Four kinds of bioagents simulants, Pantoea agglomerans (Pan), Staphylococcus aureus subsp. Aureus (Sta), Bacillus globigii (BG) and Escherichia coli (EH) were cultured and the strains growth curves were obtained, and the generation time were 0.99, 0.835, 1.07 and 1.909 h respectively. Two wavelengths at 266 and 355 nm of a designed short range fluorescence lidar were engaged for the measurements of two dimensional fluorescence spectra in the amino acid band and NADH band of the biological warfare agents, respectively. The simulants were diffused homogeneously inside a controlled fluorescence measurement chamber and interogated by the lidar. The 2 dimensional fluorescence spectra of four kinds of vegetative bacteria, BSA and OVA(as simulant of toxin agents) were obtained with resolution of 4 nm. The fluorescence spectra of Pan, Sta, EH and BG, BSA and OVA were consistent with the standard fluorescent component tryptophan in the amino acid band with FWHM of 60 nm, but the central wavelength of the fluorescence spectra of these simulants blue/purple shifted obviously as affected by the external biochemical environment, concentration and ratio of different bacterial internal fluorophores, so the energy level between the excited state and the ground state of the fluorescence molecule increased. Accordingly, weak NADH fluorescence spectra with 100 nm FWHM inside the four vegetative bacteria aerosols were also detected, but Raman scattering contribution of water and nitrogen could not be effectively extracted. The second-order derivative fluorescence spectra of the four simulants showed that the high-order processing and recognition of the fluorescence spectrum was feasible.