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Isolation and in vitro cultivation of the aphid pathogenic fungus Entomophthora planchoniana

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Abstract:

Entomophthora planchoniana is an important fungal pathogen of aphids. Although Entomophthora chromaphidis has been considered a synonym for E. planchoniana, the two species are now separated, and E. planchoniana is reported not to grow in vitro. In this paper, we describe for the first time the isolation and cultivation of this species. Entomophthora planchoniana was isolated from a population of Ovatus crataegarius (Homoptera, Aphididae), which was infected by E. planchoniana only. The isolates did not sporulate, but the sequence of the small subunit rDNA and the restriction fragment length polymorphism patterns of the first part of the large subunit rDNA and the ITS II region confirm that the isolates were E. planchoniana. The isolated fungus grew in a medium consisting of Grace's insect cell culture medium supplemented with lactalbumin hydrolysate, yeastolate, and 10% fetal bovine serum or in GLEN medium with 10% fetal bovine serum. Vegetative cells of E. planchoniana were long and club-shaped and did not stain with Calcofluor, thus suggesting that they were protoplasts.Key words: biological control, entomopathogen, Entomophthorales, protoplast, Zygomycetes.

Entomophthora planchoniana est un champignon pathogène important des pucerons. Bien qu'Entomophthora chromaphidis ait été considéré comme synonyme d'E. planchoniana, les deux espèces sont maintenant séparées et il a été rapporté qu'E. planchoniana ne pousse pas en culture in vitro. Dans cet article, nous décrivons pour la première fois l'isolement et la culture de cette espèce. Entomophthora planchoniana a été isolé d'une population d'Ovatus crataegarius (Homoptera, Aphididae) qui était infectée par E. planchoniana uniquement. Les isolats ne produisaient pas de spores, mais les séquences de la petite sous-unité de l'ADNr et les motifs de polymorphisme de la longeur des fragments de restriction de la première partie de la grosse sous-unité de l'ADNr et de ITS II suggèrent que les isolats étaient du E. planchoniana. Le champignon isolé poussait dans le milieu de culture pour cellules d'insecte de Grace enrichi d'hydrolysat de lactalbumine, de yeastolate et de 10% de sérum fœtal de bovin ou dans le milieu GLEN enrichi avec 10% de sérum fœtal de bovin. Les cellules végétatives d'E. planchoniana étaient longues et en forme de gourdins et ne se coloraient pas avec le Calcofluor, suggérant ainsi qu'il s'agissait de protoplastes.Mots clés : lutte biologique, entomopathogène, Entomophthorales, protoplaste, Zygomycetes.

Keywords: Entomophthorales; Zygomycetes; biological control; entomopathogen; entomopathogène; lutte biologique; protoplast; protoplaste

Document Type: Research Article

Publication date: December 1, 2001

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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