Transformation of the entomopathogenic fungus Paecilomyces fumosoroseus with Agrobacterium tumefaciens.
Lett Appl Microbiol
; 42(6): 631-6, 2006 Jun.
Article
en En
| MEDLINE
| ID: mdl-16706904
ABSTRACT
AIMS:
To test the suitability of the Agrobacterium tumefaciens-mediated transformation (AMT) method with Paecilomyces fumosoroseus, a fungal pathogen that causes diseases in a wide range of insects including whiteflies. METHODS ANDRESULTS:
Conidia of P. fumosoroseus were successfully transformed to hygromycin B resistance using the hph gene of Escherichia coli as the selectable marker. Transformation frequencies were 58.3 +/- 18.5, 98.3 +/- 24.8 and 169.7 +/- 35.5 (+/-SEM) transformants per 10(5), 10(6) and 10(7) target conidia respectively. After confirmation by PCR, transformants were subjected to Southern analysis, and the results revealed that 45% (four of nine) of the transformants contained single-copy integration of the T-DNA.CONCLUSIONS:
In our AMT system, we efficiently transformed conidia of P. fumosoroseus. The employment of this method circumvents time-consuming protoplast preparation and allows the isolation of transformants containing single-copy integration of the T-DNA. SIGNIFICANCE AND IMPACT OF THE STUDY Considering the efficiency of Ag. tumefaciens-mediated transformation, this method represents a useful tool for insertional mutagenesis to characterize genes that are important for the pathogenicity of P. fumosoroseus.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Transformación Genética
/
Paecilomyces
/
Agrobacterium tumefaciens
/
Insectos
Tipo de estudio:
Evaluation_studies
Límite:
Animals
Idioma:
En
Revista:
Lett Appl Microbiol
Asunto de la revista:
MICROBIOLOGIA
Año:
2006
Tipo del documento:
Article
País de afiliación:
Brasil