The experiment consists of growing white prosperity cultivar of gladiolus at four different planting times (1st fortnight of October, 2nd fortnight of October, 1st fortnight of November and 2nd fortnight of November) and with eight different treatments of bio inoculants. The experiment was laid out in randomized block design with three replications. Among the different bio inoculants treatment, T8 treatment (RDF + Azotobacter + PSB + Mycorrhiza) proved best with respect to maximum phosphatase enzyme activity when compared to other treatments and among different time of plantings, 1st fortnight of October was the best planting time for gladiolus in terms of maximum phosphatase enzyme activity.