TY  - JOUR
AU  - Blagojević, Jovana
AU  - Vukojevic, Jelena
AU  - Ivanović, Borko
AU  - Ivanović, Žarko
PY  - 2020
UR  - https://plantarum.izbis.bg.ac.rs/handle/123456789/749
AB  - Leaf spot diseases caused by Alternaria species have been reported worldwide in plants in the Brassicaceae family. However, there is little information on Alternaria species causing diseases in horseradish. In the present study, 89 Alternaria spp. isolates from Armoracia rusticana, sampled from nine districts in Serbia, were characterized based on their morphology, physiology, and molecular markers. Morphological characterization and molecular analyses based on ITS, GAPDH, Alt a 1, and PM-ATP sequences identified three distinct species associated with the disease: Alternaria brassicae, A. brassicicola, and A. alternata. For all species, growth and sporulation rates at 0, 5, 10, 15, 20, 25, 30, 35, and 40°C showed a quadratic response, with A. alternata having the widest temperature optimum (20 to 30°C) while A. brassicicola had higher optimum temperatures (20 to 25°C) than A. brassicae (15 to 20°C). To gain a better understanding of the pathogenicity of these species, the influence of leaf age, host susceptibility, and ability to infect artificially wounded and nonwounded leaves were tested. The pathogenicity test identified A. brassicicola and A. brassicae as the main causal agents of horseradish leaf spot disease. Results indicated that young and intact leaves of horseradish and cabbage were less susceptible to infection and also suggested the potential for cross-infection between these two hosts. Haplotype networks showed haplotype uniformity for A. brassicae, two haplotype groups of A. brassicicola, and eight haplotype groups of A. alternata in Serbia and suggest the possible association of some haplotypes with the geographic area. This study is the first to investigate Alternaria leaf spot disease on A. rusticana in Serbia and is the first record of A. brassicicola on horseradish in this country.
PB  - American Phytopathological Society
T2  - Plant Disease
T1  - Characterization of Alternaria Species Associated with Leaf Spot Disease of Armoracia rusticana in Serbia
EP  - 1389
IS  - 5
SP  - 1378
VL  - 104
DO  - 10.1094/pdis-02-19-0289-re
DO  - 0191-2917
ER  - 

@article{
author = "Blagojević, Jovana and Vukojevic, Jelena and Ivanović, Borko and Ivanović, Žarko",
year = "2020",
abstract = "Leaf spot diseases caused by Alternaria species have been reported worldwide in plants in the Brassicaceae family. However, there is little information on Alternaria species causing diseases in horseradish. In the present study, 89 Alternaria spp. isolates from Armoracia rusticana, sampled from nine districts in Serbia, were characterized based on their morphology, physiology, and molecular markers. Morphological characterization and molecular analyses based on ITS, GAPDH, Alt a 1, and PM-ATP sequences identified three distinct species associated with the disease: Alternaria brassicae, A. brassicicola, and A. alternata. For all species, growth and sporulation rates at 0, 5, 10, 15, 20, 25, 30, 35, and 40°C showed a quadratic response, with A. alternata having the widest temperature optimum (20 to 30°C) while A. brassicicola had higher optimum temperatures (20 to 25°C) than A. brassicae (15 to 20°C). To gain a better understanding of the pathogenicity of these species, the influence of leaf age, host susceptibility, and ability to infect artificially wounded and nonwounded leaves were tested. The pathogenicity test identified A. brassicicola and A. brassicae as the main causal agents of horseradish leaf spot disease. Results indicated that young and intact leaves of horseradish and cabbage were less susceptible to infection and also suggested the potential for cross-infection between these two hosts. Haplotype networks showed haplotype uniformity for A. brassicae, two haplotype groups of A. brassicicola, and eight haplotype groups of A. alternata in Serbia and suggest the possible association of some haplotypes with the geographic area. This study is the first to investigate Alternaria leaf spot disease on A. rusticana in Serbia and is the first record of A. brassicicola on horseradish in this country.",
publisher = "American Phytopathological Society",
journal = "Plant Disease",
title = "Characterization of Alternaria Species Associated with Leaf Spot Disease of Armoracia rusticana in Serbia",
pages = "1389-1378",
number = "5",
volume = "104",
doi = "10.1094/pdis-02-19-0289-re, 0191-2917"
}

Blagojević, J., Vukojevic, J., Ivanović, B.,& Ivanović, Ž.. (2020). Characterization of Alternaria Species Associated with Leaf Spot Disease of Armoracia rusticana in Serbia. in Plant Disease
American Phytopathological Society., 104(5), 1378-1389.
https://doi.org/10.1094/pdis-02-19-0289-re

Blagojević J, Vukojevic J, Ivanović B, Ivanović Ž. Characterization of Alternaria Species Associated with Leaf Spot Disease of Armoracia rusticana in Serbia. in Plant Disease. 2020;104(5):1378-1389.
doi:10.1094/pdis-02-19-0289-re .

Blagojević, Jovana, Vukojevic, Jelena, Ivanović, Borko, Ivanović, Žarko, "Characterization of Alternaria Species Associated with Leaf Spot Disease of Armoracia rusticana in Serbia" in Plant Disease, 104, no. 5 (2020):1378-1389,
https://doi.org/10.1094/pdis-02-19-0289-re . .

TY  - JOUR
AU  - Blagojević, Jovana
AU  - Ivanović, Žarko
AU  - Oro, Violeta
AU  - Dolovac, Nenad
AU  - Popović Milovanović, Tatjana
AU  - Ignjatov, Maja
AU  - Vukojevic, Jelena
PY  - 2015
UR  - https://plantarum.izbis.bg.ac.rs/handle/123456789/964
AB  - In July 2014, a foliar spot disease of horseradish (Armoracia rusticana) was observed in the Bačka region of Serbia (45°14′17″ N; 19°42′22″ E). The disease was observed in several small vegetable farms and disease incidence approached 20 to 40%. The symptoms first appeared on the abaxial side of older leaves as light brown concentric spots with dark margins and sometimes haloes. The spots varied in size between 0.5 and 0.9 cm. The necrotic spots gradually enlarged, tore, and dropped out. Under high humidity and temperature in 2014 in that region, development of numerous necrosis spots resulted in a rapid reduction of leaves. Small pieces taken from infected leaf tissue of diseased horseradish were treated with 1% sodium hypochlorite for 1 min, rinsed with sterile distillated water (SDW), and placed on V8 agar. Isolates were grown in pure culture at 24°C for 5 days in 12 h light/dark photoperiod. Morphological characteristics of the colony and sporulation were determined for six representative isolates. Fungal colonies were smooth, color varied from white, off white to light brown with concentric zones and intensive sporulation. Conidia were light brown, obclavate, produced singly or in short chains. Mature conidia were 16 to 38 µm wide and 73 to 158 µm long with a beak cell 79 to 120 µm long with 8 to 11 transverse septa and 0 to 4 longitudinal septa. Based on morphological characteristics, all isolates were identified as Alternaria brassicae (Berk.) Sacc (Simmons 2007). To confirm the pathogen’s identity, DNA was extracted from the fungal isolates and subjected to PCR; the internal transcribed spacer region of rDNA was amplified using primers ITS1 and ITS4 (White et al. 1990). The resulting amplicons were sequenced by Macrogen Inc. (Seoul, South Korea) and deposited in NCBI GenBank (Accession Nos. KP115599 to KP115604). BLAST analysis of revealed 100% homology with a sequence of A. brassicae strain ATCC 58169 deposited in GenBank (JX499028). Pathogenicity of six representative isolates was tested on leaves of 2-month-old horseradish seedlings by spraying with aqueous conidial suspensions (105 conidia/ml) prepared from cultures grown on V8 agar at 24°C for 5 days. Negative control plants were sprayed with SDW. Plants were covered with plastic bags and placed for 48 h in a greenhouse at 20 to 22°C. For all isolates within 10 days, inoculated plants developed brown lesions on leaves. Plants treated with SDW were symptomless. To fulfill Koch’s postulates, reisolations from all tested strains were done by streaking margins of necrotic leaf spot tissue onto V8 agar. Reisolated strains showed the same colony morphology as described above. Based on the pathogenicity test accompanied by completion of Koch’s postulates, sequence analysis, and morphological and cultural characteristics, the strains were identified as A. brassicae. According to available literature data, this is the first report of this pathogen as the causal agent of brown leaf spot of horseradish in Serbia.
PB  - American Phytopathological Society
T2  - Plant Disease
T1  - First Report of Horseradish Leaf Spot Caused by Alternaria brassicae in Serbia
IS  - 5
SP  - 730
VL  - 99
DO  - 10.1094/PDIS-11-14-1181-PDN
ER  - 

@article{
author = "Blagojević, Jovana and Ivanović, Žarko and Oro, Violeta and Dolovac, Nenad and Popović Milovanović, Tatjana and Ignjatov, Maja and Vukojevic, Jelena",
year = "2015",
abstract = "In July 2014, a foliar spot disease of horseradish (Armoracia rusticana) was observed in the Bačka region of Serbia (45°14′17″ N; 19°42′22″ E). The disease was observed in several small vegetable farms and disease incidence approached 20 to 40%. The symptoms first appeared on the abaxial side of older leaves as light brown concentric spots with dark margins and sometimes haloes. The spots varied in size between 0.5 and 0.9 cm. The necrotic spots gradually enlarged, tore, and dropped out. Under high humidity and temperature in 2014 in that region, development of numerous necrosis spots resulted in a rapid reduction of leaves. Small pieces taken from infected leaf tissue of diseased horseradish were treated with 1% sodium hypochlorite for 1 min, rinsed with sterile distillated water (SDW), and placed on V8 agar. Isolates were grown in pure culture at 24°C for 5 days in 12 h light/dark photoperiod. Morphological characteristics of the colony and sporulation were determined for six representative isolates. Fungal colonies were smooth, color varied from white, off white to light brown with concentric zones and intensive sporulation. Conidia were light brown, obclavate, produced singly or in short chains. Mature conidia were 16 to 38 µm wide and 73 to 158 µm long with a beak cell 79 to 120 µm long with 8 to 11 transverse septa and 0 to 4 longitudinal septa. Based on morphological characteristics, all isolates were identified as Alternaria brassicae (Berk.) Sacc (Simmons 2007). To confirm the pathogen’s identity, DNA was extracted from the fungal isolates and subjected to PCR; the internal transcribed spacer region of rDNA was amplified using primers ITS1 and ITS4 (White et al. 1990). The resulting amplicons were sequenced by Macrogen Inc. (Seoul, South Korea) and deposited in NCBI GenBank (Accession Nos. KP115599 to KP115604). BLAST analysis of revealed 100% homology with a sequence of A. brassicae strain ATCC 58169 deposited in GenBank (JX499028). Pathogenicity of six representative isolates was tested on leaves of 2-month-old horseradish seedlings by spraying with aqueous conidial suspensions (105 conidia/ml) prepared from cultures grown on V8 agar at 24°C for 5 days. Negative control plants were sprayed with SDW. Plants were covered with plastic bags and placed for 48 h in a greenhouse at 20 to 22°C. For all isolates within 10 days, inoculated plants developed brown lesions on leaves. Plants treated with SDW were symptomless. To fulfill Koch’s postulates, reisolations from all tested strains were done by streaking margins of necrotic leaf spot tissue onto V8 agar. Reisolated strains showed the same colony morphology as described above. Based on the pathogenicity test accompanied by completion of Koch’s postulates, sequence analysis, and morphological and cultural characteristics, the strains were identified as A. brassicae. According to available literature data, this is the first report of this pathogen as the causal agent of brown leaf spot of horseradish in Serbia.",
publisher = "American Phytopathological Society",
journal = "Plant Disease",
title = "First Report of Horseradish Leaf Spot Caused by Alternaria brassicae in Serbia",
number = "5",
pages = "730",
volume = "99",
doi = "10.1094/PDIS-11-14-1181-PDN"
}

Blagojević, J., Ivanović, Ž., Oro, V., Dolovac, N., Popović Milovanović, T., Ignjatov, M.,& Vukojevic, J.. (2015). First Report of Horseradish Leaf Spot Caused by Alternaria brassicae in Serbia. in Plant Disease
American Phytopathological Society., 99(5), 730.
https://doi.org/10.1094/PDIS-11-14-1181-PDN

Blagojević J, Ivanović Ž, Oro V, Dolovac N, Popović Milovanović T, Ignjatov M, Vukojevic J. First Report of Horseradish Leaf Spot Caused by Alternaria brassicae in Serbia. in Plant Disease. 2015;99(5):730.
doi:10.1094/PDIS-11-14-1181-PDN .

Blagojević, Jovana, Ivanović, Žarko, Oro, Violeta, Dolovac, Nenad, Popović Milovanović, Tatjana, Ignjatov, Maja, Vukojevic, Jelena, "First Report of Horseradish Leaf Spot Caused by Alternaria brassicae in Serbia" in Plant Disease, 99, no. 5 (2015):730,
https://doi.org/10.1094/PDIS-11-14-1181-PDN . .