Fig 5. Quantification of AtNPR1 activity using qPCR.

Total RNA extracted from entire sweet orange leaf (A) or specifically midrib and petioles (B) was used as template. Sequence of primers used to amplify the AtNPR1 gene is detailed in Table 1. Transgenic lines 1 to 16 containing the 35S-NPR1 cassette are ‘Hamlin’ while lines 17 to 31 are ‘Valencia’. Transgenic lines 1 to 12 containing the AtSUC2-NPR1 cassette are ‘Hamlin’ while lines 13 to 19 are ‘Valencia’. Three independent clones were tested from each transgenic line. Total RNA from a non-transgenic plant was also included to verify the accuracy of the amplification process. Transgenic plants that had a level of expression greater than indicated by the dotted line were considered to exhibit a relative high level of expression of AtNPR1.