Development of an agroinfiltration-based transient hairpin RNA expression system in pak choi leaves (Brassica rapa ssp. chinensis) for RNA interference against Liriomyza sativae.

The vegetable leafminer (Liriomyza sativae) is a devastating invasive pest of many vegetable crops and horticultural plants worldwide, causing serious economic loss. Conventional control strategy against this pest mainly relies on the synthetic chemical pesticides, but widespread use of insecticides easily causes insecticide resistance development and is harmful to beneficial organisms and environment. In this context, a more environmentally friendly pest management strategy based on RNA interference (RNAi) has emerged as a powerful tool to control of insect pests. Here we report a successful oral RNAi in L. sativae after feeding on pak choi (Brassica rapa ssp. chinensis) that transiently express hairpin RNAs targeting vital genes in this pest. First, potentially lethal genes are identified by searching an L. sativae transcriptome for orthologs of the widely used V-ATPase A and actin genes, then expression levels are assessed during different life stages and in different adult tissues. Interestingly, the highest expression levels for V-ATPase A are observed in the adult heads (males and females) and for actin in the abdomens of adult females. We also assessed expression patterns of the target hairpin RNAs in pak choi leaves and found that they reach peak levels 72 h post agroinfiltration. RNAi-mediated knockdown of each target was then assessed by letting adult L. sativae feed on agroinfiltrated pak choi leaves. Relative transcript levels of each target gene exhibit significant reductions over the feeding time, and adversely affect survival of adult L. sativae at 24 h post infestation in genetically unmodified pak choi plants. These results demonstrate that the agroinfiltration-mediated RNAi system has potential for advancing innovative environmentally safe pest management strategies for the control of leaf-mining species.

Disparities in Genetic Diversity Drive the Population Displacement of Two Invasive Cryptic Species of the Bemisia tabaci Complex in China.

Within the whitefly Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) complex, two cryptic species, namely Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED), are important invasive pests affecting global agriculture and horticulture. They were introduced into China sequentially in the mid-1990s and around 2003, respectively. Subsequently, the latter invader MED has outcompeted the earlier invader MEAM1, becoming the dominant population in the field. Although extensive studies have explored the underlying mechanisms driving this shift, the contribution of population genetics remains notably underexplored. In this study, we analyzed the genetic diversity and structure of 22 MED and 8 MEAM1 populations from various regions of China using mitochondrial DNA sequencing and microsatellite genotyping. Our results indicate low and moderate levels of genetic differentiation among geographically separate populations of MED and MEAM1, respectively. Median-joining network analysis of mtCOI gene haplotypes revealed no clear geographic structuring for either, with common haplotypes observed across provinces, although MED had more haplotypes. Comparative analyses revealed that MED presented greater genetic diversity than MEAM1 on the basis of two markers. Furthermore, analysis of molecular variance supported these findings, suggesting that while some genetic variation exists between populations, a significant amount is also present within populations. These findings reveal the population genetics of the two invasive cryptic species of the B. tabaci complex in China and suggest that the disparities in genetic diversity drive the displacement of their populations in the field. This work also provides valuable information on the genetic factors influencing the population dynamics and dominance of these invasive whitefly species.

Genome-Wide Identification, Evolution, and Female-Biased Expression Analysis of Odorant Receptors in Tuta absoluta (Lepidoptera: Gelechiidae).

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.

An effector of Phthorimaea absoluta oral secretions inhibits host plant defense.

Insects have evolved effectors to regulate host defenses for efficient feeding, yet their impact on chewing insects, like the tomato leaf miner (Phthorimaea absoluta), a significant pest, is poorly understood. We used RNAi to target the REPAT38 gene in larvae, monitoring changes at 0.5, 1, 2, and 4 h in leaf stomata, plant hormone concentrations (jasmonic acid (JA), jasmonoyl-L-isoleucine (JA-Ile), salicylic acid (SA), ethylene (ET), and abscisic acid (ABA)), and 12 hormone-responsive genes to explore the molecular mechanism of REPAT38-mediated plant-insect interactions. The results showed that the effector induced stomatal closure at 0.5 h and inhibited the synthesis of JA, ET, and ABA at 1 h. Additionally, seven plant hormone-responsive genes-AOC, MYC2, ACS1A, PAL, PR1, EIL2, and SRK2E-were inhibited at various time points. Our data suggest that REPAT38, as an effector with conserved functions, can weaken tomato host defenses and conducive to insect adaptation to host plants.

Phototactic Changes in Phthorimaea absoluta Long-Wavelength Opsin Gene Mutants (LW2-/-) and Short-Wavelength Opsin Gene Mutant (BL-/-) Strains.

Phthorimaea absoluta (Meyrick) is an invasive pest that has caused damage to tomatoes and other crops in China since 2017. Pest control is mainly based on chemical methods that pose significant threats to food safety and environmental and ecological security. Light-induced control, a green prevention and control technology, has gained attention recently. However, current light-trapping technology is non-specific, attracting targeted pests alongside natural enemies and non-target organisms. In this study, we characterized the phototactic behavior of tomato leaf miners for the development a specific light-trapping technology for pest control. In situ hybridization revealed opsin expression throughout the body. Furthermore, we investigated the tropism of pests (wild T. absoluta, Toxoptera graminum, and Bemisia tabaci) and natural enemies (Nesidiocoris tenuis and Trichogramma pintoi) using a wavelength-lamp tropism experiment. We found that 365 ± 5 nm light could accurately trap wild P. absoluta without trapping natural enemies and other insects. Finally, we analyzed the phototactic behavior of the mutant strains LW2(-/-) and BL(-/-). LW2 and BL mutants showed significant differences in phototactic behavior. The LW2(-/-) strain was attracted to light at 390 ± 5 nm and the BL(-/-) strain was unresponsive to any light. Our findings will help to develop specific light-trapping technology for controlling tomato leaf miners, providing a basis for understanding pest population dynamics and protecting crops against natural enemies.

Predicting global geographical distribution and latitudinal suitability gradient for light Brown apple moth.

Epiphyas postvittana, commonly known as the light brown apple moth (LBAM), is native to Australia and has a restricted global distribution. Its polyphagous nature and the recent surge in interceptions have emphasized the need for focused risk assessments to guide effective measures to curb the entry of this pest into new countries. This study aimed to perform a detailed global invasion risk assessment using an optimized MaxEnt model that incorporated 19 bioclimatic variables and elevation. The predictive outcomes underscored the significance of key variables, specifically the minimum temperature of the coldest month (bio6), precipitation of the driest month (bio14), and precipitation of the coldest quarter (bio19), in shaping the potential geographical distribution of LBAM. Regions beyond the existing range, including the southeastern United States, southern Brazil, eastern Argentina, Uruguay, southern Chile, and various Western European countries, were identified as susceptible to invasion and establishment by LBAM. An increase in suitability was observed above 45°N and 40°S under future climate scenario. With respect to climate change, LBAM would expand its potential range in Western Europe and the United States, especially under SSP5-8.5, in the 2050s. An upward trend in the latitudinal suitability gradient for LBAM in mid-high latitude areas implies that amid changing climate conditions, LBAM may find favorable habitats in these regions. For countries and regions with invasion risk, it is imperative to implement corresponding inspections and quarantine measures to thwart the introduction of LBAM, particularly in countries with established trade ties with invaded regions.

Temperature extremes nip invasive macrophyte Cabomba caroliniana A. Gray in the bud: potential geographic distributions and risk assessment based on future climate change and anthropogenic influences.

Cabomba caroliniana A. Gray, an ornamental submerged plant indigenous to tropical America, has been introduced to numerous countries in Europe, Asia, and Oceania, impacting native aquatic ecosystems. Given this species is a popular aquarium plant and widely traded, there is a high risk of introduction and invasion into other environments. In the current study the potential global geographic distribution of C. caroliniana was predicted under the effects of climate change and human influence in an optimised MaxEnt model. The model used rigorously screened occurrence records of C. caroliniana from hydro informatic datasets and 20 associated influencing factors. The findings indicate that temperature and human-mediated activities significantly influenced the distribution of C. caroliniana. At present, C. caroliniana covers an area of approximately 1531×104 km2 of appropriate habitat, especially in the south-eastern parts of South, central and North America, Southeast Asia, eastern Australia, and most of Europe. The suitable regions are anticipated to expand under future climate scenarios; however, the dynamics of the changes vary between different extents of climate change. For example, C. caroliniana is expected to expand to higher latitudes, following global temperature increases under SSP1-2.6 and SSP2-4.5 scenarios, however, intolerance to temperature extremes may mediate invasion at higher latitudes under future extreme climate scenarios, e.g., SSP5-8.5. Owing to the severe impacts its invasion causes, early warning and stringent border quarantine processes are required to guard against the introduction of C. caroliniana especially in the invasion hotspots such as, Peru, Italy, and South Korea.

Risk Assessment of Spodoptera exempta against Food Security: Estimating the Potential Global Overlapping Areas of Wheat, Maize, and Rice under Climate Change.

Spodoptera exempta, known as the black armyworm, has been extensively documented as an invasive agricultural pest prevalent across various crop planting regions globally. However, the potential geographical distribution and the threat it poses to host crops of remains unknown at present. Therefore, we used an optimized MaxEnt model based on 841 occurrence records and 19 bioclimatic variables to predict the potential suitable areas of S. exempta under current and future climatic conditions, and the overlap with wheat, rice, and maize planting areas was assessed. The optimized model was highly reliable in predicting potential suitable areas for this pest. The results showed that high-risk distribution areas for S. exempta were mainly in developing countries, including Latin America, central South America, central Africa, and southern Asia. Moreover, for the three major global food crops, S. exempta posed the greatest risk to maize planting areas (510.78 × 104 km2), followed by rice and wheat planting areas. Under future climate scenarios, global warming will limit the distribution of S. exempta. Overall, S. exempta had the strongest effect on global maize production areas and the least on global wheat planting areas. Our study offers a scientific basis for global prevention of S. exempta and protection of agricultural crops.

DNA methylase 1 influences temperature responses and development in the invasive pest Tuta absoluta.

DNA methylase 1 (Dnmt1) is an important regulatory factor associated with biochemical signals required for insect development. It responds to changes in the environment and triggers phenotypic plasticity. Meanwhile, Tuta absoluta Meyrick (Lepidoptera: Gelechiidae)-a destructive invasive pest-can rapidly invade and adapt to different habitats; however, the role of Dnmt1 in this organism has not been elucidated. Accordingly, this study investigates the mechanism(s) underlying the rapid adaptation of Tuta absoluta to temperature stress. Potential regulatory genes were screened via RNAi (RNA interference), and the DNA methylase in Tuta absoluta was cloned by RACE (Rapid amplification of cDNA ends). TaDnmt1 was identified as a potential regulatory gene via bioinformatics; its expression was evaluated in response to temperature stress and during different development stages using real-time polymerase chain reaction. Results revealed that TaDnmt1 participates in hot/cold tolerance, temperature preference and larval development. The full-length cDNA sequence of TaDnmt1 is 3765 bp and encodes a 1254 kDa protein with typical Dnmt1 node-conserved structural features and six conserved DNA-binding active motifs. Moreover, TaDnmt1 expression is significantly altered by temperature stress treatments and within different development stages. Hence, TaDnmt1 likely contributes to temperature responses and organismal development. Furthermore, after treating with double-stranded RNA and exposing Tuta absoluta to 35°C heat shock or -12°C cold shock for 1 h, the survival rate significantly decreases; the preferred temperature is 2°C lower than that of the control group. In addition, the epidermal segments become enlarged and irregularly folded while the surface dries up. This results in a significant increase in larval mortality (57%) and a decrease in pupation (49.3%) and eclosion (50.9%) rates. Hence, TaDnmt1 contributes to temperature stress responses and temperature perception, as well as organismal growth and development, via DNA methylation regulation. These findings suggest that the rapid geographic expansion of T absoluta has been closely associated with TaDnmt1-mediated temperature tolerance. This study advances the research on 'thermos Dnmt' and provides a potential target for RNAi-driven regulation of Tuta absoluta.

Projecting the Global Potential Geographical Distribution of Ceratitis capitata (Diptera: Tephritidae) under Current and Future Climates.

The Mediterranean fruit fly, Ceratitis capitata (Wiedemann), which is native to tropical Africa, has invaded more than 100 countries and constitutes a risk to the citrus sector. Studying its potential geographical distribution (PGD) in the context of global climate change is important for prevention and control efforts worldwide. Therefore, we used the CLIMEX model to project and assess the risk of global invasion by C. capitata under current (1981-2010) and future (2040-2059) climates. In the prevailing climatic conditions, the area of PGD for C. capitata was approximately 664.8 × 105 km2 and was concentrated in South America, southern Africa, southern North America, eastern Asia, and southern Europe. Under future climate conditions, the area of PGD for C. capitata is projected to decrease to approximately 544.1 × 105 km2 and shift to higher latitudes. Cold stress was shown to affect distribution at high latitudes, and heat stress was the main factor affecting distribution under current and future climates. According to the predicted results, countries with highly suitable habitats for C. capitata that have not yet been invaded, such as China, Myanmar, and Vietnam, must strengthen quarantine measures to prevent the introduction of this pest.