Horse mackerel egg identification using DNA methodology

Egg identification in plankton samples is usually needed for purposes of stock assessment. Until recently, only morphological characters were used for identifying the eggs of different fish species. Late developmental stages are easily distinguishable due to pigmentation as well as egg and oil globule size range. However, for early stages, these characteristics are difficult to be discriminated and may overlap with other species. European horse mackerel species (Trachurus trachurus, T. mediterraneus, T. picturatus) overlap significantly in their spawning areas in some European waters. Because of the fact that their eggs are morphologically similar, genetic methodologies are needed to identify eggs and larvae to species correctly. In the present study, formalin‐ and ethanol‐preserved eggs were tested to estimate the efficacy of genetic methodologies for species identification of eggs when different preservatives are used. A 370‐bp fragment of cytochrome b was successfully amplified followed by restriction fragment analysis with two restriction enzymes aiming to identify the eggs to species. Horse mackerel egg identification was accomplished with the maximum success in ethanol‐preserved eggs. However, it seems that various preservatives had different effects on the DNA quality of eggs as genetic identification was less successful in formalin‐preserved eggs. Preserving in ethanol a part of the eggs obtained in plankton surveys is suggested for purposes of accurate genetic identification, even if their morphological features are distorted in time.     

4种笛鲷的线粒体DNA多样性分析

取红鳍笛鲷(Lutjanus erythopterusBloch)、紫红笛鲷(Lutjanus argentimaculatusFor-skal)、勒氏笛鲷(Lutjanus russelliBleeker)和约氏笛鲷(Lutjanus johniBloch)肝脏组织,分离纯化其线粒体DNA,用限制性内切酶酶切进行了限制性片段长度多态性分析。在红鳍笛鲷共发现4种单倍型,单倍型间的平均遗传距离为0.010 1,其mtDNA多态度为0.002 9。在紫红笛鲷共发现6种单倍型,单倍型间的平均遗传距离为0.015 1,其mtDNA多态度为0.005 5。在勒氏笛鲷共发现3种单倍型,单倍型间的平均遗传距离为0.008 6,其mtDNA多态度为0.001 2。在约氏笛鲷共发现5种单倍型,单倍型间的平均遗传距离为0.008 3,其mtDNA多态度为0.002 0。     

星洲银罗非鱼线粒体DNA的RFLP研究

用EcoRⅤ、MboⅡ、NaeⅠ、PvuⅡ、StyⅠ、KpnⅠ、SacⅠ、HindⅢ、DraⅠ、HpaⅡ等10种限制性核酸内切酶对星洲银罗非鱼线粒体DNA(mtDNA)进行酶切分析,其中StyⅠ、SacⅠ和DraⅠ表现出多态性,HpaⅡ酶切后产生零碎片段,EcoRⅤ、NaeⅠ、PvuⅡ、KpnⅠ只有一个切点,HindⅢ有两个切点,MboⅡ有5个切点但有小片段丢失。在星洲银罗非鱼中共发现4种单倍型,单倍型间平均遗传距离为0.008 0,其mtDNA多态度为0.002 4,遗传多样性较丰富。     

Genetic discrimination for three gynogenetic clones of silver carp Hypophthalmichthys molitrix, based on restriction endonuclease analysis of Nd5-Nd6 region of mitochondrial DNA

1 INTRODUCTION Silver carp (Hypophthalmichthys molitrix) is an important species in freshwater fisheries and ranks the first in world fish production (Li, 1993). China has a long history of aquaculture of silver carp that is very difficult for selective breeding because it has a long life cycle. However, development of cytoge-netics and modern molecular genetics built up a strong impact to the research on selective breeding. A rapid, efficient approach for establishment of a pure line of…     

Preliminary study on mitochondrial 16S rRNA gene sequences and phylogeny of flatfishes （Pleuronectiformes）

A 605 bp section of mitochondrial 16S rRNA gene from Paralichthys olivaceus, Pseudorhombus cinnamomeus, Psetta maxima and Kareius bicoloratus, which represent 3 families of Order Pleuronectiformes was amplified by PCR and sequenced to show the molecular systematics of Pleuronectiformes for comparison with related gene sequences of other 6 flatfish downloaded from GenBank. Phylogenetic analysis based on genetic distance from related gene sequences of 10 flatfish showed that this method was ideal to explore the relationship between species, genera and families. Phylogenetic trees set-up is based on neighbor-joining, maximum parsimony and maximum likelihood methods that accords to the general rule of Pleuronectiformes evolution. But they also resulted in some confusion. Unlike data from morphological characters, P olivaceus clustered with K.bicoloratus, but P cinnamomeus did not cluster with t3. olivaceus, which is worth further studying.     

栉孔扇贝16S rRNA基因片段序列的多态性研究

采用PCR技术扩增了栉孔扇贝（Chlamys farreri)线粒体DNA的16SrRNA基因片段，PCR产物经T载体连接之后进行了克隆，测序，得到634bp的核苷酸序列；分析了该片段的大连，烟台，青岛，朝鲜4个自然群体31个个体的核苷酸序列多态性，共检测到26个多态性核苷酸位点，18种单倍型，结果表明，4个群体中以朝鲜群体遗传多样性最高，其次为烟台，青岛群体，大连群体最低；不同自然分布区的栉孔扇贝未出现明显的遗传分化。     

The genetic structure of the exotic ascidian Styela plicata (Tunicata) from Italian ports,with a re‐appraisal of its worldwide genetic pattern

The pleated ascidian Styela plicata (Lesueur, 1823) is a solitary species commonly found in ports and marinas around the world. It has been recorded in the Mediterranean region since the mid‐19th century. In the present work, the species’ genetic diversity was analysed, employing a 613‐bp portion of the mitochondrial cytochrome c oxidase subunit I (COI) gene from 149 individuals collected in 14 ports along Italian coasts at spatial scales ranging from 1 to approximately 2200 km. Haplotype and nucleotide diversity values were h = 0–0.933 (total h = 0.789) and π = 0–0.145 (total π = 0.0094), respectively. A general southward trend of increasing within‐population genetic diversity was observed. Analysis of molecular variance revealed significant genetic structuring but no significant differences were detected among basins, and no isolation by distance was found. Our data were integrated with the COI sequences available from previous studies and re‐analysed in order to investigate the possible routes of introduction of this ascidian into the Mediterranean Sea. The presence of the two COI haplogroups detected in previous molecular investigations on S. plicata at intercontinental spatial scale was confirmed in the Mediterranean Sea. The results revealed multiple introductions of S. plicata, although some locations appear to have experienced rapid expansion from few founding individuals with reduced genetic diversity. However, continuous introductions would confound the pattern deriving from single founder events and make it difficult to estimate the time needed for gene diffusion into established populations. This mixing of effects creates difficulties in understanding the past and current dynamics of this introduction, and managing this alien invasive ascidian whose genetic structure is continuously shuffled by vessel‐mediated transport.     

The invasive Asian green mussel Perna viridis in South Africa: all that is green is not viridis

The Asian green mussel Perna viridis is an invasive Indo-Pacific species recently reported from South African harbours. To verify the invasion, a phylogenetic (and morphological) analysis of green-shelled mussels (n = 39), found in six South African harbours, was conducted using the mitochondrial cytochrome c oxidase subunit I gene (COI). Estimates of genetic distances using the neighbour-joining analysis identified P. viridis only from Durban Harbour. All other green mussels were more than 3.2% divergent from P. viridis and were identified as green-shelled variants of indigenous P. perna. The only reliable morphological differences distinguishing the two species were the poorly developed mantle papillae and the wavy pallial line in P. viridis. The confirmed occurrence of P. viridis in a South African harbour suggests that there is a possible threat of the species becoming established and then spreading onto the open coast and competing with indigenous P. perna.