METHOD FOR THE MOLECULAR GENOTYPIFICATION OF AGAVE PLANTS USING ITSS NUCLEOTIDE SEQUENCES OF RIBOSOMAL DNA.

摘要

The present invention describes an analysis of the nucleotide sequences of ITSs of different species of Agave plants for establishing a clear and accurate molecular strategy useful for molecularly genotypificating Agave at a specimen level. Agave from striatae and tequilana species were collected in order to extract DNA therefrom by modifying the lithium chloride precipitation technique described by Sullivan et al., in 2000. An amplification of the ITSs regions was subsequently carried out by the PCR technology using specific oligonucleotides which were led to the 18S, 5.8S and 28S DNAr sequences; finally said sequences were cloned into the 2.1 PCR vector and sequenced for carrying out a comparative analysis therebetween. The analysis of the nucleotide sequence previously obtained was useful for achieving the genotypification of the specimens obtained from the different Agave species pertaining to the Striatae and A. tequilana series, as well as the recognition of specific markers fou nd in different region populations.