Fuel moisture content (FMC) proved to be one of the most relevant parameters for controlling fire behavior and risk, particularly at the wildland-urban interface (WUI). Data relating FMC to spectral indexes for different species are an important requirement identified by the wildfire safety community. In Valparaíso, the WUI is mainly composed of Eucalyptus Globulus and Pinus Radiata—commonly found in Mediterranean WUI areas—which represent the 97.51% of the forests plantation inventory. In this work we study the spectral signature of these species under different levels of FMC. In particular, we analyze the behavior of the spectral reflectance per each species at five dehydration stages, obtaining eighteen spectral indexes related to water content and, for Eucalyptus Globulus, the area of each leave—associated with the water content—is also computed. As the main outcome of this research, we provide a validated linear regression model associated with each spectral index and the fuel moisture content and moisture loss, per each species studied.