STANDARDIZATION PROTOCOLS FOR EXTRACTING CHROMOSOMAL DNA OF colletotrichum gloeosporioides STRAINS ISOLATED IN MANGO PLANTS (Mangifera indica L.)

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Luis Alberto Valdés
Adys Gómez
María Elena Carballo
Maricela Capote
Ingrid González
W. Rohde

Abstract

Colletotrichum gloeosporioides (Penz) Penz & Sacc. is responsible for anthracnose in mango, a disease that affects all aerial parts of the plant, causing losses of up to 12 % of production. Microbial identification and characterization is important for determining prevention and pathogen control measure in plantations. This research aimed to standardize protocols for isolation of genomic DNA in strains of Colletotrichum gloeosporioides (Penz) Penz & Sacc., to ensure quality and concentration for future molecular studies in different representatives of this fungal species. Isolation of genomic DNA was performed by three methods: CTAB Doyle y Doyle (1990); Kuramane-Izioka (1997) with modifications and CTAB California modified by Ramirez et al., (2004) combined with NucleoSpin commercial reagent kits. The quality of the DNA obtained was tested by absorbance measurement at 260 and 280nm. The first method was not effective while the other methods yielded were obtained with genomic DNA with the appropriate concentrations for molecular research on the species. Modifications to the Kuramane-Izioka (1997) method, contributed to its effectiveness. The California CTAB method modified by Ramirez et al. (2004) constitutes the first report of its use for representatives of the Fungi kingdom.
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