Control of Aculops lycopersici with the Predatory Mite Transeius montdorensis.

In this study, the predatory mite Transeius montdorensis (Acari, Phytoseiidae) was tested for the control of the tomato russet mite (TRM) Aculops lycopersici (Acari, Eriophyidae) in experiments with small plants, under semi-field and crop conditions. The releasing strategy consisted of repeatedly introducing the predator together with additional breeding prey. The predator was able to move and disperse to the upper part of the tomato plant where the TRM seeks refuge. At the crop level, significant reductions in TRM populations were observed that resulted in a significantly higher yield compared to the conventional control plot, where pesticides were used to control the pest. Caution should be taken when extreme temperatures or humidity occur as they could be deleterious to the predator population. Hence, crop practices should include the management of environmental parameters in the greenhouse to ensure the success of this TRM-control strategy. In conclusion, this biological approach seems to be an effective measure to control the pest and should be further implemented at crop level.

Development of a multiprimer metabarcoding approach to understanding trophic interactions in agroecosystems.

To understand trophic interactions and the precise ecological role of each predatory species, it is important to know which arthropod and plant resources are used by generalist predators in agroecosystems. Molecular approaches, such as the use of high-throughput sequencing (HTS), play a key role in identifying these resources. This study develops a multiprimer metabarcoding approach for screening the most common trophic interactions of two predatory arthropods with contrasting morphologies, Rhagonycha fulva (Coleoptera: Cantharidae) and Anthocoris nemoralis (Hemiptera: Anthocoridae) collected from a peach crop. To reduce the time and cost of this metabarcoding approach, we first evaluated the effect of using two different predator-pools of different size (10 and 23 individuals of the same species). We also used our system to analyze the performance of one and two primer pairs in the same library. Our results show that the analysis of 23 individuals together with the use of two primer pairs in the same library optimize the HTS analysis. Using these best-performing conditions, we then analyzed the entire bodies of field-collected predators as well as the washing solutions used to clean the insect bodies. We were able to identify both gut content (i.e., diet) and external pollen load (i.e., on the insects' bodies). This study also demonstrates the importance of washing predatory insects' bodies prior to HTS analysis when the target species have a considerable size (>10 mm) and hairy structures. This metabarcoding approach has significant potential for the study of trophic links in agriculture, revealing expected and unexpected trophic relationships.

Oviposition behavior of the mirid Macrolophus pygmaeus under risk of intraguild predation and cannibalism.

Zoophytophagous mirid species, that feed and develop either on prey or plant resources, are often found simultaneously on the same host. Hence, these species can engage in both intraguild predation and cannibalism, which can pose a threat to mirid eggs. Ovipositing females may respond to such risks of predation on their eggs by reducing the number of eggs laid or selecting safer oviposition sites. We tested the oviposition behavior of Macrolophus pygmaeus (Rambur) (Hemiptera: Miridae) females under the risk of cannibalism by M. pygmaeus males and intraguild predation by Nesidiocoris tenuis (Reuter) males (Hemiptera: Miridae) under laboratory conditions. Intraguild predators and cannibals were introduced during or after the oviposition period. The number of eggs laid (using counts of newly hatched nymphs) and their proportion on each part of a tomato plant were both measured. The results reveal that only cannibalism by M. pygmaeus males after the period of oviposition significantly decreased the number of hatched eggs. Cannibalism thus represents a greater risk to mirid eggs than intraguild predation. The M. pygmaeus female responded to the presence of potential intraguild predators (or competitors) by decreasing the number of eggs laid in the upper leaves. The results suggest that M. pygmaeus females avoid competition by N. tenuis, by laying fewer eggs on upper leaves. Cannibalism could regulate zoophytophagous predator populations under prey scarcity conditions and minimize the risk of crop damage associated with those biological control agents.

Development of a PCR-based method to monitor arthropod dispersal in agroecosystems: Macrolophus pygmaeus (Hemiptera: Miridae) from banker plants to tomato crops.

Development of conservation biological control programs requires the identification of sources that contribute to predator colonization of crops. Macrolophus pygmaeus (Rambur) (Hemiptera: Miridae) is an efficient polyphagous predator used in biological control programs in vegetable crops in Europe. We have developed a marking method based on spraying with a solution of the brine shrimp Artemia spp. (Anostraca: Artemiidae) cysts, followed by a PCR detection of Artemia DNA to monitor M. pygmaeus dispersal from banker plants to tomato crops. Experiments conducted in climatic chambers show that the topical application of this marking solution on M. pygmaeus does not significantly reduce adult longevity and that it is detected up to 6 d after the application. When this Artemia solution was applied on Calendula officinalis L. banker plants harboring M. pygmaeus and maintained outdoors, Artemia DNA was still detected on 62% of the insects after 6 d. The conducted field applications in commercial greenhouses have confirmed the usefulness of this method to monitor M. pygmaeus dispersal from banker plants to a newly planted tomato crop. This method can be used to assess arthropod movement, being an interesting molecular approach for further improving future pest management strategies.

Understanding trophic interactions of Orius spp. (Hemiptera: Anthocoridae) in lettuce crops by molecular methods.

BACKGROUND: The aphid Nasonovia ribisnigri (Mosley) (Hemiptera: Aphididae) and the thrips Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) are common pests in Mediterranean lettuce crops, where Orius spp. are common generalist predators. Predation by Orius spp. was studied in a lettuce plot by conventional polymerase chain reaction (PCR) and real-time PCR analyses using specific primers of both main pests. Also, high-throughput sequencing was used to have a wider approach of the diet of these predators in natural field conditions. RESULTS: Molecular analyses indicated a higher predation on N. ribisnigri in spring and on F. occidentalis in summer. Predation on alternative prey, like Collembola, was also found in both seasons. Real-time PCR was more sensitive than conventional PCR in showing the target trophic links, whereas high-throughput sequencing revealed predation on other natural enemies - intraguild predation (IGP), showing other trophic interactions of Orius majusculus within the studied ecosystem. CONCLUSIONS: This study gives important information about the trophic relationships present in Mediterranean lettuce crops in different periods of the year. The detected predation by Orius spp. on alternative prey, as well as on other natural enemies, should be further investigated to clarify whether it adds or detracts to the biological control of N. ribisnigri and F. occidentalis.

Comparative cytogenetic study of three Macrolophus species (Heteroptera, Miridae).

Macrolophus pygmaeus (Rambur, 1839) (Insecta, Heteroptera, Miridae) is a predator of key vegetable crop pests applied as a biocontrol agent in the Mediterranean region. Macrolophus pygmaeus and Macrolophus melanotoma (A. Costa, 1853) are cryptic species with great morphological similarity which results in their misidentification and negative consequences for the conservation of their populations on greenhouse and outdoor crops. In order to find out specific markers for their separation we studied the karyotype, male meiosis and heterochromatin composition of these species and additionally of a third species (as a reference one), Macrolophus costalis Fieber, 1858. We demonstrate here that all the three species share achiasmate male meiosis and sex chromosome pre-reduction. On the other hand, the species differ in karyotype, with 2n=28 (26+XY) in Macrolophus pygmaeus, 2n=27 (24+X1X2Y) in Macrolophus costalis, and 2n=34 (32+XY) in Macrolophus melanotoma, and heterochromatin distribution and composition. In addition, the species differ in sperm morphology: sperm cells of Macrolophus costalis are significantly longer with longer head and tail than those of Macrolophus melanotoma and Macrolophus pygmaeus, whereas sperm cells of Macrolophus melanotoma have a longer tail than those of Macrolophus pygmaeus. All these characters can be used as markers to identify the species, in particular the cryptic species Macrolophus melanotoma and Macrolophus pygmaeus.

Molecular assessment of predation by hoverflies (Diptera: Syrphidae) in Mediterranean lettuce crops.

BACKGROUND: Hoverflies (Diptera: Syrphidae) are generalist predators of a great variety of pests. Nasonovia ribisnigri (Hemiptera: Aphididae) and Frankliniella occidentalis (Thysanoptera: Thripidae) are two common pests in Mediterranean lettuce crops, where they occur alongside alternative prey (e.g., Collembola). A semi-field experiment was conducted in an experimental lettuce plot where hoverfly predation on N. ribisnigri, F. occidentalis and Collembola was studied by conventional PCR and qPCR using specific primers, as well as by next-generation sequencing (NGS) in order to reveal other potential trophic interactions. RESULTS: Trophic linkages between hoverflies and N. ribisnigri were the strongest both in spring and summer. F. occidentalis and Collembolans were also detected in both seasons, but with less frequency. qPCR detected a higher frequency of consumption than conventional PCR when both tests were run at optimal conditions. NGS analyses showed intraguild predation on other hoverfly species, as well as on anthocorids, spiders and even aphid parasitoids. CONCLUSIONS: Conventional PCR and qPCR provided important insights into Mediterranean hoverfly species predation on target pest and non-pest prey. NGS gave a complementary approach revealing a broader diet of these predators within the studied ecosystem.

Nectarivory by the plant-tissue feeding predator Macrolophus pygmaeus Rambur (Heteroptera: Miridae): nutritional redundancy or nutritional benefit?

Most predators and parasitoids feed on plant-provided food (nectar, pollen) or engage in herbivory during at least part of their life stages. Plant feeding by these insects plays an important role in driving predator-herbivore dynamics. Thus, understanding the effects of plant feeding on omnivores could be an important element in improving biological control strategies. The mirid Macrolophus pygmaeus is an omnivorous heteropteran predator of whitefly and other pests. Unlike other predators that need to seek out accessible nectar to meet their carbohydrate requirements, mirid bugs can access the plant's carbohydrate resources by feeding directly on plant tissues. Leaf and stem feeding could be seen as a nutritional surrogate that allows mirids to become independent of nectar availability. However, to date feeding experiments have not yet considered nectar feeding by these mirid predators. In this study we demonstrate that M. pygmaeus survival is prolonged on broad bean plants featuring extrafloral nectar as compared to broad bean with extrafloral nectaries removed, irrespective of the presence of cattail pollen. Survival on extrafloral nectar was comparable to the survival by individuals kept on broad bean provided with eggs of Ephestia kuehniella as prey. Also, a greater proportion of mirid females laid eggs when extrafloral nectar was available as compared to those confined on nectariless plants without supplemental food.

Impact of Macrolophus caliginosus presence on damage production by Dicyphus tamaninii (Heteroptera: Miridae) on tomato fruits.

In northeast Spain, the most common predators found in tomato fields and greenhouses are the mirids Macrolophus caliginosus Wagner and Dicyphus tamaninii Wagner. Natural colonization occurs during the growing season and both species prey on whiteflies. Because D. tamaninii can damage tomato fruits during periods of prey scarcity, a semifield experiment was carried out to evaluate whether the presence of M. caliginosus affects damage produced by D. tamaninii. In a tomato greenhouse, 60 plants were individually caged and distinct predator treatments were introduced: D. tamaninii, D. tamaninii + eggs of Ephestia kuehniella Zeller, D. tamaninii + M. caliginosus, M. caliginosus alone, and a control without any insect. Damage to tomato fruits was recorded (>25% of the fruit) in all the treatments with D. tamaninii, whereas no significant damage was detected with M. caliginosus alone. Finally, no intraguild predation was detected between both mirid species.