Genome-Wide Characterization of the PaO Gene Family and Pyramiding Effects of Superior Haplotypes on Yield-Related Traits in Sorghum

The Pheophorbide a oxygenase (PaO) is a key enzyme in chlorophyll degradation and plays an important role in plant senescence. However, the PaO gene’s function in sorghum remains underexplored. In this study, we identified five SbPaO gene family members in the sorghum genome through bioinformatics analysis. Analyses of gene structure, phylogeny, and collinearity revealed high conservation of this gene family among grass crops, suggesting similar functions. Subcellular localization and protein network predictions indicated that SbPaOs may participate in chlorophyll catabolism and regulate leaf senescence. Expression pattern analysis showed that SbPaO1, SbPaO3, SbPaO4, and SbPaO5 were highly expressed in leaves and significantly upregulated during senescence. Haplotype analysis found three SbPaO genes significantly linked to thousand-grain weight (TGW); superior haplotypes SbPaO1-hap4, SbPaO3-hap5, and SbPaO4-hap4 notably increased this trait. Single-gene improvements increased TGW by 10.57–17.20%, dual-gene aggregation by 18.78–24.75%, and three-gene aggregation by 29.09%. The study also developed Kompetitive Allele-Specific PCR (KASP) markers that identify superior haplotypes with 100% accuracy. In summary, this study’s results provide a theoretical basis and genetic resources for further exploration of haplotype pyramiding strategies to breed new high-yielding sorghum varieties and delineate a clear research direction for subsequent functional validation and breeding practices.