Aberrant splicing of a nicotinic acetylcholine receptor alpha 6 subunit is associated with spinosad tolerance in the thrips predator Orius laevigatus.

Susceptibility to insecticides is one of the limiting factors preventing wider adoption of natural enemies to control insect pest populations. Identification and selective breeding of insecticide tolerant strains of commercially used biological control agents (BCAs) is one of the approaches to overcome this constraint. Although a number of beneficial insects have been selected for increased tolerance to insecticides the molecular mechanisms underpinning these shifts in tolerance are not well characterised. Here we investigated the molecular mechanisms of enhanced tolerance of a lab selected strain of Orius laevigatus (Fieber) to the commonly used biopesticide spinosad. Transcriptomic analysis showed that spinosad tolerance is not a result of overexpressed detoxification genes. Molecular analysis of the target site for spinosyns, the nicotinic acetylcholine receptor (nAChR), revealed increased expression of truncated transcripts of the nAChR α6 subunit in the spinosad selected strain, a mechanism of resistance which was described previously in insect pest species. Collectively, our results demonstrate the mechanisms by which some beneficial biological control agents can evolve insecticide tolerance and will inform the development and deployment of insecticide-tolerant natural enemies in integrated pest management strategies.

Inheritance and Biological Characterization of an Orange-nymph Mutant in Orius laevigatus (Hemiptera: Anthocoridae).

A mutation showing a distinct orange color in the nymph stages was found in Orius laevigatus (Fieber) (Hemiptera: Anthocoridae), a key biological control agent in protected crops, used to control small pests, especially thrips. A laboratory strain carrying this body color mutation ambar was established. Genetic analysis determined that the mutation (ambar) was controlled by a single autosomal recessive allele. Some biological and ecological characteristics of this orange strain were compared to a normal population. Longevity, fecundity and fertility were similar in both populations, but immature survival, development rate, body size, starvation tolerance and predation capacity were inferior in the orange strain. The utility of the orange mutant as a visible marker for biological and ecological studies of this important biological control agent is discussed.

Effect of Parental Age and Mating Status on Reproductive Performance of Orius laevigatus (Hemiptera: Anthocoridae).

The reproductive potential of biological control agents (BCAs) is crucial for efficient mass-rearing and field performance, and it all begins with mating. Fecundity can be strongly influenced by intrinsic conditions, such as female age and, often neglected, male age and mating status. However, little is known about the impact of parental status at mating on female reproductive outcomes in BCAs. Orius laevigatus (Fieber) (Hemiptera: Anthocoridae) is widely used to control thrips in protected crops. We evaluated how many consecutive females a male could successfully mate and the effect on a female's reproductive output. In addition, we studied the effects of male and female age on mating. In the multiple mating experiment, the males showed a high capacity to fertilize females successively, not reducing fecundity until the sixth mated female. In the delayed mating experiment, copulation duration and fecundity increased with male age but decreased with female age. In contrast, fertility followed an opposite pattern, increasing with female age but decreasing with male age. However, fecundity gains outweighed fertility declines in both sexes. Therefore, reproductive capacity is increased when mating newly emerged females with males a few days old. The implications of our results for mass rearing and field performance are discussed.

Effect of the Amount of Ephestia kuehniella Eggs for Rearing on Development, Survival, and Reproduction of Orius laevigatus.

Orius laevigatus is a key tool for the success of augmentative biological control programs in protected crops. This biological control agent is mass-reared feeding on eggs of Ephestia kuehniella. However, this factitious prey is expensive, accounting for a significant percentage of the rearing costs. Therefore, there is a need to optimize the amount of Ephestia eggs needed per individual leading to a cost reduction, which in turn will favor biological control adoption. This study investigated the effect of the amount of Ephestia eggs provided on the developmental and reproductive fitness of O. laevigatus. At least a daily supply of 1 and 3 Ephestia eggs was needed for optimal development of the first two nymphal instars, respectively, although for maximum survival, 1 egg was enough for both instars. For subsequent development until adulthood, a minimum of 8 eggs per day were needed to fully support growth, but only 3 eggs for optimal survival. Similarly, male body size was also maximized by feeding 8 eggs, but for maximum female body size 10 eggs per day were required. Oviposition rate of females increased with the daily number of Ephestia eggs provided, until a plateau was reached at 8 eggs/day. Benefits and savings for industrial production of O. laevigatus are discussed.

Selection for resistance to pyrethroids in the predator Orius laevigatus.

BACKGROUND: Insecticide resistance in the natural enemies of pest species is a very desirable trait. It allows better integration of biological control with the synthetic and natural compounds applied to manage certain pests, especially secondary pest outbreaks. Insecticide resistance in predatory insects has been documented for lady beetles and lacewings, but intriguingly no cases of field-evolved resistance have been noted in any heteropteran predator. In this work, we first explored the variation in susceptibility to pyrethroids in wild and commercial populations of Orius laevigatus (Fieber). Second, we exploited this genetic variation to artificially select a strain of O. laevigatus resistant to pyrethroids. RESULTS: We found significant variation [median lethal dose (LC50 ) 1.6-77.0 mg L-1 ] in susceptibility to pyrethroids in wild populations of the heteropteran predator O. laevigatus, with a baseline LC50 value of 14.6 mg L-1 . We successfully selected a strain of O. laevigatus highly resistant to pyrethroids (LC50 = 1059.9 mg L-1 ). In addition, such resistance was expressed in every instar, particularly in the last nymphal stages, increasing the resilience of the whole population present in the crop facing pyrethroid application. CONCLUSION: The level of resistance achieved may be sufficient to allow survival of adults and nymphs of O. laevigatus exposed to the maximum field rate of several pyrethroids and natural pyrethrins, widely used to control a number of pests in organic and integrated pest management crops. Therefore, this strain resistant to pyrethrins and pyrethroids would improve the resilience of biocontrol protocols, which is an urgent requirement for wider adoption of biological control. © 2021 Society of Chemical Industry.

Challenges facing arthropod biological control: identifying traits for genetic improvement of predators in protected crops.

Biological control is an efficient pest control method but there are still limitations that are hindering its wider adoption. Genetic improvement of biological control agents (BCAs) can help to overcome these constraints, but the choice of key attributes for better performance that need to be selected is still an open question. Several characteristics have been suggested but the harsh reality is that selective breeding of BCAs has received a lot of attention but resulted in very little progress. Identifying the appropriate traits to be prioritized may be the first step to reverse this situation. In our opinion, the best way is to look at the factors limiting the performance of key BCAs, especially generalist predators (pesticide compatibility, prey-density dependence, non-suitable crops, and extreme environmental conditions), and according to these challenges, to choose the attributes that would allow BCAs to overcome those limitations. The benefits of selection for higher resistance to toxins, whether artificially applied (pesticides) or plant produced (plant defenses); increased fitness when feeding on non-prey food (supplemented or plant-derived); and better adaptation to extreme temperature and humidity are discussed. In conclusion, genetic improvement of BCAs can bring about new opportunities to biocontrol industry and users to enhance biocontrol resilience. © 2020 Society of Chemical Industry.

Comparative Genomics of Facultative Bacterial Symbionts Isolated from European Orius Species Reveals an Ancestral Symbiotic Association.

Pest control in agriculture employs diverse strategies, among which the use of predatory insects has steadily increased. The use of several species within the genus Orius in pest control is widely spread, particularly in Mediterranean Europe. Commercial mass rearing of predatory insects is costly, and research efforts have concentrated on diet manipulation and selective breeding to reduce costs and improve efficacy. The characterisation and contribution of microbial symbionts to Orius sp. fitness, behaviour, and potential impact on human health has been neglected. This paper provides the first genome sequence level description of the predominant culturable facultative bacterial symbionts associated with five Orius species (O. laevigatus, O. niger, O. pallidicornis, O. majusculus, and O. albidipennis) from several geographical locations. Two types of symbionts were broadly classified as members of the genera Serratia and Leucobacter, while a third constitutes a new genus within the Erwiniaceae. These symbionts were found to colonise all the insect specimens tested, which evidenced an ancestral symbiotic association between these bacteria and the genus Orius. Pangenome analyses of the Serratia sp. isolates offered clues linking Type VI secretion system effector-immunity proteins from the Tai4 sub-family to the symbiotic lifestyle.