*4.2. Molecular Confirmation of the Transgene Insertion*

The selected shoots were subcultured four/five times for rapid and clonal multiplication onto the PQL1 media contained the same composition present in PQL0 and enriched with 0.7 mg L−<sup>1</sup> BAP.

Based on the assumption that genes encoding for the same proteins in different species show conserved domain with a high degree of identity, divergent regions between pear *PHYB* gene and *AtPHYB*, and between pear *CRY1* and *LeCRY1* were selected to design specific primers. The selected regions were checked against bacteria genes as well and they did not match any homologous eukaryotic sequences of genes present in data banks. *AtPHYB* and *LeCRY1* sequences, expected product sizes and annealing temperatures used to detect each gene are presented in Table S1.

PCR amplification tests were conducted to test to validate the insertion of both genes on plantlets of selected lines (Figure S4c). For DNA extraction procedure, from 100 mg leafy shoot tissues, and PCRs chemicals and amplification profile have been using the procedure reported in previous work [72]. Amplification products were visualized on agarose gels (1.2%, *w*/*v*) and 10 µg mL−<sup>1</sup> of ethidium bromide (Figures S6 and S7).
