Authorship：Lead author,　Corresponding author   Language：Japanese   Publisher：Kanagawa Prefectural Museum of Natural History (Kanagawa Prefectural Museum)  

DOI： 10.32225/nkpmnh.2019.40_25

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Authorship：Lead author,　Corresponding author   Publishing type：Research paper (scientific journal)  

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Authorship：Lead author,　Corresponding author  

DOI： 10.1016/j.marpolbul.2018.07.016

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Language：Japanese   Publisher：岩波書店  

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE PUBLISHING GROUP  

Previous studies have suggested that blastomeres from the 2-, 4-, or 8-cell stage of corals have the ability to develop into normal primary polyps. However, it is still not known which developmental stage's blastomere produces which juvenile. In this study, we demonstrated that only the blastomeres with animal hemispheres have the capacity to develop into normal primary polyps. Individuality was evaluated using blastomeres isolated from the corals Acropora digitifera, A. intermedia, Dipsastraea lizardensis, and Favites chinensis. On commencement of embryo cleavage, the animal pole was marked using Neutral red staining, and at the 2-, 4-, and 8-cell stages, embryos were divided into individual blastomeres using glass needles. We found that the survival rate and percentage metamorphosis were higher in the larger-sized blastomeres with animal hemispheres. The vegetal hemisphere alone is incapable of developing into a normal primary polyp; however, a ball-shaped embryo with incomplete mesenteries and no pharynx developed in some cases. These results indicate that the animal hemisphere is needed for corals to develop into normal primary polyps, and that the individuality of corals is possibly determined by a combination of the chance physical splitting of embryos by waves and their innate developmental ability.

DOI： 10.1038/s41598-017-16273-w

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Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE PUBLISHING GROUP  

Reef-building corals form symbiotic relationships with dinoflagellates of the genus Symbiodinium. Symbiodinium are genetically and physiologically diverse, and corals may be able to adapt to different environments by altering their dominant Symbiodinium phylotype. Notably, each coral species associates only with specific Symbiodinium phylotypes, and consequently the diversity of symbionts available to the host is limited by the species specificity. Currently, it is widely presumed that species specificity is determined by the combination of cell-surface molecules on the host and symbiont. Here we show experimental evidence supporting a new model to explain at least part of the specificity in coral-Symbiodinium symbiosis. Using the laboratory model Aiptasia-Symbiodinium system, we found that symbiont infectivity is related to cell size; larger Symbiodinium phylotypes are less likely to establish a symbiotic relationship with the host Aiptasia. This size dependency is further supported by experiments where symbionts were replaced by artificial fluorescent microspheres. Finally, experiments using two different coral species demonstrate that our size-dependent-infection model can be expanded to coral-Symbiodinium symbiosis, with the acceptability of large-sized Symbiodinium phylotypes differing between two coral species. Thus the selectivity of the host for symbiont cell size can affect the diversity of symbionts in corals.

DOI： 10.1038/ismej.2017.17

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Language：Japanese   Publisher：北海道大学 高等教育推進機構 オープンエデュケーションセンター 科学技術コミュニケーション教育研究部門（CoSTEP）  

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Language：English   Publishing type：Part of collection (book)   Publisher：Edward Elgar Publishing Ltd.  

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Language：Japanese   Publisher：東京経済大学人文自然科学研究会  

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Language：Japanese   Publisher：東京経済大学人文自然科学研究会  

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Authorship：Lead author,　Corresponding author   Language：Japanese   Publisher：日本生物科学者協会 ; 1949-  

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Authorship：Lead author,　Corresponding author   Language：Japanese   Publisher：日本生物科学者協会 ; 1949-  

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：ZOOLOGICAL SOC JAPAN  

The order Scleractinia includes two distinct groups, which are termed "complex" and "robust" as indicated by the molecular phylogeny of mitochondrial 16S ribosomal gene sequences. Since this discovery, coral taxonomists have been seeking morphological characters for grouping this deep division in the order Scleractinia. Recently, morphological characteristics during embryogenesis that facilitate grouping the two clades as "complex" and "robust" were reported, thus clarifying a deep division in the Scleractinia. In the present report, I establish two new suborders, Refertina and Vacatina, on the basis of the embryogenetic morphological characteristics, molecular data, and new observations of Tubastraea coccinea and Cyphastrea serailia embryogenesis. In particular, the embryo of T. coccinea has a possible fertilization membrane that was first observed in the phylum Cnidaria. The new suborder Refertina consists of the families that belong to the "complex" clade and have no or little blastocoel. The new suborder Vacatina is composed of the families that fall into the "robust" clade and have an apparent blastocoel.

DOI： 10.2108/zs150094

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：BIOMED CENTRAL LTD  

Background: Research into various aspects of coral biology has greatly increased in recent years due to anthropogenic threats to coral health including pollution, ocean warming and acidification. However, knowledge of coral early development has lagged. The present paper describes the embryonic development of two previously uncharacterized robust corals, Favia lizardensis (a massive brain coral) and Ctenactis echinata (a solitary coral) and compares it to that of the previously characterized complex coral, Acropora millepora, both morphologically and in terms of the expression of a set of key developmental genes.
Results: Illumina sequencing of mixed age embryos was carried out, resulting in embryonic transcriptomes consisting of 40605 contigs for C.echinata (N50 = 1080 bp) and 48536 contigs for F.lizardensis (N50 = 1496 bp). The transcriptomes have been annotated against Swiss-Prot and were sufficiently complete to enable the identification of orthologs of many key genes controlling development in bilaterians. Developmental series of images of whole mounts and sections reveal that the early stages of both species contain a blastocoel, consistent with their membership of the robust clade. In situ hybridization was used to examine the expression of the developmentally important genes brachyury, chordin and forkhead. The expression of brachyury and forkhead was consistent with that previously reported for Acropora and allowed us to confirm that the pseudo-blastopore sometimes seen in robust corals such as Favia spp. is not directly associated with gastrulation. C.echinata chordin expression, however, differed from that seen in the other two corals.
Conclusions: Embryonic transcriptomes were assembled for the brain coral Favia lizardensis and the solitary coral Ctenactis echinata. Both species have a blastocoel in their early developmental stages, consistent with their phylogenetic position as members of the robust clade. Expression of the key developmental genes brachyury, chordin and forkhead was investigated, allowing comparison to that of their orthologs in Acropora, Nematostella and bilaterians and demonstrating that even within the Anthozoa there are significant differences in expression patterns.

DOI： 10.1186/s12862-016-0615-2

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Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：TAYLOR & FRANCIS LTD  

Reproductive traits of scleractinian corals are important in understanding their ecology and for maintaining coral populations; however, it has been reported for no more than 30% of species. Oulastrea crispata (Lamark, 1816) has the greatest north-south distribution in the Indo-West Pacific. This species is also known as one of the most stress-tolerant corals. Thus, most previous studies on O. crispata have examined its physiological, ecological, morphological, or genetic traits and its symbiotic dinoflagellates relative to its broad distribution and its stress tolerance. However, there have been few studies on its reproduction. In this study, the release of sperm and eggs of O. crispata was observed under laboratory rearing conditions in Shirahama, Kii Peninsula, Japan. This is the first direct evidence of sexual reproduction in this species, and the contribution of sexual reproduction to Oulastrea biology is briefly discussed.

DOI： 10.1080/07924259.2015.1006340

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER SCIENCE BV  

The deterioration of coral reefs in Japan is a serious environmental problem. Conventional conservation policies for terrestrial ecosystems are sometimes difficult to apply to coral reef protection because of the large number of stakeholders involved. In what seems to be an interesting attempt to solve this problem, tourist divers in Okinawa, Japan have begun to transplant coral fragments onto deteriorated coral reefs, by participating in a tour provided by diving shops. However, the problem here is that when the transplanted fragments have been taken out from the natural coral colonies, it tends to cause a host of potential problems such as decreasing fecundity of donor colonies, negative effects on the surrounding environment of the exploited corals and low species diversity of transplanted fragments. In this paper, we examine the merits of commercial coral transplantation in marine ecosystem conservation, and to suggest some reforms that could help to mitigate the problems encountered when using sexually propagated coral transplants. Finally, we discuss how the commercial transplantation in Okinawa could be applied to the conservation of other marine ecosystem. (C) 2014 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.ecoser.2014.07.009

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Authorship：Lead author,　Corresponding author   Language：Japanese   Publisher：環境経済・政策学会  

DOI： 10.14927/reeps.7.1_54

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：PUBLIC LIBRARY SCIENCE  

A comprehensive understanding of coral reproduction and development is needed because corals are threatened in many ways by human activity. Major threats include the loss of their photosynthetic symbionts (Symbiodinium) caused by rising temperatures (bleaching), reduced ability to calcify caused by ocean acidification, increased storm severity associated with global climate change and an increase in predators caused by runoff from human agricultural activity. In spite of these threats, detailed descriptions of embryonic development are not available for many coral species. The current consensus is that there are two major groups of stony corals, the "complex" and the "robust". In this paper we describe the embryonic development of four "complex" species, Pseudosiderastrea tayamai, Galaxea fascicularis, Montipora hispida, and Pavona Decussata, and seven "robust" species, Oulastrea crispata, Platygyra contorta, Favites abdita, Echinophyllia aspera, Goniastrea favulus, Dipsastraea speciosa (previously Favia speciosa), and Phymastrea valenciennesi (previously Montastrea valenciennesi). Data from both histologically sectioned embryos and whole mounts are presented. One apparent difference between these two major groups is that before gastrulation the cells of the complex corals thus far described (mainly Acropora species) spread and flatten to produce the so-called prawn chip, which lacks a blastocoel. Our present broad survey of robust and complex corals reveals that prawn chip formation is not a synapomorphy of complex corals, as Pavona Decussata does not form a prawn chip and has a well-developed blastocoel. Although prawn chip formation cannot be used to separate the two clades, none of the robust corals which we surveyed has such a stage. Many robust coral embryos pass through two periods of invagination, separated by a return to a spherical shape. However, only the second of these periods is associated with endoderm formation. We have therefore termed the first invagination a pseudo-blastopore.

DOI： 10.1371/journal.pone.0084115

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：MARINE BIOLOGICAL LABORATORY  

Temporal dynamics of larval survival were examined in vitro in four broadcast-spawning reef coral species, Acropora hyacinthus, A. japonica, A. solitaryensis, and Goniastrea pectinata. Larval size was treated as an important characteristic that may relate to larval lifespan. Two patterns were observed in larval survival dynamics between the three Acropora species (mean initial larval size; 0.05-0.08 mm(3)) and G. pectinata (0.02 mm(3)), based on the timing of a sharp drop in larval survival rates (ca. > 50% reduction over a 1-2 week period). Consequently, the majority of larvae of the three Acropora species had a lifespan of less than 2-3 weeks, whereas those of G. pectinata were extended a further 2-3 weeks despite the smaller larval size. No significant relationship was detected between the initial larval size and larval lifespan in any of the four reef coral species. These results suggest that (1) larval dispersal of spawning Acropora species may be on a more local scale than that of G. pectinata and most other reef coral species previously reported, and (2) larval size is not a good estimator of larval lifespan in reef coral species.

DOI： 10.1086/BBLv220n1p15

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Authorship：Lead author   Language：Japanese   Publisher：The Japanese Coral Reef Society  

The deterioration of coral reefs is a serious environmental problem. Conventional conservation policies for terrestrial ecosystems are sometimes difficult to apply to coral reefs because of the large number of stakeholders. In an interesting attempt to solve this problem, some diving tours in Japan have begun to transplant coral fragments on to deteriorated coral reefs. However, the fragments used for transplantation are taken from natural coral colonies, causing potential problems. These include decreasing the fecundity of the donor colonies; having a negative effect on the surrounding environment of the exploited corals; and contributing to low species diversity of transplanted fragments. In this paper, we suggest that seedlings can be used for transplantation. This is costly but has a lower negative environmental effect. We show how to determine the optimal ratio of fragments and seedlings to produce the best environmental effect, depending on the cost and the environmental effect of each coral transplant.

DOI： 10.3755/jcrs.12.69

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：MARINE BIOLOGICAL LABORATORY  

In the late 1990s, the once prolific populations of the coral Acropora intermedia surrounding Okinawa, Japan, dramatically declined because of thermal stress, bleaching caused by heat stress, and consequent mortality. Before the bleaching event, 72 fragments (about 15 cm in length) were collected and transferred to the Okinawa Churami Aquarium. Through growth and repeated fragmentation, these original fragments developed into about 100 colonies that spawned from 1999 to 2009. In this study, we compared gametogenesis, fertilization, survival, and O(2) consumption in cultured and wild colonies of A. intermedia and their offspring. Cultured A. intermedia had larger oocytes and higher fertilization and survival rates than samples from wild colonies. O(2) consumption of cultured embryos was similar to that of wild embryos. These results suggest that cultured A. intermedia and their offspring are as viable as wild colonies. Aquaria can play a role in the conservation of endangered corals, and their cultured colonies could be used to re-establish devastated species on the Okinawa reefs.

DOI： 10.1086/BBLv218n3p230

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：ACAD SINICA INST ZOOLOGY  

Nami Okubo, Hiroki Taniguchi, and Makoto Omori (2009) Sexual reproduction in transplanted coral fragments of Acropora nasuta. Zoological Studies 48(4): 442-447. The survival rate and sexual reproduction were examined after fragmentation and transplantation of the reef-building coral Acropora nasuta. Fragments of 2 different lengths, of approximately 5 and 10 cm, were transplanted onto a reef substrate in July 2001 (n = 85 at 5 cm and 71 at 10 cm) and Feb. 2002 (n = 66 at 5 cm and 66 at 10 cm), corresponding to the early and late vitellogenic stages of oocyte development, respectively. Oocyte development, fecundity, and spawning, were monitored over a 3 yr period. Oocyte development was influenced by both fragment size and season of transplantation. In smaller fragments, oocytes were resorbed, while development continued in larger fragments, suggesting that smaller fragments could not afford to invest in sexual reproduction and converted resources from oocytes into growth/survival. Oocytes of July-transplanted fragments (in the early vitellogenic stage) were resorbed, while oocytes of Feb.-transplanted fragments (in the late vitellogenic stage) continued to develop. This may have occurred because of the large amount of energy needed for further oocyte development in July-transplanted fragments until spawning in June of the following year. Transplanted fragments spawned in the 1st yearn none, except for 1 fragment, spawned in the 2nd year: and no gametes were produced in the 3rd year, indicating that fragments reallocated energy resources. and that infertility occurred for a certain period of time. http://zoolstud.sinica.edu.tw/Journals/48.4/442.pdf

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

RNA 3'-terminal phosphate cyclase (Rtc) is an enzyme involved in RNA splicing that converts the 3'-terminal hydroxyl group of truncated RNA to 2',3'-cyclic phosphate, which is required just before its ligation. This reaction may occur in the following two steps: (i) Rtc + ATP -> Rtc-AMP + PP(i) and (ii) RNA-N3'p + Rtc-AMP -> RNA-N>p + Rtc + AMP. In order to reveal the reaction mechanism, Rtc of Sulfolobus tokodaii (St-Rtc) overexpressed in Escherichia coli was purified and crystallized in the following states: St-Rtc, St-Rtc+Mn, St-Rtc+ATP, St-Rtc+AMP and St-Rtc-AMP. The crystals diffracted to 2.25-3.00 angstrom resolution and preliminary solutions of their structures have been obtained by molecular replacement using the structure of a selenomethionine-labelled St-Rtc crystal which was solved in advance using the MAD method as a model. These crystals grew in two different space groups (P3(1) and P4(2)), with the former space group displaying two distinct packing modes.

DOI： 10.1107/S1744309109014663

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：INTER-RESEARCH  

O-2 consumption of a single embryo/planula at each developmental stage was monitored in the reef-building coral Acropora intermedia using an optical O-2-sensing system with our original micro-chamber system (6.28 mu l). The lowest rate Of O-2 consumption was observed in unfertilized eggs. After fertilization, O-2 consumption increased and remained constant until the prawn chip blastula stage. However, O-2 consumption began to increase again during the bowl-shaped blastula stage, which involves the formation of 2 germ layers and corresponds to the beginning of gastrulation. The rate Of O-2 consumption peaked during the teardrop-shaped planula stage. During this stage planulae are able to swim actively, especially in the vertical plane, so an increase in energy consumption during this stage is to be expected. O-2 consumption began to decrease gradually 5 d after spawning. At this stage, the larvae frequently touched the substrate with their concave aboral end, which features numerous spirocysts required for substrate attachment. When the planulae began to settle, 7 d after spawning, the rate Of O-2 consumption dropped to that of unfertilized eggs, suggesting that the planulae slowly use stored energy for crawling/settlement behavior and/or post-settlement growth and survivorship.

DOI： 10.3354/meps07562

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Authorship：Lead author,　Corresponding author   Language：English   Publisher：The Japanese Coral Reef Society  

To reveal the optimal resource allocation among regeneration, growth, sexual reproduction, and survivorship in a colonial animal, a specimen of the branching coral <I>Acropora muricata</I> (<I>formosa</I>), without oocytes, was separated into fragments of three different sizes (5, 10, and 20cm long) that were then transplanted onto coral pavement. Regardless of the fragment size, a wound was fully closed within 11 days after fragmentation. The number of newly formed polyps on the new tissue layer did not differ significantly among fragment sizes. No significant difference was observed in the monthly growth rate among the three fragment sizes. These results showed that regeneration and growth were independent of fragment size. During the observation period, one 5cm fragment died, the 5cm and 10cm fragments did not spawn, but the 20cm fragments did spawn. The investment to survivorship by the 5cm fragments might have been smaller than that of the 10cm fragments, because the 5cm fragments had smaller energy stocks than did the 10cm fragments. Combined, these results suggest that the energetic investment for regeneration and growth was constant while investment for reproduction was proportional to fragment size. This finding agrees with the theoretical optimal dispersal strategy, in which the allocation to growth by mothers is constant. In conclusion, the colonial coral <I>Acropora muricata</I> generally shows size-independent growth and regeneration, and exhibits a tradeoff among regeneration, growth, survivorship, and reproduction.

DOI： 10.3755/galaxea.10.83

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：ZOOLOGICAL SOC JAPAN  

Embryogenesis in the reef building corals Acropora intermedia, A. solitaryensis, A. hyacinthus, A. digitifera, and A. tenuis was studied in detail at the morphological level, and the relationships among the animal pole, blastopore, and mouth were investigated for the first time in corals. These species showed essentially the same sequence of development. The embryo undergoes spiral-like holoblastic cleavage despite the presence of a dense isolecithal yolk. After the morula stage, the embryo enters the "prawn-chip" stage, which consists of an irregularly shaped cellular bilayer. The embryo begins to roll inward to form the bowl stage; the round shape observed during this stage suggests that it may be the beginning of gastrulation. However, the blastopore closes and the stomodeum (mouth and pharynx) is formed via invagination at a site near the closed blastopore. During the planula stage, a concavity forms in the aboral region in conjunction with numerous spirocysts, suggesting that spirocysts are used to attach to the substrate before the onset of metamorphosis.

DOI： 10.2108/zsj.24.1169

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：ZOOLOGICAL SOC JAPAN  

We examined gametogenesis and the periodicity of planula release in the brooding coral Acropora (Isopora) brueggemanni (Brook, 1893) on Akajima Island, Okinawa, Japan. We captured the moment when A. brueggemanni would be in the process of self-fertilization. Whole colonies of this species were cultured separately or together with other colonies in plastic containers. We observed no apparent periodicity of planula release in the collected colonies. A few planulae were released intermittently during the observation period, regardless of whether the colonies were cultured individually or with other colonies. Serial paraffin sections of A. brueggemanni showed follicle-like cells surrounding the oocyte during vitellogenesis. In September and October, some spermaries looked half-broken and some ova were surrounded by sperm or spermaries instead of follicle-like cells. Such ova were heart-shaped and may have been cells at the first cleavage stage. These observations suggest that the migration of spermaries and/or oocytes resulted in the close proximity of oocytes and sperm, which would allow self-fertilization. This possibility, together with the production of planula larvae by isolated colonies, suggests that this brooding coral engages in self-fertilization.

DOI： 10.2108/zsj.24.277

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER  

In order to determine competency of sexual reproduction and survival rate after fragmentation, the branching coral Acropora formosa was fragmented and fragments in three different sizes (ca. 5, 10 and 20 cm long) and three different stages of gametogenesis were transplanted on coral pavement. Their oocyte development and fecundity, as well as spawning were monitored for a 3-year period. The oocyte development was affected by both fragment size and by the developmental stage of oocytes when fragmented. In small fragments, the oocytes were resorbed while in large fragments they continued development. Oocytes in the early vitellogenic stage at the time of fragmentation were resorbed, whereas those in the late stage continued developing. Smaller fragments showed a lower survival rate and histological observations of their gonads revealed resorption of oocytes, suggesting that there was a trade-off of energy between reproduction and survival. Transplanted fragments often spawned one month earlier than the donor colonies in the first year, but spawning occurred in the same month as the donors or did not occur at all in the second year and none spawned in the third year. The risk of colony death may cause the fragments to re-allocate energy for sexual reproduction.

DOI： 10.1007/s00227-006-0490-2

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER  

In order to establish a successful method for the transplantation of branching and tabular coral fragments, we tested the effects of orientations of attachment, seasons of transplantation, and size of fragments on survival, growth, and spawning using Acropora formosa and A. hyacinthus. Vertically attached, large-sized fragments of A. formosa showed 98-100% survival rate after 18 months. The fragments transplanted in August exhibited better survival than those transplanted in November. The larger fragments had the higher percentage of spawning. The fragments that spawned had lower growth rate, while those resorbed the oocytes carried at the time of transplantation showed higher growth rate, suggesting the trade-off between growth and reproduction. Half of the fragments spawned 1 month earlier than the donor colonies. Only the vertically attached fragments of A. hyacinthus fused to the substratum, and those transplanted in February showed 100% survival rate after 14 months, indicating that this species is well suited for transplantation.

DOI： 10.1007/s00338-005-0496-0

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Authorship：Lead author,　Corresponding author   Language：Japanese   Publisher：日本サンゴ礁学会  

In order to restore disturbed coral reefs, transplantation of coral fragments or coral heads has been conducted in many parts of the world. We reviewed methods and results of the transplantation experiments described in numerous previous reports in order to determine recommendable methodologies. Problems that should be solved by future studies are discussed. For transplantation, generally, small pieces of coral are taken from the donor colony using hammer, chisel and the like. If the collection site is far from the transplantation site, fragments should be placed in bucket full of seawater. There are various methods to fix the coral fragment onto substrate, e. g. by means of epoxy cement, plastic coated wire and nails. They should be chosen according to size and shape of coral fragments, but we found fixation with nails and cable ties is easy and reliable for staghorn corals. The cost of transplantation with a density of 245, 000 fragments per hectare secured by means of nails and cable ties has been estimated at about 36, 000, 000 JPN Yen. So far, species of the genera &lt;i&gt;Acropora&lt;/i&gt;, &lt;i&gt;Porites&lt;/i&gt;, &lt;i&gt;Pavona&lt;/i&gt; and &lt;i&gt;Galaxea&lt;/i&gt; were most frequently used for transplantation as they have relatively high survival rates.

DOI： 10.3755/jcrs.2001.31

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