Estandarización de una PCR-SSP para la identificación del fitopatógeno phythophtora infestans (Oomycete) a partir de muestras de ADN en frutos inoculados en la berenjena (Solanum Melongena)

Abstract

Infection caused by Phytophtora infestans causes gout, wilt, leak, boat or late blight disease that affects most species of the family Solanaceae. In the eggplant (Solanum melongena) it is still unclear what species, or the species of the genus Phytophtora that affect the plant and that is the cause of the rot of the fruits, and, many times, becomes confused with Other genera of fungi such as Fusarium spp and Botrytis spp. Consequently, it is not possible to determine what type of control can be used against the pathogen and what type of fungicide to use. Inoculating up to 25 aubergine fruits using the clonal EC-1 lineage, DNA extractions were performed, which were then quantified by the Nanodrop 2000c team; To standardize a PCR-SPP using ITS Sequence primers or primers. The results of these extractions and the PCRs were visualized on 2% and 3% agarose gels. To test the sensitivity of the test, dilutions of up to 10-3 were performed. The Clonal lineage of Phytophtora infestans EC-1, is not reported as a causal agent of gout, wilt disease, leak, boat or late blight in eggplant, is proposed as a new pathogen for Solaum melongena. Three sets of ITS sequence primers were used, only two of them amplify the actual size of an ITS and meet its characteristics. The third set is discarded as an ITS sequence, but not discarded as a specific protein sequence as consulted with the NBCI database.