Abstract:
Eight isolates (7 species) of white rot fungi were grown on soil extract agar amended
with 0, 5 10 and 20 mg l- simazine, trifluralin and dieldrin, individually and as a
mixture, under two different water regimes (-0.7 and -2.8 MPa water potential). The
best isolates were T.versicolor (R26 and R101) and P.ostreatus, exhibiting good
tolerance to the pesticides and water stress and the ability to degrade lignin and
produce laccase in the presence of these pesticides.
As a result, the activity of those three isolates plus Phanerochaete chrysosporium
(well described for its bioremediation potential) was examined in soil extract broth in
relation to differential degradation of the pesticide mixture at different concentrations
(0-30 mg l-1) under different osmotic stress levels (-0.7 and -2.8 MPa). Enzyme
production, relevant to P and N release (phosphomonoesterase, protease), carbon
cycling (β-glucosidase, cellulase) and laccase, involved in lignin degradation was
quantified. The results suggested that the test isolates have the ability to degrade
different groups of pesticides, supported by the capacity for expression of a range of
extracellular enzymes at both -0.7 and -2.8 MPa water potential. P.chrysosporium and
T.versicolor R101, were able to degrade this mixture of pesticides independently of
laccase activity, whereas P.ostreatus and T.versicolor R26 showed higher production
of this enzyme. Complete degradation of dieldrin and trifluralin was observed, while
about 80% of the simazine was degraded regardless of osmotic stress treatment in a
nutritionally poor soil extract broth. The results with toxicity test (Toxalert®10),
suggested the pesticides were metabolised. Therefore the capacity for the degradation
of high concentrations of mixtures of pesticides and the production of a range of
enzymes, even under osmotic stress, suggested potential applications in soil.
Subsequently, microcosm studies of soil artificially contaminated with a mixture of
pesticides (simazine, trifluralin and dieldrin, 5 and 10 mg kg soil-1) inoculated with
P.ostreatus, T.versicolor R26 and P.chrysosporium, grown on wood chips and spent
mushroom compost (SMC) were examined for biodegradation capacity at 15ºC. The
three test isolates successfully grew and produced extracellular enzymes in soil.
Respiratory activity was enhanced in soil inoculated with the test isolates, and was
generally higher in the presence of the pesticide mixture, which suggested increased
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mineralization. Cellulase and dehydrogenase was also higher in inoculated soil than in
the control especially after 12 weeks incubation. Laccase was produced at very high
levels, only when T.versicolor R26 and P.ostreatus were present. Greatest
degradation for the three pesticides was achieved by T.versicolor R26, after 6 weeks
with degradation rates for simazine, trifluralin and dieldrin 46, 57, and 51% higher
than in natural soil. And by P.chrysosporium, after 12 weeks, with degradation rates
58, 74, and 70% higher than the control. The amendment of soil with SMC also
improved pesticide degradation (17, 49 and 76% increase in degradation of simazine,
trifluralin and dieldrin compared with the control).