The molecular mechanisms underlying fish tolerance to soybean meal (SBM) remain unclear. Identifying these mechanisms would be beneficial, as this trait favors growth. Two fish replicates from 19 experimental families were fed fishmeal-(100FM) or SBM-based diets supplemented with saponin (50SBM + 2SPN) from juvenile to adult stages. Individuals were selected from families with a genotype-by-environment interaction higher (HG-50SBM + 2SPN, 170 ± 18 mg) or lower (LG-50SBM + 2SPN, 76 ± 10 mg) weight gain on 50SBM + 2SPN for intestinal transcriptomic analysis. A histological evaluation confirmed middle intestinal inflammation in the LG- vs. HG-50SBM + 2SPN group. Enrichment analysis of 665 differentially expressed genes (DEGs) identified pathways associated with immunity and lipid metabolism. Genes linked to intestinal immunity were downregulated in HG fish (mpx, cxcr3.2, cftr, irg1l, itln2, sgk1, nup61l, il22), likely dampening inflammatory responses. Conversely, genes involved in retinol signaling were upregulated (rbp4, stra6, nr2f5), potentially favoring growth by suppressing insulin responses. Genes associated with lipid metabolism were upregulated, including key components of the SREBP (mbtps1, elov5l, elov6l) and cholesterol catabolism (cyp46a1), as well as the downregulation of cyp7a1. These results strongly suggest that transcriptomic changes in lipid metabolism mediate SBM tolerance. Genotypic variations in DEGs may become biomarkers for improving early selection of fish tolerant to SMB or others plant-based diets.

Chitin is an excellent material for the synthesis of nanoparticles because it is an elicitor and can form nanostructured materials. The application of chitin nanoparticles (CNPs) in plants can activate early defense responses associated with chitin. In this study, CNPs were synthesized by water in oil (W/O) emulsion using an aqueous chitin solution. The CNPs were characterized and used to evaluate the activation of genes related to early responses to chitin and the production of reactive oxygen species (ROS) on seedlings of Nicotiana benthamiana. The CNPs had an average size of 280 nm in diameter, a polydispersity of 0.299, a surface charge of 26.9 mV, and their chemical composition was corroborated by the disappearance of microaggregated CNPs treated with chitinases observed under a microscope. Seedlings treated with CNPs for one hour revealed increments in the expression of genes STZ, ATL2, and MAPK3, in contrast when they were treated with chitin oligomers, and no changes in gene CERK1 was detected in both conditions. Finally, the synthesis of ROS mediated by CNPs was detected in seedlings, which was higher than those generated by the treatment of chitin oligomers. These results demonstrated the capability to generate CNPs by emulsion, which are capable of triggering responses related to early defense in N. benthamiana more efficiently than chitin oligomers.

One of the causal agents of bacterial canker is Pseudomonas amygdali pv. morsprunorum—Pam (formerly Pseudomonas syringae pv. morsprunorum). Recently detected in Chile, Pam is known to cause lesions in the aerial parts of the plant, followed by more severe symptoms such as cankers and gummosis in the later stages of the disease. This study presents the design of PCR and LAMP detection methods for the specific and sensitive identification of Pseudomonas amygdali pv. morsprunorum (Pam) from cherry trees. Twelve Pseudomonas isolates were collected, sequenced, and later characterized by Multi-locus Sequence Analysis (MLSA) and Average Nucleotide Identity by blast (ANIb). Three of them (11116B2, S1 Pam, and S2 Pam) were identified as Pseudomonas amygdali pv. morsprunorum and were used to find specific genes through RAST server, by comparing their genome with that of other Pseudomonas, including isolates from other Pam strains. The effector gene HopAU1 was selected for the design of primers to be used for both techniques, evaluating sensitivity and specificity, and the ability to detect Pam directly from plant tissues. While the PCR detection limit was 100 pg of purified bacterial DNA per reaction, the LAMP assays were able to detect up to 1 fg of purified DNA per reaction. Similar results were observed using plant tissues, LAMP being more sensitive than PCR, including when using DNA extracted from infected plant tissues. Both detection methods were tested in the presence of 30 other bacterial genera, with LAMP being more sensitive than PCR.