Development and stability of insecticide resistance in the leafminer Liriomyza trifolii (Diptera: Agromyzidae) to cyromazine, abamectin, and spinosad

Three populations of the leafminer, Liriomyza trifolii (Burgess), were collected from commercial ornamental production greenhouses in the United States and tested for susceptibility to three commercial insecticides. A leaf dip bioassay of leaves containing young (1-2-d-old) larvae was used. Based on larval mortality and compared with a susceptible laboratory reference colony, the three strains varied in spectrum and level of resistance to the insecticides. CA-1, collected from Gerbera daisy, was moderately resistant to cyromazine (18.1-fold) and abamectin (22.0-fold), but highly resistant to spinosad (> 188-fold). CA-2, collected from chrysanthemums, was not resistant to abamectin, had a low level of resistance to cyromazine (8.2-fold), but was extremely resistant to spinosad (1,192-fold). GA-1, collected from chrysanthemums, had very low levels of resistance to cyromazine (5.4-fold) and spinosad (1.9-fold) but was moderately resistant to abamectin (30.6-fold). When reared in the absence of insecticide selection pressure, all three strains reverted to approximately the level of the reference strain. The CA-1 strain reverted in nine generations to cyromazine; however, the lowest levels of abamectin and spinosad resistance reverted to was 3.1-fold at F8 and 3.2 at the F10, respectively. The CA-2 strain reverted in five generations to both cyromazine and spinosad. GA-1 reverted in five generations to abamectin. Based on the results, resistance to these three insecticides was unstable. Additionally, there was no cross-resistance among these three insecticides.     

Phytosanitary irradiation of Liriomyza trifolii (Diptera: Agromyzidae)

Agromyzid leafminers are economic and quarantine pests of a variety of vegetables, flowers, and ornamental foliage. Methyl bromide fumigation is often used as a phytosanitary treatment when quarantined agromyzids are found in shipped commodities; alternative treatments are sought. Ionizing radiation is a viable alternative that is increasing in use worldwide. A dose of 400 Gy is accepted by USDA-APHIS for all insects (except Lepidoptera pupae and adults) on all commodities. Efforts to lower this dose and make it acceptable to other countries involve determining radiotolerance of families of major quarantine pests. Agromyzidae is one such family for which no useful information on radiotolerance exists. This research sought to determine the dose required to control a major agromyzid pest, Liriomyza trifolii (Burgess) and was performed on L. trifolii collected in Weslaco, TX, reared on Phaseolus vulgaris L. and Capsicum annuum L. and irradiated in the late puparial stage. The measure of efficacy was prevention of F1 mine formation. Puparia collected from Gossypium hirsutum L. and reared on P. vulgaris were more radiotolerant than those collected and reared on C. annuum. A dose of 214 Gy may prevent F1 mine formation of L. trifolii. This research used a variation of probit analysis where the direct response of the treated individual is not measured, but the response of the F1 generation is. This type of analysis is useful in phytosanitary irradiation research where the measure of efficacy often involves a response of the F1 generation.     

Effects of two plant extracts on larval leafminer Liriomyza trifolii (Diptera: Agromyzidae) in tomatoes

Aqueous extracts from two plants, Urginea maritima L. (Liliaceae) and Euphorbia myrsinites L. (Euphorbiaceae), were tested for their insecticidal activity against the leafminer Liriomyza trifolii (Burgess) on infested tomato, Lycopersicon esculentum Mill., plants in the laboratory and field. Two grams of plant material was extracted with 100 ml of water and then diluted 1:100, 1:50, and 1:25 with distilled water. Diluted plant extract was either applied to the infested tomato leaves or by soil drench and was compared with foliar application of cyromazine. All dilutions of both plant extracts caused significant control of the leafminer larvae and maintained populations below those of the nontreated control plants in all trials. Only at the most concentrated dilutions (1:25) were the plant extracts statistically similar to the cyromazine treatment. Furthermore, greenhouse yields from all of the foliar treatments were statistically similar to the cyromazine treatment and significantly better than the nontreated control. Four species of leafminer parasitoids were found in the greenhouse; however, the percentage of parasitism was significantly less in all treated replicates than in the nontreated control replicates. Aqueous extracts from these two plant extracts exhibited both translaminar and systemic activity and are potential candidates as new organic insecticides.     

The complete mitochondrial genome of the leafminer Liriomyza sativae (Diptera: Agromyzidae): great difference in the A+T-rich region compared to Liriomyza trifolii

The complete mitochondrial genome sequence of Liriomyza sativae Blanchard (15,551 bp) was determined and analyzed in this study. The circular genome contained 37 genes including 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes and an A + T-rich region. The initiation codons of COI and ND1 were ‘ATCA’ and ‘GTG’, respectively. ND2 gene used the truncated termination codon ‘T’. All the tRNA genes had the typical cloverleaf secondary structures except for tRNA^Ser(AGN) gene, which was found with the absence of a DHU arm. In addition, a tRNA-like secondary structure (tRNA^Met) was found in the A + T-rich region. The great difference was that the length of L. sativae A + T-rich region was 597 bp shorter than that of Liriomyza trifolii (Burgess). Meanwhile, some minor differences such as ‘TATA’ block were also observed in L. sativae in contrast to ‘TACA’ block in L. trifolii. There were also some essential structure elements such as ‘TATA’ block, ‘G(A)_nT’ block, poly-T stretch and stem-and-loop structure in the A + T-rich region of L. sativae mitochondrial genome.     

Entomopathogenic nematodes for biological control of the leafminer,Liriomyza trifolii (Dipt.: Agromyzidae)

In the laboratory, mortality rates of the agromyzid leafminer larvae,Liriomyza trifolii (Burgess), ranged from 48 to 98% by 20 strains and/or species of steinernematid and heterorhabditid nematodes. In the greenhouse, abamectin provided superior control of larval leafminers, killing 100% of them as compared withSteinernema carpocapsae (Weiser) All strain (24 to 43% leafminer mortality) orS. carpocapsae Liriomyza-selected strain (8 to 44% leafminer mortality); the maximum relative humidity (r.h.) ranged between 81 and 91% and the minimum r.h. between 50 and 70%. In the foghouse under high r.h., the commercially available All strain and the Hawaiian isolate ofS. feltiae (Filipjev) MG-14 strain caused 69 and 67% mean mortality, respectively. There was a significant correlation (P<0.01) between nematode mortality of leafminers and r.h., including the mean, standard deviation, and minimum r.h. during the 48 h after treatment. Average r.h. >92% with a standard deviation of <9% r.h. and a minimum of 72% r.h. providedS. carpocapsae All strain mortality rates of leafminers >65%. The major constraint against the use of nematodes against leafminers in the foliar environment is low r.h. The use of nematodes againstL. trifolii can be successful if the r.h. remains high and if nematodes enter leafmines before desiccation, and the nematodes should be integrated with chemical insecticides such as abamectin to manage pesticide resistance inL. trifolii.     

Polyandry and Reproduction in the Serpentine Leaf Miner Liriomyza trifolii (Diptera: Agromyzidae)

We investigated the hypotheses that Liriomyza trifolii is a polyandrous species and that multiple mating is required to maximize female fecundity. In addition, the hypothesis that male copulating frequency reduces female fecundity was examined. Using sterile and fertile males, we found that L. trifolii females mate more than once during their life span, and that multiple mating is required to maximize female fecundity. A similar amount of sperm was transferred during five successive male copulations with an equal insemination probability. However, the probability of reproduction failure was strongly correlated with the male copulating sequence. Moreover, the failure rate of sperm transfer was found to be 6.8%. In light of these results, possible selection pressures driving for polyandry in L. trifolii are discussed.     

美洲斑潜蝇室内饲养研究

筛选了室内饲养美洲斑潜蝇的适宜寄主植物,进行了饲养密度及成虫补充营养糖的浓度试验。结果表明：初生菜豆PhaseolusvulgarisL.苗是室内饲养的适宜寄主植物,用该种植物饲养,幼虫饲养密度低于1头／cm2,则幼虫成活率大于90％,饲养密度提高,则幼虫成活率降低,每平方厘米初生叶饲养1头幼虫是适宜的饲养密度,成虫补充营养糖或蜜糖的浓度以8％为宜。     

Analysis of acoustic stimulus to Liriomyza trifolii (Diptera: Agromyzidae) through Electroantennogram (EAG) technique

The study utilized the Electroantennogram (EAG) technique as biological acoustic stimulus to Liriomyza trifolii (Diptera: Agromyzidae). Pure tone sounds and courtship songs were tested in the excised head of test specimens. Significant EAG stimuli were detected when released at 20 Hz pure tone sound together with the courtship song having long pulse train patterns of Cryptonevra inquilina and Lipara japonica (Chloropidae). For their biological effectiveness, movement behaviors were analyzed with stimulus taken from EAG recordings. No attraction effect was observed even with and without acoustic stimulus. However, movement activities were transformed by information entropy and observed to be clearly different when selected acoustic stimuli were broadcast. The combination of response to EAG and behavioral observation would be useful to determine effective movement modification through acoustic stimulus in insect pests that are necessary toward the development of an acoustic pest management tool in agriculture.     

The movement and dispersal of Liriomyza trifolii (Diptera: Agromyzidae) in a chrysanthemum greenhouse

Investigation of the dispersal of Liriomyza trifolii in a 1–2 ha chrysanthemum greenhouse indicated that the average distance flown by female flies was greater than that flown by male flies. However, flies of both sexes were found at the furthest distance from the release point (102 m) which indicates that flies in the greenhouse could be considered to be a single inter-breeding population. When the data were fitted to a generalised distance-dispersal model, examination of the parameters revealed density decreased with distance more rapidly for males than females. Also, the model showed that males tended to be slightly more attracted to each other than were females. The applicability of these results to the development of pest management systems for L. trifolii is discussed.     

Egg-collection equipment for Liriomyza leafminers (Diptera: Agromyzidae)

Egg-collection equipment for Liriomyza leafminers was designed using a small glass tube with a glucose solution supported between two thin films. Adult female Liriomyza trifolii (Burgess), which were previously reared on kidney bean for 2–4 days, laid abundant eggs in 5 or 10% glucose solutions. Hatchability of the eggs in the solutions was greater than 80%. The equipment was used successfully to collect eggs from another leafminer species, Liriomyza sativae Blanchard.     

Effects of temperature on the immature development of the stone leek leafminer Liriomyza chinensis (Diptera: Agromyzidae)

The effect of nine constant temperatures (15, 17.5, 20, 22.5, 25, 27.5 30, 32.5, and 35 degrees C) on the development of the stone leek leafminer, Liriomyza chinensis (Kato), on Japanese bunching onion, Allium fistulosum L., was studied in the laboratory. Developmental times for immature stages were inversely proportional to temperature between 15 and 30 degrees C but increased at 32.5 degrees C. Total developmental times from egg to adult emergence decreased from 69.6 to 17.1 d for temperatures from 15 to 30 degrees C, with pupae requiring more time for development than the combined egg and larva stages. Both linear and nonlinear (Logan equation VI) models provided a reliable fit of development rates versus temperature for all immature stages. The lower developmental thresholds that were estimated from linear regression equations for the egg, first, second, and third instars, total larva, egg-larval, pupa, and total combined immature stages were 12.1, 10.6, 13.6, 8, 9.6, 11.3, 11.2, and 11.4 degrees C, respectively. The degree-day accumulation was calculated as 312.5 DD for development from egg to adult emergence. By fitting the nonlinear models to the data, the upper and optimal temperatures for egg, larva, pupa, and total immature stages were calculated as 37.8 and 31.7, 34.9 and 30.1, 35.8 and 30.6, and 35.0 and 30.9 degrees C, respectively. These data are useful for predicting population dynamics of L. chinensis under field conditions and determining the maximum proportion of susceptible individuals for facilitating improved timing of application of control measures.     

Feeding, oviposition and survival of Liriomyza trifolii (Diptera: Agromyzidae) on Bt and non-Bt cottons

The effects of Bt transgenic cottons (Bt-I expressing cry1Ac and Bt-II expressing cry1Ab and cry2Ab or cry1Ab and cry1Fa) and non-Bt cottons on feeding, oviposition and longevity of adults, and development and survival of Liriomyza trifolii larvae were studied under laboratory conditions; and infestation on four Bt and two non-Bt cotton traits were investigated under field conditions. Laboratory choice and no-choice tests showed that L. trifolii adults were capable of distinguishing between Bt cottons and non-Bt cottons. In a choice test on younger plants (4-5 leaves), the adults were found more often and made more feeding punctures (FP) on non-Bt cottons than on Bt cottons. On older plants (8-9 leaves), adults made the most FP on non-Bt cotton followed by those on Bt-II cottons and the least on Bt-I cotton. The females oviposited more eggs (6.7 eggs per leaf) on non-Bt cotton than on Bt-I (1.7 eggs per leaf) and Bt-II (0.8 eggs per leaf) cottons on younger plants and oviposited similar numbers of eggs (0.7-1.3 eggs per leaf) on non-Bt and Bt cottons on older plants. In a no-choice test, the females also fed more FP on non-Bt cottons than on Bt cottons on both younger and older plants. The females oviposited more eggs (15.6 eggs per leaf) on non-Bt cotton than on Bt-I (8.2 eggs per leaf) and Bt-II (6.5 eggs per leaf) cottons on younger plants and similar numbers of eggs (2.5-3.3 eggs per leaf) on non-Bt and Bt cottons on older plants. Larval and puparial survivals were not different among Bt and non-Bt cottons. The occurrence and damage of leafminers on cottons in the field showed that L. trifolii infested more plants and leaves and had more mines on non-Bt cotton than on Bt cottons.     

Polymerase chain reaction-restriction fragment-length polymorphism method to distinguish Liriomyza huidobrensis from L. Langei (Diptera: Agromyzidae) applied to three recent leafminer invasions.

A molecular method is presented for differentiating the morphologically cryptic leafminers Liriomyza langei Frick and L. huidobrensis (Blanchard). This method requires polymerase chain reaction (PCR) amplification of a 1031-bp region of mitochondrial cytochrome oxidase DNA followed by restriction fragment analysis using the restriction enzymes SpeI and EcoRV. Spel cuts the mitochondrial fragment of L. langei into two fragments, but does not cut the L. huidobrensis fragment. EcoRV cuts the L. huidobrensis fragment into two fragments, but does not cut the L. langei fragment. This PCR-restriction fragment-length polymorphism (RFLP) method is faster and less costly than DNA sequencing,which is currently the only other way to differentiate these two species. We apply the method to samples from recently introduced leafminer populations in Sri Lanka, Canada, and South Africa and find that the invasive leafminer in all three locations is L. huidobrensis.     

Abamectin, pymetrozine and azadirachtin sequence as a unique solution to control the leafminer Liriomyza trifolii (Burgess) (Diptera: Agromyzidae) infesting garden beans (Phaseolus vulgaris L.) in Egypt

Field trails were conducted to determine the performance of three different sequences as a unique solution for the control of the leaf miner Liriomyza trifolii (Burgess) (Diptera: Agromyzidae) infesting garden beans (Phaseolus vulgaris L.) during the two successive seasons of 2004 and 2005. Furthermore, during the evaluation period, the side effect against the ectoparasite Diglyphus isaea (Walker) (Hymenoptera: Eulophidae) was put into consideration. Meanwhile, the comparative evaluation of the pesticides alone showed that abamectin and azadirachtin were highly effective against Liriomyza trifolii, while carbosulfan, pymetrozine and thiamethoxam provided to be of a moderate effect. Moreover, carbosulfan showed harmful effect to the larvae of the ectoparasite Diglyphus isaea (Walker), while abamectin and azadirachtin gave a moderate effect. Thiamethoxam and the the detergent (Masrol 410) had slight effect in this respect. The highly effective sequence among the sequences was abamectin, pymetrozine and azadirachtin, against Liriomyza trifolii (Burgess), with slight harmful effect on Diglyphus isaea (Walker). However the sequence of azadirachtin, pymetrozine and abamectin had a moderate effect on Liriomyza trifolii (Burgess) and exhibited a slight toxic effect on Diglyphus isaea (Walker). In contrast, the sequence of carbosulfan, thiamethoxam and pymetrozine was the least effective and represented a slight effect on Diglyphus isaea (Walker). From this study, it was concluded that abamectin, pymetrozine and azadirachtin sequence has proved to be a unique solution for the control of the leaf miner Liriomyza trifolii (Burgess) infesting garden beans (Phaseolus vulgaris L.) in Egypt.     

Host plant-related parasitism and host feeding activities of Diglyphus isaea (Hymenoptera: Eulophidae) on Liriomyza huidobrensis, Liriomyza sativae, and Liriomyza trifolii (Diptera: Agromyzidae)

Host plant species can affect the behavior and attributes of parasitoids, such as host searching, oviposition, and offspring fitness. In this study, parasitism, host feeding, and sex ratios of Diglyphus isaea (Walker) (Hymenoptera: Eulophidae) on Liriomyza huidobrensis (Blanchard), Liriomyza sativae Blanchard, and Liriomyza trifolii (Burgess) (Diptera: Agromyzidae) larvae reared on Phaseolus vulgaris L., Pisum sativum L., Solanum lycopersicum L., and Vicia faba L. were determined. In no-choice tests, L. huidobrensis had the highest rate of parasitism when reared on P. vulgaris (46%), L. sativae when reared on V. faba (59%) and P. vulgaris (59%), and L. trifolii when reared on S. lycopersicum (68%). Host feeding in no-choice tests ranged between 2% and 36% and was highest on L. trifolii reared on V. faba. Results of choice tests showed a significant interaction effect for host plant and Liriomyza species on parasitism and host feeding. Within plant mixtures, L. sativae reared on P. vulgaris had the highest rate of parasitism (31%), followed by L. trifolii on S. lycopersicum (29%) and L. huidobrensis on V. faba (28%). Host feeding was highest on L. trifolii reared on S. lycopersicum (14%) and lowest on L. huidobrensis reared on P. sativum and S. lycopersicum (1%). In some instances, plant mixtures resulted in a higher proportion of females of D. isaea than single plant species. The highest proportion of females was obtained in plant mixtures on L. huidobrensis and L. trifolii on V. faba (71 and 72%, respectively). This study suggests that planting crop mixtures can potentially lead to higher proportions of females, thus improving parasitism and host feeding, depending on Liriomyza and host plant species.     

Parasitoids of chickpea leafminer Liriomyza cicerina (Diptera: Agromyzidae) and their parasitism rate on chickpea fields in North Tunisia

In recent years, as a consequence of climatic changes, the leafminer Liriomyza cicerina had emerged as a serious pest in chickpea crops causing considerable damage in Tunisia. Thus, the search of control methods is necessary. The aim of this research was to monitor and identify the naturally occurring parasitoids of the chickpea leafminer Liriomyza cicerina in insecticide-free chickpea-growing areas located in North-Western Tunisia. Chickpea leaves with mines were weekly sampled from Beja and Kef sites during 2016 and 2017 and kept to observe and count emerging leafminer and parasitoid adults in the laboratory. Results revealed that L. cicerina parasitoids complex species included three hymenopteran parasitoids namely Opius monilicornis (Fisher, 1962) (Braconidae); Diglyphus isaea (Walker, 1838) (Eulophidae) and Diaulinopsis arenaria (Erdös, 1951) (Eulophidae). These parasitoids were identified and recorded for the first time in Tunisia on L. cicerina with parasitism rate varied between 13% and 52.5%. The most common species was O. monilicornis. The correlation between infestation rate on chickpea plants and number of emerged three parasitoids was calculated revealing high significant and positive correlations. The diversity index (H′) was also calculated. The three parasitoids were found in two sites Beja and Kef; while the diversity differed during the developmental stage of chickpea crops.     

Kairomone attractant for the leafmining fly, Liriomyza bryoniae (Diptera, Agromyzidae)

A field test carried out in an industrial greenhouse in Lithuania revealed the attractiveness of synthetic methyl salicylate (MeSa) towards an economically important leafmining tomato pest, Liriomyza bryoniae (Kaltenbach) (Diptera, Agromyzidae). The behavioural reaction of the flies depended very much on the simultaneous presence of both olfactory and visual stimuli. The attractiveness depended on the colour of a sticky trap: MeSa attracted significantly more flies (ca. 2.2 times) when placed in yellow traps than in aluminium foil colour ones, when catches in such traps were compared to a corresponding control. L. bryoniae is the first species within the Agromyzidae family attracted by MeSa. The attractant was attributed to kairomones, as the compound is known as a plant-produced volatile. MeSa can be an effective extra-tool for increasing the attractiveness of traps. It should be evaluated in future whether such trap/bait combination is effective for the mass trapping of L. bryoniae leafminers in greenhouses (closed area).     

Molecular evidence of cryptic species within the Liriomyza huidobrensis (Diptera: Agromyzidae)

Phylogenetic relationships among populations of the polyphagous pea leafminer, Liriomyza huidobrensis (Blanchard), were investigated using DNA sequence data. Maximum parsimony analysis of 941 bp of mitochondrial cytochrome oxidase I and II genes showed that L. huidobrensis contains two well-defined monophyletic groups, one composed of specimens from California and Hawaii and one composed of specimens from South and Central America together with populations that have been recently introduced into other parts of the world. The differentiation between the two clades within L. huidobrensis is equivalent to that seen between other agromyzid species, suggesting that L. huidobrensis as currently defined contains two cryptic species. This finding is consistent with field observations of differences in pest status and insecticide resistance between L. huidobrensis populations. Until additional studies are complete, no changes in L. huidobrensis taxonomy are proposed. However, researchers and quarantine officials may wish to consider the findings of the current study in designing research, pest management, and quarantine programs for L. huidobrensis.     

福建省发现加藤斑潜蝇（双翅目：潜蝇科Diptera：Agromyzidae）

1995年在调查潜叶蝇种类时,在菊花上发现加藤斑潜蝇,属中国大陆首次报道,本文对其形态特征进行描述。