Genome-wide identification of trihelix transcription factor family genes in pear (Pyrus bretschneideri) and functional characterization of PbrGT15 in black spot resistance

Pear ( Pyrus bretschneideri ), a valuable widely cultivated fruit, faces significant economic losses due to black spot disease caused by Alternaria alternate (Fr.) Keissl. Trihelix transcription factors (TFs) are crucial in regulating plant defense and autoimmunity. This study aimed to analyze the trihelix transcription factor (GT) genes within pear through genome-wide identification, phylogenetic, gene structure, synteny, and cis- acting elements analyses. Among the 31 trihelix genes, 28 were on 12 known chromosomes, while the remaining 3 were located on unknown chromosomes. These genes were categorized into five clades: SIP1, GTγ, GT1, GT2 and SH4, containing 7, 2, 9, 11 and 2 genes, respectively. Synteny analysis indicated eight duplicated gene pairs. Based on the expression pattern of PbGT genes in seven tissues from the database, the PbGT genes of the GT2 clade were selected for further investigation. The quantitative reverse transcriptase–polymerase chain reaction confirmed that PbrGT5 , PbrGT6 , PbrGT15 and PbrGT16 correlated with black spot disease resistance. Notably, the salicylic acid (SA) treatment significantly upregulated the expression levels of PbrGT10 , PbrGT13 , PbrGT15 and PbrGT23 . Among these, PbrGT15 showed the highest induction to both SA and black spot infection. Subcellular localization demonstrated that PbrGT15 functions as a nuclear protein. Virus-induced gene silencing of PbrGT15 increased pear plants' susceptibility to black spot disease, indicating its pivotal role in enhancing resistance. These results indicated that PbrGT15 positively regulated black spot disease resistance in pears.