Fungal culture and maintenance
Cultures of filamentous fungi - Trichoderma viride (NCIM
1053) and two virulent cultures of Fusarium oxysporum
(NCIM 1072) and Alternaria alternata (NCIM 718) causing wilt and blight in legumes were obtained from National
collection of Industrial microorganisms (NCIM), National
chemical laboratory (NCL), Pune, India. These fungi were
cultured on Potato dextrose agar slants and maintained in an
environmental chamber set at 28 ± 2°C, 90% relative humidity
and 16:8 h light: dark regime. An aqueous conidial suspension
of 106 condia/ml containing 105 mg/ml Tween 80 was prepared
from 14 days old culture (Nageswara Rao Reddy, 2006).
Conidial viability was tested and used for further experiments
if the conidial germination was more than 95%. Cultures were
initiated by inoculating 1 ml of conidial suspension containing
107 conidia in 250 ml of Potato dextrose broth for further
experiments.