Fine structure and development of the retina of the grenadier anchovy Coilia nasus (Engraulididae, Clupeiformes)

A study of the morphogenesis of the grenadier anchovy retina was undertaken using light and electron microscopy. Five developmental stages from prelarvae 3 days after fertilization to adult fish were studied. In addition to the general morphology of the eye and retina, special emphasis was given to the development of the photoreceptors and pigment epithelium (PE). The earliest retinae showing structural features indicative of a functioning eye are pure cone retinae composed of rows of alternating long and short cones forming a transient, tesselated pattern. At this stage there is a conventional PE containing melanin. In older stages cone rows are separated by the newly formed rods and by PE wedges filled with diffusely reflecting guanine crystallites. The findings are compared with the retinae of other engraulidids and with the development of teleost retinae in general. Moreover, the observed structural changes are discussed with respect to the photic habitat conditions of these anadromous fish that move between coastal waters, estuary, and river.     

Digestive enzyme activity of the Japanese grenadier anchovy Coilia nasus during spawning migration: influence of the migration distance and the water temperature

In this study, we investigated the activity levels of two major digestive enzymes (pepsin and lipase) in the commercially important Japanese grenadier anchovy Coilia nasus during its upstream migration to analyse the digestive physiological responses to starvation and to analyse the influence of the water temperature on enzyme activity. Water temperature had a significant effect on pepsin activity, while long-term starvation resulted in a significant decrease in pepsin activity. As starvation continued, however, a slight increase in pepsin activity between the Wuhu (440 river km) and Anqing (620 river km) regions may indicate that C. nasus had refeeding behaviour due to its large expenditure of energy reserves. In contrast, lipase activity was not significantly affected by the water temperature but the effect of fasting increased as much as 13% of lipase activity from the Chongming region (20 river km) to Anqing region, suggesting that the stored lipids of grenadier anchovy were mobilised to meet energy requirements of upstream migration activity and gonad development. Lipid mobilisation activated lipoprotein lipase (LPL; proteins with lipase activity) to hydrolyse triacylglycerides (TAG), which is the first step of lipid assimilation and obtained energy from fatty acids under fasting conditions. Therefore, the increased lipase activity is attributed mainly to the lipase that is involved in endogenous lipid hydrolysis. Grenadier anchovy appears to adapt to long-term starvation during migration and the increased lipase activity may indicate a crucial effect on lipid metabolism. This study demonstrated that distinct alterations occur in pepsin and lipase activities during the spawning migration of grenadier anchovy due to exogenous nutrition and endogenous metabolism. Furthermore, it provides a basis for further research on the digestive physiology and energy metabolism in this species.     

Richness and diversity of helminth communities in the Japanese grenadier anchovy, Coilia nasus, during its anadromous migration in the Yangtze River, China

To determine the relationship between the species richness, diversity of helminth communities, and migration distance during upward migration from coast to freshwater, helminth communities in the anadromous fish Coilia nasus were investigated along the coast of the East China Sea, the Yangtze Estuary, and 3 localities on the Yangtze River. Six helminth species were found in 224 C. nasus . Changes in salinity usually reduced the survival time of parasites, and thus the number of helminth species and their abundance. Except for the 2 dominant helminths, the acanthocephalan Acanthosentis cheni and the nematode Contracaecum sp., mean abundance of other 4 species of helminths was rather low (<1.0) during the upward migration in the Yangtze River. Mean abundance of the 2 dominant helminths peaked in the Yangtze Estuary and showed no obvious decrease among the 3 localities on the Yangtze River. Mean species richness, Brillouin's index, and Shannon index were also highest in the estuary (1.93 ± 0.88, 0.28 ± 0.25, and 0.37 ± 0.34, respectively) and did not exhibit marked decline at the 3 localities on the Yangtze River. A significant negative correlation was not seen between the similarity and the geographical distance (R = -0.5104, P = 0.1317). The strong salinity tolerance of intestinal helminths, relatively brief stay in the Yangtze River, and large amount of feeding on small fish and shrimp when commencing spawning migration perhaps were responsible for the results.     

Elemental signature in otolith nuclei for stock discrimination of anadromous tapertail anchovy (Coilia nasus) using laser ablation ICPMS

The elemental signature in otolith nuclei was determined using laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS) for stock discrimination of adult anadromous tapertail anchovy, Coilia nasus, in five Chinese estuaries. Five elements (Na, Mg, K, Sr, and Ba) were well detected in the otolith nuclei of the adult fish. Results showed that the elemental composition in the otolith nuclei varied substantially among the estuaries. Age and fish length data showed no significant influences on the elemental concentration ratios across the sample sites. The Sr:Ca and Ba:Ca ratios were inter-site distinct and could be used as natal tags for discriminating among stocks. Discriminant function analyses (DFA) showed that these ratios can be used in discriminating the Liaohe River estuary (LD, 92.3?%), the Yangtze River estuary (CJ, 86.7?%), and the Yellow River estuary (HH, 76.9?%) samples with high classification accuracy, followed by the Haihe River estuary (BH, 58.3?%) and the Daguhe River estuary (JZ, 46.2?%) samples. An overall classification accuracy rate of 72.7?% from the discriminant functions indicated that elemental fingerprinting appeared to have the potential to discriminate between tapertail anchovy stocks in these estuaries.     

Differentiation between populations of a termite in eastern Africa: implications for biogeography

Aim  African forests are divided by an arid corridor which runs from the Horn of Africa to the Namib Desert. Several forest species occur in the forests of eastern Africa as well as in the Guineo-Congolian forest block. We evaluate the possibility that such species may have crossed the arid corridor along a route through the Kenyan Highlands and down the eastern drainages during climatologically favourable periods in the past.
Locations  Eastern Africa, Ivory Coast.
Methods  We used the termite species Schedorhinotermes lamanianus (Sjöstedt). This species occurs in lowland forests and woodland throughout Africa south of the Sahara. We sampled termites from 12 populations. We evaluated the differentiation between populations using amplified fragment length polymorphisms as well as morphometrical measurements.
Results  Genetic and morphometrical analysis demonstrated substantial differentiation between populations west and east of the arid corridor in Kenya. To the east of this corridor we found an increase of morphological distance with geographical distance. Schedorhinotermes lamanianus occurs not only along the coast but also at isolated locations (e.g. ground-water forests in foothills) within the arid hinterland.
Main conclusions  We interpret these populations as remnants of a wider distribution during wet climatic periods. At these times, populations of S. lamanianus were apparently able to establish along extensive gallery forests protruding into the arid belt of the Kenyan hinterland. There have been no connections between populations of this species east and west of the arid corridor across the Kenyan Highlands.     

Polymorphic microsatellite markers for stock identification in Japanese anchovy (Engraulis japonica)

Japanese anchovy (Engraulis japonica) is a migratory marine fish of high economic significance in Taiwan. The adult Japanese anchovies migrate from the East China sea to spawn in coastal waters of Taiwan; the larvae then drift back to the East China Sea to complete their life cycle. We developed six highly polymorphic microsatellites for E. Japonica (expected heterozygosity ranging from 0.751 to 0.971) and these microsatellites can be used as genetic markers for identifying stocks to establish regulations in fishing management. Moreover, the markers will be useful in inferring the stock origins and migration routes in the future.     

Phylogeny and biogeography of the Japanese rhinoceros beetle,Trypoxylus dichotomus (Coleoptera: Scarabaeidae) based on SNP markers

The Japanese rhinoceros beetle Trypoxylus dichotomus is one of the largest beetle species in the world and is commonly used in traditional Chinese medicine. Ten subspecies of T. dichotomus and a related Trypoxylus species (T. kanamorii) have been described throughout Asia, but their taxonomic delimitations remain problematic. To clarify issues such as taxonomy, and the degree of genetic differentiation of Trypoxylus populations, we investigated the genetic structure, genetic variability, and phylogeography of 53 specimens of Trypoxylus species from 44 locations in five Asian countries (China, Japan, Korea, Thailand, and Myanmar). Using specific‐locus amplified fragment sequencing (SLAF‐seq) techniques, we developed 330,799 SLAFs over 114.16M reads, in turn yielding 46,939 high‐resolution single nucleotide polymorphisms (SNPs) for genotyping. Phylogenetic analysis of SNPs indicated the presence of three distinct genetic groups, suggesting that the various subspecies could be treated as three groups of populations. PCA and ADMIXTURE analysis also identified three genetic clusters (North, South, West), which corresponded to their locations, suggesting that geographic factors were important in maintaining within population homogeneity and between population divergence. Analyses of SNP data confirmed the monophyly of certain subspecies on islands, while other subspecies (e.g., T. d. septentrionalis) were found to be polyphyletic and nested in more than one lineage. AMOVA demonstrated high level of differentiation among populations/groups. Also, pairwise F_ST values revealed high differentiation, particularly between South and West, as well as between North and South. Despite the differentiation, measurable gene flow was inferred between genetic clusters but at varying rates and directions. Our study demonstrated that SLAF‐seq derived markers outperformed 16S and COII sequences and provided improved resolution of the genetic differentiation of rhinoceros beetle populations from a large part of the species’ range.     

长江刀鲚mor-4k13基因的分离鉴定及表达分析

分布于长江的刀鲚(Coilia nasus)具有洄游和定居两个生态型,生殖洄游是区别两者的主要表征。为了探索嗅觉受体(OR)基因是否参与了刀鲚的生殖洄游过程,本文采用RACE技术从洄游型刀鲚中获得了mor-4k13基因,该基因全长1 098 bp,编码区长963 bp,单外显子结构,可编码320个氨基酸。预测表明,mor-4k13基因编码的蛋白质,为7个疏水性的α-螺旋跨膜结构,属G-蛋白偶联受体,有胆固醇和油酸两个配体。MOR-4K13蛋白与已报道的其他鱼类OR蛋白的同源性在40%~68%之间,其中,与近缘种大西洋鲱(Clupea harengus)嗅觉受体蛋白同源性高达68%。采用Real-time PCR方法对10个组织或器官所作的荧光定量分析显示,mor-4k13基因在定居型刀鲚嗅囊和性腺中高表达,在肌肉、眼球、胃壁、肝和鳃中低表达,心肌中几乎不表达。mor-4k13基因在洄游型刀鲚嗅囊中的表达量总体高于定居型,且洄游型雄性刀鲚嗅囊中此基因的表达量约是其雌性嗅囊中的3倍。这表明mor-4k13基因不仅与嗅觉功能和性腺发育相关,也可能与生殖洄游习性相关,不同性别的个体间也存在着嗅觉能力的差异。     

The migratory history of anadromous and non-anadromous tapertail anchovy Coilia nasus in the Yangtze River Estuary revealed by the otolith Sr:Ca ratio

The migratory history of tapertail anchovy Coilia nasus in the Yangtze River Estuary, China was investigated using otolith Sr:Ca ratios and two-dimensional images of the Sr level from an X-ray electron probe microanalyzer (EPMA). The results showed that 17 of the 22 young-of-the-year (YOY) specimens had low Sr:Ca ratios (1.2–2.4?×?10^?3;1.5?±?0.3?×?10^?3) at the central otolith area, indicating their riverine origin and initial freshwater residence. In addition, 11 of the 14 adult specimens had low Sr:Ca ratios (1.3–2.2?×?10^?3; 1.7?±?0.4?×?10^?3) at the central otolith area but showed alternating changes between high (>4.0?×?10^?3) and low (<2.5?×?10^?3) values outside of this region, reflecting their riverine origin and the migration between freshwater and estuarine habitats. These 28 specimens represented the anadromous population in this region. The other 5 YOY specimens had high Sr:Ca ratios (3.6–5.9?×?10^?3; 4.8?±?0.8?×?10^?3) throughout the life history. Similarly, the other 3 adult specimens had high Sr:Ca ratios (4.0–5.7?×?10^?3; 4.8?±?0.7?×?10^?3) at the central otolith area but showed alternating changes between low and high values outside this region, suggesting that estuarine-origin non-anadromous individuals occurred in this region. The average of the otolith Sr:Ca ratios and Sr level mapping along the life-history transects could be used as a scalar for charting the migratory history of the tapertail anchovy in the Yangtze River Estuary: <2.0?×?10^?3 for freshwater residence and 3.5–6.0?×?10^?3 for estuarine residence.     

基于线粒体控制区的序列变异分析中国东南部沿海凤鲚种群遗传结构

为了解中国东南部沿海凤鲚(Coilia mystus)的种群遗传多样性和遗传结构, 本文分析了长江口(CJ)、钱塘江口(QT)、闽江口(MJ)和九龙江口(JL)4个凤鲚地理群体的mtDNA控制区561 bp片段的序列变异。65尾样本共检测到28个单元型。4个群体总的单元型多样性和核苷酸多样性均较高(h = 0.9433 ± 0.0168, π = 0.0317 ± 0.0158), 但单个群体的核苷酸多样性水平却很低, 其中以CJ最高(π = 0.0080 ± 0.0046), MJ最低(π = 0.0015 ± 0.0013)。MJ与JL群体之间以及CJ与QT群体之间的平均K2P遗传距离很小, 分别为0.3%和0.8%; 而CJ、QT分别与MJ、JL群体之间的遗传距离均较大, 达到了6%。采用最大似然法(ML)、最大简约法(MP)和邻接法(NJ)分别构建的单元型间的系统发育树揭示, 4个凤鲚群体构成CJ-QT 和MJ-JL 2个支系, 且具有极高的支持率。单元型的网络分析也显示这两个支系间有高达28步的突变次数。AMOVA分析显示大部分的遗传变异来自这两支系群体间(90.77%), 表明凤鲚群体间存在着显著的地理分化。种群分化指数和基因流分析也表明, 支系间群体有着明显的遗传分化(F_ST > 0.9, N_m < 0.03)。所有分析结果支持所研究的凤鲚标本属于两个不同的地理种群, 且种群的分化至少已达到亚种水平。采用BEAST和TRACER软件得到凤鲚两个亚种的最近共同祖先约在0.34–0.46百万年前, 处于更新世晚期, 推测可能是第四纪晚期的气候旋回和海平面的升降导致了凤鲚的种群分化。     

Seasonal occurrence of helminths in the anadromous fish Coilia nasus (Engraulidae): parasite indicators of fish migratory movements

To understand the seasonal migration of the anadromous Coilia nasus , we attempted to identify the parasites infecting C. nasus and determine their seasonal occurrence. From June 2007 to July 2008, a survey of 775 C. nasus individuals from the estuary of the Yangtze River and the coast of the East China Sea revealed more than 7,300 parasites associated with the gills and alimentary tracts of C. nasus . The following 6 helminth taxa were identified, i.e., the monogeneans Heteromazocraes lingmueni and Helciferus tenuis, the digenean Elytrophallus coiliae, the acanthocephalan Acanthosentis cheni , and larvae of the nematodes Anisakis simplex and Contracaecum sp., all of which are marine or brackish-water parasites. The absence of freshwater helminths suggested that the parasites acquired in freshwater may have been accidentally, and easily, lost by the time the fish had reached the estuary and coast. Contrary to seasonal occurrence of the parasites' life cycles, the lowest mean abundance and prevalence of H. lingmueni and A. cheni occurred in August, which suggested the immigration of C. nasus from freshwater to the Yangtze estuary, with lower parasite burdens. The highest mean abundance and prevalence of the nematodes A. simplex and Contracaecum sp. in May and June, and the lowest in August, indicated the arrival of the fish from the coast and the Yangtze River, to the estuary, respectively. These findings suggested that a majority of the fish prepared for spawning migration in the estuary in spring and early summer and returned to the estuary after spawning in the lower and middle reaches of the Yangtze River in late summer.     

Genetic diversity analysis of Perinereis aibuhitensis based on ISSR and SRAP markers of Chinese coast populations

The objective of this study was to obtain an overview of the genetic relationships within Perinereis aibuhitensis using Inter-Simple Sequence Repeat (ISSR) and Sequence-Related Amplified Polymorphism (SRAP) markers that were derived from related populations residing in the Chinese coasts. The percentage of polymorphic bands, Nei's gene diversity and Shannon's information index revealed a high level of genetic diversity at the species level. The analysis of molecular variance revealed that 81.22% (ISSR) and 76.29% (SRAP) of variability were partitioned among individuals within populations, which indicated the coherent trend by Nei's genetic differentiation (Gst) (0.2568/0.2876). The gene flow number (Nm) was 1.4470/1.2385, which indicated that there was limited gene exchange between populations. The phylogenetic tree of the ten P. aibuhitensis populations was separated into four major clusters using the neighbor-joining (NJ) method. These results provide a simple and useful basis for P. aibuhitensis germplasm research and aquaculture breeding.     

Use of inter-simple sequence repeat (ISSR) markers for discrimination between and within species of blackflies (Diptera: Simuliidae)

The present study is the first report of fingerprinting in blackflies (Diptera: Simuliidae), using inter-simple sequence repeat (ISSR) markers. Among five primers tested, three tetranucleotide repeat primers ((GACA)₄, (ACTG)₄, (ACAG)₄) generated a high proportion of polymorphic bands. Seven species representing various genera, subgenera or species groups were compared. No similar profiles were found. Intraspecific and interspecific banding patterns were analysed for two species in the Prosimulium hirtipes (Fries, 1824) species group and four species in the Simulium variegatum (Meigen, 1818) species group. The UPGMA cluster analysis based on Jaccard’s coefficient demonstrated the intraspecific and interspecific diversity and the resolving power of the ISSR markers to differentiate blackfly species and populations. In Simulium maximum (Knoz, 1961), geographically defined populations were successfully discriminated.     

Linkage maps for the Pacific abalone (genus Haliotis) based on microsatellite DNA markers

This study presents linkage maps for the Pacific abalone (Haliotis discus hannai) based on 180 microsatellite DNA markers. Linkage mapping was performed using three F1 outbred families, and a composite linkage map for each sex was generated by incorporating map information from the multiple families. A total of 160 markers are placed on the consolidated female map and 167 markers on the male map. The numbers of linkage groups in the composite female and male maps are 19 and 18, respectively; however, by aligning the two maps, 18 linkage groups are formed, which are consistent with the haploid chromosome number of H. discus hannai. The female map spans 888.1 cM (Kosambi) with an average spacing of 6.3 cM; the male map spans 702.4 cM with an average spacing of 4.7 cM. However, we encountered several linkage groups that show a high level of heterogeneity in recombination rate between families even within the same sex, which reduces the precision of the consolidated maps. Nevertheless, we suggest that the composite maps are of significant potential use as a scaffold to further extend the coverage of the H. discus hannai genome with additional markers.     

准噶尔无叶豆片断化居群的遗传变异及克隆多样性

准噶尔无叶豆(Eremosparton songoricum)是豆科无叶豆属小半灌木, 既能开花结实进行有性繁殖, 又可以靠根茎进行无性克隆繁殖, 为国家三级保护植物, 在中国仅片断化分布于新疆古尔班通古特沙漠局部区域。本文采用ISSR分子标记对采自古尔班通古特沙漠腹地及边缘的7个准噶尔无叶豆自然居群共148个个体进行了遗传变异和克隆多样性分析。8个引物共扩增出84个位点, 其中77个为多态性位点, 物种水平上的多态位点百分比PPB为91.67％, Nei's基因多样性指数I为0.3192, Shannon信息指数H为0.3540; 居群水平上的多态位点百分比PPB为58.45％, Nei's基因多样性指数I为0.2248, Shannon信息指数H为0.3270。居群间的遗传分化系数G_ST为0.2978。AMOVA分析表明, 有31.88%的遗传变异存在于居群间, 表明居群间存在显著的遗传分化。克隆多样性分析表明, 居群水平上, 居群G的Simpson多样性指数和均匀度指数最高, 分别为0.9400和0.9885; 居群E最低, 分别为0.8457和0.9021。物种水平上, Simpson多样性指数为0.9858, 均匀度指数为0.9673。本研究结果表明, 和其他荒漠植物相比, 准噶尔无叶豆表现出较高的遗传变异水平和克隆多样性, 这主要与该物种兼性生殖的繁育方式及多克隆起源有关; 而居群间产生了显著的遗传分化则主要由于人为干扰引起的生境片断化和居群减小而导致的基因交流障碍所致。遗传变异水平和遗传结构的研究将为分析准噶尔无叶豆致濒原因及进化潜力提供重要资料, 对该物种保护具有指导意义。     

湖北野生春兰资源遗传多样性的ISSR分析

近年来,由于过度采挖和生境片断化,湖北野生春兰(Cymbidiumgoeringii)资源正面临着灭绝的危险。本文采用ISSR分子标记技术对湖北省内的11个春兰野生居群共325个个体的遗传多样性水平及居群遗传结构进行了研究。11个引物共检测到127个位点,其中112个为多态位点,占88.19%。POPGENE分析结果表明:与其他兰科植物相比,春兰具有丰富的遗传变异(在物种水平上,He=0.2628,Ho=0.4037;在居群水平上,PPL=63.06%,He=0.1945,Ho=0.2958)。Nei's遗传多样性分析和AMOVA分析表明,各居群间产生了一定程度的遗传分化(GST=0.2440,FST=0.2207)。居群间一定程度的遗传分化可能是由生境破坏和基因流障碍(Nm=0.8828)引起。UPGMA聚类分析可知,与其他居群相比,恩施地区的5个居群,即巴东(BD)、福宝山(FBS)、宣恩(XE)、毛坝(MB)、来凤(LF)优先聚成一支,而大悟(DW)居群单独聚为一支。同时本研究也表明,虽然春兰自交亲和,但在自然界中其繁育系统还是以异交为主。鉴于春兰资源的遗传多样性现状和其相应的居群遗传结构,我们建议在遗传多样性较高的来凤(LF)、京山(JS)、大悟(DW)居群设立保护点进行就地保护;而对资源破坏最为严重的毛坝(MB)和宣恩(XE)居群要实行迁地保护。     

Assessing genetic diversity of Elymus sibiricus (Poaceae: Triticeae) populations from Qinghai-Tibet Plateau by ISSR markers

Inter-simple sequence repeats (ISSR) markers were used to assess the genetic diversity and population structure in eight populations of Elymus sibiricus L. from the southeast of Qinghai-Tibet Plateau of China. Of the 100 primers screened, 13 produced highly reproducible ISSR bands. Using these primers, 193 discernible DNA fragments were generated with 149 (77.2%) being polymorphic, indicating considerable genetic variation at the species level. In contrast, there were relatively low levels of polymorphism at the population level with the percentage of polymorphic bands (PPB) ranging from 44.04 to 54.92%. The mean gene diversity (H_E) was estimated to be 0.181 within populations (range 0.164–0.200), and 0.274 at the species level. A high level of genetic differentiation among populations was detected based on Nei's genetic diversity analysis (33.1%), Shannon's index analysis (34.5%), Bayesian method (33.2%) and AMOVA analysis (42.5%). No significant statistical differences (analysis of molecular variance [AMOVA], P = 0.08) in ISSR variation were found between the sample collection regions. However, among populations (42.5% of the variance) and within populations (57.5% of the variance), there were significant differences (P < 0.001). Populations shared high levels of genetic identity. This pattern of genetic variation was different from that for most of inbreeding Triticeae species reported. In addition, a geographical pattern of population differentiation, where the populations from south and north of sampling sites were clearly separated from each other, was revealed by both the cluster and principal coordinates analyses. Generally, the result of this study indicates that E. sibiricus contains high molecular variation in its populations. The implications of these results for the conservation of the species are discussed.