Effect of liquid culture media on morphology, growth, propagule production, and pathogenic activity of the Hyphomycete, Metarhizium flavoviride.
Two isolates of Metarhizium spp. were studied for propagule production, because of their pathogenic activity towards locusts and grasshoppers (Mf189 = M. flavoviride (or M. anisopliae var. acridum) strain IMI 330189, and Mf324 = M. flavoviride strain ARSEF324). Both isolates were grown in seven different liquid media, which have been developed for mass production of various Hyphomycetes, considered as candidates for microbial control of noxious insects. Shake-flask experiments were carried out at 28 degress C in the dark. Production was quantified for 72 h and the effects of the tested media were evaluated on propagule concentration, morphology and pathogenicity. Based on preliminary experiments, all tested media were supplemented with 0.4% Tween 80 to avoid the formation of pellets and to produce unicellular propagules. Submerged propagule yields were higher with Mf189 than with Mf324 in all seven media. While high concentrations of propagules (1.4 to 2.4 x 10(8) propagules ml(-1) for MF189 and 1.4 to 8.3 x 10(7) propagules ml(-1) for Mf324) were produced in four media (Adamek, Catroux, Jackson, and Jenkins-Prior media), production of propagules was lower in the three other media (Goral, Kondryatiev, and Paris media). Both isolates produced oblong blastospore-like propagules, except in Kondryatiev medium in which they provided ovoid propagules. In this case, Mf189 submerged propagules looked like aerial conidia, but scanning observations did not demonstrate a typical conidiogenesis via phialides. In Kondryatiev medium, Mf324 submerged propagules were significantly smaller than aerial conidia. Infection potential of submerged propagules was assayed on Schistocerca gregaria. Second-instar larvae fed for 48 h on fresh wheat previously contaminated by a spraying suspension of each inoculum titrated at 10(7) propagules ml(-1). All seven media produced submerged propagules that were highly infectious for S. gregaria larvae. Shake flask culture assays permitted us to select three low-costmedia, Adamek, Jenkins-Prior, and Catroux for improving scale-up of liquid fermentation focused on mass-production of Metarhizium propagules for mycoinsecticides devoted to locust control.
Fargues
Jacques
J
Centre de Biologie et de Gestion des Populations, INRA-Montpellier, Campus International de Montferrier-Baillarguet, Montferrier-sur-Lez, France. fargues@ensam.inra.fr
Smits
Nathalie
N
Vidal
Claire
C
Vey
Alain
A
Vega
Fernando
F
Mercadier
Guy
G
Quimby
Paul
P
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D003470:Culture Media / Q000378:metabolism*; D006110:Grasshoppers / Q000254:growth & development / Q000378:metabolism / Q000382:microbiology; D007306:Insecticides / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D003904:Mitosporic Fungi / Q000254:growth & development* / Q000378:metabolism / Q000472:pathogenicity / Q000648:ultrastructure
D003470:Culture Media; D007306:Insecticides