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Revista Colombiana de Biotecnología

Print version ISSN 0123-3475

Abstract

PARRA-FUENTES, M.; REYES-PERDOMO, C.  and  HERNANDEZ-FERNANDEZ, J.. Molecular detection of potyvirus in leaves and small bulbs of garlic, Allium sativum, associated a clean seed production program. Rev. colomb. biotecnol [online]. 2014, vol.16, n.2, pp.30-36. ISSN 0123-3475.  http://dx.doi.org/10.15446/rev.colomb.biote.v16n2.47237.

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.

Keywords : Allium sativum; ELISA; micropropagation; LSYV; potyvirus; OYDV; RT-PCR.

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