Integration of full-length Iso-Seq, Illumina RNA-Seq, and flavor testing reveals potential differences in ripened fruits between two Passiflora edulis cultivars.

Abstract

Passion fruit (Passiflora edulis) is loved for its delicious flavor and nutritious juice. Although studies have delved into the cultivation and enhancement of passion fruit varieties, the underlying factors contributing to the fruit's appealing aroma remain unclear. This study analyzed the full-length transcriptomes of two passion fruit cultivars with different flavor profiles: "Tainong 1" (TN1), known for its superior fruit flavor, and "Guihan 1" (GH1), noted for its strong environmental resilience but lackluster taste. Utilizing PacBio Iso-Seq and Illumina RNA-Seq technologies, we discovered terpene synthase (TPS) genes implicated in fruit ripening that may help explain the flavor disparities. We generated 15,913 isoforms, with N50 lengths of 1,500 and 1,648 bp, and mean lengths of 1,319 and 1,463 bp for TN1 and GH1, respectively. Transcript and isoform lengths ranged from a maximum of 7,779 bp to a minimum of 200 and 209 bp. We identified 14,822 putative coding DNA sequences (CDSs) averaging 1,063 bp, classified 1,007 transcription factors (TFs) into 84 families. Additionally, differential expression analysis of ripening fruit from both cultivars revealed 314 upregulated and 43 downregulated unigenes in TN1 compared to GH1. The top 10 significantly enriched Gene Ontology (GO) terms for the differentially expressed genes (DEGs) indicated that TN1's upregulated genes were primarily involved in nutrient transport, whereas GH1's up-regulated genes were associated with resistance mechanisms. Meanwhile, 17 PeTPS genes were identified in P. edulis and 13 of them were TPS-b members. A comparative analysis when compared PeTPS with AtTPS highlighted an expansion of the PeTPS-b subfamily in P. edulis, suggesting a role in its fruit flavor profile. Our findings explain that the formation of fruit flavor is attributed to the upregulation of essential genes in synthetic pathway, in particular the expansion of TPS-b subfamily involved in terpenoid synthesis. This finding will also provide a foundational genetic basis for understanding the nuanced flavor differences in this species.