Brazilian Journal of Microbiology
On-line version ISSN 1678-4405
FREIRE, Laurineide Lopes de Carvalho; COSTA, Ana Bolena Lima da; GOES, Larissa Brandão and OLIVEIRA, Neiva Tinti de. DNA polymorphism and total protein in mutants of Metarhizium anisopliae var. Anisopliae (Metsch.) Sorokin strain E9. Braz. J. Microbiol. [online]. 2001, vol.32, n.2, pp. 93-97. ISSN 1678-4405. http://dx.doi.org/10.1590/S1517-83822001000200004.
Five mutants (MaE10, MaE27, MaE24, MaE41 e MaE49) of Metarhizium anisopliae wild strain E9 were analysed for DNA profile through the RAPD technique and for changes in total protein content by spectrophotometry, polyacrylamide gel electrophoresis and densitometry. The pattern of RAPD markers showed genetic polymorphism among the strains: out of twenty primers seven were selected, producing 113 bands. Forty seven bands were present in all strains (41.6% of monomorphic bands) and 66 showed polymorphism (58.4%). The mean coefficient of similarity among all strains was 0.75 (75%). The total protein content varied, staining in the interval of 6.0-8.0 µg/µl. The electrophoresis analysis, through zymogram and protein fraction profiles by densitometry, allowed the observation of seven bands for the wild strain E9 and five bands for the mutants MaE10, MaE27, MaE34, MaE41 and MaE49, evidence of variations in µg% among protein fractions. The RAPD technique was very sensitive to detect genetic differences between the wild type and the mutants obtained through gamma radiation. The total protein analysis also showed changes in quantity and pattern of bands after electrophoresis in the mutants compared to the wild type.
Keywords : Metarhizium anisopliae; mutants; RAPD; total protein.