Honey bee queen mandibular pheromone inhibits ovary development and fecundity in a fruit fly

A key feature of eusocial insects is their reproductive division of labour. The queen signals her fecundity to her potentially reproductive daughters via a pheromone, which renders them sterile. In contrast, solitary insects lack division in reproductive labour and there is no such social signalling or need for ovary‐regulating pheromones. Nonetheless, females from both non‐social and eusocial lineages are expected to regulate their ovaries to maximize inclusive lifetime reproductive success. It is not known, however, whether the underlying networks that regulate ovary activation are homologous between non‐social and eusocial taxa, especially when these taxa are phylogenetically distant. In this study, we provide evidence that solitary fruit flies may share a conserved ovary‐regulating pathway with a eusocial honey bee, Apis mellifera L. (Hymenoptera: Apidae). Specifically, we demonstrate that honey bee queen mandibular pheromone (QMP) inhibits fly ovaries in much the same way as it suppresses worker ovaries. Drosophila melanogaster Meigen (Diptera: Drosophilidae) exposed to sufficient doses of QMP showed a reduction in ovary size, produced fewer eggs, and generated fewer viable offspring, relative to unexposed controls. Drosophila melanogaster therefore responds to an interspecific social cue to which it would not normally be exposed. Although we cannot strictly rule out an incidental effect, this conspicuous response suggests that these two species may share an underlying mechanism for ovary regulation. Why a non‐social species of fly responds to a highly social bee's pheromone is not clear, but one possibility is that solitary and social insects share pathways associated with female reproduction, as predicted by the ‘groundplan’ hypothesis of social evolution.     

Varying degree of physiological integration among host instars and their endoparasitoid affects stress‐induced mortality

In natural populations of insect herbivores, genetic differentiation is likely to occur due to variation in host plant utilization and selection by the local community of organisms with which they interact. In parasitoids, engaging in intimate associations with their host during immature development, local variation may exist in host quality for parasitoid development. We compared the development of a gregarious endoparasitoid, Cotesia glomerata L. (Hymenoptera: Braconidae), collected in The Netherlands, in three strains and three caterpillar instars (L1–L3) of its main host, Pieris brassicae L. (Lepidoptera: Pieridae). Hosts had been collected in The Netherlands and France, and were reared in the laboratory for one generation. We also used an established Dutch laboratory strain that had not been exposed to parasitoids for at least 24 generations. Parasitoid survival to adulthood was inversely correlated with host instar at parasitism. Adult parasitoid body mass was largest when hosts were parasitized as L1 and smallest when hosts were parasitized as L3, whereas egg‐to‐adult development time was quickest on L3 hosts and slowest on L1 hosts. Higher survival and faster development of C. glomerata on French L2 hosts also showed that there is variation in host‐instar‐related suitability. Many L2 and most L3 caterpillars that were parasitized exhibited signs of pathogen infection and perished within a few days of parasitism, whereas this never happened when hosts were parasitized as L1 or in non‐parasitized control caterpillars. Our results reveal that, irrespective of the host strain, L1 hosts are optimally synchronized with C. glomerata development. By contrast, the high precocious mortality of L3 larvae may be due to stress‐induced regulation by the parasitoid in order to ‘force’ its developmental program into synchrony with the developing parasitoid larvae. Our results underscore a potentially important role played by pathogens in mediating herbivore–parasitoid interactions that are host‐instar‐dependent in their expression.     

In vitro rearing of Campoletis sonorensis, a larval endoparasitoid of Heliothis virescens from egg to third instar in an artificial medium devoid of insect sources

Campoletis sonorensis is a solitary larval endoparasitoid of several Heliothis spp. pests. This study was carried out to develop media devoid of insect sources for in vitro rearing the parasitoid. Trehalose, lysine, threonine, asparagine, glutamine, hydroxyproline, serine, bovine serum albumin, and lactalbumin were beneficial to C. sonorensis. Addition of fresh chicken egg yolk at a low level improved the artificial media. Addition of 20-hydroxyecdysone increased the molting rate, reduced the critical size at molt and decreased development time. Parasitoid development in vitro was dependent on eggs that had been in the host for at least 22 h. Luminosity was also critical for the development of C. sonorensis in vitro. Optimal development occurred in a L14:D10 photoperiod at 43 Lux light intensity. Utilizing the best media and conditions, 100% of the parasitoid larvae reached the second instar and over 37% molted to the third instar. However, no further development occurred.     

Food sources for adult Diachasmimorpha longicaudata, a parasitoid of tephritid fruit flies: effects on longevity and fecundity

We report the results of a study on potential food sources of the widely distributed Indo‐Australian braconid fruit fly parasitoid Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae). Adults sustained life on diets of fruit juice or fruit pulp, a homopteran and its associated honeydew, or extrafloral nectary secretions. Longevities on all these foods and fecundity on fruit juice were comparable to those achieved on the honey that is typically provided in mass‐rearing programs. Certain of the flower species Bidens alba (L.), Spermacoce verticillata L., Lobularia maritima (L.) Desv., Brassica nigra (L.), Lantana camara L., their nectar or pollen, provided a diet that resulted in longer maximum life spans than water alone. Unlike some tephritid flies, the braconid did not feed on fresh bird feces or leaf‐surface exudates. Feeding by D. longicaudata on wounded host fruits of tephritid flies suggests that adult parasitoids would not need separate forays for adult food and oviposition sites, as these occur in the same locations. We conclude that an inventory of adult foods may help target inundative releases of D. longicaudata and lead to improvements in diets used for mass rearing.     

Effects of age,sex, and genotype on high‐sensitivity metabolomic profiles in the fruit fly,Drosophila melanogaster

Researchers have used whole‐genome sequencing and gene expression profiling to identify genes associated with age, in the hope of understanding the underlying mechanisms of senescence. But there is a substantial gap from variation in gene sequences and expression levels to variation in age or life expectancy. In an attempt to bridge this gap, here we describe the effects of age, sex, genotype, and their interactions on high‐sensitivity metabolomic profiles in the fruit fly, Drosophila melanogaster. Among the 6800 features analyzed, we found that over one‐quarter of all metabolites were significantly associated with age, sex, genotype, or their interactions, and multivariate analysis shows that individual metabolomic profiles are highly predictive of these traits. Using a metabolomic equivalent of gene set enrichment analysis, we identified numerous metabolic pathways that were enriched among metabolites associated with age, sex, and genotype, including pathways involving sugar and glycerophospholipid metabolism, neurotransmitters, amino acids, and the carnitine shuttle. Our results suggest that high‐sensitivity metabolomic studies have excellent potential not only to reveal mechanisms that lead to senescence, but also to help us understand differences in patterns of aging among genotypes and between males and females.     

Effects of quantitative and qualitative differences in volatiles from host‐ and non‐host‐infested maize on the attraction of the larval parasitoid Cotesia kariyai

Cotesia kariyai Watanabe (Hymenoptera: Braconidae) is a specialist larval parasitoid of Mythimna separata Walker (Lepidoptera: Noctuidae). Cotesia kariyai wasps use herbivore‐induced plant volatiles (HIPVs) to locate hosts. However, complex natural habitats are full of volatiles released by both herbivorous host‐ and non‐host‐infested plants at various levels of intensity. Therefore, the presence of non‐hosts may affect parasitoid decisions while foraging. Here, the host‐finding efficiency of naive C. kariyai from HIPVs influenced by host‐ and non‐host‐infested maize [Zea mays L. (Poaceae)] plants was investigated with a four‐arm olfactometer. Ostrinia furnacalis Guenée (Lepidoptera: Crambidae) was selected as a non‐host species. One unit (1 U) of host‐ or non‐host‐infested plant was prepared by infesting a potted plant with five host or seven non‐host larvae. In two‐choice bioassays, host‐infested plants fed upon by different numbers of larvae, and various units of host‐ and non‐host‐infested plants (infestation units; 1 U, 2 U, and 3 U) were arranged to examine the effects of differences in volatile quantity and quality on the olfactory responses of C. kariyai with the assumption that volatile quantity and quality changes with differences in numbers of insects and plants. Cotesia kariyai was found to perceive quantitative differences in volatiles from host‐infested plants, preferring larger quantities of volatiles from larger numbers of larvae or plants. Also, the parasitoids discriminated between healthy plants, host‐infested plants, and non‐host‐infested plants by recognising volatiles released from those plants. Cotesia kariyai showed a reduced preference for host‐induced volatiles, when larger numbers of non‐host‐infested plants were present. Therefore, quantitative and qualitative differences in volatiles from host‐ and non‐host‐infested plants appear to affect the decision of C. kariyai during host‐habitat searching in multiple tritrophic systems.     

Salicylic and jasmonic acid pathways are necessary for defence against Dickeya solani as revealed by a novel method for Blackleg disease screening of in vitro grown potato

Potato is major crop ensuring food security in Europe, and blackleg disease is increasingly causing losses in yield and during storage. Recently, one blackleg pathogen, Dickeya solani has been shown to be spreading in Northern Europe that causes aggressive disease development. Currently, identification of tolerant commercial potato varieties has been unsuccessful; this is confounded by the complicated etiology of the disease and a strong environmental influence on disease development. There is currently a lack of efficient testing systems. Here, we describe a system for quantification of blackleg symptoms on shoots of sterile in vitro potato plants, which saves time and space compared to greenhouse and existing field assays. We found no evidence for differences in infection between the described in vitro‐based screening method and existing greenhouse assays. This system facilitates efficient screening of blackleg disease response of potato plants independent of other microorganisms and variable environmental conditions. We therefore used the in vitro screening method to increase understanding of plant mechanisms involved in blackleg disease development by analysing disease response of hormone‐ related (salicylic and jasmonic acid) transgenic potato plants. We show that both jasmonic (JA) and salicylic (SA) acid pathways regulate tolerance to blackleg disease in potato, a result unlike previous findings in Arabidopsis defence response to necrotrophic bacteria. We confirm this by showing induction of a SA marker, pathogenesis‐related protein 1 (StPR1), and a JA marker, lipoxygenase (StLOX), in Dickeya solani infected in vitro potato plants. We also observed that tubers of transgenic potato plants were more susceptible to soft rot compared to wild type, suggesting a role for SA and JA pathways in general tolerance to Dickeya.     

Characterization of a novel Drosophila melanogaster cis‐regulatory module that drives gene expression to the larval tracheal system and adult thoracic musculature

In a previous bioinformatics analysis we identified 10 conserved Drosophila melanogaster sequences that reside upstream from protein coding genes (CGs). Here we characterize one of these genomic regions, which constitutes a Drosophila melanogaster cis‐regulatory module (CRM) that we denominate TT‐CRM. The TT‐CRM is 646 bp long and is located in one of the introns of CG32239 and resides about 3,500 bp upstream of CG13711 and about 620 bp upstream of CG12493. Analysis of 646 bp‐lacZ lines revealed that TT‐CRM drives gene expression not only to the larval, prepupal, and pupal tracheal system but also to the adult dorsal longitudinal muscles. The patterns of mRNA expression of the transgene and of the CGs that lie in the vicinity of TT‐CRM were investigated both in dissected trachea and in adult thoraces. Through RT‐qPCR we observed that in the tracheal system the pattern of expression of 646 bp‐lacZ is similar to the pattern of expression of CG32239 and CG13711, whereas in the thoracic muscles 646 bp‐lacZ expression accompanies the expression of CG12493. Together, these results suggest new functions for two previously characterized D. melanogaster genes and also contribute to the initial characterization of a novel CRM that drives a dynamic pattern of expression throughout development.     

Growth,development, and behaviour of the parasitised and unparasitised larvae of a shelter‐building moth and consequences for the resulting koinobiont parasitoid

Most attention to size‐time trade‐offs of insects has focused on herbivore risk, with considerably less attention paid to parasitoids. Here, we focus on parasitoid risk, comparing the fates of unparasitised herbivore hosts and parasitised hosts that protect the parasitoids. Success of a koinobiont parasitoid (host grows after parasitisation) depends on maintaining a delicate balance with its host, thereby ensuring its own survival while the host grows. To evaluate growth rate–mortality rate relationships of host and parasitoid, we compared several aspects of the growth, phenology, and behaviour of unparasitised fern moth [Herpetogramma theseusalis (Walker) (Lepidoptera: Crambidae)] larvae and larvae parasitised by Alabagrus texanus (Cresson) (Hymenoptera: Braconidae), a solitary koinobiont (one parasitoid per host) wasp. Host larvae feed and construct shelters on sensitive fern, Onoclea sensibilis L. (Dryopteridaceae). Alabagrus texanus parasitise early‐instar moths in late summer, which overwinter in their host, emerging in mid‐summer to pupate and eclose. During the autumn following hatching and the immediately following spring, parasitised and unparasitised moth larvae did not differ in size, took similar time to choose between satisfactory and unsatisfactory foods, and built similar shelters. Prior to any other changes noted, more parasitised than unparasitised larvae also died when severely starved. Parasitised larvae subsequently grew less and pupated later than unparasitised ones (small size, slow growth), but consumed similar amounts of food. Although the numerically dominant parasitoid of fern moths, we concluded that A. texanus do not efficiently exploit their hosts.     

In the absence of a “landscape of fear”: How lions,hyenas, and cheetahs coexist

Aggression by top predators can create a “landscape of fear” in which subordinate predators restrict their activity to low‐risk areas or times of day. At large spatial or temporal scales, this can result in the costly loss of access to resources. However, fine‐scale reactive avoidance may minimize the risk of aggressive encounters for subordinate predators while maintaining access to resources, thereby providing a mechanism for coexistence. We investigated fine‐scale spatiotemporal avoidance in a guild of African predators characterized by intense interference competition. Vulnerable to food stealing and direct killing, cheetahs are expected to avoid both larger predators; hyenas are expected to avoid lions. We deployed a grid of 225 camera traps across 1,125 km² in Serengeti National Park, Tanzania, to evaluate concurrent patterns of habitat use by lions, hyenas, cheetahs, and their primary prey. We used hurdle models to evaluate whether smaller species avoided areas preferred by larger species, and we used time‐to‐event models to evaluate fine‐scale temporal avoidance in the hours immediately surrounding top predator activity. We found no evidence of long‐term displacement of subordinate species, even at fine spatial scales. Instead, hyenas and cheetahs were positively associated with lions except in areas with exceptionally high lion use. Hyenas and lions appeared to actively track each, while cheetahs appear to maintain long‐term access to sites with high lion use by actively avoiding those areas just in the hours immediately following lion activity. Our results suggest that cheetahs are able to use patches of preferred habitat by avoiding lions on a moment‐to‐moment basis. Such fine‐scale temporal avoidance is likely to be less costly than long‐term avoidance of preferred areas: This may help explain why cheetahs are able to coexist with lions despite high rates of lion‐inflicted mortality, and highlights reactive avoidance as a general mechanism for predator coexistence.     

Chemical Composition and In vitro Anthelmintic Activity of Extracts of Tagetes patula Against a Multidrug‐Resistant Isolate of Haemonchus contortus

Sheep breeding has suffered economic losses due to parasitism by gastrointestinal nematodes, particularly Haemonchus contortus. The use of natural products, specifically Tagetes patula, has been suggested as an alternative method of combatting this issue. Chemical analyses of the extracts of this species described in the literature report the presence of important classes of secondary metabolites such as thiophenes, flavonoids, alkaloids, and benzofurans, some of which were identified and isolated in this study. The aim of this work was to test the effect of the essential oil (EO) and the ethanolic extract of the aerial parts (Tp_EtOH) of T. patula on eggs and larvae of H. contortus, through an egg hatch test (EHT) and a larval development test (LDT). In the EHT, the EO showed 100% inhibition at 0.75 mg mL^?1 (LC₅₀ = 0.0780 mg mL^?1), and the Tp_EtOH showed 100% inhibition at 100 mg mL^?1 (LC₅₀ = 12.8 mg mL^?1). In the LDT, the EO showed 100% inhibition at 0.375 mg mL^?1 (LC₅₀ = 0.0400 mg mL^?1), and the Tp_EtOH showed 100% inhibition at 1.56 mg mL^?1 (LC₅₀ = 0.340 mg mL^?1). Compared to available literature data, the results presented here suggest that the crude extracts of T. patula have substantial potential for controlling this nematode by interrupting its life cycle and/or preventing it from reaching the infective stage.     

The plant Pontin and Reptin homologues,RuvBL1 and RuvBL2a,colocalize with TERT and TRB proteins in vivo,and participate in telomerase biogenesis

Telomerase maturation and recruitment to telomeres is regulated by several telomerase‐ and telomere‐associated proteins. Among a number of proteins, human Pontin and Reptin play critical roles in telomerase biogenesis. Here we characterized plant orthologues of Pontin and Reptin, RuvBL1 and RuvBL2a, respectively, and show association of Arabidopsis thaliana RuvBL1 (AtRuvBL1) with the catalytic subunit of telomerase (AtTERT) in the nucleolus in vivo. In contrast to mammals, interactions between AtTERT and AtRuvBL proteins in A. thaliana are not direct and they are rather mediated by one of the Arabidopsis thaliana Telomere Repeat Binding (AtTRB) proteins. We further show that plant orthologue of dyskerin, named AtCBF5, is indirectly associated with AtTRB proteins but not with the AtRuvBL proteins in the plant nucleus/nucleolus, and interacts with the Protection of telomere 1 (AtPOT1a) in the nucleolus or cytoplasmic foci. Our genome‐wide phylogenetic analyses identify orthologues in RuvBL protein family within the plant kingdom. Dysfunction of AtRuvBL genes in heterozygous T‐DNA insertion A. thaliana mutants results in reduced telomerase activity and indicate the involvement of AtRuvBL in plant telomerase biogenesis.     

Effect of different diets on digestive enzyme activities,in vitro digestibility,and midgut gland structure in juvenile crayfish,Cherax quadricarinatus

This study investigated the effects of food quality on digestive enzyme activities, in vitro protein digestibility and histological traits of the midgut gland in juvenile crayfish Cherax quadricarinatus. Animals of a wide weight range were fed different diets: two commercial diets with high or low lipid content (high lipid and low lipid, respectively) and were compared with a reference diet (RF) previously formulated for this species. Proteinase, lipase and amylase activities were significantly influenced by diet and weight. Specific trypsin activity was significantly higher for crayfish fed with the HL diet. Trypsin activity depended on diet and weight. Protein digestibility showed that HL was the most digestible diet and RF the least. The weight of the animals did not affect protein digestibility. Structural disorganization, hypertrophy of B‐cells and presence of large vacuoles in R‐cells were mainly observed in juveniles fed with HL, indicative of malnutrition. Thus, our data suggest that the HL diet would not be the most appropriate for C. quadricarinatus, while RF diet would be more convenient for culture of this species.     

Structural,mutagenic and in silico studies of xyloglucan fucosylation in Arabidopsis thaliana suggest a water‐mediated mechanism

The mechanistic underpinnings of the complex process of plant polysaccharide biosynthesis are poorly understood, largely because of the resistance of glycosyltransferase (GT) enzymes to structural characterization. In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2‐fucosyl residues to xyloglucan side chains – a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. We unravel the mechanistic basis for fucosylation by AtFUT1 with a multipronged approach involving protein expression, X‐ray crystallography, mutagenesis experiments and molecular simulations. Mammalian cell culture expressions enable the sufficient production of the enzyme for X‐ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs. The lack of an appropriately positioned active site residue as a catalytic base leads us to propose an atypical water‐mediated fucosylation mechanism facilitated by an H‐bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations.