Skuhrovec, J., O. Douda, M. Zouhar, M. Maňasová, M. Božik, and P. Klouček. Journal of Economic Entomology, toz279, https://doi.org/10.1093/jee/toz279

The Colorado potato beetle ranks as one of the most important potato pests, mainly due to its high feeding rate during all developmental stages, particularly third and fourth larval instar, and high fecundity. The effect of essential oil (EO) from anise (Pimpinella anisum L. [Apiales: Apiaceae]) prepared as conventional and encapsulated (EN) formulations on the mortality and antifeedant responses of young larvae of Colorado potato beetles was studied to evaluate the insecticidal and antifeedant effects of five concentrations of this EO and to assess the persistence of both formulations on potato plants. The EN formulation had a significantly higher residual amount compared with that of the conventionally formulated EO. Significantly different values of LC50 and LC90 (ppm) were established for the EO (LC50 = 1,700 and LC90 = 9500) and EN (LC50 = 3,100 and LC90 = 14,300) formulations. The effects of both P. anisum formulations (EO and EN) applied topically to Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae) larvae were distinctly different from those observed with the contact treatment. At the highest concentration of 20,000 ppm, the mortality of the second instars of the L. decemlineata larvae did not exceed 25%. On the other hand, both tested formulations of P. anisum were highly effective when administered orally. The encapsulated EO formulation achieved a distinctly higher biological activity. Our results confirm that the EO from P. anisum, especially the encapsulated formulation, has high insecticidal properties that may lead to the development of new organic products for the control of Colorado potato beetles.

Dumas, P. , M. Sambou , J. D. Gaudet , M. D. Morin , C. E. Moffat , S. Boquel , P. Morin. Archives of Insect Biochemistry and Physiology https://doi.org/10.1002/arch.21642

The Colorado potato beetle (Leptinotarsa decemlineata [Say]) is an insect pest that can significantly harm potato plants worldwide. Control of this insect relies heavily on chemical insecticides such as chlorantraniliprole. Nevertheless, the complete molecular signature associated with response to this compound is lacking in L. decemlineata. In this study, amplification and quantification by qRT‐PCR (quantitative reverse transcription‐polymerase chain reaction) of targets relevant to chlorantraniliprole were undertaken in insects exposed to this chemical. This approach showed modulation of numerous cytochrome P450s, such as CYP350D1 and CYP4Q3, as well as upregulation of microRNAs (miRNAs), including miR‐1‐3p and miR‐305‐5p, in chlorantraniliprole‐exposed insects. Functional assessment of transcript targets predicted to be regulated by these miRNAs was performed and revealed their likely impact on transcriptional regulation. RNAi‐based targeting of CYP350D1 notably provided preliminary evidence of its underlying implication for chlorantraniliprole response in L. decemlineata. Overall, this study strengthens the current knowledge of the molecular changes linked to chlorantraniliprole response in L. decemlineata and provides novel targets with potential relevance to chlorantraniliprole susceptibility in this insect pest of global relevance.

Tomilova, O. G., O. N. Yaroslavtseva, M. D. Ganina, M. V. Tyurin, E. I. Chernyak, I. V. Senderskiy, Y. A. Noskov, O. V. Polenogova, Y. B. Akhanaev, V. Yu. Kryukov, V. V. Glupov, S. V. Morozov. Journal of Insect Physiology 116: 106-117. https://doi.org/10.1016/j.jinsphys.2019.05.003

Susceptibility to the fungus Metarhizium robertsii and changes in host defences were evaluated in different stages of the intermoult period (4–6 h, 34–36 h and 84–86 h post moult in IV larval instars) of the Colorado potato beetle. A significant thickening of the cuticle during larval growth was accompanied by decreases in cuticle melanization, phenoloxidase activity and epicuticular hydrocarbon contents (C28-C32). At the same time, a decrease in the conidial adhesion rate and an increase in resistance to the fungus were observed. In addition, we recorded significant elevation of the encapsulation rate and total haemocyte counts in the haemolymph during the specified period. The activity of detoxification enzymes decreased in the haemolymph but increased in the fat body during larval growth. No significant differences in the fatty acid content in the epicuticle were observed. The role of developmental disorders in susceptibility to entomopathogenic fungi is also discussed.

Margus, A., Piiroinen, S., Lehmann, P. et al. Scientific Reports 9, 11320 (2019) doi:10.1038/s41598-019-47473-1

Stress tolerance and adaptation to stress are known to facilitate species invasions. Many invasive species are also pests and insecticides are used to control them, which could shape their overall tolerance to stress. It is well-known that heavy insecticide usage leads to selection of resistant genotypes but less is known about potential effects of mild sublethal insecticide usage. We studied whether stressful, sublethal pyrethroid insecticide exposure has within-generational and/or maternal transgenerational effects on fitness-related traits in the Colorado potato beetle (Leptinotarsa decemlineata) and whether maternal insecticide exposure affects insecticide tolerance of offspring. Sublethal insecticide stress exposure had positive within-and transgenerational effects. Insecticide-stressed larvae had higher adult survival and higher adult body mass than those not exposed to stress. Furthermore, offspring whose mothers were exposed to insecticide stress had higher larval and pupal survival and were heavier as adults (only females) than those descending from control mothers. Maternal insecticide stress did not explain differences in lipid content of the offspring. To conclude, stressful insecticide exposure has positive transgenerational fitness effects in the offspring. Therefore, unsuccessful insecticide control of invasive pest species may lead to undesired side effects since survival and higher body mass are known to facilitate population growth and invasion success.

Xu, Q.-Y., Q.-W. Meng, P. Deng, K.-Y. Fu, W.-C. Guo, and G.-Q. Li. Bulletin of Entomological Research https://doi.org/10.1017/S0007485318001050

Dietary delivery of bacterially expressed double-stranded RNA (dsRNA) has a great potential for management of Leptinotarsa decemlineata. An important first step is to discover possible RNA-interference (RNAi)-target genes effective against larvae, especially the old larvae. In the present paper, five putative Broad-Complex (BrC) cDNAs (Z1-Z4, and Z6) were identified in L. decemlineata. The expression of the five LdBrC isoforms was suppressed by juvenile hormone signaling, whereas the transcription was upregulated by 20-hydroxyecdysone signaling at the fourth (final) instar larval stage. Feeding of bacterially expressed dsBrC (derived from a common fragment of the five LdBrC variants) in the third- and fourth-instar larvae successfully knocked down the target mRNAs. For the fourth-instar LdBrC RNAi hypomorphs, they had a higher larval mortality compared with the controls. Moreover, most dsBrC-fed beetles did not pupate normally. After removal of the apolysed larval cuticle, a miniature adult was found. The adult head, compound eyes, prothorax, mesothorax, metathorax were found on the dorsal view. Distinct adult cuticle pigmentation was seen on the prothorax. The mouthparts, forelegs, midlegs, and hindlegs could be observed on the ventral view of the miniature adults. For the third-instar LdBrC RNAi specimens, around 20% moribund beetles remained as prepupae and finally died. Therefore, LdBrC is among the most attractive candidate genes for RNAi to control the fourth-instar larvae in L. decemlineata.

Kryukov, V. Y., M. R. Kabilov, N. Smirnova, O. G. Tomilova, M. V. Tyurin, Y. B. Akhanaev, O. V. Polenogova, V. P. Danilov, S. K. Zhangissina, T. Alikina, O. N. Yaroslavtseva, V. V. Glupov, Fungal Biology https://doi.org/10.1016/j.funbio.2019.09.012

Strains of entomopathogenic fungi may have substantial differences in their final stages of mycosis. Insect cadavers are usually overgrown with mycelium after colonization of the insect body, but in many cases, bacterial decomposition of the colonized hosts occurs. We used two Metarhizium robertsii strains in the work: Mak-1 (cadavers become overgrown with mycelium and conidia) and P-72 (cadavers decay after fungal colonization). We conducted a comparative analysis of gut and cadaver microbiota in Colorado potato beetle larvae using 16S rRNA gene sequencing after infection with these strains. In addition, we estimated the content of different forms of nitrogen in cadavers and the influence of cadavers on the growth of Solanum lycopersicum on sand substrates under laboratory conditions. It was shown that infections did not lead to a significant shift in the midgut bacterial communities of infected insects compared to those of untreated insects. Importantly, bacterial communities were similar in both types of cadaver, with predominantly enterobacteria. Decomposing cadavers (P-72) were characterized by increased nitrate and ammonium, and they had a stronger growth-promoting effect on plants compared to cadavers overgrown with mycelium and conidia (Mak-1). We also estimated the colonization and growth of plants after treatment with conidia of both strains cultivated on artificial medium. Both cultures successfully colonized plants, but strain P-72 showed stronger growth promotion than Mak-1. We propose that the use of deviant strains that are unable to sporulate on cadavers leads to a faster (though only passive) flow of nitrogen from killed insects to plants.

Krey, K. L., C. K. Blubaugh, J. T. Van Leuven, and W. E. Snyder. Environmental Entomology, nvz123, https://doi.org/10.1093/ee/nvz123

Soil chemistry and microbial diversity can impact the vigor and nutritive qualities of plants, as well as plants’ ability to deploy anti-herbivore defenses. Soil qualities often vary dramatically on organic versus conventional farms, reflecting the many differences in soil management practices between these farming systems. We examined soil-mediated effects on herbivore performance by growing potato plants (Solanum tuberosum L.) in soils collected from organic or conventional commercial farm fields, and then exposing these plants to herbivory by green peach aphids (Myzus persicae Sulzer, Hemiptera: Aphididae) and/or Colorado potato beetles (Leptinotarsa decemlineata Say, Coleoptera: Chrysomelidae). Responses of the two potato pests varied dramatically. Survivorship of Colorado potato beetles was almost 3× higher on plants grown in organic than in conventional soils, but was unaffected by the presence of aphids. In contrast, aphid colony growth was twice as rapid when aphids were reared alone rather than with Colorado potato beetles, but was unaffected by soil type. We saw no obvious differences in soil nutrients when comparing organic and conventional soils. However, we saw a higher diversity of bacteria in organic soils, and potato plants grown in this soil had a lower carbon concentration in foliar tissue. In summary, the herbivore species differed in their susceptibility to soil- versus competitor-mediated effects, and these differences may be driven by microbe-mediated changes in host plant quality. Our results suggest that soil-mediated effects on pest growth can depend on herbivore species and community composition, and that soil management strategies that promote plant health may also increase host quality for pests.

Naqqash, M. N., A. Gökçe, E. Aksoy, A. Bakhsh. Chemosphere, https://doi.org/10.1016/j.chemosphere.2019.124857

Colorado potato beetle, Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae), is the important pest of potato all over the world. This insect pest is resistant to more than 50 active compounds belonging to various chemical groups. Potential of RNA interference (RNAi) was explored to knock down transcript levels of imidacloprid resistant genes in Colorado potato beetle (CPB) under laboratory conditions. Three important genes belonging to cuticular protein (CP), cytochrome P450 monoxygenases (P450) and glutathione synthetase (GSS) families encoding imidacloprid resistance were targeted. Feeding bio-assays were conducted on various stages of imidacloprid resistant CPB lab population by applying HT115 expressing dsRNA on potato leaflets. Survival rate of insects exposed to CP-dsRNA decreased to 4.23%, 15.32% and 47.35% in 2nd, 3rd and 4th instar larvae respectively. Larval weight and pre-adult duration were also affected due to dsRNAs feeding. Synergism of RNAi with imidacloprid conducted on the 2nd instar larvae, exhibited 100% mortality of larvae when subjected to reduced doses of GSS and CP dsRNAs along with imidacloprid. Utilization of three different dsRNAs against imidacloprid resistant CPB population reveal that dsRNAs targeting CP, P450 and GSS enzymes could be useful tool in management of imidacloprid resistant CPB populations.

García-Robles, I., De Loma, J., Capilla, M., Roger, I., Boix-Montesinos, P., Carrión, P., Vicente, M., López-Galiano, MJ, Real, MD, Rausell, C. Developmental & Comparative Immunology DOI: 10.1016/j.dci.2019.103525

Bacillus thuringiensis (Bt) toxins constitute effective, environmentally safe biopesticides. Nevertheless, insects' tolerance to Bt is influenced by environmental factors affecting immunity. To understand larval immune response in the devastating coleopteran insect pest Colorado potato beetle (CPB), we undertook a proteomic analysis of hemolymph of non-treated control larvae and larvae consuming non-lethal doses of spore-crystal mixtures containing the coleopteran-active Cry3Aa toxin. Results revealed lower amount of proteins involved in insect growth and higher amount of immune response-related proteins in challenged insects, sustaining the larval weight loss observed. Additionally, we found a potential regulatory role of the evolutionary conserved miR-8 in the insect's immune response relying on antimicrobial peptides (AMPs) production. Upon toxin challenge, different patterns of hemolymph AMPs expression and phenoloxidase activity were observed in CPB larvae reared on different Solanaceae plants. This suggests that diet and diet-associated insect midgut microbiota might modulate this insects' tolerance to non-lethal doses of Bt.

Nguyen D, Poeschl Y, Lortzing T, et al. International Journal of Molecular Sciences. 2018;19(12):3845.

In nature, plants are frequently subjected to multiple biotic and abiotic stresses, resulting in a convergence of adaptive responses. We hypothesised that hormonal signalling regulating defences to different herbivores may interact with drought responses, causing distinct resistance phenotypes. To test this, we studied the hormonal and transcriptomic responses of Solanum dulcamara subjected to drought and herbivory by the generalist Spodoptera exigua (beet armyworm; BAW) or the specialist Leptinotarsa decemlineata (Colorado potato beetle; CPB). Bioassays showed that the performance of BAW, but not CPB, decreased on plants under drought compared to controls. While drought did not alter BAW-induced hormonal responses, it enhanced the CPB-induced accumulation of jasmonic acid and salicylic acid (SA), and suppressed ethylene (ET) emission. Microarray analyses showed that under drought, BAW herbivory enhanced several herbivore-induced responses, including cell-wall remodelling and the metabolism of carbohydrates, lipids, and secondary metabolites. In contrast, CPB herbivory enhanced several photosynthesis-related and pathogen responses in drought-stressed plants. This may divert resources away from defence production and increase leaf nutritive value. In conclusion, while BAW suffers from the drought-enhanced defences, CPB may benefit from the effects of enhanced SA and reduced ET signalling. This suggests that the fine-tuned interaction between the plant and its specialist herbivore is sustained under drought.