Insights into the origin of the invasive populations of Trioza erytreae in Europe using microsatellite markers and mtDNA barcoding approaches.

The African citrus psyllid Trioza erytreae is one of the major threats to citrus industry as the vector of the incurable disease known as huanglongbing (HLB) or citrus greening. The psyllid invaded the northwest of the Iberian Peninsula 6 years ago. The invasion alarmed citrus growers in the Mediterranean basin, the largest citrus producing area in Europe, which is still free of HLB. Before our study, no research had been carried out on the genetic diversity of T. erytreae populations that have invaded the Iberian Peninsula and the archipelagos of the Macaronesia (Madeira and the Canary Islands). In this study, combining microsatellites markers and mtDNA barcoding analysis, we characterize the genetic diversity, structure and maternal relationship of these new invasive populations of T. erytreae and those from Africa. Our results suggest that the outbreaks of T. erytreae in the Iberian Peninsula may have derived from the Canary Islands. The populations of T. erytreae that invaded Macaronesia and the Iberian Peninsula are likely to have originated from southern Africa. We anticipate our results to be a starting point for tracking the spread of this invasive pest outside of Africa and to be important for optimizing contingency and eradication plans in newly invaded and free areas.

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.

Sulfoxaflor efficacy in the laboratory against imidacloprid-resistant and susceptible populations of the green peach aphid, Myzus persicae: Impact of the R81T mutation in the nicotinic acetylcholine receptor.

A key to effective insect pest management and insecticide resistance management is to provide growers with a range of new tools as potential alternatives to existing compounds or approaches. Sulfoxaflor (Isoclast™ active) is a new sulfoximine insecticide which is active on a broad range of sap-feeding insects, including species that have reduced susceptibility to currently used insecticides, such as imidacloprid from the neonicotinoid class. Sulfoxaflor (SFX) and imidacloprid (IMI) were tested in laboratory bioassays to compare the susceptibility of field populations of green peach aphid, Myzus persicae (Sulzer), exhibiting varying degrees of resistance involving an alteration (R81T) to the insect nicotinic acetylcholine receptor. The LC50 values for M. persicae exposed to SFX ranged from 0.09 to 1.31 (mg litre-1), whereas when the same populations were exposed to IMI the LC50 values ranged from 0.6 to 76.2 (mg litre-1). M. persicae were significantly more sensitive to SFX as compared to IMI for nine of the 13 populations tested. For M. persicae populations confirmed to be homozygous susceptible (ss) or heterozygous rs) for the R81T point mutation, there was no significant differences in the observed LC50 values for either SFX or IMI relative to the susceptible reference population (15LP1). However, in all M persicae populations that were homozygous (rr) for the R81T point mutation, susceptibility was significantly less to IMI as compared to the reference population with resistance ratios ranging from 22.1 to 63.5-fold. In contrast, only one homozygous resistant population (15MP9) exhibited a statistically significant change in susceptibility (RR = 10-fold) to SFX as compared to the reference population, which was far less than the 56-fold observed for imidacloprid in that same population. Thus, this study indicates there is no specific correlation between the laboratory efficacy of SFX and IMI in field collected populations in Spain displaying varying degrees of resistance to IMI. Furthermore, the presence of target site resistance in M. persicae to IMI, in the form of the R81T mutation, does not a priori translate to a reduction in sensitivity to sulfoxaflor. Consequently, SFX can be an effective tool for use in insect pest management programs for green peach aphid. These data also serve as a baseline reference for green peach aphid sensitivity to SFX prior to commercial uses in Spain.

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.

Identification of mutations associated with pyrethroid resistance in the voltage-gated sodium channel of the tomato leaf miner (Tuta absoluta).

The tomato leaf miner, Tuta absoluta (Lepidoptera) is a significant pest of tomatoes that has undergone a rapid expansion in its range during the past six years and is now present across Europe, North Africa and parts of Asia. One of the main means of controlling this pest is through the use of chemical insecticides. In the current study insecticide bioassays were used to determine the susceptibility of five T. absoluta strains established from field collections from Europe and Brazil to pyrethroids. High levels of resistance to λ cyhalothrin and tau fluvalinate were observed in all five strains tested. To investigate whether pyrethroid resistance was mediated by mutation of the para-type sodium channel in T. absoluta the IIS4-IIS6 region of the para gene, which contains many of the mutation sites previously shown to confer knock down (kdr)-type resistance to pyrethroids across a range of different arthropod species, was cloned and sequenced. This revealed that three kdr/super-kdr-type mutations (M918T, T929I and L1014F), were present at high frequencies within all five resistant strains at known resistance 'hot-spots'. This is the first description of these mutations together in any insect population. High-throughput DNA-based diagnostic assays were developed and used to assess the prevalence of these mutations in 27 field strains from 12 countries. Overall mutant allele frequencies were high (L1014F 0.98, M918T 0.35, T929I 0.60) and remarkably no individual was observed that did not carry kdr in combination with either M918T or T929I. The presence of these mutations at high frequency in T. absoluta populations across much of its range suggests pyrethroids are likely to be ineffective for control and supports the idea that the rapid expansion of this species over the last six years may be in part mediated by the resistance of this pest to chemical insecticides.

Genetic study of Mediterranean and South American populations of tomato leafminer Tuta absoluta (Povolny, 1994) (Lepidoptera: Gelechiidae) using ribosomal and mitochondrial markers.

BACKGROUND: Before its introduction into Europe at the end of 2006, Tuta absoluta (Povolny, 1994) was confined solely to South America. Currently, this invasive pest is well established in various European and Mediterranean countries, causing important economic losses to tomato (Lycopersicon esculentum Mill.) crops. In order to study the genetic variability of this pest, 23 Mediterranean and ten native South American populations were analysed with nuclear ribosomal DNA (rDNA) and mitochondrial DNA (mtDNA) markers. RESULTS: The internal transcribed spacers 1 (ITS1) and 2 (ITS2) of rDNA and a fragment in the mtDNA gene encoding cytochrome oxidase I (COI) were PCR amplified and sequenced in T. absoluta. Sequence analyses consistently revealed neither intrapopulation nor interpopulation variation in either genomic region. CONCLUSIONS: High genetic homogeneity was detected in T. absoluta populations from the Mediterranean Basin and South America, based on mtCOI and ITS rDNA sequence analysis. A single genetic type was identified in this pest.

Insecticide resistance status of Bemisia tabaci Q-biotype in south-eastern Spain.

BACKGROUND: Bemisia tabaci Gennadius Q-biotype has readily developed resistance to numerous insecticide classes. Studies in the Mediterranean area are needed to clarify the resistance status and cross-resistance patterns in this invasive whitefly biotype. The levels of resistance in nymphs of seven strains of B. tabaci Q-biotype from south-eastern Spain to representative insecticides were determined. RESULTS: Six populations had low to moderate levels of resistance to azadirachtin (0.2- to 7-fold), buprofezin (11- to 59-fold), imidacloprid (1- to 15-fold), methomyl (3- to 55-fold), pyridaben (0.9- to 9-fold), pyriproxyfen (0.7- to 15-fold) and spiromesifen (1- to 7-fold), when compared with a contemporary Spanish Q-biotype reference population (LC(50) = 2.7, 8.7, 15.2, 19.9, 0.34, 20.9 and 1.1 mg L(-1) respectively). A single population collected from a greenhouse subject to intensive insecticide use exhibited generally higher resistance levels to the same array of compounds (31-, 1164-, 3-, 52-, 9-, 19- and 3-fold respectively). Pyridaben and spiromesifen were extremely effective against nymphs of all strains, with LC(50) values significantly below recommended application rates. CONCLUSION: In contrast to previous reports, high rates of efficacy exist for numerous insecticide classes against B. tabaci Q-biotype populations in these intensive agricultural regions of south-eastern Spain. This probably reflects the recent and significant reductions in exposure that have resulted from a wider uptake of IPM technologies and strategies. However, the continued presence of resistance genes also suggests that a reversion to levels of high insecticide exposure could result in a rapid selection for resistance.

Inheritance of resistance to acrinathrin in Frankliniella occidentalis (Thysanoptera: Thripidae).

BACKGROUND: The western flower thrips (WFT), Frankliniella occidentalis (Pergande), is an economically important pest. The genetic basis of acrinathrin resistance was investigated in WFT. RESULTS: The resistant strain, selected in the laboratory for acrinathrin resistance from a pool of thrips populations collected in Almeria (south-eastern Spain), showed a high resistance to acrinathrin (43-fold based on LC(50) values) compared with the laboratory susceptible strain. Mortality data from reciprocal crosses of resistant and susceptible thrips indicated that resistance was autosomal and not influenced by maternal effects. Analysis of probit lines from the parental strains and reciprocal crosses showed that resistance was expressed as a codominant trait. To determine the number of genes involved, a direct test of monogenic inheritance based on the backcrosses suggested that resistance to acrinathrin was probably controlled by one locus. Another approach, which was based on phenotypic variances, showed n(E), or the minimum number of freely segregating genetic factors for the resistant strain, to be 0.79. CONCLUSION: The results showed that acrinathrin resistance in WFT was autosomal and not influenced by maternal effects, and was expressed as a codominant trait, probably controlled by one locus.