Noctuid Species Research Articles

A sperm-activating trypsin-like protease from the male reproductive tract of Spodoptera litura: Proteomic identification, sequence characterization, gene expression profile, RNAi and the effects of ionizing radiation

Like other lepidopteran insects, males of the tobacco cutworm moth, Spodoptera litura produce two kinds of spermatozoa, eupyrene (nucleate) and apyrene (anucleate) sperm. Formed in the testis, both kinds of sperm are released into the male reproductive tract in an immature form and are stored in the duplex region of the tract. Neither type of sperm is motile at this stage. When stored apyrene sperm from the duplex are treated in vitro with an extract of the prostatic region of the male tract, or with mammalian trypsin, they become motile; activation is greater and achieved more rapidly with increasing concentration of extract or enzyme. The activating effect of prostatic extract is blocked by soybean trypsin inhibitor (SBTI), also in a dose-dependent way. These results suggest that the normal sperm-activating process is due to an endogenous trypsin-like protease produced in the prostatic region.Proteomic analysis of S. litura prostatic extracts revealed a Trypsin-Like Serine Protease, TLSP, molecular weight 27 kDa, whose 199-residue amino acid sequence is identical to that of a predicted protein from the S. litura genome and is highly similar to predicted proteins encoded by genes in the genomes of several other noctuid moth species. Surprisingly, TLSP is only distantly related to Serine Protease 2 (initiatorin) of the silkmoth, Bombyx mori, the only identified lepidopteran protein so far shown to activate sperm. TLSP has features typical of secreted proteins, probably being synthesized as an inactive precursor zymogen, which is later activated by proteolytic cleavage.cDNA was synthesized from total RNA extracted from the prostatic region and was used to examine TLSP expression using qPCR. tlsp mRNA was expressed in both the prostatic region and the accessory glands of the male tract. Injection of TLSP-specific dsRNA into adult males caused a significant reduction after 24 h in tlsp mRNA levels in both locations. The number of eggs laid by females mated to adult males that were given TLSP dsRNA in 10 % honey solution, and the fertility (% hatched) of the eggs were reduced. Injecting pupae with TLSP dsRNA caused the later activation of apyrene sperm motility by adult male prostatic extracts to be significantly reduced compared to controls.Exposure of S. litura pupae to ionizing radiation significantly reduced expression of tlsp mRNA in the prostatic part and accessory gland of irradiated males in both the irradiated generation and also in their (unirradiated) F1 progeny. The implications of these findings for the use of the inherited sterility technique for the control of S. litura and other pest Lepidoptera are discussed.

The novel function of an orphan pheromone receptor reveals the sensory specializations of two potential distinct types of sex pheromones in noctuid moth

Sex pheromones play crucial role in mating behavior of moths, involving intricate recognition mechanisms. While insect chemical biology has extensively studied type I pheromones, type II pheromones remain largely unexplored. This study focused on Helicoverpa armigera, a representative species of noctuid moth, aiming to reassess its sex pheromone composition. Our research unveiled two previously unidentified candidate type II sex pheromones—3Z,6Z,9Z-21:H and 3Z,6Z,9Z-23:H—in H. armigera. Furthermore, we identified HarmOR11 as an orphan pheromone receptor of 3Z,6Z,9Z-21:H. Through AlphaFold2 structural prediction, molecular docking, and molecular dynamics simulations, we elucidated the structural basis and key residues governing the sensory nuances of both type I and type II pheromone receptors, particularly HarmOR11 and HarmOR13. This study not only reveals the presence and recognition of candidate type II pheromones in a noctuid moth, but also establishes a comprehensive structural framework for PRs, contributing to the understanding of connections between evolutionary adaptations and the emergence of new pheromone types.

A new noctuid genus and species (Lepidoptera, Noctuidae, Amphipyrinae, Psaphidini, Triocnemidina) from New Mexico and Texas, United States of America.

Pooleagen. nov. is described for two noctuid species from southwestern United States: Pooleagrandimacula Barnes & McDunnough, comb. nov., previously in Oxycnemis Grote, and Pooleapsaphidoidessp. nov.Poolea is compared to Oxycnemis (Amphipyrinae, Psaphidini, Triocnemidina) and is retained in the same subtribe. Adult moths and male and female genitalia of Poolea species are illustrated along with those of Oxycnemisadvena Grote, the genus type species. Pertinent recent taxonomic changes to Amphipyrinae classification are reviewed.

A new species of winter noctuid moth (Lepidoptera, Noctuidae, Xyleninae) from Zhejiang, China, with a key to species of the genus.

The tribe Xylenini is one of the main large taxonomic groups known as Winter Noctuidae. A new species of the genus Antivaleria Sugi, 1980, Antivaleriaronkayorum Zhang & Wang, sp. nov. is described and illustrated from Zhejiang Prov., China. The species resembles Antivaleriaperegovitsi Ronkay et al., 2010, but differs in wing pattern, foot-like cucullus and special shape of harpe. A key to the Antivaleria species is presented. The holotype is deposited in the Department of Entomology, College of Plant Protection, South China Agricultural University, Guangzhou.

An odorant receptor tuned to an attractive plant volatile vanillin in Spodoptera litura

The insect olfaction plays crucial roles in many important behaviors, in which ORs are key determinants for signal transduction and the olfactory specificity. Spodoptera litura is a typical polyphagous pest, possessing a large repertoire of ORs tuning to broad range of plant odorants. However, the specific functions of those ORs remain mostly unknown. In this study, we functionally characterized one S. litura OR (OR51) that was highly expressed in the adult antennae. First, by using Xenopus oocyte expression and two-electrode voltage clamp recording system (XOE-TEVC), OR51 was found to be strongly and specifically responsive to vanillin (a volatile of S. litura host plants) among 77 tested odorants. Second, electroantennogram (EAG) and Y-tube behavioral experiment showed that vanillin elicited significant EAG response and attraction behavior especially of female adults. This female attraction was further confirmed by the oviposition experiment, in which the soybean plants treated with vanillin were significantly preferred by females for egg-laying. Third, 3D structural modelling and molecular docking were conducted to explore the interaction between OR51 and vanillin, which showed a high affinity (−4.46 kcal/mol) and three residues (Gln163, Phe164 and Ala305) forming hydrogen bonds with vanillin, supporting the specific binding of OR51 to vanillin. In addition, OR51 and its homologs from other seven noctuid species shared high amino acid identities (78–97%) and the same three hydrogen bond forming residues, suggesting a conserved function of the OR in these insects. Taken together, our study provides some new insights into the olfactory mechanisms of host plant finding and suggests potential applications of vanillin in S. litura control.

New records of two noctuid species (Lepidoptera, Noctuidae) from Iran

New records of two noctuid species (Lepidoptera, Noctuidae) from Iran

No interaction of fluctuating or constant temperature and virulence of entomopathogenic fungi in two noctuid species

AbstractThe potential interaction of fluctuating temperature and the virulence of entomopathogens has implications for biological control. The objective of this study was to investigate the effect of entomopathogenic fungi (Metarhizium brunneum, Beauveria bassiana) on noctuid pest caterpillars (Spodoptera littoralis, Heliothis virescens) under constant and fluctuating temperature regimes. The results revealed similar virulence of entomopathogenic fungi under fluctuating temperature (20–30 °C or 15–35 °C) compared with constant temperature (25 °C). Experiments with cotton leaves as food confirmed that S. littoralis was less susceptible to entomopathogenic fungi (M. brunneum) than H. virescens. Results of additional experiments with H. virescens larvae, B. bassiana, and artificial diet were comparable to experiments with M. brunneum and cotton leaves, despite that susceptibility to B. bassiana was three orders of magnitude lower than to M. brunneum. The fact that both fungus species showed reduced growth on medium under fluctuating temperatures when compared to constant temperatures did not translate to an interaction of temperature regime and the virulence against noctuid larvae. Our study implies that virulence studies with noctuid larvae under constant temperatures using plant material or artificial diet might be fair models also for environments with fluctuating temperatures.

Transposable Elements Shape the Genome Diversity and the Evolution of Noctuidae Species.

Noctuidae is known to have high species diversity, although the genomic diversity of Noctuidae species has yet to be studied extensively. Investigation of transposable elements (TEs) in this family can improve our understanding of the genomic diversity of Noctuidae. In this study, we annotated and characterized genome-wide TEs in ten noctuid species belonging to seven genera. With multiple annotation pipelines, we constructed a consensus sequence library containing 1038-2826 TE consensus. The genome content of TEs showed high variation in the ten Noctuidae genomes, ranging from 11.3% to 45.0%. The relatedness analysis indicated that the TE content, especially the content of LINEs and DNA transposons, is positively correlated with the genome size (r = 0.86, p-value = 0.001). We identified SINE/B2 as a lineage-specific subfamily in Trichoplusia ni, a species-specific expansion of the LTR/Gypsy subfamily in Spodoptera exigua, and a recent expansion of SINE/5S subfamily in Busseola fusca. We further revealed that of the four TE classes, only LINEs showed phylogenetic signals with high confidence. We also examined how the expansion of TEs contributed to the evolution of noctuid genomes. Moreover, we identified 56 horizontal transfer TE (HTT) events among the ten noctuid species and at least three HTT events between the nine Noctuidae species and 11 non-noctuid arthropods. One of the HTT events of a Gypsy transposon might have caused the recent expansion of the Gypsy subfamily in the S. exigua genome. By determining the TE content, dynamics, and HTT events in the Noctuidae genomes, our study emphasized that TE activities and HTT events substantially impacted the Noctuidae genome evolution.

Renewing and updating of the scientific and common names of the most noctuid species (Lepidoptera: Noctuidae) in Egypt

Renewing and updating of the scientific and common names of the most noctuid species (Lepidoptera: Noctuidae) in Egypt

Potential host range of Cotesia vanessae (Hymenoptera: Braconidae), a parasitoid new to North America and a possible biological control agent of noctuid pest species.

The likelihood of parasitoids establishing in new geographic regions depends upon the availability of suitable host species. Identifying these hosts and the degree of their suitability is particularly important when they include species that are economically important as pests. In laboratory studies, we examined the suitability of 47 species of Lepidoptera as potential hosts of a parthenogenetic strain of the gregarious parasitoid Cotesia vanessae (Hymenoptera: Braconidae). Previously known from Eurasia and northern Africa, the first known recovery of C. vanessae in North America was in 2009. C. vanessae completed development in 34 species, of which three were known hosts (Noctuidae) and 31 (30 Noctuidae, 1 Nymphalidae) were not. Many of these noctuid species are economic pests. Parasitoid fitness was generally highest on species of Plusiinae (Noctuidae), measured as either percentage of successful parasitism, developmental time, or number and mass of F1 progeny. Closely related species were generally similar in their suitability as hosts. In some cases, parasitoid eggs or larvae were killed by the immune system of the parasitized host, but the host eventually failed to excrete food waste, did not pupate, and ultimately died. Such cases reached up to 100% mortality depending upon the lepidopteran species. The suitability of many species of noctuid pests as hosts for C. vanessae suggests that this parasitoid will become established widely throughout North America and may help to suppress populations of some pest species.

Pollen and nectar have different effects on the development and reproduction of noctuid moths

Although many noctuid insects are agricultural pests that threaten food production, they are also the major nocturnal pollinators of flowering plants. Larval foods of noctuid pest insects have been well studied for developing control strategies, but knowledge on host plants for the adults is rather scarce. Here, the impact of plant-derived foods on adult survival, fecundity and reproductive physiology of four global species of noctuid pests (Mythimna separata Walker, Mythimna loreyi Duponchel, Athetis lepigone Möschler, and Hadula trifolii Hufnagel) was assessed in laboratory experiments. Our results indicated that nectar slowed testis decay and prolonged the oviposition period and lifespan, increasing fecundity. Acacia nectar increased the longevity of male and female adults by 3.2∼10.9 and 2.4∼5.0 days, respectively, and fecundity of females by 1.22∼3.34 times compared to water-fed individuals. The fitness among the different species of noctuid moths differed on specific pollen diets. On pine pollen, the fecundity of female moths of M. separata, A. lepigone and H. trifolii was 10.06, 33.52, and 28.61%, respectively, lower than those of the water-fed females, but the fecundity of female moths of M. loreyi on pine pollen was 2.11 times greater than for the water-fed individuals. This work provides valuable information on the nutritional ecology for noctuid moths, which can aid the development and design of nutritional attractants within noctuid pests-infected cropping systems and provide a basis for effective and targeted management of global noctuid pests.

Identification of the Sex Pheromone of the Pink Grass Worm, Tmetolophota atristriga, Reveals Possible Population Differences in Male Response to Sex Pheromone

The pink grass worm, Tmetolophota atristriga (Walker), is an endemic New Zealand noctuid moth species that is abundant throughout the North and South Islands. The larvae are minor defoliators of agricultural pasture. We investigated the sex pheromone of this species. Analysis of extract of the female sex pheromone gland identified six compounds: two monounsaturated compounds, (Z)-11-hexadecenal (Z11-16:Ald) and (Z)-11-hexadecenyl acetate (Z11-16:Ac), three saturated compounds, hexadecanal (16:Ald), hexadecyl acetate (16:Ac) and octadecan-1-ol (18:OH), and a triene hydrocarbon, (3Z,6Z,9Z)-tricosatriene (Z3Z6Z9-23:Hy). Several field-trapping experiments testing combinations of the six compounds were conducted. Results suggested that males of two different populations of T. atristriga responded differently to different blends of the compounds. Males of one population responded equally to a two-component blend as to other blends, including the one with all six compounds. By contrast, males of the second population responded only to the six-component blend or a ternary blend of Z11-16:Ald, Z11-16:Ac and Z3Z6Z9-23:Hy. In experiments testing different doses of Z11-16:Ald and Z11-16:Ac in a binary or a six-component blend, a 1 mg dose of the binary blend gave the greatest male catch for both populations. This is the second sex pheromone identification of a New Zealand species of Noctuidae and is the first reported occurrence of Z3Z6Z9-23:Hy as a sex pheromone component of any noctuid species.

Validation of reference-assisted assembly using existing and novel Heliothine genomes

Chloridea subflexa and Chloridea virescens are a pair of closely related noctuid species exhibiting pheromone-based sexual isolation and divergent host plant preferences. We produced a novel Illumina short read C. subflexa genome assembly and an improved C. virescens genome assembly, which offer opportunities to study the genomic basis for evolutionarily important traits in this lepidopteran family with few genomic resources. We then examined the feasibility of reference-assisted assembly, an approach that leverages existing high quality genomic resources for genome improvement in closely related taxa and applied it to our Heliothine genomes. Our work demonstrates that reference-assisted assembly has the potential to enhance contiguity and completeness of existing insect genomic resources with minimal additional laboratory costs. We conclude by discussing both the potential and pitfalls of reference-assisted assembly according to the intended downstream assembly application.

Spodoptera littoralis genome mining brings insights on the dynamic of expansion of gustatory receptors in polyphagous noctuidae.

The bitter taste, triggered via gustatory receptors, serves as an important natural defense against the ingestion of poisonous foods in animals, and the increased host breadth is usually linked to an increase in the number of gustatory receptor genes. This has been especially observed in polyphagous insect species, such as noctuid species from the Spodoptera genus. However, the dynamic and physical mechanisms leading to these gene expansions and the evolutionary pressures behind them remain elusive. Among major drivers of genome dynamics are the transposable elements but, surprisingly, their potential role in insect gustatory receptor expansion has not been considered yet. In this work, we hypothesized that transposable elements and possibly positive selection would be involved in the highly dynamic evolution of gustatory receptor in Spodoptera spp. We first sequenced de novo the full 465 Mb genome of S. littoralis, and manually annotated the main chemosensory genes, including a large repertoire of 373 gustatory receptor genes (including 19 pseudogenes). We also improved the completeness of S. frugiperda and S. litura gustatory receptor gene repertoires. Then, we annotated transposable elements and revealed that a particular category of class I retrotransposons, the SINE transposons, was significantly enriched in the vicinity of gustatory receptor gene clusters, suggesting a transposon-mediated mechanism for the formation of these clusters. Selection pressure analyses indicated that positive selection within the gustatory receptor gene family is cryptic, only 7 receptors being identified as positively selected. Altogether, our data provide a new good quality Spodoptera genome, pinpoint interesting gustatory receptor candidates for further functional studies and bring valuable genomic information on the mechanisms of gustatory receptor expansions in polyphagous insect species.

Development of Multiplex PCR-based Protocols for Simultaneous Caterpillar Diagnosis of Three Spodoptera and One Mamestra Species (Lepidoptera: Noctuidae).

Since many noctuid moth species are highly destructive crop pests, it is essential to establish proper management strategies, which primarily require accurate and rapid species identification. However, diagnosis of noctuid species in the field, particularly at the larval stage, is very difficult due to their morphological similarity and individual color variation. In particular, caterpillars of Spodoptera exigua (Hübner), Spodoptera litura (Fabricius), Spodoptera frugiperda (Smith), and Mamestra brassicae (L.) (Lepidoptera: Noctuidae) are hard to be identified by morphology and frequently found on the same host crops in the same season, thus requiring a reliable species diagnosis method. To efficiently diagnose these species, we identified species-specific internal transcribed spacer 1 (ITS1) sequences and developed two molecular species diagnosis protocols using ITS1 markers. The first protocol was multiplex conventional PCR in conjunction with subsequent gel electrophoresis for species identification based on amplicon size. The second protocol was based on multiplex real-time PCR using fluorescent dye-labeled primers for single-step diagnosis. Template genomic DNA (gDNA) prepared by the DNA release method was also suitable for both protocols as the template prepared by DNA extraction. The two protocols enabled rapid and robust species diagnosis using a single multiplex PCR step. Depending on laboratory instrumentation, one of the two protocols can be easily adapted for species diagnosis of the four noctuid caterpillars in the field, which is essential for establishing proper management strategies. The multiplex real-time PCR protocol, in particular, will facilitate accurate diagnosis of the four species in a single step regardless of template gDNA quality.

New and rare noctuid species (Lepidoptera, Noctuidae) in the fauna of the Republic of Moldova

Data on 17 rare species of noctuids (Noctuidae) in the fauna of the Republic of Moldova are given in the present paper. The investigations were carried out during 2012-2020 in 26 sites throughout the Republic of Moldova. The genus Actebia Stephens, 1829 and the species Actebia praecox (Linnaeus, 1758) and Xylena solidaginis (Hübner, 1803) are reported as new for the fauna of the Republic of Moldova. Thus, the number of noctuid species in the fauna of the Republic of Moldova reached 427. The species: Acontia titania (Esper, 1798), Aedophron rhodites (Eversmann, 1851), Periphanes delphinii (Linnaeus, 1758), Eucarta amethystina (Hübner, 1803), Dasypolia temples (Thunberg, 1792), Oxytripia orbiculosa (Esper, 1799), Euxoa cos (Hübner, 1824), Gortyna cervago (Eversmann, 1844), Meganephria bimaculosa (Linnaeus, 1758) and Cucullia argentea (Hufnagel, 1766) needs protection and conservation, so it is recommended according to IUCN criteria to be included in the next edition of the Red Book of the Republic of Moldova and the elaboration of concrete management measures.

Modern groups of insecticides: diamides and meta-diamides

The review summarizes and analyzes the data of foreign authors on the practical efficacy, mechanisms of action and insect resistance concerning insecticides of the groups of diamides and meta-diamides. The prospects of their application in Russia in agriculture and medical disinsection are considered. Insects resistant to OР’s, carbamates, pyrethroids remain the susceptibility to diamides and meta-diamides. Broflanilide, a pro-insecticide, which, due to its transformation into desmethylbroflanilide in the body of arthropods, acquires physicochemical properties leading to improved lipophilicity, water solubility, stability, affecting systemic activity, slowing down the action on harmful insects and increased selectivity for non-target species, is considered in detail. In several countries of the world where diamides are widely used in the controlling agricultural pests, a high resistance of several species of noctuids, diamondback moth, pyralid moths, tomato leafminer, etc. has been established. The key factors that determined the resistance to diamide in Thailand were the lack of insecticide rotation, minimal crop rotation, insufficient insecticide dosage, and irrigation. The necessity of introducing diamides and meta-diamides into the insecticide rotation schemes in order to control the insects resistant to traditionally used active substances was revealed.

Pheromone antagonism in Plu tella xylostella (Linnaeus) by sex pheromones of two sympatric noctuid moths.

Responses to sex pheromones are commonly antagonized by pheromone components of closely related species. Pheromone antagonism has not been widely explored for phylogenetically distant species that have completely different pheromone components. Yet, pheromone components of sympatrically occurring species may also interfere with each other even if these species are distantly related. Here, the effects of heterospecific pheromones on electrophysiology (electroantennogram, EAG) and behavioral responses were tested on the diamondback moth Plutella xyloslella (Plutellidae) and two sympatric noctuid moth species, Spodoptera litura and Spodoptera exigua, whose larvae also feed on Brassica crops. The sex pheromone blend of P. xyloslella, and its components, did not elicit EAG responses in males of the two noctuid species, while sex pheromone components of the noctuid moths elicited significant EAG responses in P. xyloslella males. In wind tunnel bioassays, both (Z, E)-9, 12-tetradecadienyl acetate (ZE-9,12-14:OAc) and (Z, E)-9, 11-tetradecadienyl acetate (ZE-9,11-14:OAc), sex pheromone components from the noctuid moths, inhibited the upwind flight behavior of P. xyloslella males toward an intraspecific pheromone odor source. In Brassica fields, sex pheromone lures of P. xyloslella did not influence trap catches of the noctuid moths, while P. xyloslella pheromone lures baited with either ZE-9,11-14:OAc or ZE-9,12-14:OAc decreased trap catches of P. xyloslella males in a dose-dependent manner. Trap catches of P. xylostella males were also affected by the proximity of ZE-9,11-14:OAc or ZE-9,12-14:OAc to P. xylostella lures. The uni-directional pheromone antagonism by ZE-9,11-14:OAc and ZE-9,12-14:OAc suggests innovative semiochemical-based strategies for the management of P. xyloslella and other economically important pests in Brassica fields. © 2021 Society of Chemical Industry.

A Guide for Using Flight Simulators to Study the Sensory Basis of Long-Distance Migration in Insects.

Studying the routes flown by long-distance migratory insects comes with the obvious challenge that the animal’s body size and weight is comparably low. This makes it difficult to attach relatively heavy transmitters to these insects in order to monitor their migratory routes (as has been done for instance in several species of migratory birds. However, the rather delicate anatomy of insects can be advantageous for testing their capacity to orient with respect to putative compass cues during indoor experiments under controlled conditions. Almost 20 years ago, Barrie Frost and Henrik Mouritsen developed a flight simulator which enabled them to monitor the heading directions of tethered migratory Monarch butterflies, both indoors and outdoors. The design described in the original paper has been used in many follow-up studies to describe the orientation capacities of mainly diurnal lepidopteran species. Here we present a modification of this flight simulator design that enables studies of nocturnal long-distance migration in moths while allowing controlled magnetic, visual and mechanosensory stimulation. This modified flight simulator has so far been successfully used to study the sensory basis of migration in two European and one Australian migratory noctuid species.

Alypiodes flavilinguis Grote, 1883 as a Valid Species (Noctuidae, Agaristinae)

Alypiodes Grote currently includes four recognized species of day-flying agaristine noctuids from Mexico and the American Southwest. Here we add a fifth, elevating the name Alypiodes flavilinguisGrote 1883rev. status from synonymy. DNA barcodes, larval characters, and adult markings that reliably differentiate Alypiodes flavilinguis from Alypiodes bimaculata and other species in the genus are enumerated. Adults, including their male and female genitalia, and the larvae are illustrated for both species.