Molecular detection of potyvirus in leaves and small bulbs of garlic, Allium sativum, associated a clean seed production program
Garlic (Allium sativum L), reproduces vegetatively using bulbils, condition that favors the spread of diseases, especially bacteria, fungi and viruses, which affect the quality and crop yield. For this reason, the molecular identification by RT-PCR of potyvirus: LYSV and OYDV in the production system of clean seed garlic of three national clones were implemented. In the production phase of clean seed was establishing garlic meristems micropropagation. Potyvirus presence in 586 seedlings was analyzed by ELISA and for RT-PCR in 70. RNA was extracted from leaves and small bulbs, yielding 1.7 to 226 ng/μl, and with this RNA, between 35 to 50 ng of cDNA. The results showed that the disinfection protocol produced a 73.6% viability of plants. ELISA analysis showed 96% sanitation of seedling to potyvirus, whereas, Leek Yellow Strip Virus, LYSV was identified in 8.6% of samples used RT-PCR methodology. Onion yellow dwarf virus (OYDV) was not detected in any sample. The results show that the in vitro culture of meristem of garlic, is an excellent alternative for seed production, showing a 92% efficiency. Moreover, efficient diagnostics of LYSV potyvirus was validated in leaves and small bulbs of garlic.Key words: Allium sativum, ELISA, micropropagation, LSYV, potyvirus, OYDV, RT-PCR.
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Universidad Nacional de Colombia - Sede Bogotá - Instituto de Biotecnología
2014
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Parra Fuentes, M. Reyes Perdomo, C. Hernandez Fernandez, Javier |
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Parra Fuentes, M. Reyes Perdomo, C. Hernandez Fernandez, Javier Molecular detection of potyvirus in leaves and small bulbs of garlic, Allium sativum, associated a clean seed production program |
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Parra Fuentes, M. Reyes Perdomo, C. Hernandez Fernandez, Javier |
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Parra Fuentes, M. |
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Molecular detection of potyvirus in leaves and small bulbs of garlic, Allium sativum, associated a clean seed production program |
| title_short |
Molecular detection of potyvirus in leaves and small bulbs of garlic, Allium sativum, associated a clean seed production program |
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Molecular detection of potyvirus in leaves and small bulbs of garlic, Allium sativum, associated a clean seed production program |
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Molecular detection of potyvirus in leaves and small bulbs of garlic, Allium sativum, associated a clean seed production program |
| title_full_unstemmed |
Molecular detection of potyvirus in leaves and small bulbs of garlic, Allium sativum, associated a clean seed production program |
| title_sort |
molecular detection of potyvirus in leaves and small bulbs of garlic, allium sativum, associated a clean seed production program |
| description |
Garlic (Allium sativum L), reproduces vegetatively using bulbils, condition that favors the spread of diseases, especially bacteria, fungi and viruses, which affect the quality and crop yield. For this reason, the molecular identification by RT-PCR of potyvirus: LYSV and OYDV in the production system of clean seed garlic of three national clones were implemented. In the production phase of clean seed was establishing garlic meristems micropropagation. Potyvirus presence in 586 seedlings was analyzed by ELISA and for RT-PCR in 70. RNA was extracted from leaves and small bulbs, yielding 1.7 to 226 ng/μl, and with this RNA, between 35 to 50 ng of cDNA. The results showed that the disinfection protocol produced a 73.6% viability of plants. ELISA analysis showed 96% sanitation of seedling to potyvirus, whereas, Leek Yellow Strip Virus, LYSV was identified in 8.6% of samples used RT-PCR methodology. Onion yellow dwarf virus (OYDV) was not detected in any sample. The results show that the in vitro culture of meristem of garlic, is an excellent alternative for seed production, showing a 92% efficiency. Moreover, efficient diagnostics of LYSV potyvirus was validated in leaves and small bulbs of garlic.Key words: Allium sativum, ELISA, micropropagation, LSYV, potyvirus, OYDV, RT-PCR. |
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Universidad Nacional de Colombia - Sede Bogotá - Instituto de Biotecnología |
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2014 |
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https://revistas.unal.edu.co/index.php/biotecnologia/article/view/47237 |
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AT parrafuentesm moleculardetectionofpotyvirusinleavesandsmallbulbsofgarlicalliumsativumassociatedacleanseedproductionprogram AT reyesperdomoc moleculardetectionofpotyvirusinleavesandsmallbulbsofgarlicalliumsativumassociatedacleanseedproductionprogram AT hernandezfernandezjavier moleculardetectionofpotyvirusinleavesandsmallbulbsofgarlicalliumsativumassociatedacleanseedproductionprogram AT parrafuentesm deteccionmoleculardepotyvirusenhojasyminibulbillosdeajoalliumsativumasociadosaunprogramadeproducciondesemillalimpia AT reyesperdomoc deteccionmoleculardepotyvirusenhojasyminibulbillosdeajoalliumsativumasociadosaunprogramadeproducciondesemillalimpia AT hernandezfernandezjavier deteccionmoleculardepotyvirusenhojasyminibulbillosdeajoalliumsativumasociadosaunprogramadeproducciondesemillalimpia |
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oai:www.revistas.unal.edu.co:article-472372016-09-09T14:55:02Z Molecular detection of potyvirus in leaves and small bulbs of garlic, Allium sativum, associated a clean seed production program Detección molecular de potyvirus en hojas y minibulbillos de ajo, Allium sativum, asociados a un programa de producción de semilla limpia Parra Fuentes, M. Reyes Perdomo, C. Hernandez Fernandez, Javier Allium sativum ELISA micropropagación LSYV potyvirus OYDV RT-PCR Garlic (Allium sativum L), reproduces vegetatively using bulbils, condition that favors the spread of diseases, especially bacteria, fungi and viruses, which affect the quality and crop yield. For this reason, the molecular identification by RT-PCR of potyvirus: LYSV and OYDV in the production system of clean seed garlic of three national clones were implemented. In the production phase of clean seed was establishing garlic meristems micropropagation. Potyvirus presence in 586 seedlings was analyzed by ELISA and for RT-PCR in 70. RNA was extracted from leaves and small bulbs, yielding 1.7 to 226 ng/μl, and with this RNA, between 35 to 50 ng of cDNA. The results showed that the disinfection protocol produced a 73.6% viability of plants. ELISA analysis showed 96% sanitation of seedling to potyvirus, whereas, Leek Yellow Strip Virus, LYSV was identified in 8.6% of samples used RT-PCR methodology. Onion yellow dwarf virus (OYDV) was not detected in any sample. The results show that the in vitro culture of meristem of garlic, is an excellent alternative for seed production, showing a 92% efficiency. Moreover, efficient diagnostics of LYSV potyvirus was validated in leaves and small bulbs of garlic.Key words: Allium sativum, ELISA, micropropagation, LSYV, potyvirus, OYDV, RT-PCR. Título en español: Detección molecular de potyvirus en hojas y minibulbillos de ajo, Allium sativum, asociados a un programa de producción de semilla limpiaTítulo en ingles: Molecular detection of potyvirus in leaves and small bulbs of garlic, Allium sativum, associated a clean seed production programTitulo corto: Detección de potyvirus en plantas de ajoResumen: El ajo (Allium sativum L) se reproduce vegetativamente utilizando bulbillos, condición que favorece la propagación de enfermedades, especialmente bacterias, hongos y virus que afectan la calidad y el rendimiento del cultivo. Por este motivo se implementó la identificación molecular por RT-PCR de los potyvirus LYSV y OYDV en el sistema de producción de semilla limpia de ajo en tres clones nacionales. En la fase de producción de semilla limpia mediante micropropagación, se estandarizó el establecimiento de meristemos de ajo. La presencia de potyvirus se analizó en 586 plántulas mediante ELISA y en 70 por RT-PCR. Para la RT-PCR se extrajo ARN a partir de microbulbillos y hojas de plántulas, obteniéndose 1.7 a 226 ng/ml de ARN y se sintetizó entre 35 a 50 ng de cADN. Los resultados obtenidos mostraron que el protocolo de desinfección produjo una viabilidad del 73.6%. El análisis ELISA presentó un saneamiento del 96.1% de las plántulas a potyvirus, mientras que con RT-PCR se identificó la presencia de LYSV en el 8.6% de las muestras evaluadas. El virus del enanismo amarrillo de la cebolla (OYDV) no fue detectado en ninguna de las muestras. Los resultados muestran que el cultivo in vitro de meristemos de ajo es una excelente alternativa para la producción de semilla, mostrando un 92% de eficiencia. Además, validan el diagnóstico eficiente del potyvirus LYSV en hojas y microbulbillos de ajo.Palabras clave: Allium sativum, ELISA, micropropagación, LSYV, potyvirus, OYDV, RT-PCR.Abstract: Garlic (Allium sativum L), reproduces vegetatively using bulbils, condition that favors the spread of diseases, especially bacteria, fungi and viruses, which affect the quality and crop yield. For this reason, the molecular identification by RT-PCR of potyvirus: LYSV and OYDV in the production system of clean seed garlic of three national clones were implemented. In the production phase of clean seed was establishing garlic meristems micropropagation. Potyvirus presence in 586 seedlings was analyzed by ELISA and for RT-PCR in 70. RNA was extracted from leaves and small bulbs, yielding 1.7 to 226 ng/µl, and with this RNA, between 35 to 50 ng of cDNA. The results showed that the disinfection protocol produced a 73.6% viability of plants. ELISA analysis showed 96% sanitation of seedling to potyvirus, whereas, Leek Yellow Strip Virus, LYSV was identified in 8.6% of samples used RT-PCR methodology. Onion yellow dwarf virus (OYDV) was not detected in any sample. The results show that the in vitro culture of meristem of garlic, is an excellent alternative for seed production, showing a 92% efficiency. Moreover, efficient diagnostics of LYSV potyvirus was validated in leaves and small bulbs of garlic.Key words: Allium sativum, ELISA, micropropagation, LSYV, potyvirus, OYDV, RT-PCR.Recibido: octubre 18 de 2013 Aprobado: octubre 1 de 2014 Universidad Nacional de Colombia - Sede Bogotá - Instituto de Biotecnología 2014-07-01 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Artículo revisado por pares application/zip application/pdf text/html https://revistas.unal.edu.co/index.php/biotecnologia/article/view/47237 10.15446/rev.colomb.biote.v16n2.47237 Revista Colombiana de Biotecnología; Vol. 16 No. 2 (2014); 30-36 Revista Colombiana de Biotecnología; Vol. 16 Núm. 2 (2014); 30-36 1909-8758 0123-3475 spa https://revistas.unal.edu.co/index.php/biotecnologia/article/view/47237/48805 https://revistas.unal.edu.co/index.php/biotecnologia/article/view/47237/48806 https://revistas.unal.edu.co/index.php/biotecnologia/article/view/47237/49652 Derechos de autor 2014 Revista Colombiana de Biotecnología https://creativecommons.org/licenses/by/4.0 |