TY  - JOUR
AU  - Pavlović, Snežana
AU  - Starović, Mira
AU  - Stojanović, S.
AU  - Aleksić, G.
AU  - Kojić, S.
AU  - Zdravković, Milan
AU  - Jošić, Dragana
PY  - 2014
UR  - http://RIVeC.institut-palanka.rs/handle/123456789/162
AB  - Pot marigold (Calendula officinalis L.) is native to southern Europe. Compounds of marigold flowers exhibit anti-inflammatory, anti-tumor-promoting, and cytotoxic activities (4). In Serbia, pot marigold is cultivated as an important medicinal and ornamental plant. Typical phyllody, virescence, proliferation of axillary buds, and witches' broom symptoms were sporadically observed in 2011 in Pancevo plantation, Serbia (44°51′49″ N, 20°39′33″ E, 80 m above sea level). Until 2013, the number of uniformly distributed affected pot marigold plants reached 20% in the field. Due to the lack of seed production, profitability of the cultivation was seriously affected. Leaf samples from 10 symptomatic and 4 symptomless marigold plants were collected and total nucleic acid was extracted from midrib tissue (3). Direct PCR and nested PCR were carried out with primer pairs P1/16S-SR and R16F2n/R16R2n, respectively (3). Amplicons 1.5 and 1.2 kb in length, specific for the 16S rRNA gene, were amplified in all symptomatic plants. No PCR products were obtained when DNA isolated from symptomless plants was used. Restriction fragment length polymorphism (RFLP) patterns of the 1.2-kb fragments of 16S rDNA were determined by digestion with four endonucleases separately (TruI1, AluI, HpaII, and HhaI) and compared with those of Stolbur (Stol), Aster Yellows (AY), Flavescence dorée-C (FD-C), Poinsettia Branch-Inducing (PoiBI), and Clover Yellow Edge (CYE) phytoplasmas (2). RFLP patterns from all symptomatic pot marigold plants were identical to the Stol pattern, indicating Stolbur phytoplasma presence in affected plants. The 1.2-kb amplicon of representative Nv8 strain was sequenced and the data were submitted to GenBank (accession no. KJ174507). BLASTn analysis of the sequence was compared with sequences available in GenBank, showing 100% identity with 16S rRNA gene of strains from Paeonia tenuifolia (KF614623) and corn (JQ730750) from Serbia, and peach (KF263684) from Iran. All of these are members of the 16SrXII ‘Candidatus Phytoplasma solani’ group, subgroup A (Stolbur). Phytoplasmas belonging to aster yellows (16SrI) (Italy and Canada) and peanut witches' broom related phytoplasma (16SrII) group (Iran) have been identified in diseased pot marigold plants (1). To our knowledge, this is the first report of natural infection of pot marigold by Stolbur phytoplasma in Serbia.
PB  - Amer Phytopathological Soc, St Paul
T2  - Plant Disease
T1  - The First Report of Stolbur Phytoplasma Associated with Phyllody of Calendula officinalis in Serbia.
EP  - 1152
IS  - 8
SP  - 1152
VL  - 98
DO  - 10.1094/PDIS-01-14-0085-PDN
ER  - 

@article{
author = "Pavlović, Snežana and Starović, Mira and Stojanović, S. and Aleksić, G. and Kojić, S. and Zdravković, Milan and Jošić, Dragana",
year = "2014",
abstract = "Pot marigold (Calendula officinalis L.) is native to southern Europe. Compounds of marigold flowers exhibit anti-inflammatory, anti-tumor-promoting, and cytotoxic activities (4). In Serbia, pot marigold is cultivated as an important medicinal and ornamental plant. Typical phyllody, virescence, proliferation of axillary buds, and witches' broom symptoms were sporadically observed in 2011 in Pancevo plantation, Serbia (44°51′49″ N, 20°39′33″ E, 80 m above sea level). Until 2013, the number of uniformly distributed affected pot marigold plants reached 20% in the field. Due to the lack of seed production, profitability of the cultivation was seriously affected. Leaf samples from 10 symptomatic and 4 symptomless marigold plants were collected and total nucleic acid was extracted from midrib tissue (3). Direct PCR and nested PCR were carried out with primer pairs P1/16S-SR and R16F2n/R16R2n, respectively (3). Amplicons 1.5 and 1.2 kb in length, specific for the 16S rRNA gene, were amplified in all symptomatic plants. No PCR products were obtained when DNA isolated from symptomless plants was used. Restriction fragment length polymorphism (RFLP) patterns of the 1.2-kb fragments of 16S rDNA were determined by digestion with four endonucleases separately (TruI1, AluI, HpaII, and HhaI) and compared with those of Stolbur (Stol), Aster Yellows (AY), Flavescence dorée-C (FD-C), Poinsettia Branch-Inducing (PoiBI), and Clover Yellow Edge (CYE) phytoplasmas (2). RFLP patterns from all symptomatic pot marigold plants were identical to the Stol pattern, indicating Stolbur phytoplasma presence in affected plants. The 1.2-kb amplicon of representative Nv8 strain was sequenced and the data were submitted to GenBank (accession no. KJ174507). BLASTn analysis of the sequence was compared with sequences available in GenBank, showing 100% identity with 16S rRNA gene of strains from Paeonia tenuifolia (KF614623) and corn (JQ730750) from Serbia, and peach (KF263684) from Iran. All of these are members of the 16SrXII ‘Candidatus Phytoplasma solani’ group, subgroup A (Stolbur). Phytoplasmas belonging to aster yellows (16SrI) (Italy and Canada) and peanut witches' broom related phytoplasma (16SrII) group (Iran) have been identified in diseased pot marigold plants (1). To our knowledge, this is the first report of natural infection of pot marigold by Stolbur phytoplasma in Serbia.",
publisher = "Amer Phytopathological Soc, St Paul",
journal = "Plant Disease",
title = "The First Report of Stolbur Phytoplasma Associated with Phyllody of Calendula officinalis in Serbia.",
pages = "1152-1152",
number = "8",
volume = "98",
doi = "10.1094/PDIS-01-14-0085-PDN"
}

Pavlović, S., Starović, M., Stojanović, S., Aleksić, G., Kojić, S., Zdravković, M.,& Jošić, D.. (2014). The First Report of Stolbur Phytoplasma Associated with Phyllody of Calendula officinalis in Serbia.. in Plant Disease
Amer Phytopathological Soc, St Paul., 98(8), 1152-1152.
https://doi.org/10.1094/PDIS-01-14-0085-PDN

Pavlović S, Starović M, Stojanović S, Aleksić G, Kojić S, Zdravković M, Jošić D. The First Report of Stolbur Phytoplasma Associated with Phyllody of Calendula officinalis in Serbia.. in Plant Disease. 2014;98(8):1152-1152.
doi:10.1094/PDIS-01-14-0085-PDN .

Pavlović, Snežana, Starović, Mira, Stojanović, S., Aleksić, G., Kojić, S., Zdravković, Milan, Jošić, Dragana, "The First Report of Stolbur Phytoplasma Associated with Phyllody of Calendula officinalis in Serbia." in Plant Disease, 98, no. 8 (2014):1152-1152,
https://doi.org/10.1094/PDIS-01-14-0085-PDN . .

TY  - JOUR
AU  - Jošić, Dragana
AU  - Starović, Mira
AU  - Stojanović, S.
AU  - Popović, Tatjana
AU  - Dolovac, Nenad
AU  - Zdravković, Jasmina
AU  - Pavlović, Snežana
PY  - 2013
UR  - http://RIVeC.institut-palanka.rs/handle/123456789/146
AB  - Saponaria officinalis L. (Caryophyllaceae; also known as bouncingbet or soapwort) is a perennial medicinal plant important for the pharmaceutical industry and used as an expectorant, alterative, laxative, and ointment for some skin diseases and arthritic conditions. S. officinalis plants with typical symptoms (23% in 2011 and 47% in 2012) of phytoplasma infection were observed in Pancevo plantation, Serbia. The symptoms appeared in May with leaves changing color from green to brown with severe reddening and necrosis. Severely diseased plants died. The infected plants had a significant reduction in biomass and quality. To investigate the presence of phytoplasma, total DNA was extracted from 10 symptomatic and four asymptomatic plants by a CTAB method. The nested PCR was carried out using phytoplasma-specific primer set P1/16S-SR followed by R16F2n/R16R2, targeting the 16S rRNA gene sequence of 1.5 and 1.2 kb in length, respectively. The amplicons of expected size were obtained from the symptomatic plants, but not from the asymptomatic plants. To obtain restriction fragment length polymorphism (RFLP) patterns, the R16F2n/R2 amplicons were digested with AluI, TruI1, HpaII, and HhaI endonucleases. The resulting patterns indicated that seven plants were infected by a Stolbur phytoplasma belonging to the 16SrXII-A subgroup, since it had the identical RFLP pattern as the STOL reference strain. The 1.2 kb nested PCR products of representative isolate Sap7 were purified using PCR purification kit (Fermentas, Vilnius, Lithuania) according to the recommended protocol and sequenced using facilities of IMGGI SeqService, Belgrade, Serbia. The obtained sequence was deposited in the NCBI database (GenBank Accession No. JX866951). The phytoplasma 16S rRNA gene sequence from Sap7 had a sequence identity of 97% with GenBank accessions GQ273961.1 (‘Euonymus japonicus’ phytoplasma), JX311953.1 (Candidatus Phytoplasma solani clone 5043), JQ412100.1 (Iranian alfalfa phytoplasma M21), and JN561702.1 (‘Convolvulus arvensis’ stolbur phytoplasma clone P1/P7-Conv2/2010-Bg). To our knowledge, this is the first report of a natural infection of S. officinalis by 16SrXII-A subgroup (Stolbur) phytoplasma in Serbia. As cited by Lee et al. (1), the 16SrI-M subgroup phytoplasma in S. officinalis sample was already detected in Lithuania by Valiunas (2). The identification of phytoplasma in the Pancevo plantation caused the intensification of our biological control tests and efforts to reduce the ecological and economic impacts of these phytoplasmas.
PB  - Amer Phytopathological Soc, St Paul
T2  - Plant Disease
T1  - First Report of Group 16SrXII-A Phytoplasma Causing Stolbur Disease in Saponaria officinalis Plants in Serbia
EP  - 420
EP  - Saponaria officinalis L.
EP  - Caryophyllaceae
EP  - bouncingbet
EP  - soapwort
IS  - 3
SP  - 420
VL  - 97
DO  - 10.1094/PDIS-09-12-0885-PDN
ER  - 

@article{
author = "Jošić, Dragana and Starović, Mira and Stojanović, S. and Popović, Tatjana and Dolovac, Nenad and Zdravković, Jasmina and Pavlović, Snežana",
year = "2013",
abstract = "Saponaria officinalis L. (Caryophyllaceae; also known as bouncingbet or soapwort) is a perennial medicinal plant important for the pharmaceutical industry and used as an expectorant, alterative, laxative, and ointment for some skin diseases and arthritic conditions. S. officinalis plants with typical symptoms (23% in 2011 and 47% in 2012) of phytoplasma infection were observed in Pancevo plantation, Serbia. The symptoms appeared in May with leaves changing color from green to brown with severe reddening and necrosis. Severely diseased plants died. The infected plants had a significant reduction in biomass and quality. To investigate the presence of phytoplasma, total DNA was extracted from 10 symptomatic and four asymptomatic plants by a CTAB method. The nested PCR was carried out using phytoplasma-specific primer set P1/16S-SR followed by R16F2n/R16R2, targeting the 16S rRNA gene sequence of 1.5 and 1.2 kb in length, respectively. The amplicons of expected size were obtained from the symptomatic plants, but not from the asymptomatic plants. To obtain restriction fragment length polymorphism (RFLP) patterns, the R16F2n/R2 amplicons were digested with AluI, TruI1, HpaII, and HhaI endonucleases. The resulting patterns indicated that seven plants were infected by a Stolbur phytoplasma belonging to the 16SrXII-A subgroup, since it had the identical RFLP pattern as the STOL reference strain. The 1.2 kb nested PCR products of representative isolate Sap7 were purified using PCR purification kit (Fermentas, Vilnius, Lithuania) according to the recommended protocol and sequenced using facilities of IMGGI SeqService, Belgrade, Serbia. The obtained sequence was deposited in the NCBI database (GenBank Accession No. JX866951). The phytoplasma 16S rRNA gene sequence from Sap7 had a sequence identity of 97% with GenBank accessions GQ273961.1 (‘Euonymus japonicus’ phytoplasma), JX311953.1 (Candidatus Phytoplasma solani clone 5043), JQ412100.1 (Iranian alfalfa phytoplasma M21), and JN561702.1 (‘Convolvulus arvensis’ stolbur phytoplasma clone P1/P7-Conv2/2010-Bg). To our knowledge, this is the first report of a natural infection of S. officinalis by 16SrXII-A subgroup (Stolbur) phytoplasma in Serbia. As cited by Lee et al. (1), the 16SrI-M subgroup phytoplasma in S. officinalis sample was already detected in Lithuania by Valiunas (2). The identification of phytoplasma in the Pancevo plantation caused the intensification of our biological control tests and efforts to reduce the ecological and economic impacts of these phytoplasmas.",
publisher = "Amer Phytopathological Soc, St Paul",
journal = "Plant Disease",
title = "First Report of Group 16SrXII-A Phytoplasma Causing Stolbur Disease in Saponaria officinalis Plants in Serbia",
pages = "420-Saponaria officinalis L.-Caryophyllaceae-bouncingbet-soapwort-420",
number = "3",
volume = "97",
doi = "10.1094/PDIS-09-12-0885-PDN"
}

Jošić, D., Starović, M., Stojanović, S., Popović, T., Dolovac, N., Zdravković, J.,& Pavlović, S.. (2013). First Report of Group 16SrXII-A Phytoplasma Causing Stolbur Disease in Saponaria officinalis Plants in Serbia. in Plant Disease
Amer Phytopathological Soc, St Paul., 97(3), 420-420.
https://doi.org/10.1094/PDIS-09-12-0885-PDN

Jošić D, Starović M, Stojanović S, Popović T, Dolovac N, Zdravković J, Pavlović S. First Report of Group 16SrXII-A Phytoplasma Causing Stolbur Disease in Saponaria officinalis Plants in Serbia. in Plant Disease. 2013;97(3):420-420.
doi:10.1094/PDIS-09-12-0885-PDN .

Jošić, Dragana, Starović, Mira, Stojanović, S., Popović, Tatjana, Dolovac, Nenad, Zdravković, Jasmina, Pavlović, Snežana, "First Report of Group 16SrXII-A Phytoplasma Causing Stolbur Disease in Saponaria officinalis Plants in Serbia" in Plant Disease, 97, no. 3 (2013):420-420,
https://doi.org/10.1094/PDIS-09-12-0885-PDN . .