Acid phosphatases of Metarhizium anisopliae during infection of the tobacco hornworm Manduca sexta.
Three acid phosphatase (AcP) isozymes, pI 8.1, 8.0 and 7.8, were isolated, purified and partially characterised from optimised cultures of the entomopathogenic fungus Metarhizium anisopliae. The enzymes had similar molecular masses (approximately 44.0 kDa), and could degrade sugar phosphates found in the haemolymph of a host insect, the tobacco hornworm Manduca sexta. The AcP activity in haemolymph of mycosed insects increased significantly over controls, and some new isozymes were present. The infection-related isoforms were similar in molecular mass and pI to some of the in vitro AcP isozymes of M. anisopliae. Results of dot blot and Western blot analyses using anti-AcP antibodies suggested that at least one Metarhizium phosphatase isoform was present in haemolymph of infected caterpillars. Antibodies did not cross-react with immune (chemically stimulated) or non-immune haemolymph from Manduca sexta. Consistent with the appearance of highly active fungal phosphatase in caterpillar blood, free phosphate concentration increased dramatically during the late stages of infection to a level two to five times that of controls. Phosphate was limiting to growth of the fungus at the concentration found in control haemolymph and supplementation of phosphate significantly increased fungal growth in vitro in haemolymph. These results suggest that Metarhizium AcP may play a key role in providing phosphorus for fungal growth at the expense of the insect.
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Y
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Department of Biology and Biochemistry, University of Bath, Claverton Down, UK.
Clarkson
J M
JM
Charnley
A K
AK
D000135:Acid Phosphatase / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D006458:Hemolymph / Q000737:chemistry; D015151:Immunoblotting; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism; D018402:Manduca / Q000382:microbiology*; D003904:Mitosporic Fungi / Q000201:enzymology* / Q000254:growth & development; D013997:Time Factors; D014026:Nicotiana / Q000469:parasitology*
D007527:Isoenzymes; D000135:Acid Phosphatase