Colloidal chitin was prepared following the method
of Roberts and Selintrenikoff (1988) with certain
modifications. The per cent transmission of the
standard and the plant samples was recorded against
reagent blank which was adjusted to 100% T at 540
nm and reducing sugars present per gram of the
sample were calculated based on the NAG standard
graph. Further, for plant samples, 100 µl of crude
protein, 100 µl of McILvaine (1920) buffer and 100 µl
of colloidal chitin were added and standard protocol
(Katany et al. 2000) for chitinase assay was followed.
In order to identify the chitinase activity, reducing
sugars released by 100 µl of leaf extract in 30 min
was converted into pico moles of reducing sugars
released per microgram of crude protein per min.
Further glucanase enzyme activity was assayed by
using colorimetric method described by Katany et al
(2000).