Phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) catalyzes the first step in the phenylpropanoid pathway, and is considered an important regulation point between primary and secondary metabolism. In the present work we analyzed expression of the PAL genes in leaves of Arabidopsis thaliana rosette-stage plants in response to nitrogen depletion at temperatures ranging from 5 to 30 degrees C. Only PAL1 and PAL2 responded strongly to both environmental factors, nitrogen and temperature. Regardless of nitrogen treatments, PAL1 and 2 transcript levels increased at 5 and 10 degrees C. Averaged across all temperatures, nitrogen depletion led to a two-fold increase in PAL1 and PAL2 transcripts. PAL activity was correlated with PAL transcript levels (R=0.94). Accumulation of major soluble phenylpropanoids, sinapic acid esters and flavonoids, increased in response to lowering temperature. The flavonoids, kaempferols, quercetins and anthocyanins, showed significantly increased levels as a result of nitrogen depletion (two-, five- and six-fold increases, respectively) when averaged across all temperatures. PAL1, PAL2 and PAL4 have previously been shown to be related with tissue-specific lignin synthesis, and the present work shows that PAL1 and PAL2 also have functional specialization in abiotic environmental-triggered flavonoid synthesis.