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Öğe Additional description and genome analyses of Caenorhabditis auriculariae representing the basal lineage of genus Caenorhabditis(Nature Research, 2021) Dayi, Mehmet; Kanzaki, Natsumi; Sun, Simo; Ide, Tatsuya; Tanaka, Ryusei; Masuya, Hayato; Kikuchi, TaiseiCaenorhabditis auriculariae, which was morphologically described in 1999, was re-isolated from a Platydema mushroom-associated beetle. Based on the re-isolated materials, some morphological characteristics were re-examined and ascribed to the species. In addition, to clarify phylogenetic relationships with other Caenorhabditis species and biological features of the nematode, the whole genome was sequenced and assembled into 109.5 Mb with 16,279 predicted protein-coding genes. Molecular phylogenetic analyses based on ribosomal RNA and 269 single-copy genes revealed the species is closely related to C. sonorae and C. monodelphis placing them at the most basal clade of the genus. C. auriculariae has morphological characteristics clearly differed from those two species and harbours a number of species-specific gene families, indicating its usefulness as a new outgroup species for Caenorhabditis evolutionary studies. A comparison of carbohydrate-active enzyme (CAZy) repertoires in genomes, which we found useful to speculate about the lifestyle of Caenorhabditis nematodes, suggested that C. auriculariae likely has a life-cycle with tight-association with insects.Öğe Assessment of the behaviour and survival of nematodes under low oxygen concentrations(Public Library Science, 2018) Kitazume, Hiromi; Dayı, Mehmet; Tanaka, Ryusei; Kikuchi, TaiseiOxygen is required for the completion of almost all known metazoan lifecycles, but many metazoans harbour abilities to withstand varying degrees and periods of hypoxia. Caenorhabditis elegans, one of the most popular model organism is extensively used as a model for the study of hypoxia and anoxia biology and it has been found that this nematode is capable of tolerance to varying degrees of hypoxia. Considering the extremely high diversity of nematodes, the effects of low oxygen concentration and mechanisms of adaptation to oxygen depletion differ among species. In this study, we used a simple assay to examine anoxia tolerance in four nematode species, including three free-living and one plant parasitic nematode. We found that the plant parasitic nematode Bursaphelenchus xylophilus can survive more than 14 days under anoxic conditions. Comparisons of behaviour during anoxia induction and the repertoire of oxygen sensation genes among the tested species suggested the existence of different oxygen sensation systems between B. xylophilus and C. elegans, which quickly introduce suspended animation in response to oxygen depletion to survive long-term anoxia.Öğe The compact genome of Caenorhabditis niphades n. sp., isolated from a wood-boring weevil, Niphades variegatus(Bmc, 2022) Sun, Simo; Kanzaki, Natsumi; Dayı, Mehmet; Maeda, Yasunobu; Yoshida, Akemi; Tanaka, Ryusei; Kikuchi, TaiseiBackground The first metazoan genome sequenced, that of Caenorhabditis elegans, has motivated animal genome evolution studies. To date > 50 species from the genus Caenorhabditis have been sequenced, allowing research on genome variation. Results In the present study, we describe a new gonochoristic species, Caenorhabditis niphades n. sp., previously referred as C. sp. 36, isolated from adult weevils (Niphades variegatus), with whom they appear to be tightly associated during its life cycle. Along with a species description, we sequenced the genome of C. niphades n. sp. and produced a chromosome-level assembly. A genome comparison highlighted that C. niphades n. sp. has the smallest genome (59 Mbp) so far sequenced in the Elegans supergroup, despite being closely related to a species with an exceptionally large genome, C. japonica. Conclusions The compact genome of C. niphades n. sp. can serve as a key resource for comparative evolutionary studies of genome and gene number expansions in Caenorhabditis species.Öğe Development and validation of SSR markers for the plant-parasitic nematode Subanguina moxae using genome assembly of Illumina pair-end reads(Brill Academic Publishers, 2015) Takeuchi, Tomoaki; Yamaguchi, Madoka; Tanaka, Ryusei; Dayı, Mehmet; Ogura, Nobuo; Kikuchi, TaiseiSubanguina moxae, belonging to the subfamily Anguininae, is an obligate parasite of Artemisia plants, which are widely used as cooking herbs and in traditional medicine in East Asia. Because the nematode is distributed throughout East Russia and East Asia, there is concern about the potential for significant damage to commercial farming; however, details about its biology remain unclear. To investigate the genetic diversity of S. moxae, we developed 2243 simple sequence repeat (SSR) markers using Illumina short reads of the genomic DNA. We validated 100 randomly selected markers indicating their robustness and examined polymorphisms among nematode populations sampled from four different locations in Japan. These SSR markers will be a useful tool for understanding the population structure and transmission patterns of this parasitic nematode.Öğe Differentially Expressed Genes Associated with Body Size Changes and Transposable Element Insertions between Caenorhabditis elegans and Its Sister Species, Caenorhabditis inopinata(Oxford Univ Press, 2023) Kawahara, Kazuma; Inada, Taruho; Tanaka, Ryusei; Dayi, Mehmet; Makino, Takashi; Maruyama, Shinichiro; Kikuchi, TaiseiWhy the recently discovered nematode Caenorhabditis inopinata differs so greatly from its sibling species Caenorhabditis elegans remains unknown. A previous study showed that C. inopinata has more transposable elements (TEs), sequences that replicate and move autonomously throughout the genome, potentially altering the expression of neighboring genes. In this study, we focused on how the body size of this species has evolved and whether TEs could affect the expression of genes related to species-specific traits such as body size. First, we compared gene expression levels between C. inopinata and C. elegans in the L4 larval and young adult stages-when growth rates differ most prominently between these species-to identify candidate genes contributing to their differences. The results showed that the expression levels of collagen genes were consistently higher in C. inopinata than in C. elegans and that some genes related to cell size were differentially expressed between the species. Then, we examined whether genes with TE insertions are differentially expressed between species. Indeed, the genes featuring C. inopinata-specific TE insertions had higher expression levels in C. inopinata than in C. elegans. These upregulated genes included those related to body size, suggesting that these genes could be candidates for artificial TE insertion to examine the role of TEs in the body size evolution of C. inopinata.Öğe Genome of the fatal tapeworm Sparganum proliferum uncovers mechanisms for cryptic life cycle and aberrant larval proliferation(Nature Research, 2021) Kikuchi, Taisei; Dayi, Mehmet; Hunt, Vicky L.; Ishiwata, Kenji; Toyoda, Atsushi; Kounosu, Asuka; Kojima, SomeiThe cryptic parasite Sparganum proliferum proliferates in humans and invades tissues and organs. Only scattered cases have been reported, but S. proliferum infection is always fatal. However, S. proliferum's phylogeny and life cycle remain enigmatic. To investigate the phylogenetic relationships between S. proliferum and other cestode species, and to examine the mechanisms underlying pathogenicity, we sequenced the entire genomes of S. proliferum and a closely related non-life-threatening tapeworm Spirometra erinaceieuropaei. Additionally, we performed larvae transcriptome analyses of S. proliferum plerocercoid to identify genes involved in asexual reproduction in the host. The genome sequences confirmed that the S. proliferum has experienced a clearly distinct evolutionary history from S. erinaceieuropaei. Moreover, we found that nonordinal extracellular matrix coordination allows asexual reproduction in the host, and loss of sexual maturity in S. proliferum are responsible for its fatal pathogenicity to humans. Our high-quality reference genome sequences should be valuable for future studies of pseudophyllidean tapeworm biology and parasitism. Kikuchi et al. sequence the genome of the fatal tapeworm Sparganum proliferum and a closely related non-life-threatening tapeworm Spirometra erinaceieuropaei, and describe its genomic features suggesting the natural history and molecular mechanisms underlying pathogenicity. Their findings indicate that nonordinal extracellular matrix coordination is important for its asexual reproduction, and suggest that loss of sexual maturity contributes to the fatal pathogenicity of S. proliferum to humans.Öğe Nearly Complete Genome Assembly of the Pinewood Nematode Bursaphelenchus xylophilus Strain Ka4C1(Amer Soc Microbiology, 2020) Dayi, Mehmet; Sun, Simo; Maeda, Yasunobu; Tanaka, Ryusei; Yoshida, Akemi; Tsai, Isheng Jason; Kikuchi, TaiseiBursaphelenchus xylophilus has been destroying pine forests in East Asia and western Europe. Here, we report its nearly complete genomic sequence containing five similar to 12-Mb scaffolds and one similar to 15-Mb scaffold representing six chromosomes. Large repeat regions that were previously unidentified are now reasonably integrated, particularly in the similar to 15-Mb scaffold.Öğe Reduced Genome of the Gut Symbiotic Bacterium Candidatus Benitsuchiphilus tojoi Provides Insight Into Its Possible Roles in Ecology and Adaptation of the Host Insect(Frontiers Media Sa, 2020) Mondal, Shakhinur Islam; Akter, Arzuba; Koga, Ryuichi; Hosokawa, Takahiro; Dayi, Mehmet; Murase, Kazunori; Kikuchi, TaiseiDiverse animals, including insects, harbor microbial symbionts within their gut, body cavity, or cells. The subsocial parastrachiid stinkbug Parastrachia japonensis is well-known for its peculiar ecological and behavioral traits, including its prolonged non-feeding diapause period and maternal care of eggs/nymphs in an underground nest. P. japonensis harbors a specific bacterial symbiont within the gut cavity extracellularly, which is vertically inherited through maternal excretion of symbiont-containing white mucus. Thus far, biological roles of the symbiont in the host lifecycle has been little understood. Here we sequenced the genome of the uncultivable gut symbiont Candidatus Benitsuchiphilus tojoi. The symbiont has an 804 kb circular chromosome encoding 606 proteins and a 14.5 kb plasmid encoding 13 proteins. Phylogenetic analysis indicated that the bacterium is closely related to other obligate insect symbionts belonging to the Gammaproteobacteria, including Buchnera of aphids and Blochmannia of ants, and the most closely related to Ishikawaella, an extracellular gut symbiont of plataspid stinkbugs. These data suggested that the symbiont genome has evolved like highly reduced gamma-proteobacterial symbiont genomes reported from a variety of insects. The presence of genes involved in biosynthesis pathways for amino acids, vitamins, and cofactors in the genome implicated the symbiont as a nutritional mutualist, supplementing essential nutrients to the host. Interestingly, the symbiont's plasmid encoded genes for thiamine and carotenoid synthesis pathways, suggesting the possibility of additional functions of the symbiont for protecting the host against oxidative stress and DNA damage. Finally, possible involvement of the symbiont in uric acid metabolism during diapause is discussed.Öğe Stage-specific transcriptome of Bursaphelenchus xylophilus reveals temporal regulation of effector genes and roles of the dauer-like stages in the lifecycle(Nature Publishing Group, 2019) Tanaka, Suguru E.; Dayı, Mehmet; Maeda, Yasunobu; Tsai, Isheng J.; Tanaka, Ryusei; Bligh, Mark; Kikuchi, TaiseiThe pine wood nematode Bursaphelenchus xylophilus is the causal agent of pine wilt disease, one of the most devastating forest diseases in East Asian and West European countries. The lifecycle of B. xylophilus includes four propagative larval stages and gonochoristic adults which are involved in the pathogenicity, and two stages of dispersal larvae involved in the spread of the disease. To elucidate the ecological roles of each developmental stage in the pathogenic life cycle, we performed a comprehensive transcriptome analysis using RNA-seq generated from all developmental stages of B. xylophilus and compared transcriptomes between stages. We found more than 9000 genes are differentially expressed in at least one stage of the life cycle including genes involved in general nematode biology such as reproduction and moulting but also effector genes likely to be involved in parasitism. The dispersal-stage transcriptome revealed its analogy to C. elegans dauer and the distinct roles of the two larval stages from each other regarding survival and transmission. This study provides important insights and resources to understand B. xylophilus parasitic biology.Öğe Telomere-to-Telomere Genome Assembly of Bursaphelenchus okinawaensis Strain SH1(Amer Soc Microbiology, 2020) Sun, Simo; Shinya, Ryoji; Dayi, Mehmet; Yoshida, Akemi; Sternberg, Paul W.; Kikuchi, TaiseiBursaphelenchus okinawaensis is a self-fertilizing, hermaphroditic, fungus-feeding nematode used as a laboratory model for the genus Bursaphelenchus, which includes the important pathogen Bursaphelenchus xylophilus. Here, we report the nearly complete genome sequence of B. okinawaensis. The 70-Mbp assembly contained six scaffolds (>11 Mbp each) with telomere repeats on their ends, indicating complete chromosomes.
