Online measurement of soil moisture is significant for agricultural production. The abundance and deficiency of leaf water shortage has been able to effect measured by fluorescence, but the research of correlation between soil moisture and laser-induced fluorescence spectroscopy has not been carried out yet. In this paper, the relationship between laser-induced plant chlorophyll fluorescence and soil moisture was studied. In the experiment with rice as the research object, the chlorophyll fluorescence induced by a 450 nm LED light source was measured by a Y-shaped fiber probe, and the soil moisture was obtained by TDR sensors. The changes in the peak intensity near 743 nm of chlorophyll fluorescence under continuous water stress and intermittent water stress were studied. The result showed that the intensity decreased with the soil moisture content. Finally, the model between the soil water content and the intensity of chlorophyll fluorescence under continuous water stress was created by using Lorentzian equation, and the coefficient of determination (R2) of the model was high. The result indicated that the method mentioned in paper can be applied to measure soil moisture in agricultural production.