AcbHLH144 transcription factor negatively regulates phenolic biosynthesis to modulate pineapple internal browning.

Internal browning (IB), a major physiological disorder of pineapples, usually happens in postharvest processes, but the underlying mechanism remains elusive. The bHLH transcription factors are involved in regulating various biological processes, but whether they could regulate tissue browning in fruit during storage remains unknown. Here we showed that the phenolic biosynthesis pathway was activated in pineapples showing IB following 9 days of storage. expression was the highest of the 180 transcription factors identified, downregulated in pineapple with IB, and negatively correlated with the major phenolic biosynthetic genes. AcbHLH144 was shown to be localized in the nucleus and its transient overexpression in pineapples and overexpression in decreased phenolic biosynthesis. The yeast one-hybrid assay and electrophoretic mobility shift assay showed that AcbHLH144 directly bound to the promoter and the dual-luciferase reporter assay showed that it inactivated transcription. These results strongly suggest AcbHLH144 as a repressor for phenolic biosynthesis. Abscisic acid (ABA) alleviated IB, reduced phenolic accumulation, and downregulated phenolic biosynthetic genes, including . Transcriptomic analysis showed that was the most upregulated of all 39 bHLHs in response to ABA. ABA enhanced expression, reduced phenolic contents, and downregulated phenolic biosynthetic genes in pineapples transiently overexpressing Moreover, ABA enhanced enzyme activity of GUS driven by the promoter. These results showed that AcbHLH144 as a repressor for phenolic biosynthesis could be activated by ABA. Collectively, the work demonstrated that AcbHLH144 negatively regulated phenolic biosynthesis via inactivating transcription to modulate pineapple IB. The findings provide novel insight into the role of AcbHLH144 in modulating pineapple IB during postharvest processes.