The utility, advantages, and disadvantages of several spectrofluorometric and in vivo spectrophotometric methods in plant science are reviewed, with emphasis on their use in weed science. Examples of the use of in vivo, dual-wavelength spectrophotometry to probe the effects of herbicides on cytochrome f oxidation/reduction, the P515 chromatic shift, phytochrome synthesis, and other processes are discussed. Use of in vivo spectrophotometry to examine effects of herbicides on phytylation of chlorophyllide, protochlorophyllide photoconversion, porphyrin accumulation, and other processes that result in spectral changes is discussed and examples are provided. Spectrofluorometric methods for pigment identification and quantification and for enzyme assays are illustrated with examples. Spectrofluorometric methods used in conjunction with high-pressure liquid chromatography (HPLC) and thin-layer chromatography (TLC) are discussed.