Non-targeted metabolomics characterization of Annona derivatives with lethal and sublethal toxicities to coffee leaf miner

Abstract

The bioactivity of ethanolic extracts from seeds of Neotropical Annona species (Annona montana, Annona mucosa, Annona muricata, Annona reticulata, and Annona sylvatica) was initially assessed for their potential to control the coffee leaf miner (CLM), Leucoptera coffeella (Lepidoptera: Lyonetiidae), a primary threat to coffee production in Brazil and elsewhere. At the diagnostic concentration (12 g L−1), the ethanolic seed extract from A. mucosa (ESAM) was the most promising treatment, resulting in a mortality rate (80%) of second- to third-instar larvae of CLM that was superior to a cyantraniliprole-based synthetic insecticide and an annonin-based botanical insecticide used as positive controls. Furthermore, ESAM also demonstrated significant oviposition deterrence and a notable reduction in the number of leaf mines and surviving larvae. The laboratory results were corroborated in a greenhouse trial, which also demonstrated the absence of phytotoxicity to coffee seedlings. Following liquid–liquid partitioning, the resulting fractions were subjected to untargeted metabolomic profiling based on mass spectrometry. The molecular network of the methanolic fraction enabled the annotation of several compounds, including the isoquinoline alkaloids roemerine, reticuline, and annonaine; the cinnamic acid-derived alkaloid moupinamide; the tryptamine-derived alkaloids N-stearoyltryptamine, N-lignoceroyltryptamine, and behenic acid tryptamide; as well as acetogenins and cyclic peptides. In contrast, the GC–MS data obtained from the hexane fraction facilitated the annotation of one aldehyde, five esters, four fatty acids, three phytosterols, one diterpenoid, one monoacylglycerol, and one diacylglycerol. These findings broaden the spectrum of action for A. mucosa derivatives in managing leaf-mining pest in perennial crops, including CLM in coffee production systems.