Opsin mutants alter host plant selection by color vision in the nocturnal invasive pest Tuta absoluta.

In insects, vision is crucial in finding host plants, but its role in nocturnal insects is largely unknown. Vision involves responses to specific spectra of photon wavelengths and opsins plays an important role in this process. Long-wavelength sensitive opsin (LW opsin) and blue-sensitive opsin (BL opsin) are main visual opsin proteins and play important in behavior regulation.We used CRISPR/Cas9 technology to mutate the long-wavelength-sensitive and blue wavelength-sensitive genes and explored the role of vision in the nocturnal invasive pest Tuta absoluta. Light wave experiments revealed that LW2(-/-) and BL(-/-) mutants showed abnormal wavelength tropism. Both LW2 and BL mutations affected the preference of T. absoluta for the green environment. Mutations in LW2 and BL are necessary to inhibit visual attraction. The elimination of LW2 and BL affected the preference of leaf moths for green plants, and mutations in both induced a preference in moths for white plants. Behavioral changes resulting from LW2(-/-) and BL(-/-) mutants were not affected by sense of smell, further supporting the regulatory role of vision in insect behavior. To the best of our knowledge, this is the first study to reveal that vision, not smell, plays an important role in the host-seeking behavior of nocturnal insects at night, of which LW2 and BL opsins are key regulatory factors. These study findings will drive the development of the "vision-ecology" theory.

Early warning and management of invasive crop pests under global warming: estimating the global geographical distribution patterns and ecological niche overlap of three Diabrotica beetles.

Invasive alien pests (IAPs) pose a major threat to global agriculture and food production. When multiple IAPs coexist in the same habitat and use the same resources, the economic loss to local agricultural production increases. Many species of the Diabrotica genus, such as Diabrotica barberi, Diabrotica undecimpunctata, and Diabrotica virgifera, originating from the USA and Mexico, seriously damaged maize production in North America and Europe. However, the potential geographic distributions (PGDs) and degree of ecological niche overlap among the three Diabrotica beetles remain unclear; thus, the potential coexistence zone is unknown. Based on environmental and species occurrence data, we used an ensemble model (EM) to predict the PGDs and overlapping PGD of the three Diabrotica beetles. The n-dimensional hypervolumes concept was used to explore the degree of niche overlap among the three species. The EM showed better reliability than the individual models. According to the EM results, the PGDs and overlapping PGD of the three Diabrotica beetles were mainly distributed in North America, Europe, and Asia. Under the current scenario, D. virgifera has the largest PGD ranges (1615 × 104 km2). In the future, the PGD of this species will expand further and reach a maximum under the SSP5-8.5 scenario in the 2050s (2499 × 104 km2). Diabrotica virgifera showed the highest potential for invasion under the current and future global warming scenarios. Among the three studied species, the degree of ecological niche overlap was the highest for D. undecimpunctata and D. virgifera, with the highest similarity in the PGD patterns and maximum coexistence range. Under global warming, the PGDs of the three Diabrotica beetles are expected to expand to high latitudes. Identifying the PGDs of the three Diabrotica beetles provides an important reference for quarantine authorities in countries at risk of invasion worldwide to develop specific preventive measures against pests.

Spatial-temporal shifting patterns and in situ conservation of spotted seal (Phoca largha) populations in the Yellow Sea ecoregion.

Understanding the habitat shifting pattern is a prerequisite for implementing in situ conservation of migratory species. Spotted seals (Phoca largha) inhabiting the Yellow Sea ecoregion (YSE) comprise a small population with independent genes and represent a charismatic flagship species in this region. However, this population has declined by 80% since the 1940s, and increased support from the countries around the YSE is urgently needed to address the potential local extinction risk. A time-series niche model and life-history weighted systematic conservation planning were designed on the basis of a satellite beacon tracking survey (2010-2020) of the YSE population. The results showed clustering and spreading shifting patterns during the breeding and migratory seasons, respectively. The closed-loop migration route formed in the YSE indicated that this population might be geographically isolated from populations in other breeding areas around the world. The conservation priority area (CPA), with an area of 19 632 km2 (3.58% of the total YSE area), was the most effective response to the potential in situ risk. However, nearly 80% of the CPA was exposed outside the existing marine protected areas (MPAs). Future establishment of MPAs in China should strategically consider the conservation gap identified herein, and it is recommended for Korea's closed fishing season to be spatially set in the western Korean Peninsula from May to August. This study also exemplified that the lack of temporal information would lead to the dislocation of niche modeling for migratory species represented by spotted seals. Attention should be paid to protecting small and migratory populations in marine biodiversity conservation planning.

Bacterial Communities of the Internal Reproductive and Digestive Tracts of Virgin and Mated Tuta absoluta.

Microorganisms can affect host reproduction, defense, and immunity through sexual or opportunistic transmission; however, there are few studies on insect reproductive organs and intestinal bacterial communities and their effects on mating. Tuta absoluta is a worldwide quarantine pest that seriously threatens the production of Solanaceae crops, and the microbial community within tomato leafminers remains unclear. In this study, 16s rRNA sequencing was used to analyze bacterial communities related to the reproductive organs and intestinal tracts of tomato leafminers (the sample accession numbers are from CNS0856533 to CNS0856577). Different bacterial communities were found in the reproductive organs and intestinal tracts of females and males. Community ecological analysis revealed three potential signs of bacterial sexual transmission: (1) Mating increased the similarity between male and female sex organs and intestinal communities. (2) The bacteria carried by mated individuals were found in unmated individuals of the opposite sex but not in unmated individuals of the same sex. (3) The bacteria carried by unmated individuals were lost after mating. In addition, the abundances of bacterial communities carried by eggs were significantly higher than those of adult worms. Our results confirm that mating leads to the transfer of bacterial communities in the reproductive organs and gut of tomato leafminers, and suggest that this community strongly influences the reproductive process.

Characteristics of the Accessible Chromatin Landscape and Transcriptome under Different Temperature Stresses in Bemisia tabaci.

Bemisia tabaci is an important invasive pest with worldwide distribution and strong temperature tolerance. Previous studies have shown that temperature tolerance varies significantly between the different invasive populations. Several key factors involved in epigenetic regulation have been identified and verified in B. tabaci; therefore, epigenetic adaptation mechanisms may also exist. This study aimed to detect changes in the chromatin accessibility landscape and genome-wide transcriptome under different temperature stresses in B. tabaci. Assay for transposase-accessible chromatin with high-throughput sequencing and RNA-seq analyses indicated that transcriptional activity of the genes strongly correlates with chromatin accessibility. Chromatin transcription-activated gene expression regulation is dominant during high-temperature stress in B. tabaci, mainly through the transcriptional repression of genes related to low-temperature stress resistance. Furthermore, B. tabaci resists low-temperature stress by regulating enzyme activities and withstands high-temperature stress by regulating metabolism and synthesis of organic substances, both achieved by altering chromatin accessibility. In summary, this study provides a theoretical basis for exploring changes in gene expression and chromatin accessibility under different temperature stresses, offering a new approach to unravelling regulatory mechanisms underlying the onset of molecular regulation in response to various temperature stress conditions.

Suitable habitat of Lepidochelys olivacea and the changes under climate change.

As a vulnerable species identified by the International Union for Conservation of Nature (IUCN), Lepidochelys olivacea has attracted extensive attention in recent years. To examine its current distribution and that under future climate change scenarios, we compiled the occurrence data of L. olivacea. With eight predictor variables, including depth, offshore distance, mean primary productivity, minimum primary productivity, mean sea surface temperature, minimum sea surface temperature, mean sea surface salinity, and minimum sea surface salinity, we predicted its distribution in an ensemble species distribution model. The accuracy of the model was evaluated with the parameters of areas under curves (AUC) and true skill statistics (TSS). The results showed that the AUC and TSS values were 0.96 and 0.81, respectively, indicating a good predictive performance of the ensemble model. Sea surface temperature and salinity were the two most important variables determining the distribution of L. olivacea, with the suitable temperature ranging from 23 to 29 ℃ and salinity below 34. The current distribution range of L. olivacea was between 30° N-25° S. Under future climate scenarios, its distribution range would decrease, especially under the RCP85 scenario in the 2100s (with a 28% reduction in the suitable survival range). The results of model validation showed that it had high accuracy and could make accurate predictions of the distribution. This study would provide references for the development of more rational conservation measures and management strategies.

Krüppel-homologue 1 regulates the development of Tuta absoluta and its cascade regulation pattern in the juvenile hormone signalling pathway.

Tomato leaf miner, Tuta absoluta (Meyrick), is one of the most destructive quarantine pests globally. It has been confirmed that Krüppel-homologue 1 (kr-h1) plays a key role in the regulation of juvenile hormone (JH). However, it is unclear how kr-h1 regulates the synthesis of JH and its cascade regulation pattern in tomato leaf miner. Here, we obtained the six JH signalling genes (kr-h1, Methoprene-tolerant, Forkhead box O, Juvenile acid methyltransferase, Juvenile hormone esterase and Fatty acid synthase 2), and applied RNA interference to explore the role of kr-h1 and the seven genes (plus Vitellogenin) regulation relationship in T. absoluta. Bioinformatics analysis revealed the structural characteristics of kr-h1 protein and JH receptor Met, which contained eight C2H2 zinc finger structures and three typical domains of the bHLH-PAS family, respectively. The expression levels of Met and Vg were upregulated after RNAi of kr-h1 gene, while the gene levels of JHAMT and FAS2 were downregulated. Furthermore, topical application of JH analogue to second instar larvae could induce the expression of kr-h1 and inhibit the expression of Met. Our study reveals the mechanism by which kr-h1 regulates JH pathway genes, which could be applied to control the growth of tomato leaf miners.

Gene Expression Analysis Reveals Potential Regulatory Factors Response to Temperature Stress in Bemisia tabaci Mediterranean.

Exposure to extreme temperatures can hinder the development of insects and even reduce their survival rate. However, the invasive species Bemisia tabaci exhibits an impressive response to different temperatures. This study aims to identify important transcriptional changes of B. tabaci occupying different temperature habitats by performing RNA sequencing on populations originating from three regions of China. The results showed that the gene expression of B. tabaci populations inhabiting regions with different temperatures was altered and identified 23 potential candidate genes that respond to temperature stress. Furthermore, three potential regulatory factors' (the glucuronidation pathway, alternative splicing, and changes in the chromatin structure) response to different environmental temperatures were identified. Among these, the glucuronidation pathway is a notable regulatory pathway. A total of 12 UDP-glucuronosyltransferase genes were found in the transcriptome database of B. tabaci obtained in this study. The results of DEGs analysis suggest that UDP-glucuronosyltransferases with a signal peptide may help B. tabaci resist temperature stress by sensing external signals, such as BtUGT2C1 and BtUGT2B13, which are particularly important in responding to temperature changes. These results will provide a valuable baseline for further research on the thermoregulatory mechanisms of B. tabaci that contributes to its ability to effectively colonize regions with considerable temperature differences.

Trophic transfer of polycyclic aromatic hydrocarbons in marine mammals based on isotopic determination.

The tissue distribution (liver, kidney, heart, lung, and muscle), source, and trophic transfer of 15 polycyclic aromatic hydrocarbons (PAHs) were studied on 14 stranded East Asian finless porpoises (Neophocaena asiaeorientalis sunameri), 14 spotted seals (Phoca largha), and 9 stranded minke whales (Balaenoptera acutorostrata) from Yellow Sea and Liaodong Bay. The PAHs levels ranged from below the limit of detection to 459.22 ng g-1 dry weight in the tissues of the three marine mammals, and light molecular weight PAHs were the primary pollutants. Although the PAHs levels were relatively higher in internal organs of the three marine mammals, generally no tissue-specific distribution of the PAHs congeners was found, either for gender-specific distribution of PAHs in the East Asian finless porpoises. However, species specific PAHs concentration distribution were obtained. The PAHs were mainly originated from petroleum and biomass combustion in the East Asian finless porpoises, while those for the spotted seals and minke whales were complex. Trophic level associated biomagnification was found for phenanthrene, fluoranthene, and pyrene in the minke whale. Benzo(b)fluoranthene exhibited a significant biodilution with increasing trophic levels in the spotted seals, but the total concentration of the PAHs showed a significant biomagnification with increasing trophic levels. Trophic level-associated biomagnification of acenaphthene, phenanthrene, anthracene, and ∑PAHs were found in the East Asian finless porpoise, while pyrene exhibited obvious biodilution with increasing trophic levels. Our current study filled knowledge gaps on tissue distribution and trophic transfer of the PAHs in the investigated three marine mammals.

Uncovering the role of wheat magnesium transporter family genes in abiotic responses.

Background: The CorA / MGT / MRS2 family proteins are an important group of magnesium transporter proteins that maintain magnesium ion homeostasis in plant cells. However, little is known about the MGT functions in wheat. Methods: The known MGT sequences were used as queries to BlastP against wheat genome IWGSC RefSeq v2.1 assembly (E-value <10-5). Chromosome localization information for each TaMGT gene was obtained from the GFF3 file of the wheat genome data (IWGSCv2.1).The sequence of 1500 bp upstream of the TaMGT genes was extracted from the wheat genome data. The cis-elements were analyzed using PlantCARE online tool. Result: A total of 24 MGT genes were identified on 18 chromosomes of wheat. After functional domain analysis, only TaMGT1A, TaMGT1B, and TaMGT1D had GMN mutations to AMN, while all the other genes had conserved GMN tripeptide motifs. Expression profiling showed that the TaMGT genes were differentially expressed under different stresses and at different growth and development stages. The expression levels of TaMGT4B and TaMGT4A were significantly up-regulated in cold damage. In addition, qRT-PCR results also confirmed that these TaMGT genes are involved in the wheat abiotic stress responses. Conclusion: In conclusion, The results of our research provide a theoretical basis for further research on the function of TaMGT gene family in wheat.

Tissue distribution and trophic magnification of trace elements in typical marine mammals in Bohai and north Yellow Seas.

A total of 20 stranded spotted seals (Phoca largha) and 9 stranded minke whales (Balaenoptera acutorostrata) were collected from Liaodong Bay and the northern part of the Yellow Sea to investigate the tissue distribution (liver, kidney, heart, lung, and muscle), risk, and trophic magnification of 13 trace elements (TEs, Hg, As, Cd, Co, Cr, Cu, Mn, Ni, Pb, Se, Sn, V, Zn). The 13 TEs were all detected in all spotted seal and minke whale tissue samples, with mean concentrations ranging from 0.041 to 136.3 mg kg-1 dry weight (dw) and 0.022 to 152.6 mg kg-1 dw, respectively. Zn was the dominant contaminant in all tissues for both spotted seals and minke whales. There was tissue-specific distribution of TEs in both marine mammals, and the TEs tended to accumulate in internal organs. Significant positive correlations were found in the body length of the spotted seals and minke whales among some of the TEs, especially for Cd in the internal organs. Gender-dependent distribution of the TEs was not obtained for the spotted seal. Ecological risk evaluation for spotted seals and minke whales suggested that greater concern should be given to Hg, As, and Se. Based on the TE concentrations detected in this study and trophic levels determined by stable carbon and nitrogen isotopes, trophic level-associated biodilution was obtained for As, Cd, Co, Cu, Mn, Pb, Se, Sn, and V in the spotted seal, while Zn displayed a significant biomagnification trend with increasing trophic levels. In the case of the minke whale, As, Cd, Co, Mn, Pb, Se, and V displayed significant biomagnification trends with increasing trophic levels.

First Report on CRISPR/Cas9-Based Genome Editing in the Destructive Invasive Pest Tuta Absoluta (Meyrick) (Lepidoptera: Gelechiidae).

The tomato leaf miner Tuta absoluta (Meyrick) is one of the world's most destructive pests of tomato, and because of its severe economic impacts, as well as the development of pesticide resistance, the species has been intensively studied, especially in regard to the identification of targets for T. absoluta control. However, functional genomic studies of T. absoluta have been constrained by a lack of effective genetic tools. Therefore, the aim of the present study was to develop a CRISPR/Cas9 zygote microinjection protocol for generating heritable mutations in T. absoluta, using the ommochrome synthesis gene cinnabar as an easily evaluated target gene. The injection of fertilised eggs with Cas9 protein and four sgRNAs, which targeted cinnabar exon 3, resulted in a mutagenesis rate of 31.9% for eggs reaching adulthood, and cinnabar mutagenesis resulted in either red or mosaic eye colour phenotypes. As such, this study is the first to report a complete and detailed CRISPR/Cas9 workflow for the efficient genome editing of the globally important invasive pest T. absoluta. The application of this robust genome-editing tool to T. absoluta will greatly facilitate the discovery of suitable RNAi control targets and the subsequent development of novel control strategies.

Characterization of Chromatin Remodeling Genes Involved in Thermal Tolerance of Biologically Invasive Bemisia tabaci.

As an invasive species, Bemisia tabaci Mediterranean (MED) has notable potential to adapt to a wide range of environmental temperatures, which enables it to successfully spread after invasion and occupy habitats over a wide latitude range. It has been postulated that chromatin remodeling mechanisms are related to the rapid acquisition of adaptive traits and thermal resistance in invasive species; however, relevant experimental evidence is scarce. To identify the molecular characteristics and assess the role of chromatin remodelers in thermal stress within invasive MED and native Asia II 1 of the B. tabaci species complex, we identified 13 switching defective/sucrose non-fermenting (SWI/SNF) and 10 imitation switch (ISWI) family members in the B. tabaci genome, analyzed their molecular characteristics and structures, and identified key mutation sites between MED and Asia II 1, then cloned the catalytic subunits, and revealed the difference in thermal tolerance function. The results showed that the expression levels of Bt-BRM-1 and Bt-BRM-2 were significantly higher in MED than in Asia II 1 during heat stress, and Bt-BRM-2 expression was significantly higher during cold stress. In addition, RNA interference results indicated that the two target genes had similar temperature tolerance function in the both two cryptic species. This study is the first to identify and analyze the molecular characteristics of SWI/SNF and ISWI family members and reveal their potential key roles in temperature tolerance in poikilothermic ectotherms. The results will assist in understanding the underlying temperature adaptation mechanism of invasive insects and will enrich stress adaptation research systems from an epigenetic perspective.

Spotted seals (Phoca largha) harbor unique gut microbiota shaped by their host habitat.

Assessing the structure and composition of gut microbiota of sentinel species such as spotted seals (Phoca largha) is a potential tool for assessing the health of the marine mammals and their habitats. However, the link between the host microbiome and their habitat is poorly understood. In this study, microbial communities in the habitat (sea ice and water) and marine mammalian host (fecal matter from P. largha) were evaluated in samples obtained from the Liaodong Bay, China during population aggregation period. Results from high-throughput sequencing showed that the bacterial communities in P. largha fecal matter were less rich and diverse compared to those from the water and ice samples. Significant differences in the composition and function of bacterial communities were also found among the water, ice, and fecal samples, in which sample type and sampling site had the greatest impact on composition and function variations, respectively. Several potential pathogenic bacteria and bacteria with functions associated with human disease were significantly enhanced in the communities of P. largha feces compared to those of surrounding environments. The ratios of environmental microorganisms sourced from the P. largha fecal matter were estimated. The results showed that certain bacteria in P. largha-inhabited fecal matter were associated with sea ice and had specific antibiotic resistance and infectious capacity. These findings provide critical data for monitoring the health of marine mammals and their habitats, which is essential for predicting the impact of anthropogenic disturbances on marine ecosystems.

Chromatin-Remodelling ATPases ISWI and BRM Are Essential for Reproduction in the Destructive Pest Tuta absoluta.

The tomato leaf miner (Tuta absoluta) is one of the top 20 plant pests worldwide. We cloned and identified the chromatin-remodelling ATPase genes ISWI and BRM by RACE and bioinformatic analysis, respectively; used RT-qPCR to examine their expression patterns during different life cycle stages; and elucidated their roles in insect reproduction using double-stranded RNA injections. The full-length cDNA of TaISWI was 3428 bp and it encoded a 1025-aa polypeptide. The partial-length cDNA of TaBRM was 3457 bp and it encoded a 1030-aa polypeptide. TaISWI and TaBRM were upregulated at the egg stage. Injection of TaISWI or TaBRM dsRNA at the late pupa stage significantly inhibited adult ovary development and reduced fecundity, hatchability, and longevity in the adult females. To the best of our knowledge, the present study was the first to perform molecular characterisations of two chromatin-remodelling ATPase genes and clarify their roles in T. absoluta fecundity. Chromatin-remodelling ATPases are potential RNAi targets for the control of T. absoluta and other insect pests. The present study was also the first to demonstrate the feasibility of reproductive inhibitory RNAi as a putative approach for the suppression of T. absoluta and other Lepidopteran insect populations.

Integrated proteomics and metabolomics reveal the variations in the physiological state of spotted seal (Phoca largha) pups following artificial rescue.

Spotted seals (Phoca largha) are a critically endangered pinniped in China. Artificial rescue of newborn pups is a conventional method to enhance their survival and maintain the population. However, little is known about the variations in the physiological state of spotted seal pups following artificial rescue. Here, an integrated proteomics and metabolomics study was performed on spotted seal pups by using whole blood samples to characterize the molecular response to artificial rescue. The proteome was characterized as having 1165 proteins that were predominantly associated with the metabolic pathways, and the complement and coagulation cascades. Remarkable variation was found in spotted seal pup blood following artificial rescue, whereby the levels of 193 proteins and 32 metabolites significantly varied in some metabolic pathways, including glycosylphosphatidylinositol-anchor biosynthesis, focal adhesion, cardiac muscle contraction, and fatty acid beta-oxidation. After rescue, spotted seal pups had a higher risk to mild hemolytic disorder due to the energy metabolism in the red blood cells was possibly suppressed. Moreover, spotted seal pups after rescue could have stronger anaerobic exercise abilities, while their capacity for long-term high-intensity exercise was weaker.

Chromosome-scale genome assembly of brown-spotted flathead Platycephalus sp.1 provides insights into demersal adaptation in flathead fish.

Flatheads are valuable commercial fish species endemic to the Indo-West Pacific. Due to their economic value and unique biological traits, such as metamorphosis and camouflage, they serve as ideal marine organisms for studies on demersal adaptation and evolution. The brown-spotted flathead (Platycephalus sp.1) is the most widely distributed in the northwestern Pacific. Despite the lack of a valid scientific name, it has been long recognized and exploited in the marine fisheries of China, Japan, and Korea. In the current study, we applied Illumina, PacBio, and Hi-C sequencing to assemble a chromosome-scale genome for this species. The assembled genome was 660.63 Mb long with a scaffold N50 of 28.65 Mb and 100% of the contigs were anchored onto 24 chromosomes. We predicted 22 743 protein-coding genes, 94.8% of which were functionally annotated. Comparative genomic analyses suggested that Platycephalus sp.1 diverged from its common ancestor with Gasterosteus aculeatus ~88.4 million years ago. The expanded gene families were significantly enriched in immune, biosynthetic, and metabolic pathways. Furthermore, three shared Gene Ontology terms and 377 common positively selected genes were identified between flathead and flatfish species, suggesting that these genes may contribute to demersal adaptation in flatheads. The assembled genomic data provide a valuable molecular resource for further research on the biological and adaptive evolution of flathead species.

Tissue distribution and health risk of trace elements in East Asian finless porpoises.

We investigated the tissue distribution, trophic transfer, and ecological risk of 13 trace elements in 26 East Asian finless porpoises (Neophocaena asiaeorientalis sunameri), an endangered species found in the Liaodong Bay and the north Yellow Sea. All the investigated trace elements were detected in the tissue and food web of the East Asian finless porpoises. The concentrations of the potentially toxic elements were 2.37 × 10-5 - 754 mg kg-1 dry weight (dw) in stranded porpoises and 0.01-159 mg kg-1 dw in their food web. Tissue-specific distribution of the trace elements generally ranked as: liver > kidney > heart > lung > muscle. Zn was the dominant contaminant in the five investigated tissues. Significant positive correlations were found between body length or age and some trace elements, especially Cd. Adults (≥2 years old) presented higher concentrations of most of the trace elements than juveniles (<2 years old). Sex-dependent distribution of the trace elements was insignificant except for Mn, Ni, and Zn in muscle and renal tissue. As, Cu, Mn, Ni, Pb, and V biodiluted across the East Asian finless porpoise food web while Zn biomagnified. However, Hg, Cd, Co, Cr, Se, and Sn did not exhibit apparent trophic transfer trends. Overall, ecological risk assessment of trace elements in East Asian finless porpoises suggested that greater attention should be given to Hg, As, Cd, and Se.

Molecular Characterization of TRPA Subfamily Genes and Function in Temperature Preference in Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae).

To reveal the mechanism of temperature preference in Tuta absoluta, one of the top 20 plant pests in the world, we cloned and identified TaTRPA1, TaPain, and TaPyx genes by RACE and bioinformatic analysis, and clarified their expression profiles during different development stages using real-time PCR, and revealed their function in preference temperature by RNAi. The full-length cDNA of TaPain was 3136 bp, with a 2865-bp open reading frame encoding a 259.89-kDa protein; and the partial length cDNA of TaPyx was 2326-bp, with a 2025-bp open reading frame encoding a 193.16-kDa protein. In addition, the expression of TaTRPA1 and TaPyx was significantly lower in larvae than other stages, and it was significantly higher in pupae and newly emerging males for TaPain. After feeding target double-stranded RNA (dsRNA), the preferred temperature decreased 2 °C more than the control group. In conclusion, the results firstly indicated the molecular characterization of TRPA subfamily genes and their key role in temperature perception in T. absoluta, and the study will help us to understand the temperature-sensing mechanism in the pest, and will provide some basis for study of other Lepidoptera insects' temperature preference. Moreover, it is of great significance in enriching the research progress of "thermos TRP".