A fluorescent probe NSS of benzothiazole derivatives was synthesized using 2-mercaptobenzothiazole as raw material and the structure was characterized by IR, MS, 1H NMR, 13C NMR. The fluorescence spectroscopy studies demonstrated that the probe NSS showed fast-response, high selectivity and good anti-interference ability towards Zn2+ ion through the significant fluorescence enhancement effect in the DMSO solution. The fluorescence intensity of probe NSS had a good linear relationship with the concentration of Zn2+ (0～11 μmol/L), and the detection limit could reach 19.1 nmol/L. The bingding stoichiometry of probe NSS with Zn2+ was 1:1, which was confirmed by Job's plot analysis and MS. Meanwhile, complex NSS-Zn2+ had obviously fluorescence intensity quenched response to glyphosate while suffering from a slight interference in the presence of other organophosphorus pesticides. The quenching efficiency reached a maximum of 99.4%, and the detection limit of complex NSS-Zn2+ to glyphosate was 16.0 nmol/L (2.68 ng/mL). In addition, probe NSS was succesfully used in the detection of Zn2+ and glyphosate in the actual water samples, the method had potential application in the field of environmental detection.