*2.6. Plasmid Construction and Plant Transformation*

The *Put2*-overexpression vector (full-length coding sequence of *Put2*) was constructed as previously described [30]. Gene loss-function of *put2* lines was generated through gene editing approaches. To generate the CRISPR/Cas9 vector, the two target sequences for *put2* were designed using the online software CRISPR-GE (http://skl.scau.edu.cn/ targetdesign/, accessed on 22 January 2022), which were inserted into two single guide RNA (sgRNA) expression cassettes through overlap PCR, followed by cloning into the pYLCRISPR/Cas9Pubi-H vector via Golden Gate ligation method [31]. The confirmed pFGC1008-*Put2*-3HA vector and pYLCRISPR/Cas9Pubi-H-*put2* binary vector were transformed into *Agrobacterium tumefaciens* strain GV3101 by electroporation after transgenic plants were generated with *Agrobacterium*-mediated cotyledon transformation of *Solanum lycopersicum* cv. Ailsa Craig via a method previously described [32]. Two separate homozygous T2 lines from mutation and overexpression lines were confirmed with Sanger sequencing and qRT-PCR. The *put2* mutants and *Put2*-OE plants were used in this study. The primers for vector construction are listed in Supplemental data S3 (Table S1).
