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Fitopatologia Brasileira

Print version ISSN 0100-4158

Abstract

FALEIRO, Fábio G. et al. Mycelial production of Crinipellis perniciosa on four culture media for DNA extraction. Fitopatol. bras. [online]. 2004, vol.29, n.3, pp. 312-315. ISSN 0100-4158.  http://dx.doi.org/10.1590/S0100-41582004000300013.

Mycelial production is the first step in getting good quality and quantity of DNA samples for molecular analysis. The objective of this work was to analyse the quantity and quality of DNA extracted from Crinipellis perniciosa mycelial mass obtained on four culture media. The lyophilized mycelial mass weight was evaluated in the following culture media: 1. yeast extract (2.5%); 2. Malt extract (2.5%); 3. PD (potato 20% and dextrose 2%) and 4. malt extract + PD (50% of each culture media 2 and 3). Mycelial growth was produced by depositing of a mycelial disk in the center of 90 mm Petri dishes containing each culture medium. The dishes were kept in a BOD incubator at 25 ºC under a photoperiod of 12 h, for ten days. The experiment was organized in complete random design with ten repetitions. The DNA was extracted from 250mg of lyophilizated mycelial mass using the SDS method with desproteinization carried out either with or without phenol. The DNA purity based on the absorbance A260/A280 and the DNA integrity in agarose gel 0,8% were analyzed. The quantity of mycelial mass-produced on culture media 1 and 4 was larger than on culture media 2 and 3. The DNA extracted from the mycelial mass produced in culture media 2 and 3 demonstrated greater integrity, yielding good amplification products. The DNA purity was higher after desproteinization with phenol. However, good quality and abundant DNA could be extracted from the mycelial mass produced in PD, even without desproteinization with phenol.

Keywords : vassoura-de-bruxa; cacau; metodologia.

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