Abstract:
The
aim of this work was the production of
high
numbers of propagules of the
entomopathogenic fungus Metarhizium
anisopliae with
high
germination capacity under
low
water availability, good storage stability and enhanced pathogenicity.
To this end
modifications of cultural conditions were employed.
Water-stress in
solid substrate
fermentation
reduced the conidial production
but
resulted
in
enhancement of
germination capacity.
Conidial
production and germination was also considerably
influenced by the type of solid substrate (bulgar
wheat, millet grain).
Intracellular
accumulation of endogenous reserves of selected polyols, which
have been linked
with
enhancement of germination under conditions of restricted water availability, was
altered by
such substrate modifications.
Imposed water-stress in liquid
culture resulted
in increased blastospore production
in the 0.998 to 0.96
water activity
(a. )
range when
ionic
solutes (NaCl, KCI)
were used for
medium modification
but
when
PEG 200
was
used, this range was narrower
(0.998-0.98 aw). Water-stress resulted in the production of
modified
blastospores
with
increased levels
of the low
molecular weight polyol
erythritol.
Higher
amounts of endogenous polyols were retained
intracellularly
when
modified
blastospores were
harvested in isotonic
solutions compared to those subjected
to hypo-osmotic
shock by
washing with water.
Intracellular
polyol patterns of
blastospores were also affected
by
nitrogen source, pH and aW-modifying solute as well
as
by interaction between
these factors. Optimum
conditions
for increased erythritol
accumulation occurred when
ionic
solutes
(NaCl, KCl)
were used
for
aW modification
and
in the pH range between 6.8-8. Total
endogenous protein was also enhanced under
the same conditions. Germination
of
blastospores produced under these conditions was
between 62-89%
under conditions of water-stress (0.96
a, ). In
contrast,
blastospores
with
lower
amounts of erythritol and total protein content had decreased germination
(8-
67%). Osmoprotection
of such modified
blastospores
resulted
in increased
storage
stability as wet pastes and as freeze-dried preparations. Fluidised bed dried blastospores
had
either
decreased viability
(<25%)
or good viability
(>40%) but high
conent of
moisture
(>30%). Destruxin A
production was not affected
by
osmoprotection whereas
protease activity was enhanced by isotonic
washing with
PEG 200
solution but
not
by.
ionic (NaCl)
or carbohydrate (glucose)
solutions. Modified blastospores
were not more
infective
to aphids than unmodified ones under 100% R. H.
regimes.