Leaf Segment Size and Culture Media influence the recovery and Diversity of Fungal Endophytes isolated from Madhuca longifolia with their Enzymes Activity

This study was performed to analyse the impact of leaf segment size and culture media on isolation and diversity of endophytic fungi from Madhuca longifolia with enzyme production potential. Total 14 taxa of endophytic fungi were recovered, dominated by Colletotrichum gloeosporioides, Corynespora and Macrophoma respectively. Reduction in leaf segment size showed increased isolation rate and significantly changed the diversity to all culture media. On potato dextrose agar 153 isolates were recovered with minimum isolation rate and diversity (1.20 isolates/cm², Shannon index 1.43) to larger size segments (T₁), followed by medium size segments (T₂) (1.95 isolates/cm², Shannon index 1.78) and maximum to small size segments (T₃) (4.50 isolates/cm², Shannon 1.89). On malt extract agar 145 isolates were recovered in which large size segments showed minimum isolation rate and diversity (1.30 isolates/cm², Shannon of 1.33) followed by medium size (isolation rate 1.95 isolates/cm², Shannon of 1.55) and maximum to small size segments (isolation rate 4.0 isolates/cm², Shannon of 1.96). On water agar, 108 isolates were recovered with similar trends of isolation rate and diversity to large size (0.65 isolates/cm², Shannon of 0.27) followed by medium size (isolation rate 1.40 isolates/cm², Shannon of 1.42) and maximum to small size segments (3.35 isolates/cm², Shannon of 1.59). This study revealed significantly that smaller leaf segment was most suitable size that gives real measures of isolation rate and diversity. Interestingly, of 24 endophytic fungi evaluated for enzyme activity, 18 were found positive to amylase, 9 to cellulase, 5 to pectinase and 14 to urease, respectively.