This work investigated the application of two
selected beneficial microorganisms (T. harzianum
and T. viride) to corn seed and their subsequent
survival and establishment in the rhizosphere once
the seed was planted. Briefly this involved storing
agar plugs taken from the periphery of an actively
growing colony in a 10% (v:v) aqueous solution of
glycerol in straw ampoules. Fungi recovered from
this system were subsequently grown on potato
dextrose agar (PDA; Merck) [21] slopes for storage
for 7 days at 30 °C in a termostated incubator
chamber with air circulation. This period showed to
be sufficient for fungi sporulation. The substrates
used were wheat bran. Wheat bran showed to be the
most suitable substrate to produce Trichoderma
spores for all strains that were evaluated. The
substrates were sterilized at 121 °C for 15 min in a
Phoenix autoclave model AV 50 and cooled down to
room temperature before the inoculation, which was
done until 24 h after sterilization. The dosages of two
funguses applied were 5 and 10g per pot. Control
plants were also available. A control plot in each
block was left untreated and only water without the
antagonistic fungus was provided. Corn seeds were
sown in plastic pots on July 6, 2009. Availability of
water in the soil plays an important role in
facilitating establishment and effectiveness of
Trichoderma in the soil, Therefore irrigation was
done so on a regular basis