Heterospecific sperm reduction in interspecific crosses between two closely related phytophagous ladybird beetles,Henosepilachna vigintioctomaculata and H. pustulosa (Coleoptera: Coccinellidae)

In interspecific crosses, a mismatch in internal physiological conditions between two species can reduce sperm viability in the interval from insemination to fertilization, leading to gametic isolation. Two closely related Japanese phytophagous ladybird beetles, Henosepilachna vigintioctomaculata and H. pustulosa, show several isolating barriers, including reduction in the number of heterospecific sperm in the female reproductive tract and low egg‐hatching rates in interspecific matings. However, the mechanisms of these two potential isolating barriers and the association between them are unknown. Here we investigated temporal changes in the number of sperm stored in the female reproductive tract and egg‐hatching rates in inter‐ and intraspecific crosses between these species. Although the number of sperm decreased after both inter‐ and intraspecific crosses, the reduction was more drastic in inter‐ than in intraspecific crosses for females of both species. Most of the sperm reduction occurred early on, during sperm transfer from the bursa copulatrix to the paired ampullae of the common oviduct (the sperm storage organs). These two species also demonstrated stably low egg‐hatching rates in interspecific crosses. Since the degree and timing of the sperm reduction did not correlate with egg‐hatching rates, the reduction in heterospecific sperm in interspecific crosses may not directly cause the low hatching rates. These two isolating barriers could be different expressions of the physiological mismatch and/or genetic incompatibility between gametes of these species.     

Molecular population genetics of a host‐associated sibling species complex of phytophagous ladybird beetles (Coleoptera: Coccinellidae: Epilachninae)

To clarify evolutionary relationships and assess the intensity and persistence of reproductive isolation because of host fidelity in three closely related host‐associated phytophagous ladybird beetles (Henosepilachna pustulosa, Henosepilachna niponica and Henosepilachna yasutomii), we determined sequences of part of the nuclear DNA internal transcribed spacer‐II (ITS‐2) region (388 bp long) for 70 individuals from seven populations, and part of the mitochondrial cytochrome c oxidase subunit I (COI) gene (985 bp long) for 280 individuals from 28 populations across the three species. We failed to detect any polymorphic sites in the ITS‐2 region. We detected 59 COI haplotypes, among which two were shared by different species. While a haplotype network and a phylogenetic gene tree of the COI haplotypes did not support monophyly for any of the three species, the results of AMOVA did not invalidate the classification of these beetles into three species. The isolation‐with‐migration analytic (IMa) test suggested that gene flow between allopatric species (i.e., H. pustulosa versus H. niponica; H. pustulosa versus H. yasutomii) was well prevented, probably because of their limited dispersal power. On the other hand, the IMa test detected a low level of unidirectional gene flow between the sympatric species H. niponica and H. yasutomii, from the former to the latter. This result was consistent with a survival rate of F1 hybrids between the two species that is higher on the host plant of H. yasutomii than on the host plant of H. niponica. High F_ST values among two sympatric and three nearly sympatric population pairs of H. niponica and H. yasutomii, however, indicated that gene flow between the two species has been quite restricted even if it has occurred. Because no reproductive barriers other than the difference in host plants is known for sympatric H. niponica and H. yasutomii, our results suggest that host differentiation and associated ecological divergence function as an effective reproductive barrier between the two species.     

Differentiation in the ability to utilize Pterostyrax hispida (Ebenales: Styracaceae) among four local populations of the phytophagous ladybird beetle Henosepilachna yasutomii (Coleoptera: Coccinellidae)

The oligophagous ladybird beetle Henosepilachna yasutomii Katakura (Coleoptera: Coccinellidae) shows inter-population differences in its host-use. In this beetle, populations using the deciduous tree Pterostyrax hispida Sieb. et Zucc. (Styracaceae) were recently rediscovered in the Kanto districts of central Honshu, Japan. In the present study, the ability to utilize P. hispida and other host plants was compared among four populations of H. yasutomii occurring on P. hispida and Scopolia japonica Maxim. (Solanaceae), Chelidonium japonicum Thunb. (Papaveraceae), and Solanum tuberosum L. (Solanaceae). As regards the feeding habits of adult beetles, only the populations occurring on P. hispida accepted this plants leaves, which differed distinctly from the feeding habits of the other populations, although the differences among the four populations were not readily apparent with respect to the leaf amounts consumed. The larvae from the populations occurring on P. hispida showed significantly higher survivorship on this plant than did the larvae from the other populations. Considering the host use patterns and the life cycles of beetles under natural conditions, the large abundance of P. hispida leaves throughout the season may have played an important role in selection for the ability to utilize P. hispida observed in the H. yasutomii populations occurring on this woody host.     

Variable correspondence of female host preference and larval performance in a phytophagous ladybird beetle Epilachna pustulosa (Coleoptera: Coccinellidae)

Genetic and environmental factors causing intraspecific variation of the thistle Cirsium kamtschaticum Ledeb. as a host plant of the phytophagous ladybird Epilachna pustulosa Kôno were investigated through simple food-choice tests and rearing of larvae. Two thistle clones (T1U₁ and T4H₂) were used, originally growing approximately 12 km apart. A previous study showed that adult female ladybirds preferred T1U₁ to T4H₂, and that larval performance was better on T1U₁, when leaves from the clones in situ were examined. The two clones retained their characteristics with respect to beetle preference after transplantation into a common garden. However, the difference between T1U₁ and T4H₂ with respect to larval performance was reduced after the transplantation. When leaves from shoots of T1U₁ exposed to different sunlight intensities were offered, adult female ladybirds did not show obvious preferences. Larval eclosion rates increased significantly with the increase in leaf sunlight intensity exposure. These results suggest strongly that both genetic and environmental factors are involved in interclonal variation of thistle quality in beetle preference and/or performance. It is suggested that the quality of thistle leaves for larval performance is largely affected by environmental factors, while leaf quality for beetle preference may be determined strictly by genetic factors. Under certain conditions, E. pustulosa females may behave maladaptively, preferring plants not appropriate for larval growth, or not choosing plants appropriate for the larval growth.     

Asymmetric local adaptation to host plants (Cirsium spp.) by the phytophagous ladybird beetle Henosepilachna niponica and its implications

Divergent host specialization by phytophagous insects is often detected as local adaptation and is thought to have played an important role in their diversification even within an ecological specialist. The phytophagous ladybird beetle Henosepilachna niponica Lewis (Coleoptera: Coccinellidae) predominantly depends on thistles (Cirsium spp., Asteraceae). The distribution of H. niponica occupies multiple areas dominated by different thistle species. This implies the possibility of the occurrence of host-associated divergent specialization of H. niponica. In this study, we investigated the pattern of host-use ability of three allopatric H. niponica populations (Aomori, Iwate, and Yamagata) on three thistle species – Cirsium alpicola Nakai, Cirsium nipponicum (Maxim.) Makino, and Cirsium tonense Nakai – under laboratory conditions. The results displayed asymmetric local adaptation by the beetles. The adults and larvae of the Aomori population showed sufficient acceptance and performance on C. nipponicum and C. tonense, the hosts of the Iwate and Yamagata populations, respectively. On the other hand, the Iwate and Yamagata populations fed small amounts of and performed poorly on C. alpicola, the host of the Aomori population. In contrast, the adults from all the populations clearly preferred feeding on C. nipponicum or C. tonense to C. alpicola. We concluded that the small but significant population differentiation and asymmetric local adaptation by beetles to congeneric host plant species could be a sign of the earliest stage of population divergence by divergent natural selection, given that these divergences will act as ‘immigrant inviability’ and ‘habitat isolation’ at least asymmetrically when these populations come into contact.     

C-value estimates for 31 species of ladybird beetles (Coleoptera: Coccinellidae)

This study provides C-value (haploid nuclear DNA content) estimates for 31 species of ladybird beetles (representing 6 subfamilies and 8 tribes), the first such data for the family Coccinellidae. Despite their unparalleled diversity, the Coleoptera have been very poorly studied in terms of genome size variation, such that even this relatively modest sample of species makes the Coccinellidae the third best studied family of beetles, behind the Tenebrionidae and Chrysomelidae. The present study provides a comparison of patterns of genome size variation with these two relatively well-studied families. No correlation was found between genome size and body size in the ladybirds, in contrast to some other invertebrate groups but in keeping with findings for other beetle families. However, there is some indication that developmental time and/or feeding ecology is related to genome size in this group. Some phylogenetic patterns and possible associations with subgenomic features are also discussed.     

A host shift from wild blue cohosh to cultivated potato by the phytophagous ladybird beetle, Epilachna yasutomii (Coleoptera, Coccinellidae)

Life history traits of the phytophagous ladybird beetle Epilachna yasutomii were compared between a nonpest population feeding on wild blue cohosh and a pest population feeding on cultivated solanaceous crops, mainly potato. Newly emerged adults of the nonpest population entered diapause early in midsummer when blue cohosh withered, while adults of the pest population were found in tomato and eggplant fields until late autumn. The pest population had larger females, a higher population growth rate, a shorter larval developmental period, and reduced longevity of overwintered females, compared with the nonpest population. ANOVA indicated that all these life history traits were influenced by the food plant, and that the number of eggs laid per female and the longevity of overwintered females were also affected by the population type. These findings suggest that the life history pattern of E. yasutomii changed to high fecundity with a short life span from low fecundity with a long life span as a result of the host shift from wild blue cohosh to cultivated solanaceous crops. Received: May 22, 1998 / Accepted: January 13, 1999     

Host plant suitability of a recently naturalized thistle Cirsium vulgare (Asteraceae) for a phytophagous ladybird beetle,Epilachna pustulosa (Coleoptera: Coccinellidae)

The phytophagous ladybird beetle, Epilachna pustulosa Kôno occurs mainly on the thistle Cirsium kamtschaticum Ledeb. In recent years, we have confirmed feeding and oviposition by E.pustulosa, and some degree of larval development of this beetle, on a naturalized congeneric thistle, Cirsium vulgare (Savi) Tenore. In this study, we assessed the host suitability of C.vulgare for E.pustulosa, using larval rearing and simple food choice tests, by comparing the performance on, and preference for, their legitimate host C.kamtschaticum. All experiments were initiated in July, when the utilization of C.vulgare by E.pustulosa became obvious. For larval performance, E.pustulosa showed a lower eclosion rate, shorter developmental duration and smaller body size when reared on C.vulgare than on C.kamtschaticum. Cirsium vulgare appeared to be a less adequate larval food than C.kamtschaticum. Adult feeding preference was altered seasonally; females preferred C.kamtschaticum to C.vulgare in a food choice test conducted in early July, whereas they showed the opposite preference in tests conducted in mid and late July. The utilization of C.vulgare by E.pustulosa appeared to be induced by qualitative deterioration of C.kamtschaticum leaves in July, which resulted in increased dispersion of adult beetles.     

Genome size evolution and polyploidy in the Daphnia pulex complex (Cladocera: Daphniidae)

Genome size was estimated in 49 clones of the Daphnia pulex complex from temperate and subarctic locations using flow cytometry and microsatellite DNA analyses. Significant genome size differences were found in diploid species belonging to the two genetically distinct groups (the pulicaria and the tenebrosa groups), with clones from the tenebrosa group having genome sizes 22% larger than those in the pulicaria group. Combined flow cytometry and microsatellite DNA analyses revealed that nearly all polyploid clones in the D. pulex complex are triploid and not tetraploid, as was previously suggested. Sequencing analyses of the ND5 gene to position clones in their respective clades within the D. pulex complex have uncovered three triploid clones of Daphnia middendorffiana with a D. pulex maternal parent. This result was unexpected because Daphnia pulicaria has always been identified as the maternal parent of these hybrid polyploid clones. Triploid clones likely owe their origins to interactions between sexual and asexual populations. Further interactions in the tenebrosa group have generated tetraploid clones but these events have been rare.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 97 , 68–79.     

Individual variation in two host plants of the ladybird beetle,Epilachna pustulosa (Coleoptera: Coccinellidae)

Individual variation in two species of host plants (thistle,Cirsium kamtschaticum, and blue cohosh,Caulophyllum robustum) of the herbivorous ladybird beetleEpilachna pustulosa was examined under laboratory conditions for their acceptability to adult beetles as a food resource, for adult preference and for larval performance. When clones of these plants were subjected to non-choice tests using posthibernating female beetles, there was found to be significant intraspecific variation among clones in terms of their acceptability, but interspecific variation was not detected. Significant intraspecific as well as interspecific variation were frequently detected in the two host plants when clones of these plants were subjected to choice tests using posthibernating female beetles; the magnitude of interspecific plant variation for beetle preference is not necessarily larger than that of intraspecific plant variation. Individual variation across plant species with respect to beetle larval performance was also significant. A positive correlation between adult preference and larval performance is suggested across the two taxonomically remote host plant species, thistle and blue cohosh, although this needs further investigation.     

Extremely divergent host plant acceptance with gene flow between sympatric host races of the phytophagous ladybird beetle Henosepilachna diekei

Ecological divergence alone can prevent the majority of gene flow in the absence of other forms of reproductive isolation. Although the importance of ecological divergence in promoting reproductive isolation has been broadly recognized, its net impact on speciation has rarely been estimated in the wild. The phytophagous ladybird beetle Henosepilachna diekei Jadwiszczak & Wegrzynowicz includes two sympatric host races that are reproductively isolated solely by extreme specialization to either of the host plants Mikania micrantha Kunth (Asteraceae) or Leucas lavandulifolia Sm. (Lamiaceae) in West Java, Indonesia. To investigate the impact of differential host use as an isolating barrier, we carried out adult host acceptance tests and molecular population genetic analyses based on mitochondrial ND2 and nucleic ITS2 gene sequences using 13 wild populations of the host races, including four sympatric population pairs. Almost all individuals of these host races persistently accepted only the original host plant. We detected restricted but a degree of gene flow between these host races. A migration event occurred only in very recent time compared to their divergence time, indicating recent secondary contact of these host races in the surveyed area. These results reveal the remarkably large impact of host‐plant shift over almost the entire process of speciation and illustrate that ecological divergence has been maintained even under the presence of a certain degree of gene flow.     

Identification of Henosepilachna vigintioctomaculata (Coleoptera: Coccinellidae) midgut putative receptor for Bacillus thuringiensis insecticidal Cry7Ab3 toxin

Bt WZ-9 strain, containing a single Cry7Ab3 toxin, had effective insecticidal activity against larvae of Henosepilachna vigintioctomaculata. By incubation with larvae midgut homogenate and trypsin in vitro, 130 kDa Cry7Ab3 protoxin was degraded into the ～75 kDa proteinase-resistant fragments. In vivo analysis, 130 kDa Cry7Ab3 protoxin was also processed into ～75 kDa fragment. Histopathological observations indicated that Cry7Ab3 ingestion by H. vigintioctomaculata larvae causes acceleration in the blebbing of the midgut epithelium cells into the gut lumen and eventual lysis of the epithelium cells resulting in larval death. A ligand blotting experiment demonstrated that Cry7Ab3 toxin bound a 220 kDa BBMV protein. This receptor protein was identified as cadherin by matrix assisted laser desorption-time of flight-mass spectrometry (MALDI-TOF-MS). The cadherin protein may be the receptor of Cry7Ab3. The data obtained may contribute to a better understanding of the mechanism of Cry7Ab3 toxin against H. vigintioctomaculata larvae.     

Interference competition between ladybird beetle adults (Coleoptera: Coccinellidae): effects on growth and reproductive capacity

Cage experiments were used to determine the minimum number of Aphis fabae Scopoli that females of the Azorean native ladybird beetle Coccinella undecimpunctata L. require per day to achieve maximum fecundity and to assess the effects of interference competition from conspecific and heterospecific [Harmonia axyridis (Pallas)] adult ladybirds on its growth and reproduction. The number of A. fabae at which females of C. undecimpunctata ate most aphids, achieved maximum reproduction and maintained their body weight, was 200. The presence of H. axyridis adults, unlike the presence of C. undecimpunctata, significantly affected the reproductive numerical response of C. undecimpunctata even though the resource was not in short supply. The results suggest that interference competition with heterospecifics may adversely affect the reproductive capacity of C. undecimpunctata. Based on previous results and those reported here, we suggest that the introduction of H. axyridis could endanger the population abundance and/or result in the competitive displacement of C. undecimpunctata from the Azorean ecosystem.     

Genome size and chromosome number in the New Zealand species of Schoenus (Cyperaceae)

Schoenus (Cyperaceae) has holocentric chromosomes. Chromosome numbers were counted and nuclear DNA amounts were measured for all the New Zealand species of the genus. Chromosome numbers ranged from 2n = 8 to c. 2n = 90. Two chromosome races, with 2n = 28 and 2n = 56, were found in S. pauciflorus. Flow cytometry using propidium iodide‐stained nuclei was used to measure genome size. A 14.8‐fold variation in 2C DNA content was found, with values ranging from 1.33 to 19.71 pg/2C nucleus. Phylogenetic trees based on sequence variation in the internal transcribed spacer (ITS) region of the 45S ribosomal DNA locus were constructed using several phylogenetic models to reveal possible evolutionary relationships among the New Zealand Schoenus spp. and a sample of Australian Schoenus spp. Analysis revealed heterogeneity of chromosome number, size and DNA C value within clades. Meiosis in four species showed only bivalent formation at metaphase I. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 169 , 555–564.     

Antennal sensilla and sexual dimorphism of the adult ladybird beetle Semiadalia undecimnotata Schn. (Coleoptera: Coccinellidae)

The antennal receptors of the adult male and female ladybird beetle, Semiadalia undecimnotata (Coleoptera: Coccinellidae), were examined by scanning and transmission electron microscopy. Twelve types of receptors were characterized and grouped into 5 morphological classes: Böhm, trichoid, coeloconic, basiconic, and chetiform sensilla. Sensory function was determined on the basis of sensillar ultrastructure and electrophysiological response. Olfactory sensilla are confined in both sexes to the 2 terminal antennal segments. In contrast, gustatory and mechanosensitive organs are present along the entire length of the antennae. Sexual dimorphism of antennal receptors is limited to the latter 2 functional classes. The principal characteristics of this dimorphism are the following: a) males possess 540 sensilla (all types), while females possess only 500; b) males exhibit 2 types of taste receptors not present in females; c) females exhibit one type of mechanoreceptor absent in males; d) the 3 sex-specific types of sensilla, which occupy the same position in males and females, are confined to the inner side of the antennae. The possible role of male-specific sensilla in intersexual communication is discussed.     

成虫体型大小驱动六斑月瓢虫性间相互选择(英文)

【目的】配偶选择是性选择的中心过程,当繁殖成本发生波动和配偶质量存在差异时,传统的性别作用也随之不同。为了验证在某些条件下雄性也可以像雌性一样进行选择,我们研究了雄性和雌性六斑月瓢虫Menochilus sexmaculatus(Fabricius)的交配成功率与作为适合度指标的成虫体型大小之间的关系。【方法】从初始种群中随机选择体型小和体型大的两性成虫,进行雄性和雌性配偶选择试验。并对成功交配的各个配对组的后代也进行配偶选择试验,测试其交配成功率。【结果】配偶选择试验结果表明,在雄性和雌性交配试验中交配的成功取决于体型大小。雄性根据其品质即体型大小这一繁殖力指标对雌性进行区分。同样,在雌性交配试验中体型大的雄虫比体型小的雄虫交配成功率高。无论亲本体型如何,体型大的雌虫都比体型小的雌虫产卵量高;不过,体型大小不影响卵的存活力。体型大的亲本的后代比体型小的亲本的后代发育更快,寿命更长,但是将其与来自初始种群的后代进行配偶选择时,它们在性合意(sexual desirability)或竞争力方面没有表现任何优势。【结论】六斑月瓢虫中存在以体型大小为目的性状的相互配偶选择。     

Phylogeny and evolution of phytophagous ladybird beetles (Coleoptera: Coccinellidae: Epilachnini), with recognition of new genera

Phytophagous ladybird beetles of the tribe Epilachnini are a cosmopolitan, species‐rich group of significant economic importance as pests of agricultural crops. The tribe is well characterized morphologically and clearly monophyletic, but very little is known about its internal phylogenetic relationships and their genus‐level taxonomy. In order to infer the evolutionary history of Epilachnini, test its monophyly and provide a phylogeny‐based classification, we assembled a comprehensive dataset, consisting of four DNA markers (18S and 28S rRNA and 16S, COI mtDNA) and a matrix of 104 morphological characters for 153 species of Epilachnini representing all previously recognised genera, ～11% of the known species, and 14 outgroup taxa. Molecular, morphological and combined datasets were analysed using maximum likelihood, parsimony and Bayesian inference. Bayes factors and Approximately Unbiased tests (AU) were used to compare alternative phylogenetic hypotheses of unconstrained and backbone‐constrained analysis. Only 14 of the 25 included genera were recovered monophyletic, as originally defined. Afidentula Kapur, Afidenta Dieke, Afissula Kapur, Epilachna Chevrolat, Henosepilachna Li Toxotoma Weise and Mada Mulsant are shown to be poly‐ or paraphyletic; Chnootriba Chevrolat, Subafissa Bielawski, Lalokia Szawaryn & Tomaszewska and Papuaepilachna Szawaryn & Tomaszewska form monophyletic groups within larger clades of genus level. All of these genera are redefined here. The two largest genera of Epilachnini, Epilachna Chevrolat and Henosepilachna Li were represented by multiple monophyletic clades, which we described as new genera: Chazeauiana Tomaszewska & Szawaryn gen.n. ; Diekeana Tomaszewska & Szawaryn gen.n .; Fuerschia Tomaszewska & Szawaryn gen.n. and Ryszardia Tomaszewska & Szawaryn gen.n . The following new synonyms are proposed: Afissa Dieke (=Afissula Kapur); Henosepilachna Li in Li & Cook (=Subafissa Bielawski); Papuaepilachna Szawaryn & Tomaszewska (=Lalokia Szawaryn & Tomaszewska). This published work has been registered in ZooBank, http://zoobank.org/urn:lsid:zoobank.org:pub:440E7FA4‐C859‐47E0‐8335‐30D478CBA8FA .     

Use of prey and non-prey food by the ladybird beetle Eriopis connexa (Coleoptera: Coccinellidae) under laboratory-rearing conditions

Prey for predators can fluctuate in abundance and in quality over time requiring predator strategies to cope with food shortage. Coccinellinae are often associated with sap-sucking pests that exhibit high population unpredictability such as aphids and psyllids. Eriopis connexa (Germar) (Coleoptera: Coccinellidae) is a predator with potential for biological control, especially a well-studied population which is resistant to pyrethroids used to control insect defoliators. Both larvae and adult E. connexa were provided ad libitum prey and non-prey foods (pollen and honey water solution) at increasing intervals from 1 to 10 days. Neonate larvae of E. connexa required eating prey daily to develop into adults. However, non-prey food such as honey water solution did prolong larval and adult survival but neither fulfilled larval development nor adult reproduction. Honey water solution promoted 100% adult survival up to 25 days in the adult stage without prey with oviposition returning after daily feeding on prey. Females subjected to increased feeding intervals over four days reduced oviposition and lived longer, but 10-day feeding intervals correlated with risk to adult survival. These results indicate the importance of non-food sources in E. connexa maintenance and the ability of larvae and adult females to compensate for prey scarcity.     

Age specific mating incidence and reproductive behavior of the ladybird beetle,Anegleis cardoni (Weise) (Coleoptera: Coccinellidae)

Age specific mating incidence, mating behavior, mate recognition cues and effect of sexual status on mating of ladybird beetle, Anegleis cardoni (Weise) (Coleoptera: Coccinellidae) were studied in detail. The mating duration was very short (14.40 ± 0.72 min) in comparison to other ladybirds. Courtship was male dominated and was displayed in steps, viz. approach, watch, examine, sharp turns, mount, attempt and mate. Sharp turns is an additional behavior observed during courtship in this ladybird, and is characterized by sharp turning movements towards the mate, and is undertaken by both males and females. Active mating of A. cardoni involved multiple strokes of the aedeagus. Mating duration and the number of strokes were higher in unmated males than in mated ones. Results reveal that physical cues probably play a role in the initiation of courtship, while chemical signals are probably involved in the continuation of courtship, culminating in intromission. The removal of male antennae and obscurity of elytral patterns significantly decreased mating success, whereas female antennal ablation and obscuring of the pronotum did not affect mate recognition.