Morphology of spines and spine joint in the crown-of-thorns starfish Acanthaster planci (Echinodermata,Asteroida)

Summary Morphology and movement of the spines of Acanthaster planci were studied. All surfaces of the animal are covered with spines. The spines on the aboral surface are cylindrical with sharp tips. The spines on the oral surface are flat; they bend over to cover the mouth and the ambulacral grooves when these soft parts are stimulated. Those on the side of the animal make a barrier of crossed spines. Thus the structure and movement of the spines are well-adapted for defense.The junction between the primary aboral spine and its pedicel makes a movable joint. The ultrastructure of the connective tissue at the joint was studied. The connective tissue is mainly composed of collagen fibers. Presumed neurosecretory cells with processes which are filled with electron-dense granules of 0.2 m diameter were found between collagen fibers. Muscle fibers are mainly found in the connective tissue at the central holes. These observations support the view that the joint connective tissue has catch properties.     

Evaluation of a crown-of-thorns starfish (Acanthaster planci) control program at Grub Reef (central Great Barrier Reef)

A crown-of-thorns starfish control program was conducted at Grub Reef (central Great Barrier Reef) in an area (0.64 km²) which encompassed 53 individual patch reefs. During a two week period, 15 divers injected 3175 starfish with copper sulphate. The program was considered unsuccessful. Although starfish abundance had declined significantly after the control efforts, biological surveys indicated that a relatively large number of starfish remained. The surveys also indicated a general decline in the number of starfish along the reef perimeter, outside the control area. The total cost of the control program was $35 per starfish. These results have important implications for the implementation of future control programs and highlight the need to undertake before and after biological surveys to assess the effectiveness of the control efforts.     

Population genetic studies of the crown-of-thorns starfish,Acanthaster planci (L.), in the Great Barrier Reef region

Seven populations of the crown-of-thorns starfish, Acanthaster planci, were compared genetically using starch gel electrophoresis in order to investigate the extent of genetic exchange throughout the Great Barrier Reef (GBR) region. These populations extended from Lizard Island in the north to One Tree Island in the south, a distance of approximately 1300 km. Thirteen of 36 enzymes assayed were genetically interpretable, and 10 (77%) of these were polymorphic. Mean heterozygosity over all loci was 0.225. An analysis of the gene frequency heterogeneity between the populations using Wright's F _ST statistic gave an overall F _ST of 0.019. The mean unbiased value of Nei's genetic distance between the populations was 0.009. These values indicate a homogeneous genetic composition throughout the range, and are consistent with the hypothesis that gene flow between these populations is high, and that A. planci throughout the GBR region are members of a single, effectively panmictic population. Within this group, the Green Island population was most distinct genetically because of differences in allele frequencies at the MDH-1 locus. Although there is no rigorous method for determining the selective basis for such differences, it is argued that the differences observed in the Green Island population were the result of selection. The basis for selective differences was possibly food availability since, at the time of sampling, the Green Island A. planci were the remnants of a large, high-density population that caused extensive coral mortality, and suffered severe population decline as food became scarce. These findings are consistent with observations of a relatively ordered sequence of outbreaks from north to south along the GBR, suggesting that all outbreaks but the first are secondary. Control measures, both on the GBR and elsewhere, have been unsuccessful except on a very small scale. Unless a vulnerable part of the A. planci life cycle can be identified, it would seem that the greatest chance for successful control would be to identify and control the causes of the primary population outbreak.     

History and dynamics of the crown-of-thorns starfish Acanthaster planci (L.) in the Suva area,Fiji

Annual recruitment of Acanthaster planci over a 13 year period in a known juvenile refuge substratum on Suva Reef was very patchy in time and space. Macroscale recruitment events resulted in outbreaks over many thousands of hectares over 100 km of coastline whereas localised but intensive mesoscale events resulted in concenrated aggregations over areas of several tens of hectares. A very low level of recruitment occurred in most years but three intensive, macroscale recruitment events occurred in 1977, 1984 and 1987, and two localised but intensive recruitment events occurred in 1982 and 1983. Annual recruitment of A. planci between 1975–89 had no overt relationship with rainfall events, as proposed in the terrestrial run-off hypothesis on the cause of primary out-breaks. Growth rates within monitored cohorts were highly variable, but mean growth rates of two cohorts 1984 and 1987 were relatively similar. Mortality rates of three monitored cohorts were variable. Longevity of the 1977 cohort was 7–8 years, while that of the 1984 cohort was 2–3 years. Mortality of the 1987 cohort has been low to date (i.e. 4 years of age). Disease, attributed to an undescribed sporozoan pathogen, was responsible for two mass mortalities in the 1984 cohort, in 1984 and in 1986, and eventually resulted in the extinction of this cohort. Anecdotal reports indicate that outbreaks of A. planci are increasing in frequency and severity. According to Fijian reef fishermen, the starfish were low to moderate in abundance in the Suva area from the 1920s to the 1960s. A progressive build up occurred on some reefs in 1963–67 leading to a major outbreak episode from 1967–1971. The outbreaks were in a chronic phase between 1977–1989. Our findings demonstrate that there is no invariable pattern or process involved in the population dynamics of A. planci and that a single explanation of the A. planci phenomenon is therefore unlikely.     

Lipolytic enzymes of the digestive organs of the crown-of-thorns starfish (Acanthaster planci): comparison of the stomach and pyloric caeca

1. Stomach and pyloric caeca homogenates from the crown-of-thorns starfish hydrolysed p-nitrophenyl esters, alpha-naphthyl esters, cholesteryl oleate and tributyrin. The pyloric caeca contained the highest activities. 2. The p-nitrophenyl acetate hydrolytic activity eluted at 0.23 M NaCl on ion exchange chromatography while the p-nitrophenyl palmitate hydrolytic activity eluted between 0.2 and 1.0 M NaCl. 3. Polyacrylamide gel zymograms for alpha-naphthyl acetate hydrolytic activity revealed one major band and several minor bands of activity for both tissues. 4. Isoelectric focusing zymograms revealed one major band with a pI = 4.2 for both tissues, with an additional band at pI = 3.5 for pyloric caeca. 5. The pyloric caeca contained twice as much lipid as the stomach. Lipid extracts contained mixtures of steroids and steroid-esters; a cholesterol-like sterol was tentatively identified.     

Genetic diversity and phylogenetic analysis of blackbuck (Antilope cervicapra) in southern India

Blackbuck (Antilope cervicapra) is a threatened species endemic to the Indian subcontinent. Many populations of blackbuck are found in southern India. Populations of blackbuck are negatively affected in many places for various reasons, such as habitat destruction and poaching. Their range decreased sharply during the 20th century. There is very limited information available on the population dynamics of blackbuck in southern India. For the phylogenetic and genetic diversity analyses of blackbuck populations among different distribution ranges in southern India, we sequenced mt DNA of cytochrome b (Cyt b) for 120, cytochrome c oxidase subunit-1 (COI) for 137 and the control region (CR) for 137 fecal pellets from eleven different locations in southern India. We analyzed the genetic structure of three mitochondrial markers, the CR, Cyt b and the COI region, separately and in a combined dataset. The haplotype diversity and nucleotide diversity of CR were 0.969 and 0.047, respectively, and were higher than those of Cyt b and COI. A Bayesian phylogeny and an MJ network based on the CR and combined dataset (105 sequences) signified several distinct haplotype clusters within blackbuck, whereas no clusters were identified with the Cyt b and COI phylogenetic analyses. The analysis of molecular variance of the combined data set revealed 52.46% genetic variation within the population. Mismatch distribution analysis revealed that blackbuck populations underwent complex changes with analysis of the combined dataset in each population and analysis of each marker separately in the overall population. The results provide evidence that blackbuck in different geographic locations has a distinct population structure due to habitat fragmentation after the formation of the Western and Eastern Ghats.

     

Genetic polymorphism of thyroxin-binding globulin (TBG) in the Pacific area.

Human plasma samples, radiolabeled with [125I]thyroxin, from the Asian, Pacific, and Australian area have been subjected to isoelectric focusing to reveal genetic variation in thyroxin-binding globulin (TBG). A genetically determined electrophoretic slow variant, TBG S, indistinguishable from the variant found in black Africans, has been observed with a frequency of 1%-10% in all Melanesian and Polynesian populations studied. The TBG S variant is present also with low frequency in Micronesians and in some Indonesian populations. However, East Asians, Indians, and Australian Aborigines were found to be monomorphic.     

Genetic diversity and population structure of spottedtail goby (Synechogobius ommaturus) based on AFLP analysis

Larval dispersal may have an important impact on genetic structure of benthic fishes. To examine population genetic structure of spottedtail goby Synechogobius ommaturus, samples from five different locations of China and Kunsan population in Korea were analyzed by using amplified fragment length polymorphism (AFLP) technology. A total of 253 bands were identified from 91 individuals by 5 primer combinations and the percentage of polymorphic bands was 43.87%. The average gene diversity was 0.0794 ± 0.1470 and Shannon’s information index was 0.1279 ± 0.2138. The pairwise F_st values ranged from 0.022 to 0.201. The results of AMOVA analysis indicated that 90.54% of the genetic variation contained within populations and 9.46% occurred among populations. The gene flow estimates (Nm) demonstrated that different gene flow existed among populations from 0.994 to 11.114. No significant genealogical branches or clusters were recognized on the UPGMA tree. The results support the hypothesis that larval dispersal ability can be responsible for the genetic diversity and population structuring.     

Genetic structure of the asiatic black bear in Japan using mitochondrial DNA analysis

The genetic structure of the Asiatic black bear (Ursus thibetanus) in Japan was studied to understand the events that occurred during its evolution. The left domain of the mitochondrial control region (about 240 bp) was sequenced, defining 27 haplotypes that consisted of 23 haplotypes from 333 bears in Japan and 22 bears in the Asian continent. The network tree of the control region indicated that the Japanese population formed a distinct clade from the continental population. The phylogeographic analysis of the haplotypes indicated that the Shikoku and Kii Hanto populations had diverged during the initial phase from the ancestral population. After the 3 dominant haplotypes were rapidly distributed throughout Japan in the early stage of the population dispersal, the Japanese population diverged into eastern and western populations. Using the entire mitochondrial cytochrome b sequence, divergence time between the Japanese and the Continental populations suggested that the Japanese population might have colonized into Japan through the land bridge from the Korean Peninsula around 500 ka, which is consistent with paleontological evidence. Our finding that bears in western Japan exhibit lower genetic diversity and higher levels of genetic differentiation than bears in eastern Japan provides a vital contribution to conservation policy for these isolated populations.     

Genetic population structure of the swimming crab Callinectes danae (Crustacea: Decapoda) in southern Brazil

The population genetic structure of the swimming crab Callinectes danae (Crustacea, Portunidae) was studied by allozyme electrophoresis along the southern coast of Brazil (Santa Catarina and Rio Grande do Sul States). The biology of C. danae is poorly known, but some studies suggest that this species depends on estuaries for reproduction and for completing its life cycle, using them as recruitment sites and nursery grounds. If estuarine retention is an important process acting in favour of the recruitment of local populations, we should expect restriction of gene flow among populations inhabiting different estuaries. Therefore, our aim was to establish whether gene flow between populations of C. danaefrom different estuaries was restricted. Samples were collected in four estuaries: São Francisco do Sul, Laguna (Santa Catarina), Patos Lagoon and Chuí Stream (Rio Grande do Sul). Eleven loci were resolved. Contingency table tests (² and G) showed significant differences (P0.05) between pair-wise subpopulation comparisons. The estimated F _ST was =0.065±0.019, suggesting a moderate structuring of C. danae populations. No relationship was found for the number of migrants between pairs of subpopulations and the geographic distance separating them (P= 0.292; r ²= 0.269). Nevertheless, UPGMA analysis clustered together those estuaries separated by less than 250 km. Low adult migration, the requirement of estuaries as reproductive areas, recruitment sites and nursery grounds for juveniles, together with larval and post-larval retention processes, are discussed in the context of their importance in preventing panmixia among subpopulations inhabiting different estuaries.     

Genetic diversity analysis of Jatropha curcas L. (Euphorbiaceae) based on methylation-sensitive amplification polymorphism

Genetic analysis of 56 samples of Jatropha curcas L. collected from Thailand and other countries was performed using the methylation-sensitive amplification polymorphism (MSAP) technique. Nine primer combinations were used to generate MSAP fingerprints. When the data were interpreted as amplified fragment length polymorphism (AFLP) markers, 471 markers were scored. All 56 samples were classified into three major groups: γ-irradiated, non-toxic and toxic accessions. Genetic similarity among the samples was extremely high, ranging from 0.95 to 1.00, which indicated very low genetic diversity in this species. The MSAP fingerprint was further analyzed for DNA methylation polymorphisms. The results revealed differences in the DNA methylation level among the samples. However, the samples collected from saline areas and some species hybrids showed specific DNA methylation patterns. AFLP data were used, together with methylation-sensitive AFLP (MS-AFLP) data, to construct a phylogenetic tree, resulting in higher efficiency to distinguish the samples. This combined analysis separated samples previously grouped in the AFLP analysis. This analysis also distinguished some hybrids. Principal component analysis was also performed; the results confirmed the separation in the phylogenetic tree. Some polymorphic bands, involving both nucleotide and DNA methylation polymorphism, that differed between toxic and non-toxic samples were identified, cloned and sequenced. BLAST analysis of these fragments revealed differences in DNA methylation in some known genes and nucleotide polymorphism in chloroplast DNA. We conclude that MSAP is a powerful technique for the study of genetic diversity for organisms that have a narrow genetic base.     

Genetic diversity among Zygophyllum (Zygophyllaceae) populations based on RAPD analysis

Zygophyllum species are succulent plants that are drought resistant and/or salt tolerant, growing under severe, dry climatic conditions. Despite their importance and abundance in the Mediterranean and Middle East regions, there is little information concerning molecular variations among species of this genus. Genetic diversity was assessed, using RAPD primers, of 12 populations of Z. coccineum, Z. album and Z. aegyptium collected from various locations in Egypt and Saudi Arabia. Yong leaves were used for DNA extraction. Genetic distances were calculated using Nei's method. A dendrogram was constructed based on the similarity data matrix by unweighted pair group method using arithmetic averages cluster analysis. Analysis with RAPD markers revealed genetic variation between and within populations of Zygophyllum. Zygophyllum coccineum showed higher levels of genetic variation and more unique alleles than the other species.     

Genetic structure of Euclea schimperi (Ebenaceae) populations in monsoonal fog oases of the southern Arabian Peninsula

Euclea schimperi, a widespread Afromontane shrub or tree, occurs in the Arabian Peninsula in fragmented, semi-evergreen or evergreen woodland refugia in wet escarpment localities of the western and southern mountain chains. In the southern coastal mountains, the (semi-) evergreen woodland with E. schimperi is close to its ecological limit and consequently today very rare, with the exception of the monsoonal fog oases of east Yemen and south Oman in the central south coast of the Arabian Peninsula. Due to the steep precipitation gradient from the centre to the western edge in this monsoon affected area, E. schimperi is found in two different habitat types: in continuous woodland belts in the Hawf and Dhofar mountains, and in isolated, scattered woodland patches in the Fartak Mountains. Ten populations (138 individuals) from across the southern Arabian distribution area of the species were analysed using chloroplast microsatellites and AFLP fingerprinting to a) reconstruct the phylogeographical pattern of E. schimperi on the southern Arabian Peninsula and b) to evaluate the consequences of population fragmentation on the genetic diversity harboured in isolated patches vs cohering stands. Phylogeographical reconstructions show that the distribution area of E. schimperi in the southern Arabian Peninsula is characterised by a geographical split that separates the southwestern populations (representated by material from Jabal Eraf and Jabal Uthmar), from the southcentral populations, which themselves are split from each other into a western (Ras Fartak) and an eastern refugium (Hawf/Dhofar). The analysis of the within-population genetic diversity in E. schimperi populations resulted in a slightly, but not significantly higher genetic variation in small and isolated woodland patches (H_S=0.302) compared to larger, cohering stands (H_S=0.291).     

Genetic structure of the critically endangered plant Tricyrtis ishiiana (Convallariaceae) in relict populations of Japan

Tricyrtis ishiiana is a relic endemic plant taxon of the Convallariaceae that inhabits two nearby gorges in Kanagawa Prefecture, Japan. The distribution range and number of populations are thought to have been reduced to the present refugial populations during the Quaternary climatic oscillations. Because of its showy flowers, this plant has faced illegal removal from its natural habitats for horticultural use and has been designated a critically endangered species (class IA). In this study, we analyzed the genetic structure of the relict populations of T. ishiiana in order to contribute to the conservation strategies of the prefectural government. Our analyses of nine nuclear microsatellite loci detected high genetic diversity (H _E = 0.704 and H _O = 0.541) for the two populations. The two populations were slightly differentiated (R _ST = 0.032), accompanied by faint substructure across the populations (K = 3). In addition, each population exhibited spatial genetic structuring. The relatively low inbreeding coefficient for both populations together (F _IS = 0.233) and each population separately (F _IS = 0.217?C0.246) may be attributable to crossing among descendants within a population along with occasional gene flow between the populations. These results suggested that the extant populations have not experienced a severe bottleneck. The two extant populations were genetically differentiated at a very low level, accompanied by occasional pollen flow via pollinators and/or seed dispersal by gravity in the mountainous environment. Occasional gene exchange between the populations has allowed T. ishiiana to harbor high genetic diversity despite being a relic plant confined to two small refugial populations.     

Genetic structure within a Japanese stone pine (Pinus pumila regel) population on Mt. Aino-dake in central Honshu, Japan

In previous investigations, natural layering of Japanese stone pine (Pinus pumila) was suggested by the occurrence of adventitious roots. However, there is no genetic evidence so far that this species actually produces offspring by natural layering. We, therefore, investigated clonal structure and spatial genetic structure within a 38×18 m plot on Mt. Aino-dake, using allozyme, random amplified polymorphic DNA (RAPD), and inter-simple sequence repeat (ISSR) analyses. We found 24 genets, with stems found to be genetically identical in multiple tests, which extended later-ally against the direction of the slope, indicating that there were clonal structures originating from elongation of ramified stems and subsequent natural layering. The results suggest, however, that less than one third of the 200 stems analyzed from this site were clonaly propagated. We also analyzed spatial genetic structure by spatial autocorrelation. Many of the spatial autocorrelation coefficients were significantly positive in short distance classes. We concluded that the species has genetic structures which largely originate from clonal propagation and avian seed dispersal.     

Genetic architecture of aluminum tolerance in rice (Oryza sativa) determined through genome-wide association analysis and QTL mapping

Aluminum (Al) toxicity is a primary limitation to crop productivity on acid soils, and rice has been demonstrated to be significantly more Al tolerant than other cereal crops. However, the mechanisms of rice Al tolerance are largely unknown, and no genes underlying natural variation have been reported. We screened 383 diverse rice accessions, conducted a genome-wide association (GWA) study, and conducted QTL mapping in two bi-parental populations using three estimates of Al tolerance based on root growth. Subpopulation structure explained 57% of the phenotypic variation, and the mean Al tolerance in Japonica was twice that of Indica. Forty-eight regions associated with Al tolerance were identified by GWA analysis, most of which were subpopulation-specific. Four of these regions co-localized with a priori candidate genes, and two highly significant regions co-localized with previously identified QTLs. Three regions corresponding to induced Al-sensitive rice mutants (ART1, STAR2, Nrat1) were identified through bi-parental QTL mapping or GWA to be involved in natural variation for Al tolerance. Haplotype analysis around the Nrat1 gene identified susceptible and tolerant haplotypes explaining 40% of the Al tolerance variation within the aus subpopulation, and sequence analysis of Nrat1 identified a trio of non-synonymous mutations predictive of Al sensitivity in our diversity panel. GWA analysis discovered more phenotype-genotype associations and provided higher resolution, but QTL mapping identified critical rare and/or subpopulation-specific alleles not detected by GWA analysis. Mapping using Indica/Japonica populations identified QTLs associated with transgressive variation where alleles from a susceptible aus or indica parent enhanced Al tolerance in a tolerant Japonica background. This work supports the hypothesis that selectively introgressing alleles across subpopulations is an efficient approach for trait enhancement in plant breeding programs and demonstrates the fundamental importance of subpopulation in interpreting and manipulating the genetics of complex traits in rice.     

First record of the association between Malmgreniella cf. variegata (Polychaeta, Polynoidae) and Ophionereis annulata (Echinodermata, Ophionereididae) in the Mexican Pacific

An association between the scale-worm Malmgreniella cf. variegata and the ophiuroid Ophionereis annulata is reported for first time in the Mexican Pacific. The relationship was found in five localities across a total of 22 sites where O. annulata was collected. The polynoids collected were similar to the Western Atlantic scale-worm Malmgreniella variegata, which was previously recorded as commensal of O. annulata on the Pacific side of Panama, although they differ in colour pattern and the presence of notochaetae with a unidentate tip. The prevalence of infestation was 5 % and the intensity was 1.20. Experimental observations suggest that the behaviour of the scale-worm on O. annulata could be affected by light conditions, and that the commensal had a preference for its host over other ophiuroid species. More studies should be carried out to determine the taxonomic status of M. cf. variegata in the Eastern Pacific and to further elucidate the commensal–host relationship between this scale-worm and O. annulata.     

Genetic structure and demographic history of brown trout (Salmo trutta) populations from the southern Balkans

1. The present study was designed to characterize the genetic structure of brown trout ( Salmo trutta ) populations from the southern Balkans and to assess the spread of non-native strains and their introgression into native trout gene pools. We analysed polymorphism at nine microsatellite loci in seven supposedly non-admixed and three stocked brown trout populations.
2. The analyses confirmed the absence of immigration and extraordinarily strong genetic differentiation among the seven non-introgressed populations in parallel with low levels of intrapopulation genetic variability. In contrast, analyses of the stocked populations revealed that the genetic integrity of the local populations had been substantially changed, and the populations must be characterized as hybrid swarms. The pattern of population differentiation observed at microsatellites contrasted to that depicted previously by mtDNA variation. However, the close relationships between populations from the Danube and Axios river systems proposed solely by microsatellites could be explained by palaeogeographic events.
3. Our research showed that most of the populations examined represent unique gene pools, whose existence is critically compromised. Therefore, appropriate management and conservation strategies should be developed urgently in order to protect the subspecific biodiversity and to reverse currently negative trends.