Improvement of the specific detection of Xanthomonas phaseoli pv. manihotis based on the pthB gene.
Modifications were made in the PCR conditions aiming to overcome the problem of non-amplification of the Xanthomonas phaseoli pv. manihotis (Xpm) fragment, using the primer pair XV / XK described in the literature. The objective of this study was to propose changes in the primers already described (XV / XK_MOD) and validate the use of these new primers in identifying Xpm. The validation procedure was carried out with the primer pair XV and XK_MOD, using different strains of Xpm, other plant pathogenic and endophytic bacteria genera and one isolate of X. phaseoli pv. passiflorae. As a control, additional reactions were conducted in multiplex with the universal primers for the 16S rRNA gene of the bacteria together with XV / XK and XV / XK_MOD. Using the forward primer (XV) described in the literature together with the modified reverse primer (XK_MOD), it was possible to achieve amplification from DNA extracted from in vitro cultures and from infected tissue, but no amplification was noticed for the primer pair described in the literature, confirming the effectiveness of the proposed modification.
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Format: | Artigo de periódico biblioteca |
Language: | English eng |
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2019-10-15
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Subjects: | Mandioca, Doença de Planta, Plant diseases and disorders, Cassava, |
Online Access: | http://www.alice.cnptia.embrapa.br/alice/handle/doc/1113116 |
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dig-alice-doc-11131162019-10-15T18:08:38Z Improvement of the specific detection of Xanthomonas phaseoli pv. manihotis based on the pthB gene. MELO, R. de C. C. BRAGANÇA, C. A. D. PESTANA, K. N. SILVA, H. S. A. FERREIRA, C. F. OLIVEIRA, S. A. S. de RITA DE CÁSSIA CERQUEIRA MELO, UFRB; CARLOS AUGUSTO DÓREA BRAGANÇA, UFRB; KÁTIA NOGUEIRA PESTANA; HARLLEN SANDRO ALVES SILVA, CNPMF; CLAUDIA FORTES FERREIRA, CNPMF; SAULO ALVES SANTOS DE OLIVEIRA, CNPMF. Mandioca Doença de Planta Plant diseases and disorders Cassava Modifications were made in the PCR conditions aiming to overcome the problem of non-amplification of the Xanthomonas phaseoli pv. manihotis (Xpm) fragment, using the primer pair XV / XK described in the literature. The objective of this study was to propose changes in the primers already described (XV / XK_MOD) and validate the use of these new primers in identifying Xpm. The validation procedure was carried out with the primer pair XV and XK_MOD, using different strains of Xpm, other plant pathogenic and endophytic bacteria genera and one isolate of X. phaseoli pv. passiflorae. As a control, additional reactions were conducted in multiplex with the universal primers for the 16S rRNA gene of the bacteria together with XV / XK and XV / XK_MOD. Using the forward primer (XV) described in the literature together with the modified reverse primer (XK_MOD), it was possible to achieve amplification from DNA extracted from in vitro cultures and from infected tissue, but no amplification was noticed for the primer pair described in the literature, confirming the effectiveness of the proposed modification. 2019-10-15T18:08:32Z 2019-10-15T18:08:32Z 2019-10-15 2019 2019-10-15T18:08:32Z Artigo de periódico Acta Scientiarum. Agronomy, v. 41, e42708, 2019. 1807-8621 http://www.alice.cnptia.embrapa.br/alice/handle/doc/1113116 en eng openAccess |
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Mandioca Doença de Planta Plant diseases and disorders Cassava Mandioca Doença de Planta Plant diseases and disorders Cassava |
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Mandioca Doença de Planta Plant diseases and disorders Cassava Mandioca Doença de Planta Plant diseases and disorders Cassava MELO, R. de C. C. BRAGANÇA, C. A. D. PESTANA, K. N. SILVA, H. S. A. FERREIRA, C. F. OLIVEIRA, S. A. S. de Improvement of the specific detection of Xanthomonas phaseoli pv. manihotis based on the pthB gene. |
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Modifications were made in the PCR conditions aiming to overcome the problem of non-amplification of the Xanthomonas phaseoli pv. manihotis (Xpm) fragment, using the primer pair XV / XK described in the literature. The objective of this study was to propose changes in the primers already described (XV / XK_MOD) and validate the use of these new primers in identifying Xpm. The validation procedure was carried out with the primer pair XV and XK_MOD, using different strains of Xpm, other plant pathogenic and endophytic bacteria genera and one isolate of X. phaseoli pv. passiflorae. As a control, additional reactions were conducted in multiplex with the universal primers for the 16S rRNA gene of the bacteria together with XV / XK and XV / XK_MOD. Using the forward primer (XV) described in the literature together with the modified reverse primer (XK_MOD), it was possible to achieve amplification from DNA extracted from in vitro cultures and from infected tissue, but no amplification was noticed for the primer pair described in the literature, confirming the effectiveness of the proposed modification. |
author2 |
RITA DE CÁSSIA CERQUEIRA MELO, UFRB; CARLOS AUGUSTO DÓREA BRAGANÇA, UFRB; KÁTIA NOGUEIRA PESTANA; HARLLEN SANDRO ALVES SILVA, CNPMF; CLAUDIA FORTES FERREIRA, CNPMF; SAULO ALVES SANTOS DE OLIVEIRA, CNPMF. |
author_facet |
RITA DE CÁSSIA CERQUEIRA MELO, UFRB; CARLOS AUGUSTO DÓREA BRAGANÇA, UFRB; KÁTIA NOGUEIRA PESTANA; HARLLEN SANDRO ALVES SILVA, CNPMF; CLAUDIA FORTES FERREIRA, CNPMF; SAULO ALVES SANTOS DE OLIVEIRA, CNPMF. MELO, R. de C. C. BRAGANÇA, C. A. D. PESTANA, K. N. SILVA, H. S. A. FERREIRA, C. F. OLIVEIRA, S. A. S. de |
format |
Artigo de periódico |
topic_facet |
Mandioca Doença de Planta Plant diseases and disorders Cassava |
author |
MELO, R. de C. C. BRAGANÇA, C. A. D. PESTANA, K. N. SILVA, H. S. A. FERREIRA, C. F. OLIVEIRA, S. A. S. de |
author_sort |
MELO, R. de C. C. |
title |
Improvement of the specific detection of Xanthomonas phaseoli pv. manihotis based on the pthB gene. |
title_short |
Improvement of the specific detection of Xanthomonas phaseoli pv. manihotis based on the pthB gene. |
title_full |
Improvement of the specific detection of Xanthomonas phaseoli pv. manihotis based on the pthB gene. |
title_fullStr |
Improvement of the specific detection of Xanthomonas phaseoli pv. manihotis based on the pthB gene. |
title_full_unstemmed |
Improvement of the specific detection of Xanthomonas phaseoli pv. manihotis based on the pthB gene. |
title_sort |
improvement of the specific detection of xanthomonas phaseoli pv. manihotis based on the pthb gene. |
publishDate |
2019-10-15 |
url |
http://www.alice.cnptia.embrapa.br/alice/handle/doc/1113116 |
work_keys_str_mv |
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