Tuta absoluta is one of the most significant invasive pests affecting tomato plants worldwide. RT-qPCR has emerged as one of the most sensitive and accurate methods for detecting gene expression data. The screening of stable internal reference genes is the most critical step for studying the molecular mechanisms of environmental adaptability. The stable reference genes expressed in T. absoluta under specific experimental conditions have not yet been clarified. In this study, seven candidate reference genes (RPL27, RPS13, RPS15, EF1-α, TUB, TBP, and β-actin) and their optimal numbers were evaluated under biotic (developmental stages and adult tissues) and abiotic (insecticide, temperature, and plant VOC) conditions using four software programs. Our results identified the following reference genes and numbers as optimal: three genes (EF1-α, RPS13, and RPL27) for different developmental stages (egg, larva, pupa, unmated adult), two genes (RPS13 and TBP) for adult tissues (antenna, head, thorax, abdomen, leg), two genes (TBP and RPS13) for insecticides (Bacillus thuringiensis, chlorpyrifos, abamectin-aminomethyl, and chlorantraniliprole), two genes (RPL27 and TUB) for temperature-induced stresses (0, 25, and 40 °C), and two genes (RPS13 and TUB) for VOC-induced stresses (nonanal, α-phellandrene, and tomato leaves). Our results provide a reference for selecting appropriate reference genes for further study of the functional genes of T. absoluta under different experimental conditions.

The tomato leafminer, Tuta absoluta (Lepidoptera: Gelechiidae), is a highly destructive invasive pest targeting Solanaceae crops. Its olfactory system plays a crucial role in host location, mate finding, and other behavioral activities. However, there is a notable gap in the literature regarding the characterization of its chemosensory genes. In this study, we conducted a genome-wide identification of 58 odorant receptors (ORs) of T. absoluta. The identified ORs exhibit coding sequence (CDS) lengths ranging from 1062 bp to 1419 bp, encoding proteins of 354 to 473 amino acids. Gene structure analysis showed that the majority of these ORs consist of five, seven, eight, or nine exons, collectively representing 67% of the total ORs identified. Through chromosomal mapping, we identified several tandemly duplicate genes, including TabsOR12a, TabsOR12b, TabsOR12c, TabsOR21a, TabsOR21b, TabsOR34a, TabsOR34b, TabsOR34c, TabsOR62a, and TabsOR62b. The phylogenetic analysis indicated that six TabsORs were clustered within the lepidopteran sex pheromone receptor clade, while an expansion clade containing ten TabsORs resulted from tandem duplication events. Additionally, five TabsORs were classified into a specific OR clade in T. absoluta. Furthermore, through RNA-Seq and RT-qPCR analyses, we identified five TabsORs (TabsOR21a, TabsOR26a, TabsOR34a, TabsOR34c, and TabsOR36) exhibiting female-antennae-biased expression. Our study provides a valuable foundation to further investigations into the molecular and ecological functions of TabsORs, particularly in relation to oviposition behavior. These findings provide foundational data for the future exploration of the functions of female-biased expression OR genes in T. absoluta, thereby facilitating the further development of eco-friendly attract-and-kill techniques for the prevention and control of T. absoluta.

Tuta absoluta (Meyrick), a leafminer that damages tomato leaves, terminal buds, flowers, and fruits, is a destructive tomato pest and is responsible for 80–100% of tomato yield losses globally. Different insect species have different courtship responses and phototropic flight rhythms. Improving the trapping effects of the sex pheromone and light traps is important for constructing an IPM system for T. absoluta. The present study explored the hourly distribution of T. absoluta adults caught by the sex pheromone (on the ground) and UV light (380 nm) traps in greenhouses over 24 h. The responses of males to sex pheromone (false female) lures were detected at dawn and early morning. The responses lasted for 3 h, from 05:30 (1 h before sunrise) to 08:30 (2 h after sunrise), and 95.8% of the males were caught during this period. The peak of the male responses to the sex pheromone was detected at 07:30 (from 06:30 to 07:30, 1 h after sunrise), and 80.8% of the males were caught during this period. The flight of male (proportion of 54.3%) and female (45.7%) adults toward the UV light traps occurred from 19:30 (time of sunset) to 06:30 (time of sunrise), lasted for 11 h, and exhibited a scotophase rhythm; 97.4% of the adults were caught during this period. The peak of adults flying toward the UV light traps occurred at 21:30 (from 20:30 to 21:30, 2 h after sunset). The rhythms of males’ responses to the sex pheromone and of the adults’ flight toward the UV lights can help to reveal the mechanisms of chemotactic and phototactic responses and may play a significant role in constructing an IPM system for this pest.

The globally invasive cassava mealybug Phenacoccus manihoti Matile-Ferrero is a pernicious pest of cassava, and its recent introduction into Asia has raised considerable alarm. To slow or prevent further invasion, an accurate, simple, and developmental-stage-independent detection method for P. manihoti is required. In the present study, a PCR method based on a species-specific mitochondrial DNA cytochrome oxidase I (SS-COI) marker was developed for rapid identification of P. manihoti. One pair of SS-COI primers (PMSSZW-1F and PMSSZW-1R) was designed based on sequence variations in the COI gene among P. manihoti and related mealybug species. Specificity of the primer pair was validated on 21 closely related species. Sensitivity tests were performed on four immature developmental stages and female adults. Efficacy tests demonstrated that at the relatively low concentration of (135.2±14.7) pg μL⁻¹ resuspended DNA, the specific fragment was detected in all replicates. Furthermore, the SS-COI primer pair was assayed on three populations of P. manihoti from major exporting countries of cassava. The PCR assay was proved to be a rapid, simple, and reliable molecular measure for the identification of P. manihoti. This tool will be useful for quarantine, monitoring, and management of this invasive pest.

Leptinotarsa decemlineata Say 1824, an invasive and globally devastating beetle, inflicts great damage to potato crops worldwide. The complete mitogenome of L. decemlineata is described in this study. It is a 16,741 bp long circular DNA molecule with a high A + T content of 76.9%, containing a typical 37 gene pattern. All PCGs (protein-coding genes) initiate with typical ATN codons. Most PCGs use TAN as a stop codon, whereas ND4 and COX3 use the incomplete codon TA as the stop codon. The lengths of rrnL and rrnS genes are 1,337 bp and 811 bp, respectively. All 22 tRNAs ranged from 62 to 77 bp. Phylogenetic analysis of Chrysomelidae indicated that L. decemlineata clusteres with three other Chrysomelinae species, which is consistent with previous analyses.