Pgadt7

The ORF of ShATL78L was amplified using specific primers ATL78L-BD (Supplementary Table S1) containing NdeI/PstI restriction sites, thus corresponding to the yeast expression vector pGBKT7 (Clontech, USA), to construct pGBKT7-ShATL78L. The pGADT7-CSN5B was combined by the homologous recombination method (ClonExpress^TM II One Step Cloning Kit, Vazyme, China). According to the manufacturer’s protocol, the plasmids of pGBKT7-ShATL78L+pGADT7-CSN5B, pGBKT7-Lam+pGADT7-RecT (negative control), and pGBKT7-53+pGADT7-RecT (positive control) were co-transformed into yeast Saccharomyces cerevisiae strain AH109. The positive and negative controls were provided with the BD Matchmaker library construction and screening kits (Clontech, USA). The transformants were tested on the SD/-Leu/-Trp and the SD/-Ade/-His/-Leu/-Trp media. CSN5B accession numbers in the SGN database are Solyc06g073150 and Solyc11g017300.

AH109 (Clontech, Japan) was used for host strain in yeast two-hybrid interaction assays. Parent plasmids pGADT7 and pGBKT7 (Clontech, Japan) acted as activation domain (AD) and DNA binding domain (BD), respectively. Target gene fragments, amplified from the genome of PLY01, were fused to pGADT7 or pGBKT7 through ClonExpress Ultra one-step cloning kit (Vazyme, China). The premiers for plasmids construction are listed in Table S3. Bait and prey plasmids were simultaneously introduced into AH109 using the lithium acetate method as mentioned and SD medium lacking tryptophan and leucine (SD-TL) for the selection of transformants. Yeast plates were incubated at 30°C for 3 to 5 days. The yeast strain AH-AMcm2-BMcm10 was employed as a positive control (PC), whereas AH-AHog1-B and AH-A-BSet5 were employed as negative control (NC).