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Intraspecific variation can promote or inhibit species coexistence, both by increasing species’ competitive abilities, and by altering the relative strengths of intraspecific and interspecific competition. Effects of intraspecific variation on coexistence can occur via complementarity of different variants, and via a selection effect: initially-variable populations are more likely to contain highly competitive variants that might determine the ability of the population as a whole to both invade and resist invasion. We tested the effects of intraspecific variation and composition on coexistence by assaying the mutual invasibility of populations of two competing bean weevil species (Callosobruchus maculatus and C. chinensis) when each was initiated with one, three, or five genetically- and phenotypically- distinct lineages. Our results reveal that intraspecific variation is a double-edged sword for species coexistence. Increasing intraspecific variation increased species’ abilities to invade, and to resist invasion, via selection effects and intraspecific niche complementarity among conspecific lineages, thereby creating the potential for exclusion among mismatched competitors. But intraspecific variation also increased the scope for resource partitioning, creating the potential for stable coexistence. Stable coexistence occurred only when intraspecific variation caused species to exhibit both relatively evenly-matched competitive abilities and sufficiently-strong resource partitioning. Our work explains the conflicting results of previous studies.

Traumatic mating (or copulatory wounding) is an extreme form of sexual conflict whereby male genitalia physically harm females during mating. In such species females are expected to evolve counter-adaptations to reduce male-induced harm. Importantly, female counter-adaptations may include both genital and non-genital traits. In this study, we examine evolutionary associations between harmful male genital morphology and female reproductive tract morphology and immune function across 13 populations of the seed beetle Callosobruchus maculatus. We detected positive correlated evolution between the injuriousness of male genitalia and putative female resistance adaptations across populations. Moreover, we found evidence for a negative relationship between female immunity and population productivity, which suggests that investment in female resistance may be costly due to the resource trade-offs that are predicted between immunity and reproduction. Finally, the degree of female tract scarring (harm to females) was greater in those populations with both longer aedeagal spines and a thinner female tract lining. Our results are thus consistent with a sexual arms race, which is only apparent when both male and female traits are taken into account. Importantly, our study provides rare evidence for sexually antagonistic coevolution of male and female traits at the within-species level.

Two bruchid species, Callosobruchus maculatus and Callosobruchus chinensis, are the most significant stored insect pests of tropical legume crops. Previously, we identified a major QTL, qBr6.1, controlling seed resistance to these bruchids in the cultivated zombi pea (Vigna vexillata) accession ‘TVNu 240’. In this study, we have narrowed down the qBr6.1 region and identified a candidate gene conferring this resistance. Fine mapping using F2 and F2:3 populations derived from a cross between TVNu 240 and TVNu 1623 (susceptible) revealed the existence of two tightly linked QTLs, designated qBr6.1-A and qBr6.1-B, within the qBr6.1. The QTLs qBr6.1-A and qBr6.1-B explained 37.46% and 10.63% of bruchid resistance variation, respectively. qBr6.1-A was mapped to a 28.24 kb region containing four genes, from which the gene VvTaXI encoding a xylanase inhibitor was selected as a candidate gene responsible for the resistance associated with the qBr6.1-A. Sequencing and sequence alignment of VvTaXI from TVNu 240 and TVNu 1623 revealed a 1-base-pair insertion/deletion and five single-nucleotide polymorphisms (SNPs) in the 5′ UTR and 11 SNPs in the exon. Alignment of the VvTAXI protein sequences showed five amino acid changes between the TVNu 240 and TVNu 1623 sequences. Altogether, these results demonstrated that the VvTaXI encoding xylanase inhibitor is the candidate gene conferring bruchid resistance in the zombi pea accession TVNu 240. The gene VvTaXI will be useful for the molecular breeding of bruchid resistance in the zombi pea.

1. Reproductive interference is a negative interspecific sexual interaction that adversely affects the fitness of males and females during reproductive process. Theoretical studies suggest that because reproductive interference is characterized by positive frequency dependence it is far more likely to cause species exclusion than the density dependence of resource competition. However, the respective contributions of resource competition and reproductive interference to species exclusion, which have been frequently observed in many competition studies, remain unclear. 2. We show that reproductive interference is a far more critical cause of species exclusion than resource competition in the competition between Callosobruchus bean weevil species. In competition experiments over several generations, we manipulated the initial relative abundance of the adzuki bean beetle, Callosobruchus chinensis, and the southern cowpea beetle, Callosobruchus maculatus. When the initial adult ratio of C. chinensis : C. maculatus were 6 : 2 and 4 : 4, C. chinensis excluded C. maculatus. However, when C. maculatus was four times more abundant than C. chinensis at the start, we observed the opposite outcome. 3. A behavioural experiment using adults of the two species revealed asymmetric reproductive interference. The fecundity and longevity of C. maculatus females, but not those of C. chinensis females, decreased when the females were kept with heterospecific males. Fecundities of females of both species decreased as the number of heterospecific males increased. In contrast, resource competition at the larval stage resulted in higher survival of C. maculatus than of C. chinensis. 4. These results suggest that the positive frequency-dependent effect of reproductive interference resulted in species exclusion, depending on the initial population ratio of the two species, and the asymmetry of the interference resulted in C. chinensis being dominant in this study, as in previous studies. Classical competition studies should be reviewed in light of this evidence for reproductive interference.

Storage pests and the food spoilage they cause are problems of great concern. Using essential oil obtained from different plants as an insecticide against these storage pests can be considered an environmentally friendly pest management option. Lantana camara Linn. (family Verbenaceae) is a flowering species, and is also a noxious weed that can proliferate well in nearly all geographical habitats. A biopesticide derived from the essential oil extracted from this plant can offer an effective solution for controlling storage pests. The goal of this study is to extract and analyse the chemical composition of essential oil obtained from L. camara leaves, and assess its effectiveness as a bioactive substance against three storage pests: Tribolium castaneum, Lasioderma serricorne, and Callosobruchus chinensis. The yield of essential oil extracted from L. camara leaves was about 0.24 ± 0.014%. By employing the GC-MS technique, the major phytochemicals contained in L. camara leaf essential oil were identified as caryophyllene (69.96%), isoledene (12%), and ɑ-copaene (4.11%). The essential oil exhibited excellent fumigant toxicity (LC50 of 16.70 mg/L air for T. castaneum, 4.141 mg/L air for L. serricorne and 6.245 mg/L air for C. chinensis at 24 h), contact toxicity (LC50 of 8.93 mg/cm2 for T. castaneum, 4.82 mg/cm2 for L. serricorne and 6.24 mg/cm2 for C. chinensis after 24 h) along with effective repellent activity towards the test insects. In addition, the oil showed no significant phytotoxicity on the germination of paddy seeds. This presents the potential to utilize a weed in developing a biopesticide for effectively managing stored product insects because of its strong bioactivity.

Key message The Br locus confers bruchid resistance in mungbean; VrPGIP2 (encoding a polygalacturonase inhibitor) is a strong candidate gene for this resistance. The VrPGIP2 sequence differs between resistant and susceptible lines. Azuki bean weevil ( Callosobruchus chinensis ) and cowpea weevil ( Callosobruchus maculatus ) are serious insect pests of mungbean during storage. Bruchid resistance in mungbean is controlled by a single dominant locus, Br . Although the Br locus has been located on a genetic map, molecular basis and function of the gene remain unknown. In this study, high-resolution mapping using a BC 11 F 2 population of 418 plants derived from a cross between ‘Kamphaeng Saen 1’ (KPS1; susceptible) and ‘V2802’ (resistant) using simple sequence repeat (SSR) markers delimited the Br locus to a genomic region of 38 Kb of chromosome 5 containing two annotated genes. EST-SSR marker DMB-SSR158 co-segregated perfectly with the Br locus. Bioinformatics analyses revealed that DMB-SSR158 corresponds to a gene encoding a polygalacturonase inhibitor (polygalacturonase-inhibiting protein PGIP) and was designated as VrPGIP2 . Comparison of VrPGIP2 coding sequences between four bruchid-resistant (V2802, V1128, V2817 and TC1966) and four bruchid-susceptible (KPS1, Sulu-1, CM and an unknown accession) mungbean lines revealed six single nucleotide polymorphisms (SNPs) between the resistant and susceptible groups. Three of the six SNPs resulted in amino acid changes; namely, alanine (A) to serine (S) at position 320, leucine (L) to proline (P) at position 332, and threonine (T) to P at position 335 of the VrPGIP2 sequence in resistant lines, compared with that in susceptible lines. The A to S change at position 320 may affect the interaction between PGIP and polygalacuronase. These results indicate that VrPGIP2 is very likely the gene at the Br locus responsible for bruchid resistance in mungbean.

ABSTRACT Nowadays, one of the main pests affecting stored seeds is Callosobruchus maculatus, which drastically lowers their quality and marketability. Especially for grains intended for human or animal feed, the conventional chemical insecticides used to manage this pest can carry serious risks. Consequently, the quest for substitute pest management strategies, such as the application of plant extracts and bioactive substances, has received a lot of attention lately. Multiple insect pests can be successfully controlled by bioactive substances found in particular plant components, according to recent research. The purpose of this work was to assess how well-known insecticidal plants Achillea millefolium and Heliotropium bacciferum affected life of C. maculatus cycle and to extract their bioactive components. Part of the components taken from the leaves of A. millefolium and H. bacciferum were identified by GC-Mass spectrometry. Three replicates were examined using ethanol, ethyl acetate, hexane extracts, cold water, hot water, and ethanol at four concentrations: 10, 7.5, 5, and 2.5 mg.mL-1 over 24-, 48-, 72-, and 96-hour periods. The most notable effect on beetle life metrics was found to be shown by H. bacciferum extracts. Mortality rates from adult and egg extracts were especially high at 10% concentration over 96 hours. Egg mortality from a 10% ethanol extract of H. bacciferum, for example, was 74%; egg mortality from ethyl acetate and hexane extracts was 69% and 65.07%, respectively; egg mortality from hot and cold-water extracts was (51%) and (47%). Adult death rates were (84%) for the 10% ethanol extract, (79%), (75%), (59%), and (55%) for the ethyl acetate, hexane, hot water, and cold-water extracts, in that order. Keywords: A. millefolium, H. bacciferum, Callosobruchus maculatus, plant extraction, phytochemical.

Genetic differentiation can be promoted allopatrically by geographic isolation of populations due to limited dispersal ability and diversification over time or sympatrically through, for example, host-race formation. In crop pests, the trading of crops across the world can lead to intermixing of genetically distinct pest populations. However, our understanding of the importance of allopatric and sympatric genetic differentiation in the face of anthropogenic genetic intermixing is limited. Here, we examined global sequence variation in two mitochondrial and one nuclear genes in the seed beetle Callosobruchus maculatus that uses different legumes as hosts. We analyzed 180 samples from 42 populations of this stored bean pest from tropical and subtropical continents and archipelagos: Africa, the Middle East, South and Southeast Asia, Oceania and South America. For the mitochondrial genes, there was weak but significant genetic differentiation across continents/archipelagos. Further, we found pronounced differentiation among subregions within continents/archipelagos both globally and within Africa but not within Asia. We suggest that multiple introductions into Asia and subsequent intermixing within Asia have generated this pattern. The isolation by distance hypothesis was supported globally (with or without continents controlled) but not when host species was restricted to cowpeas Vigna unguiculata, the ancestral host of C. maculatus. We also document significant among-host differentiation both globally and within Asia, but not within Africa. We failed to reject a scenario of a constant population size in the recent past combined with selective neutrality for the mitochondrial genes. We conclude that mitochondrial DNA differentiation is primarily due to geographic isolation within Africa and to multiple invasions by different alleles, followed by host shifts, within Asia. The weak inter-continental differentiation is most likely due to frequent inter-continental gene flow mediated by human crop trade.

Aim: The seed beetle Callosobruchus maculatus is an important tropical and subtropical pest of legumes distributed world-wide. Archaeological evidence suggests an African origin with later world-wide invasion facilitated by the last centuries' legume trading and exchange. To date, no studies could identify the routes or timing of dispersal of the species. Here, we investigate the global phylogeography of this pest to shed light on the main inter-continental dispersal routes that led to it becoming a cosmopolitan pest. Location: World-wide. Methods: We sampled seed beetles over a large fraction of the species' range and sequenced one nuclear and three mitochondrial loci. Using this data, we estimated spatio-temporal phylogeographical reconstructions, and the demographic history of the species. We also used our dataset to evaluate the effect of panmixia on Bayesian demographic estimations. Results: Callosobruchus maculatus exhibited regional and continental genetic structure, with the highest genetic diversity found in Africa. Our discrete Bayesian phylogeographical approach indicated that the species first dispersed to Asia and then colonized the pantropical belt. The three methods used for inferring the demographic history of C. maculatus indicated a recent demographic expansion in the world-wide dataset, as well as in the subset restricted to African samples. Such a signal was, however, not observed in the subset composed of Asian specimens. This demographic expansion occurred in the Holocene and is likely explained by the spread of cowpea and other host legumes across and out of Africa. Main conclusions: The inferred dispersal routes support the idea that the evolutionary history of C. maculatus relates to the trade of its main host plant, Vigna unguiculata. Human-mediated processes appear to have shaped the global genetic structure of this pest. As a methodological implication, we demonstrate that coalescent-based demographic reconstructions can be erroneous if the dataset violates the assumption of panmixia.

Reproductive interference (RI) is a negative interspecific interaction caused by reproductive activities. Previous studies have implied that male-male competition increases RI capability, consisting of the male harm-inflicting capability of RI by males (RI infliction) and the female susceptibility to heterospecific male harm (RI resistance). Geographical populations show variations in the intensity of male-male competition and sexual conflict due to the influence of biotic/abiotic factors. We made two predictions: (1) RI capability would vary among conspecific populations, and (2) the degree of RI infliction by males would match that of female RI resistance in geographical strains. To test these predictions, we measured the fecundity of once-mated females in two scenarios: housed with heterospecific males and without males. We performed experiments using six strains from each of two bean weevil species, Callosobruchus maculatus and C. chinensis. The results demonstrated intraspecific variation in RI capability within both Callosobruchus species, but the RI infliction of males and RI resistance of females within a geographical strain did not consistently coincide. Our results suggest that RI varies depending on geographical strains, even within species, but we did not obtain evidence that sexual conflict molds RI capability, indicating traits driving RI do not follow the sexual conflict in both Callosobruchus species.