Figure 7.

ORCA3 overexpression does not increase ZCT1 expression, and ZCT1 is not significantly induced under the EASI conditions using Agrobacterium tumefaciens strain GV3101. Catharanthus roseus seedlings were transiently transformed, as described in Mortensen et al., (2019), with Agrobacterium tumefaciens containing either a GUS (Addgene ID #123197) or ORCA3 (Addgene ID #123196) overexpression construct (Figure S4). The “No Agro.” condition was treated identical to the other infiltrations but with no Agrobacteria present. Transcript levels were normalized to the housekeeping gene, SAND (Pollier et al., 2014), and fold changes were calculated according to the 2^−∆∆Ct method relative to the GUS control (Livak & Schmittgen, 2001). Data were analyzed using a one‐way ANOVA, and significant differences, compared to the GUS control, were determined using the Dunnett's method. p‐values < .001 are indicated with three stars (***)