Changes in biochemical constituents and physiological alteration were studied in various intensities (1-5%, 6-15%, 16-30%, 31-50% and ＞ 50%) of leaf spot (Myrothecium roridum) on mulberry leaves and compared with healthy leaves. Chlorophyll, total soluble sugar and total protein were decreased (P ＜ 0.01), but total phenol increased due to pathogen infection. Changes in biochemical constituents showed significant correlation with intensity of disease. Chlorophyll ($r^2$= 0.92), and protein (($r^2$= 0.83) possessed negative while phenol (($r^2$= 0.61) possessed positive correlation. Photosynthetic rate, transpiration rate, stomatal conductance, moisture content (%) and physiological water use efficiency (pWUE) were decreased, but stomatal resistance increased in the infected leaves. Physiological parameters also possessed significant (P ＜ 0.01) correlation with disease intensity. Photosynthetic rate (($r^2$= 0.96), transpiration rate ($r^2$=0.88), stomatal conductance (($r^2$= = 0.65), physiological water use efficiency (($r^2$= 0.88) and moisture content (r = 0.85) were negatively but stomatal resistance (($r^2$= 0.75) was positively correlated to disease intensities.