roots using extraction buffer (200 mM Tris-HCl pH 8.5, 250 mM NaCl, 25 mM EDTA, 0.5% SDS), purified by multi spin separation columns [Thistle Scientific (Axygen)] or in 24:1 mixture of chlorophorm-isoamyl alcohol and amplified by nested PCR (ITS 4 and DC 6 for first round, DC 1 and DC 5 for second round). Diluted DNA extracts were also amplified by conventional PCR with modified 'universal' Phytophthora primers (ITS 6, ITS 7 and ITS 8, Cooke et al., 2000) and digested with Msp1. Digestion patterns of the universal primers PCR product from infected roots matched those of Scottish strains. P. fragariae var. rubi occurred on 8 out of 14 sites. Our results indicate that nested PCR (ITS 4 and DC 6 for first round, DC 1 and DC 5 for second round) or digestion of the 'universal' Phytophthora primers PCR product for detection of P. fragariae var. rubi are more sensitive and less time-consuming and therefore recommended for use., Phytophthora fragariae var. rubi (Wilcox & Duncan), prouzrokovač truleži korena maline, je veoma značajan zemljišni patogen A2 karantinskog statusa u EPPO. Uzorci korena su sakupljeni sa jako zaraženih biljaka maline koje su ispoljavale čitav niz karakterističnih i često dramatičnih simptoma tokom pregleda koji su vršeni u zapadnoj Srbiji 2002. godine. Identifikacija prouzrokovača je izvršena u saradnji sa Scottish Crop Research Institute (S.C.R.I.), Dundee, UK. Nekrotični korenčići su unošeni u selektivnu French bean podlogu, koja je sadržala ampicilin, rifamicin, bavistin i himeksazol. Detekcija dobijenih izolata je vršena posmatranjem kulture i morfoloških karakteristika i poređenjem sa autentičnim kulturama. DNK je ekstrahovana direktno iz uzorkovanih korenova pomoću ekstrakcionog pufera (200 mM Tris-HCl pH 8,5, 250 mM NaCl, 25 mM EDTA, 0,5% SDS), prečišćena Multi spin separacionim kolonama (Thistle Scientific (Axygen) ili 24:1 mešavinom hloroforma i izoamil alkohola i amplifikovana nested PCR (ITS 4 i DC 6 za prvi krug, DC 1 i DC 5 za drugi krug). Rastvoreni DNK ekstrakti su takođe amplifikovani putem konvencionalnog PCR sa modifikovanim 'univerzalnim' Phytophthora prajmerima (ITS 6, ITS 7 i ITS 8, Cooke i sar., 2000) i di gestirani sa Msp1. Digestioni paterni proizvoda univerzalnih prajmera iz zaraženih korenova su se poklapali sa škotskim izolatima. P. fragariae var. rubi se javila na 8 od 14 lokaliteta. Naši rezultati ukazuju da su nested PCR (ITS 4 i DC 6 za prvi krug, DC 1 i DC 5 za drugi krug) i digestija produkta univerzalnih Phytophthora prajmera u cilju detekcije P. fragariae var. rubi senzitivnije i zahtevaju manje vremena, pa ih stoga preporučujemo za korišćenje.",
publisher = "Institut za pesticide i zaštitu životne sredine, Beograd i Društvo za zaštitu bilja Srbije, Beograd",
journal = "Pesticidi i fitomedicina",
title = "Methods for detection of Phytophthora fragariae var. rubi on raspberry, Metode za detekciju Phytophthora fragariae var. rubi na malini",
pages = "184-177",
number = "3",
volume = "24",
doi = "10.2298/PIF0903177K"
}
Koprivica, M., Dulić-Marković, I., Jevtić, R.,& Cooke, D. E.L.. (2009). Methods for detection of Phytophthora fragariae var. rubi on raspberry. in Pesticidi i fitomedicina
Institut za pesticide i zaštitu životne sredine, Beograd i Društvo za zaštitu bilja Srbije, Beograd., 24(3), 177-184.
https://doi.org/10.2298/PIF0903177K
Koprivica M, Dulić-Marković I, Jevtić R, Cooke DE. Methods for detection of Phytophthora fragariae var. rubi on raspberry. in Pesticidi i fitomedicina. 2009;24(3):177-184.
doi:10.2298/PIF0903177K .
Koprivica, Mirjana, Dulić-Marković, Ivana, Jevtić, Radivoje, Cooke, Dave E.L., "Methods for detection of Phytophthora fragariae var. rubi on raspberry" in Pesticidi i fitomedicina, 24, no. 3 (2009):177-184,
https://doi.org/10.2298/PIF0903177K . .
3
4

The influence of Tilletia spp. inoculum source and environmental conditions on the frequency of infected wheat spikes

Koprivica, Mirjana; Jevtić, Radivoje; Dulić-Marković, Ivana

(Institut za pesticide i zaštitu životne sredine, Beograd i Društvo za zaštitu bilja Srbije, Beograd, 2009)

TY  - JOUR
AU  - Koprivica, Mirjana
AU  - Jevtić, Radivoje
AU  - Dulić-Marković, Ivana
PY  - 2009
UR  - http://fiver.ifvcns.rs/handle/123456789/695
AB  - The influence of inoculum source on the incidence of common bunt, caused by fungi from the genus Tilletia, was estimated based on the frequency of bunt infected wheat spikes in our agroecological conditions. The cultivar Novosadska rana 5 was sown in a random split plot design with four replicates at Rimski Šančevi on three sawing dates in 1999/2000 and 2000/2001. The following variables were evaluated: I - control, II - soilborne inoculum (4 g teliospores/1 l soil), III - seedborne inoculum (2 g teliospores/1 kg seeds), IV - seedborne inoculum + soilborne inoculum (4 g teliospores/1 l soil + 2 g teliospores/1 kg seeds). Correlation and regression analysis were used to evaluate the effect of temperature and precipitation on the frequency of infected spikes. The frequency of bunt infected spikes depended on the source of Tilletia spp. inoculum, and difference in infection frequencies between variables II and III, as well as III and IV, were determined for the assessed infection parameters. When teliospores are the only source of inoculum in soil, 60 days after sawing (r>+0.52) is a critical period in which temperature influences the development of infection. The highest number of plants was infected in the first, while less were infected in the second ten days (decade) after sawing (r>0.41), when temperature was the optimal 5.0-6.0ºC. The initial 60 days after sawing were also critical for disease development when teliospores on seeds were the only source of inoculum (r>+0.50). The highest number of plants was infected in the third and fewer in the fifth decade after sawing (r>0.41), when temperature was the optimal 5.0-6.0ºC. When infection was caused by teliospores on seeds and in soil, the critical period lasted 120 days after sawing (r>0.42), with a maximum frequency of infection found at the optimal temperatures for the period of 4.0- 5.0ºC.
AB  - Uticaj izvora inokuluma na pojavu glavnice pšenice koju prouzrokuju gljive roda Tilletia u našim agroekološkim uslovima praćen je, između ostalih, ispitivanjem parametra infekcije: stepen zaraženosti klasova. Sorta Novosadska rana 5 je sejana po split plot metodu u tri roka setve sa 4 ponavljanja na lokalitetu Rimski Šančevi tokom 1999/2000. i 2000/2001. godine. Ispitivane su sledeće varijante: I - apsolutna kontrola, II - zemlja infestirana teleutosporama (4 g teleutospora/1 l zemlje), III - seme infestirano teleutosporama (2 g teleutospora/1 kg semena) i IV - zemlja infestirana teleutosporama + seme infestirano teleutosporama (4 g teleutospora/1 l zemlje + 2 g teleutospora/1 kg semena). Izvršena je korelaciona i regresiona analiza ispitivanog parametara infekcije u odnosu na temperaturu i padavine. Stepen zaraženosti klasova je zavisio od izvora inokuluma Tilletia spp., a uočene su razlike u nivou infekcije između varijanti II i III, kao i III i IV. Kasniji rokovi setve (drugi i treći) su uticali značajno na ispitivani parametar infekcije, ali između njih nisu utvrđene značajne razlike. Kritični period u kom temperature utiču na ostvarivanje infekcije u uslovima kad su teleutospore u zemlji jedini izvor inokuluma traje 60 dana nakon setve (r>+0,52). Najveći broj biljaka je zaražen u prvoj i nešto manje u drugoj dekadi posle setve (r>0,41), pri čemu je optimalna temperatura u tom periodu iznosila 5,0-6,0ºC. Prvih 60 dana nakon setve je, takođe, kritičan period za ostvarivanje infekcije u uslovima kad su teleutospore na semenu jedini izvor inokuluma (r>+0,50). Najveći broj biljaka je zaražen u trećoj i nešto manje u petoj dekadi posle setve (r lt -0,45), pri čemu je optimalna temperatura u tim dekadama iznosila 6,0-7,0ºC. U uslovima kad infekciju pšenice ostvaruju teleutospore prisutne na semenu i u zemlji kritičan period traje 120 dana posle setve (r>+0,42), pri čemu je maksimalna infekcija ostvarena pri optimalnoj temperaturi 4,0-5,0ºC.
PB  - Institut za pesticide i zaštitu životne sredine, Beograd i Društvo za zaštitu bilja Srbije, Beograd
T2  - Pesticidi i fitomedicina
T1  - The influence of Tilletia spp. inoculum source and environmental conditions on the frequency of infected wheat spikes
T1  - Uticaj izvora inokuluma Tilletia spp. i uslova sredine na učestalost zaraženih klasova
EP  - 196
IS  - 3
SP  - 185
VL  - 24
DO  - 10.2298/PIF0903185K
ER  - 
@article{
author = "Koprivica, Mirjana and Jevtić, Radivoje and Dulić-Marković, Ivana",
year = "2009",
abstract = "The influence of inoculum source on the incidence of common bunt, caused by fungi from the genus Tilletia, was estimated based on the frequency of bunt infected wheat spikes in our agroecological conditions. The cultivar Novosadska rana 5 was sown in a random split plot design with four replicates at Rimski Šančevi on three sawing dates in 1999/2000 and 2000/2001. The following variables were evaluated: I - control, II - soilborne inoculum (4 g teliospores/1 l soil), III - seedborne inoculum (2 g teliospores/1 kg seeds), IV - seedborne inoculum + soilborne inoculum (4 g teliospores/1 l soil + 2 g teliospores/1 kg seeds). Correlation and regression analysis were used to evaluate the effect of temperature and precipitation on the frequency of infected spikes. The frequency of bunt infected spikes depended on the source of Tilletia spp. inoculum, and difference in infection frequencies between variables II and III, as well as III and IV, were determined for the assessed infection parameters. When teliospores are the only source of inoculum in soil, 60 days after sawing (r>+0.52) is a critical period in which temperature influences the development of infection. The highest number of plants was infected in the first, while less were infected in the second ten days (decade) after sawing (r>0.41), when temperature was the optimal 5.0-6.0ºC. The initial 60 days after sawing were also critical for disease development when teliospores on seeds were the only source of inoculum (r>+0.50). The highest number of plants was infected in the third and fewer in the fifth decade after sawing (r>0.41), when temperature was the optimal 5.0-6.0ºC. When infection was caused by teliospores on seeds and in soil, the critical period lasted 120 days after sawing (r>0.42), with a maximum frequency of infection found at the optimal temperatures for the period of 4.0- 5.0ºC., Uticaj izvora inokuluma na pojavu glavnice pšenice koju prouzrokuju gljive roda Tilletia u našim agroekološkim uslovima praćen je, između ostalih, ispitivanjem parametra infekcije: stepen zaraženosti klasova. Sorta Novosadska rana 5 je sejana po split plot metodu u tri roka setve sa 4 ponavljanja na lokalitetu Rimski Šančevi tokom 1999/2000. i 2000/2001. godine. Ispitivane su sledeće varijante: I - apsolutna kontrola, II - zemlja infestirana teleutosporama (4 g teleutospora/1 l zemlje), III - seme infestirano teleutosporama (2 g teleutospora/1 kg semena) i IV - zemlja infestirana teleutosporama + seme infestirano teleutosporama (4 g teleutospora/1 l zemlje + 2 g teleutospora/1 kg semena). Izvršena je korelaciona i regresiona analiza ispitivanog parametara infekcije u odnosu na temperaturu i padavine. Stepen zaraženosti klasova je zavisio od izvora inokuluma Tilletia spp., a uočene su razlike u nivou infekcije između varijanti II i III, kao i III i IV. Kasniji rokovi setve (drugi i treći) su uticali značajno na ispitivani parametar infekcije, ali između njih nisu utvrđene značajne razlike. Kritični period u kom temperature utiču na ostvarivanje infekcije u uslovima kad su teleutospore u zemlji jedini izvor inokuluma traje 60 dana nakon setve (r>+0,52). Najveći broj biljaka je zaražen u prvoj i nešto manje u drugoj dekadi posle setve (r>0,41), pri čemu je optimalna temperatura u tom periodu iznosila 5,0-6,0ºC. Prvih 60 dana nakon setve je, takođe, kritičan period za ostvarivanje infekcije u uslovima kad su teleutospore na semenu jedini izvor inokuluma (r>+0,50). Najveći broj biljaka je zaražen u trećoj i nešto manje u petoj dekadi posle setve (r lt -0,45), pri čemu je optimalna temperatura u tim dekadama iznosila 6,0-7,0ºC. U uslovima kad infekciju pšenice ostvaruju teleutospore prisutne na semenu i u zemlji kritičan period traje 120 dana posle setve (r>+0,42), pri čemu je maksimalna infekcija ostvarena pri optimalnoj temperaturi 4,0-5,0ºC.",
publisher = "Institut za pesticide i zaštitu životne sredine, Beograd i Društvo za zaštitu bilja Srbije, Beograd",
journal = "Pesticidi i fitomedicina",
title = "The influence of Tilletia spp. inoculum source and environmental conditions on the frequency of infected wheat spikes, Uticaj izvora inokuluma Tilletia spp. i uslova sredine na učestalost zaraženih klasova",
pages = "196-185",
number = "3",
volume = "24",
doi = "10.2298/PIF0903185K"
}
Koprivica, M., Jevtić, R.,& Dulić-Marković, I.. (2009). The influence of Tilletia spp. inoculum source and environmental conditions on the frequency of infected wheat spikes. in Pesticidi i fitomedicina
Institut za pesticide i zaštitu životne sredine, Beograd i Društvo za zaštitu bilja Srbije, Beograd., 24(3), 185-196.
https://doi.org/10.2298/PIF0903185K
Koprivica M, Jevtić R, Dulić-Marković I. The influence of Tilletia spp. inoculum source and environmental conditions on the frequency of infected wheat spikes. in Pesticidi i fitomedicina. 2009;24(3):185-196.
doi:10.2298/PIF0903185K .
Koprivica, Mirjana, Jevtić, Radivoje, Dulić-Marković, Ivana, "The influence of Tilletia spp. inoculum source and environmental conditions on the frequency of infected wheat spikes" in Pesticidi i fitomedicina, 24, no. 3 (2009):185-196,
https://doi.org/10.2298/PIF0903185K . .
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