Maternal effect of a hybrid inviability gene in Tribolium castaneum

The Tribolium castaneum hybrid inviability gene, H, was selectively introgressed into a genetic background lacking H through serial paternal backcrosses. This revealed a poor viability phenotype (partial paralysis and poor control of the limbs, referred to as tremor) not present in the parent strains. Tremor cosegregated with H, but was expressed only when transmitted paternally and only when H was not also present maternally. The inferred maternal, self‐suppressive effect of H may explain nonreciprocal incompatibilty previously observed between H and H‐incompatible strains. This revised version was published online in July 2006 with corrections to the Cover Date.     

赤拟谷盗和杂拟谷盗的生殖隔离研究

赤拟谷盗和杂拟谷盗是两个同域发生的近缘储粮害虫。为明确它们的生殖隔离程度和机制,本研究比较分析了赤拟谷盗和杂拟谷盗雄虫对同种和异种雌虫的交配选择;赤拟谷盗与杂拟谷盗交配后,测定了精子在异种雌体内的存活情况;将赤拟谷盗和杂拟谷盗进行正反杂交,研究其F1代、F2代和回交代杂种是否产生及其雌雄比。结果显示,赤拟谷盗和杂拟谷盗雄虫对同种雌虫的爬跨率和交配率高于其对异种雌虫的爬跨率和交配率,表明赤拟谷盗和杂拟谷盗种间性隔离不完全;赤拟谷盗和杂拟谷盗交配后精子在异种雌体内是存活的,杂交所产的F1代卵为受精卵,说明交配后完成了精子传送和受精过程,表明种间机械隔离和配子隔离不完全;赤拟谷盗和杂拟谷盗的杂种F1代自交和回交产生了F2代和回交代,表明种间不存在杂种不育;一些杂交组合产生的F2代和回交代数量少且存在雌雄性比偏离,表明种间存在部分杂种不活和杂种衰败。本研究明确了赤拟谷盗和杂拟谷盗的生殖隔离机制,这对于揭示它们的物种进化关系有着重要意义。     

Genetic variation and co-variation for fitness between intra-population and inter-population backgrounds in the red flour beetle, Tribolium castaneum

Hybrids from crosses between populations of the flour beetle, Tribolium castaneum, express varying degrees of inviability and morphological abnormalities. The proportion of allopatric population hybrids exhibiting these negative hybrid phenotypes varies widely, from 3% to 100%, depending upon the pair of populations crossed. We crossed three populations and measured two fitness components, fertility and adult offspring numbers from successful crosses, to determine how genes segregating within populations interact in inter-population hybrids to cause the negative phenotypes. With data from crosses of 40 sires from each of three populations to groups of five dams from their own and two divergent populations, we estimated the genetic variance and covariance for breeding value of fitness between the intra- and inter-population backgrounds and the sire × dam population interaction variance. The latter component of the variance in breeding values estimates the change in genic effects between backgrounds owing to epistasis. Interacting genes with a positive effect, prior to fixation, in the sympatric background but a negative effect in the hybrid background cause reproductive incompatibility in the Dobzhansky-Muller speciation model. Thus, the sire × dam population interaction provides a way to measure the progress towards speciation of genetically differentiating populations on a trait by trait basis using inter-population hybrids.     

Response of Tribolium castaneum larvae to synthetic aggregation pheromone

Responses of larval Tribolium castaneum Herbst. to the synthetic adult aggregation pheromone (4,8-dimethyldecanal) were studied. Larvae of T. castaneum were attracted both by contact and vapour of 4,8-dimethyldecanal at concentrations of 1, 5 and 10 g. g^-1 of medium.

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Reaktion von Tribolium castaneum Larven auf synthetisches Aggregationspheromon

Zusammenfassung Die Reaktionen der Larven von T. castaneum auf das kunstliche Aggregations pheromon (4,8-dimethyldecanal) in Versuchen in Petrischalen untersucht. Die Larven wurden angezogen, wenn sie mit Pheromon auf Filterpapier oder mit verdampftem Pheromon in Kontakt kamen. Diese Reaktion wurde bei Konzentrationen von 1, 5 und 10 g · g^-1 Medium festgestellt. Die höchste Konzentration rief die grosste Reaktion hervor. Fruhe Larvenstadien zeigten stärkere Reaktionen als spatere Stadien.

     

An unusual barrier to gene flow: perpetually immature larvae from inter-population crosses in the flour beetle, Tribolium castaneum

We genetically characterize an unusual hybrid incompatibility phenotype manifest in F₁ offspring of crosses between two populations of Tribolium castaneum. Hybrid larvae cease development at the third larval instar, persisting as ‘perpetually immature larvae’ thereafter. Although unable to produce viable adult hybrid offspring with one another, each population produces abundant, fertile hybrids with other populations, indicating a recent origin of the incompatibility and facilitating genetic studies. We mapped the paternal component of the hybrid phenotype to a single region, which exhibits two characteristics common to hybrid incompatibility: marker transmission ratio distortion within crosses and elevated genetic divergence between populations. The incompatible variation and an elevation in between‐population genetic divergence is associated with a region containing the T. castaneum ecdysone receptor homologue, a major regulatory switch, controlling larval moults, pupation and metamorphosis. This contributes to understanding the genetics of speciation in the Coleoptera, one of the most speciose of all arthropod taxa.     

Identification of microsatellite markers in the red flour beetle,Tribolium castaneum

We isolated and characterized microsatellite markers for the red flour beetle Tribolium castaneum, an important model species for studies in various areas of evolutionary biology and ecology. A microsatellite‐enriched genomic library was constructed and screened with single stranded oligonucleotide probes [(CCT)₁₇, (AAT)₁₇ and (CAG)₁₇]. Forty‐five primer pairs were designed of which 19 pairs produced successful amplification. Polymorphism screening involved beetles from five beetle strains and revealed 15 polymorphic and four monomorphic markers. The development of polymorphic microsatellite markers will facilitate future ecological and genetic studies involving T. castaneum beetles.     

Developmental O-glycan profile analysis shows pentasaccharide mucin-type O-glycans are linked with pupation of Tribolium castaneum

Eukaryotic cells can decorate their proteins with carbohydrate structures or glycans, significantly affecting the properties and activities of these proteins. Despite the importance of protein glycosylation in numerous biological processes, our knowledge of this modification in insects is far from complete. While N-glycosylation is the most studied, the study of O-glycans in insects is still very fragmentary and these studies are limited to a specific developmental stage or a specific tissue. In this article, matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS) technology was used to analyze the O-glycan profile for the different developmental stages of egg, larva, pupa, and adult of the red flour beetle Tribolium castaneum, an important insect model and pest worldwide. The results on the O-glycan profile showed that the mucin-type glycans dominate the O-glycome of the red flour beetle. Interestingly, some of the more complex mucin-type O-glycans, such as a tetra- (O-GalNAcGalGlcAGalNAc) and pentasaccharide O-glycan (O-GalNAc(GalGlcA)GalNAcGlcA), were highly abundant during the pupa stage, the intermediate stage between larval and adult stage in holometabolous insects, demonstrating that insect metamorphosis is accompanied with a change in the insect O-glycan profile. Together with the N-glycan profile, the current data are a foundation to better understand the role of protein glycosylation in the development of insects.     

First evidence of DNA methylation in insect Tribolium castaneum

DNA methylation has been studied in many eukaryotic organisms, in particular vertebrates, and was implicated in developmental and phenotypic variations. Little is known about the role of DNA methylation in invertebrates, although insects are considered as excellent models for studying the evolution of DNA methylation. In the red flour beetle, Tribolium castaneum (Tenebrionidae, Coleoptera), no evidence of DNA methylation has been found till now. In this paper, a cytosine methylation in Tribolium castaneum embryos was detected by methylation sensitive restriction endonucleases and immuno-dot blot assay. DNA methylation in embryos is followed by a global demethylation in larvae, pupae and adults. DNA demethylation seems to proceed actively through 5-hydroxymethylcytosine, most probably by the action of TET enzyme. Bisulfite sequencing of a highly abundant satellite DNA located in pericentromeric heterochromatin revealed similar profile of cytosine methylation in adults and embryos. Cytosine methylation was not only restricted to CpG sites but was found at CpA, CpT and CpC sites. In addition, complete cytosine demethylation of heterochromatic satellite DNA was induced by heat stress. The results reveal existence of DNA methylation cycling in T. castaneum ranging from strong overall cytosine methylation in embryos to a weak DNA methylation in other developmental stages. Nevertheless, DNA methylation is preserved within heterochromatin during development, indicating its role in heterochromatin formation and maintenance. It is, however, strongly affected by heat stress, suggesting a role for DNA methylation in heterochromatin structure modulation during heat stress response.     

磷化氢对赤拟谷盗成虫体内CAT和SOD活性的影响

用赤拟谷盗Tribolium castaneum(Herbst)成虫,研究PH3对其体内超氧化物岐化酶(SOD)和过氧化氢酶(CAT)活性的影响。结果表明PH3熏蒸后,4种不同地理种群的成虫体内,CAT酶活性都有所降低。种群NGD1降低最多,接近57%,而NGD4则只降低了19%,NGD2和NGD3的降低都在40%左右。不同地理种群的成虫体内,SOD酶活性在熏蒸后都有所增加,种群NGD1增加幅度最大,NGD2增加幅度最小。SOD和CAT活性的变化幅度与成虫对PH3产生的抗性有关,抗性越大,酶活性变化越大。     

Crinkled employs wingless pathway for wing development in Tribolium castaneum

Crinkled is associated with embryonic denticle formation and auditory organ development in Drosophila melanogaster. However, the functions of Crinkled have not been fully investigated. Additionally, the genes that participate in the Crinkled pathway are unknown. Phylogenetic analysis indicates that crinkled exhibits a one‐to‐one orthologous relationship in insects. In Tribolium castaneum, the crinkled gene is 6,498 bp in length and consists of six exons. Crinkled expression peaked during two phases in Tribolium: late embryonic and pupal stages. High levels of crinkled mRNA were detected in the fat body, head, epidermis, ovary, and accessory gland of late adults. Knockdown of crinkled using RNA interference (RNAi) severely affected wing morphogenesis in T. castaneum. We further showed that crinkled silencing reduced forked expression through wingless and shaven‐baby, and RNAi of forked phenocopied the effects of crinkled knockdown in T. castaneum. This study investigated the development role of crinkled in postembryonic stages and indicated that forked mediates the functions of crinkled during wing morphogenesis in T. castaneum.     

Expression of an endoglucanase from Tribolium castaneum (TcEG1) in Saccharomyces cerevisiae

Insects are a largely unexploited resource in prospecting for novel cellulolytic enzymes to improve the production of ethanol fuel from lignocellulosic biomass. The cost of lignocellulosic ethanol production is expected to decrease by the combination of cellulose degradation (saccharification) and fermentation of the resulting glucose to ethanol in a single process, catalyzed by the yeast Saccharomyces cerevisiae transformed to express efficient cellulases. While S. cerevisiae is an established heterologous expression system, there are no available data on the functional expression of insect cellulolytic enzymes for this species. To address this knowledge gap, S. cerevisiae was transformed to express the full‐length cDNA encoding an endoglucanase from the red flour beetle, Tribolium castaneum (TcEG1), and evaluated the activity of the transgenic product (rTcEG1). Expression of the TcEG1 cDNA in S. cerevisiae was under control of the strong glyceraldehyde‐3 phosphate dehydrogenase promoter. Cultured transformed yeast secreted rTcEG1 protein as a functional β‐1,4‐endoglucanase, which allowed transformants to survive on selective media containing cellulose as the only available carbon source. Evaluation of substrate specificity for secreted rTcEG1 demonstrated endoglucanase activity, although some activity was also detected against complex cellulose substrates. Potentially relevant to uses in biofuel production rTcEG1 activity increased with pH conditions, with the highest activity detected at pH 12. Our results demonstrate the potential for functional production of an insect cellulase in S. cerevisiae and confirm the stability of rTcEG1 activity in strong alkaline environments.     

敲减Wingless / Wnt1基因表达对赤拟谷盗发育中的影响

Wnt信号通路是进化中高度保守的一条信号转导途径,在调控动物的胚胎轴向正常发育、胚胎分化、决定细胞极性、维持成体动态平衡等方面发挥重要作用. 该信号通路的异常激活还与肿瘤的发生密切相关. 本实验将体外人工合成的Wingless(Wg)/Wnt1基因dsRNA显微注射入赤拟谷盗晚期幼虫体内,研究Wingless/Wnt1蛋白在赤拟谷盗发育过程中发挥的作用. 实验结果显示,注射 Wingless(wg)/Wnt1基因dsRNA后,赤拟谷盗发育形成的蛹,翅膀宽度减小,翅间距明显增大,且羽化过程也受到严重影响. 此外,qPCR结果表明,赤拟谷盗Wingless(Wg)/Wnt1基因被沉默后,Cadherin-like 和 Smoothened (Smo)基因的表达显著上调,Armadillo-2基因略上调. 这些结果揭示,Wnt-1 信号通路和赤拟谷盗翅膀发育以及成虫羽化过程密切相关. 蛹翅宽减小,翅间距增大,可能是由于调控细胞粘连及细胞形态的Cadherin-like 和Armadillo-2基因的上调所引起.更重要的是,Smo基因的上调,表明了Wnt信号通路和Hedgehog信号通路在赤拟谷盗发育过程中有交互作用.     

Female mate choice in relation to heterozygosity in Tribolium castaneum

Abstract Female mate choice, both before and after copulation, is pervasive among insect species. It is often hypothesized that females would preferentially mate with males that are genetically dissimilar to promote the genetic variability of the offspring. We used various strains of red flour beetle, Tribolium castaneum, and tested the effect of male and female genetic backgrounds on precopulatory and post‐copulatory female mate choice. Simultaneous mate choice experiments using previously well established pheromone assays did not detect female preference for males of different strains. Post‐copulatory female mate choice was examined through paternity analysis. Two parameters were used to measure post‐copulatory female mate choice, including male defence capacity (P₁, proportion of offspring sired by the first male when a female mated with two males consecutively) and offence capacity (P₂, proportion of offspring sired by the second of two males to mate with a female). When female and male beetle strains were same, defence capacity was significantly higher than when female and male strains were different. However, such a pattern was not observed for offence capacity. The results suggest that female precopulatory mate choice is not affected by genetic background, but the outcome of post‐copulatory processes depends on the genetic background of male and female beetles.     

赤拟谷盗功能基因组学研究进展

赤拟谷盗Tribolium castaneum是一种重要的模式生物,在遗传、发育、生化与免疫等研究领域均取得了重要的研究进展。同时它也是一种危害极大的鞘翅目类储粮害虫,在世界各地都有分布,每年给储藏物造成了数十亿美元的经济损失。其全基因组测序的完成、遗传操作体系的构建及系统RNAi方法的应用都极大地促进了其功能基因组学的研究。本文综述了近年来赤拟谷盗基因组计划及功能基因组学的研究进展,拟为赤拟谷盗的生物学研究和防治奠定基础。     

Two orthodenticle-related genes in the short-germ beetle Tribolium castaneum

 To investigate the molecular basis of head evolution, we searched for genes related to the Drosophila orthodenticle (otd) homeobox gene in the short-germ beetle Tribolium castaneum. Unexpectedly, we found that there are two otd-related genes in Tribolium, with predicted homeodomains highly similar to that of the single Drosophila gene. One of the two genes (Tc otd-1) is more related in both amino acid sequence and expression pattern to fruitfly otd. Tc otd-1 is expressed in a broad anterior stripe in the blastoderm embryo, suggesting a role in early head segmentation similar to that of the Drosophila gene. The second gene (Tc otd-2) is more similar in sequence to the otd-related genes isolated from different vertebrate species (the Otx gene family). Tc otd-2 is not transcribed in the blastoderm, but is expressed later in more limited subsets of cells in the anterior brain. Both Tribolium genes and the Drosophila gene are, unlike the vertebrate genes, also expressed at the developing ventral midline of the embryo. Our results are consistent with the idea that an otd/Otx gene specified anterior head structures in the last ancestor common to arthropods and vertebrates. Within the arthropod lineage, we propose that this gene acquired a function in cells at the developing midline prior to the duplication that generated the two Tribolium genes. Received: 16 February 1996 / Accepted: 1 March 1996 Edited by C. Desplan