Tissue of 10-week-old S. pennellii LA0716 and S. lycopersicum M82 were harvested as follows: stems were flash-frozen in liquid nitrogen, and trichomes were shaved into 1.5-ml microcentrifuge tubes with a clean razor blade. Trichomes and denuded stems were kept in liquid nitrogen and ground with plastic micropestles in 1.5-ml microcentrifuge tubes. RNA was extracted from ground trichomes and stems (six biological replicates for each species and tissue type) using the RNeasy Plant Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions. For each sample, 250 ng of RNA as quantified using a NanoDrop 2000c (Thermo Fisher Scientific, Waltham, MA) was used to synthesize cDNA using SuperScript III Reverse Transcriptase (Invitrogen, Carlsbad, CA). qRT-PCR was carried out using SYBR Green PCR Master Mix on a QuantStudio 7 Flex Real-Time PCR System (Applied Biosystems, Warrington, UK) using the following cycling conditions: 48°C for 30 min, 95°C for 10 min, 40 cycles of 95°C for 15 s and 60°C for 1 min, followed by melt curve analysis. RT_ASFF_F and RT_ASFF_R primers were used to detect the ASFF1 transcript; RT_EF-1a_F/R, RT_actin_F/R, and RT_ ubiquitin_F/R primers were used to detect transcripts of the EF-1α, actin, and ubiquitin genes, respectively (table S3). For each biological replicate, relative levels of the ASFF1 transcript were determined using the ΔΔCt method (53) and normalized to the geometric mean of EF-1α, actin, and ubiquitin transcript levels.

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