Yeast one-hybrid (Y1H) assay
PCR products of CsHSFA1b and CsHSFA2 were amplified from
tea leaves (Longjing 43) and inserted into the pGAD vector with EcoR I
and BamH I restriction sites, and full-length and truncated derivatives
of the CsJAZ6 promoter were inserted into the pABAi vector with
Pst I and Nco I sites. The CsJAZ6pro -pABAi construct was then
linearized by digestion with BstB I, introduced into the Gold Y1H
strain, and plated on SD/-Ura for three days. Colonies harboring theCsJAZ6 promoter were used to prepare competent cells that were
transformed with the CsHSFA1b/A2-pGAD constructs (or pGAD plasmid as the
control) and plated on SD/Leu. Then colonies were moved to SD/Leu with
supplemented 200 ng aureobasidin A (AbA).