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. 2019 Apr 30;10:925. doi: 10.3389/fmicb.2019.00925

FIGURE 4.

FIGURE 4

Effect of endophytes inoculation on the expression of genes involved in reticuline biosynthesis. Total RNA was isolated from the green capsule of 90 days old P. somniferum plants inoculated with endophytes SM1B and SM3B individually, and in the form of consortium (SM1B+SM3B), reverse transcribed and used as a template for RT-qPCR with SYBR Green detection. The capsules of non-inoculated endophyte free plants were used as a control. Expression of TyrAT (A), TYDC (B), NCS (C), 6OMT (D), CNMT (E), NMCH (F) was analyzed. Results were normalized to actin (reference transcript) and are shown relative to the level in non-inoculated endophyte free control plants (calibrator). qRT-PCR was performed on triplicate technical replicates of triplicate biological samples (n = 3). Data are means ± SE (n = 3 biological replicates) and Y-axis represents relative quantity (RQ). RQ was calculated using the equation; RQ = 2−ΔΔCt. Different letters (a–d) indicate statistically significant differences between treatments (Duncan’s multiple range test P < 0.05).