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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.

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BACKGROUND: Cotton is an important economic crop and a host of Liriomyza sativae. Pectin methylesterase (PME)-mediated pectin metabolism plays an indispensable role in multiple biological processes in planta. However, the pleiotropic functions of PME often lead to unpredictable effects on crop resistance to pests. Additionally, whether and how PME affects susceptibility to Liriomyza sativae remain unclear. RESULTS: Here, we isolated GhPME36, which is located in the cell wall, from upland cotton (Gossypium hirsutum L.). Interestingly, the overexpression of GhPME36 in cotton caused severe susceptibility to Liriomyza sativae but increased leaf biomass in Arabidopsis. Cytological observations revealed that the cell wall was thinner with more demethylesterified pectins in GhPME36-OE cotton leaves than in WT leaves, whereas the soluble sugar content of GhPME36-OE cotton leaf cell walls was accordingly higher; both factors attracted Liriomyza sativae to feed on GhPME36-OE cotton leaves. Metabolomic analysis demonstrated that glucose was significantly differentially accumulated. Transcriptomic analysis further revealed DEGs enriched in glucose metabolic pathways when GhPME36 was overexpressed, suggesting that GhPME36 aggravates susceptibility to Liriomyza sativae by affecting both the structure and components of cell wall biosynthesis. Moreover, GhPME36 interacts with another pectin-modifying enzyme, GhC/VIF1, to maintain the dynamic stability of pectin methyl esterification. CONCLUSIONS: Taken together, our results reveal the cytological and molecular mechanisms by which GhPME36 aggravates susceptibility to Liriomyza sativae. This study broadens the knowledge of PME function and provides new insights into plant resistance to pests and the safety of genetically modified plants.

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Our aim was to develop an electrostatic apparatus to lure and capture silverleaf whiteflies (Bemisia tabaci), vegetable leafminers (Liriomyza sativae), and western flower thrips (Frankliniella occidentalis) that invade tomato greenhouses. A double-charged dipolar electric field producer (DD-EFP) was constructed by filling water in two identical transparent soft polyvinyl chloride tubes arrayed in parallel with fixed separation, and then, inserting the probes of grounded negative and positive voltage generators into the water of the two tubes to generate negatively and positively charged waters, respectively. These charged waters electrified the outer surfaces of the opposite tubes via dielectric polarization. An electric field formed between the oppositely charged tubes. To lure these phototactic insects, the water was colored yellow using watercolor paste, then introduced into the transparent insulator tubes to construct the yellow-colored DD-EFP. This apparatus lured insects in a manner similar to commercially available yellow sticky traps. The yellow-colored DD-EFP was easily placed as a movable upright screen along the plants, such that invading pests were preferentially attracted to the trap before reaching the plants. Furthermore, pests settling on the plants were attracted to the apparatus, which used a plant-tapping method to drive them off the plants. Our study provided an experimental basis for developing an electrostatic device to attract and capture insects that enter greenhouses.

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Factors limiting distribution range for most species are generally unknown regardless of whether they are native or invasive. We studied factors that could enable or restrict the distribution of two cosmopolitan invasive leafminer fly species, Liriomyza huidobrensis (Blanchard) and Liriomyza sativae (Blanchard) in their native niche. In order to test which ecological and environmental factors affect leafminer distribution we conducted thermal tolerance assays, sampled along elevation gradients and modeled species distribution. Findings from the field and rearing chambers showed a physiological restriction due to high temperatures for L. huidobrensis at 28-29 °C, above which adult emergence is compromised. We also found that maximum temperatures below 22 °C, typical of tropical highlands, favored L. huidobrensis. L. sativae was found across a wider temperature range (i.e., from 21 to 36 °C) in Guatemala. Our finding of a physiological threshold in temperature for L. huidobrensis may enable us to predict its invasive risk when combined with the environmental conditions at horticultural ports of entry and the global agricultural landscape. Further, it strengthens our predictions on shifts in distribution of the leafminer fly under future climate. We also found a temperature mediated competitive exclusion interaction between the two herbivore species, where L. sativae occurred at temperatures < 22 °C only in the absence of L. huidobrensis. We show that parasitoids had a negative effect on the leafminer flies, which varied with host plant. Finally, we show the importance of taking a multiaspect approach when investigating what limits distribution and invasiveness of a species.

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ABSTRACT Melon is one of the most important vegetable crops in the world. With short cycle in a system of phased planting, phytosanitary control is compromised, and a great volume of agricultural chemicals is used to control vegetable leafminer. Genetic control is an ideal alternative to avoid the damage caused by this insect. Thus, the aim of this study was to evaluate Cucumis accessions in regard to resistance to leafminer and correlate the variables analyzed. Fifty-four accessions and four commercial hybrids of melon were tested. The study was divided into two experiments: with and with no choice. The following characteristics were evaluated: with choice, in field - subjective score based on the infestation and the number of mines per leaf; and with no choice, in cage - number of mines per leaf, chlorophyll content, and leaf colorimetry. The results showed variability among the accessions and some genotypes showed favorable results for resistance in both experiments. There was correlation between the two variables in the experiment in the field. The accessions CNPH 11-282, CNPH 06-1047, and CNPH 11-1077 are the most recommended for future breeding programs with aim on introgression of resistance to vegetable leafminer in melon.

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In this study, partial sequences of the mitochondrial cytochrome oxidase subunit I (COI) gene of four Liriomyza sativae (Blanchard) geographic populations in China were sequenced. As for the 784 bp mtDNA-COI gene obtained, six variable sites were found which were all transitions and no base composition was insertions or deletions. Six haplotypes were identified in all the sequences, with five showing polymorphism and one was exclusive. Nucleotide diversity (π) and haplotype diversity (h) ranged from 0.00068 to 0.00300 and 0.53571 to 0.82857, respectively. The phylogenetic trees suggested that six haplotypes constructed two clades. Molecular variance analysis (AMOVA) demonstrated that the genetic variation was not obvious and mainly occurred within geographic populations (94.8%). Most molecular variance within the species was due to the difference of haplotypes among different geographic populations. The genetic characters of the four populations were analyzed by FST value and gene flow (Nm), and the FST and Nm values were 0.174-0.464 and 0.577-2.367, respectively. All results showed that not only the gene flow presented among the four populations but also the genetic differences did. The main reason causing the genetic differences among the four populations was supposed to be related to geographic isolation and host plants aggravated the differences.

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While Liriomyza sativae (Diptera: Agromyzidae), an important invasive pest of ornamentals and vegetables has been found in China for the past two decades, few studies have focused on its genetics or route of invasive. In this study, we collected 288 L. sativae individuals across 12 provinces to explore its population genetic structure and migration patterns in China using seven microsatellites. We found relatively low levels of genetic diversity but moderate population genetic structure (0.05 < F ST < 0.15) in L. sativae from China. All populations deviated significantly from the Hardy-Weinberg equilibrium due to heterozygote deficiency. Molecular variance analysis revealed that more than 89% of variation was among samples within populations. A UPGMA dendrogram revealed that SH and GXNN populations formed one cluster separate from the other populations, which is in accordance with STRUCTURE and GENELAND analyses. A Mantel test indicated that genetic distance was not correlated to geographic distance (r = -0.0814, P = 0.7610), coupled with high levels of gene flow (M = 40.1-817.7), suggesting a possible anthropogenic influence on the spread of L. sativae in China and on the effect of hosts. The trend of asymmetrical gene flow was from southern to northern populations in general and did not exhibit a Bridgehead effect during the course of invasion, as can be seen by the low genetic diversity of southern populations.

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A longitudinal study to identify the species of Liriomyza leafminer, their distribution, relative abundance, and seasonal variation, including their host range, was conducted in vegetable fields at three altitudes in Kenya from November 2011 to November 2012. Three main species were identified: Liriomyza huidobrensis (Blanchard), Liriomyza sativae Blanchard, and Liriomyza trifolii (Burgess), of which L. huidobrensis was the most abundant across all altitudes irrespective of the cropping season and accounting for over 90% of the total Liriomyza specimens collected. Liriomyza species were collected from all infested incubated leaves of 20 crops surveyed belonging to seven families: Fabaceae, Solanaceae, Cucurbitaceae, Malvaceae, Brassicaceae, Amaranthaceae, and Amaryllidaceae. However, more than 87.5% of the Liriomyza species were obtained from only four of these crops: Pisum sativum L., Phaseolus vulgaris L., Solanum lycopersicum L., and Solanum tuberosum, thereby demonstrating that Fabaceae and Solonaceae crops are the most important hosts with regard to Liriomyza species richness and relative abundance. L. huidobrensis had the widest host range (20 crops), followed by L. sativae (18 crops) and L. trifolii (12 crops). Although L. trifolii has been considered the dominant Liriomyza leafminer in Kenya, this study suggests that this may not be the case anymore, as L. huidobrensis dominates at all altitudes.

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Leafmining flies (Diptera: Agromyzidae) can be serious economic pests of horticultural crops. Some genera such as Liriomyza are particularly problematic with numerous species, some of which are highly polyphagous (wide host range), which can only be confidently identified morphologically from adult males. In our study, DNA barcoding was employed to establish new locality records of the vegetable leafminer fly, Liriomyza sativae, from the islands of Torres Strait (Queensland, Australia) and the central highlands of Papua New Guinea (PNG). These records represent significant range extensions of this highly invasive plant pest. Specimens of immature leafminers (from leaf mines) were collected over a 5-year period during routine plant health surveys in ethanol or on FTA® filter paper cards, both methods proved effective at preserving and transporting insect DNA under tropical conditions, with FTA cards possessing some additional logistical benefits. Specimens were identified through sequencing two sections of the cytochrome oxidase I gene and the utility of each was assessed for the identification of species and intra-specific genetic lineages. Our study indicates that multiple haplotypes of L. sativae occur in PNG, while a different haplotype is present in the Torres Strait, with genetic regionalization between these areas apart from a single possible instance - one haplotype 'S.7' appears to be common between these two regions - interestingly this has also been the most common haplotype detected in previous studies of invasive L. sativae populations. The DNA barcoding methods employed here not only identified multiple introductions of L. sativae, but also appear generally applicable to the identification of other agromyzid leafminers (Phytomyzinae and Agromyzinae) and should decrease the likelihood of potentially co-amplifying internal hymenopteran parasitoids. Currently, L. sativae is still not recorded from the Australian mainland; however, further sampling of leafminer flies from Northern Australia and surrounding areas is required, as surveillance for possible Liriomyza incursions, as well as to characterize endemic species with which Liriomyza species might be confused.

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Abamectin resistance was selected in the vegetable leafminer, Liriomyza sativae (Blanchard) (Diptera: Agromyzidae) under laboratory conditions, and cross-resistance patterns and possible resistance mechanisms in the abamectin-resistant strains (AL-R, AF-R) were investigated. Compared with the susceptible strain (SS), strain AL-R displayed 39-fold resistance to abamectin after 20 selection cycles during 25 generations, and strain AF-R exhibited 59-fold resistance to abamectin after 16 selection cycles during 22 generations. No cross-resistance to cyromazine was found in both abamectin-resistant strains. However, we failed to select for cyromazine resistance in L. sativae under laboratory conditions by conducting 17 selection cycles during 22 generations. However, moderate levels of cross-resistance to abamectin (6-9 fold) were observed in strains which received cyromazine treatments. Biochemical analysis showed that glutathione S-transferase (GST) activity in both abamectin-resistant strains (AL-R, AF-R) was significantly higher than in the susceptible strain (SS), suggesting metabolically driven resistance to abamectinin L. sativae. Recommendations of mixtures or rotation of cyromazine and abamectin should be considered carefully, as consecutive cyromazine treatments may select for low-level cross-resistance to abamectin.

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