Identification of Species in Genus Platycephalus from Seas of China

Platycephalus in Chinese sea area has a high commercial value.However,there were mis-identifications in previous records.In this study,we fully distinguished and diagnosed all five species,Platycephalus indicus,P.cultellatus,Platycephalus sp.,Platycephalus sp.1 and Platycephalus sp.2.The results revealed that P.cultellatus was overlooked by previous ichthyologists.Platycephalus sp.1 was misidentified as P.indicus in reality,and Platycephalus sp.2 only existed in the seas of Japan.Furthermore,morphological,especially phylogenetic analysis indicated that Platycephalus sp.from South China Sea differs from all former known species,which might be a new species.We identified all Platycephalus species in China seas for the first time,which will contribute to local species identification,biodiversity conservation and sustainable exploitation of Platycephalus species.     

中国野鲮亚科鱼类一新属新种记述

描记采于贵州省茂兰自然保护区境内一洞穴的中国野鲮亚科（Labeoninae）一新属长臀鲮属Longanalus．sp nov和一个新种大鳍长臀鲮Longanalus macrochirous sp nov。新属以臀鳍分枝鳍条为7根等特征．区别于该亚科所有属；新种以偶鳍发达．胸鳍后伸达到腹鳍基，腹鳍后伸超过臀鳍起点等显著特征命名大鳍长臀鲮。     

中国野鲮亚科鱼类一新属新种记述

描记采于贵州省茂兰自然保护区境内一洞穴的中国野鲮亚科(Labeoninae)一新属长臀鲮属Longanalus,spnov和一个新种大鳍长臀鲮Longanalus macrochiroussp nov。新属以臀鳍分枝鳍条为7根等特征,区别于该亚科所有属;新种以偶鳍发达,胸鳍后伸达到腹鳍基,腹鳍后伸超过臀鳍起点等显著特征命名大鳍长臀鲮。     

巨蛎属牡蛎DNA序列的比较及鉴定

【目的】了解巨砺属牡蛎的系统关系和种类区分。【方法】对我国沿海分布的7种巨砺属牡蛎(香港牡蛎、有明牡蛎、熊本牡蛎、岩牡蛎、艾氏牡蛎、太平洋牡蛎、葡萄牙牡蛎)的线粒体DNA片段(COI、12S、16S和tRNAs),以及核糖体RNA基因转录间隔子序列(ITS-1、ITS-2和ITS)进行分子系统学研究。【结果】无论种间或种内,tRNAs序列差异最大,亚种内TS1序列差异最大;tRNAs和COI序列较其它序列变异更快,种间变异与种内变异之间有明显的条形码间隙;12S、16S、ITS和ITS2的种间变异与种内变异无重叠及条形码间隙;ITS1的种间变异与种内变异出现重叠。在亚种层面,12S、ITS、ITS1和ITS2的亚种间变异与亚种内变异出现重叠;16S的亚种间变异与亚种内变异无重叠及条形码间隙;tRNAs与COI序列亚种间变异与亚种内变异之间有明显的条形码间隙,有利于区分亚种。【结论】tRNAs和COI序列可用于种及亚种的鉴定。     

Identification of a Long-Chain Fatty Acid Elongase from Nannochloropsis sp. Involved in the Biosynthesis of Fatty Acids by Heterologous Expression in Saccharomyces cerevisiae

The marine microalga Nannochloropsis sp. contains various elongases and desaturases that are critical for biosynthesis of polyunsaturated fatty acids. A full-length cDNA encoding a long-chain fatty acid elongase, named Ns FAE, was cloned from Nannochloropsis sp.. The open reading frame of Ns FAE(GenBank accession no. MF680548) consisted of 1068 bp and encoded a predicted protein of 355 amino acids with molecular mass 38.8 k Da. The deduced polypeptide showed 43%–44% identity to fatty acyl elongases from other algae. RT-PCR experiments indicated that the Ns FAE gene exhibited the highest expression in Nannochloropsis sp. at 72 h(i.e., during the third growth stage) and the expression was significantly lower in the other four growth stages. Plasmid p Ns FAE-CRISPR and a recombinant DNA fragment(ADH1p-Ns FAE-CYCt) were transformed into Saccharomyces cerevisiae strain BY4742 using the CRISPR-Cas system. Yeast transformants containing Ns FAE produced three fatty acids not normally present in wild-type BY4742-linoleic acid, linolenic acid and eicosadienoic acid-indicating that Ns FAE encodes a functional elongase enzyme.     

Preliminary Identification of Three New Isolates in Genus Alexandrium (Dinophyceae) from China Sea Area

1 Introduction Dinoflagellate Alexandrium has received intensive at-tention in recent years, as it is a valuable bioactive com-pound resource and related to harmful algal bloom (HAB)which has negative impact on marine ecology systemaquiculture, human health and tourism (Hallegraeff andBolch, 1992; Smayda, 1997; Vale and Antonia de MSampayo, 2001). However, not all of the species in genusAlexandrium produce toxin (Wendy et al., 2001), so rapididentification and clarification of the phyloge…     

Cloning and characterization of a delta-6 desaturase encoding gene from Nannochloropsis oculata

A gene(NANOC-D6D) encoding a desaturase that removes two hydrogen atoms from fatty acids at delta 6 position was isolated from a cDNA library of Nannochloropsis oculata(Droop) D.J.Hibberd(Eustigmatophyceae).The unicellular marine microalga N.oculata synthesizes rich long chain polyunsaturated fatty acids(LCPUFAs),including eicosapentaenoic acid(20:5n-3,EPA).The deduced protein contains 474 amino acids that fold into 4 trans-membrane domains.The neighbor-joining phylogenetic tree indicates that NANOC-D6D is ...     

Construction and characterization of a normalized cDNA library of Nannochloropsis oculata(Eustigmatophyceae)

We constructed and characterized a normalized cDNA library of Nannochloropsis oculata CS-179,and obtained 905 nonredundant sequences(NRSs) ranging from 431-1 756 bp in length.Among them,496 were very similar to nonredundant ones in the GenBank(E ≤1.0e-05),and 349 ESTs had significant hits with the clusters of eukaryotic orthologous groups(KOG).Bases G and/or C at the third position of codons of 14 amino acid residues suggested a strong bias in the conserved domain of 362 NRSs(60%).We also identified the unigenes encoding phosphorus and nitrogen transporters,suggesting that N.oculata could efficiently transport and metabolize phosphorus and nitrogen,and recognized the unigenes that involved in biosynthesis and storage of both fatty acids and polyunsaturated fatty acids(PUFAs),which will facilitate the demonstration of eicosapentaenoic acid(EPA) biosynthesis pathway of N.oculata.In comparison with the original cDNA library,the normalized library significantly increased the efficiencies of random sequencing and rarely expressed genes discovering,and decreased the frequency of abundant gene sequences.     

Construction and characterization of a normalized cDNA library of Nannochloropsis oculata (Eustigmatophyceae)

We constructed and characterized a normalized cDNA library of Nannochloropsis oculata CS-179, and obtained 905 nonredundant sequences (NRSs) ranging from 431–1 756 bp in length. Among them, 496 were very similar to nonredundant ones in the GenBank (E ≤1.0e-05), and 349 ESTs had significant hits with the clusters of eukaryotic orthologous groups (KOG). Bases G and/or C at the third position of codons of 14 amino acid residues suggested a strong bias in the conserved domain of 362 NRSs (>60%). We also identified the unigenes encoding phosphorus and nitrogen transporters, suggesting that N. oculata could efficiently transport and metabolize phosphorus and nitrogen, and recognized the unigenes that involved in biosynthesis and storage of both fatty acids and polyunsaturated fatty acids (PUFAs), which will facilitate the demonstration of eicosapentaenoic acid (EPA) biosynthesis pathway of N. oculata. In comparison with the original cDNA library, the normalized library significantly increased the efficiencies of random sequencing and rarely expressed genes discovering, and decreased the frequency of abundant gene sequences.     

Genetic Transformation of Nannochloropsis oculata with a Bacterial Phleomycin Resistance Gene as Dominant Selective Marker

The gene ble from Streptoalloteichus hindustanus is widely used as a selective antibiotic marker.It can control the phleomycin resistance,and significantly increase the tolerance of hosts to zeocin.The unicellular marine microalga Nannochloropsis oculata is extremely sensitive to zeocin.We selected ble as the selective marker for the genetic transformation of N.oculata.After the algal cells at a density of 2×10~7 cells mL~(-1) was digested with 4% hemicellulase and 2% driselase for 1 h,the protoplasts accounted for 90% of the total.The ble was placed at the downstream of promoter HSP70A-RUBS2 isolated from Chlamydomonas reinhardtii,yielding a recombinant expression construct p MS188.The construct was transferred into the protoplasts through electroporation(1kV,15 μS).The transformed protoplasts were cultured in fresh f/2 liquid medium,and selected on solid f/2 medium supplemented with 500 ngmL~(-1) zeocin.The PCR result proved that ble existed in the transformants.Three transformants had been cultured for at least 5 generations without losing ble.Southern blotting analysis showed that the ble has been integrated into the genome of N.oculata.The ble will serve as a new dominant selective marker in genetic engineering N.oculata.     

Cloning and phylogenetic analysis of a fatty acid elongase gene from Nannochloropsis oculata CS179

Nannochloropsis oculata CS179, a unicellular marine microalga, is rich in long-chain polyunsaturated fatty acids (LCPUFAs). Elongase and desaturase play a key role in the biosynthesis of PUFAs. A new elongase gene, which encodes 322 amino acids, was identified via RT-PCR and 5′ and 3′ RACE. The sequence of the elongase gene was blast-searched in the NCBI GenBank and showed a similarity to those of the cryptosporidium. But the NJ-tree revealed that the N. oculata CS179 elongase clustered with those of the microalgae Phaeodactylum tricornutum, Ostreococcus tauri and Thalassiosira pseudonana.     

Using Different Standardized Methods for Species Identification: A Case Study Using Beaks from Three Ommastrephid Species

The cephalopod beak is a vital hard structure with a stable configuration and has been widely used for the identification of cephalopod species. This study was conducted to determine the best standardization method for identifying different species by measuring 12 morphological variables of the beaks of Illex argentinus, Ommastrephes bartramii, and Dosidicus gigas that were collected by Chinese jigging vessels. To remove the effects of size, these morphometric variables were standardized using three methods. The average ratios of the upper beak morphological variables and upper crest length of O. bartramii and D. gigas were found to be greater than those of I. argentinus. However, for lower beaks, only the average of LRL (lower rostrum length)/LCL (lower crest length), LRW (lower rostrum width)/LCL, and LLWL (lower lateral wall length)/LCL of O. bartramii and D. gigas were greater than those of I. argentinus. The ratios of beak morphological variables and crest length were found to be all significantly different among the three species (P < 0.001). Among the three standardization methods, the correct classification rate of stepwise discriminant analysis (SDA) was the highest using the ratios of beak morphological variables and crest length. Compared with hood length, the correct classification rate was slightly higher when using beak variables standardized by crest length using an allometric model. The correct classification rate of the lower beak was also found to be greater than that of the upper beak. This study indicates that the ratios of beak morphological variables to crest length could be used for interspecies and intraspecies identification. Meanwhile, the lower beak variables were found to be more effective than upper beak variables in classifying beaks found in the stomachs of predators.     

基于cox1条形码的鱼肚DNA分子鉴定

为鉴定鱼肚的正品来源,运用一对特异性引物扩增并测定10 个不同价格（700～1 500 元/kg）鱼肚样本的线粒体DNA 细胞色素c 氧化酶Ⅰ（cox1）约660 bp 的序列.通过BLASTN 比对DNA 序列同源度,判定鱼肚所属种类分别为鲈形目石首鱼科（Sciaenidae）、马鲅科（Eleutheronema）、笛鲷科（Lutjanidae）、带鱼科（Trichiuridae）.cox1 条码序列获取便捷,基于此条码序列的物种鉴定技术可用于鱼肚种类鉴别.     

隐丝藻科一新属──管形藻属

中空蜈蚣藻（Grateloupiaporracea（Mert．）W．X．LietZ．F．Ding）在幼苗期（0．2cm）已中空,成体的主枝与分枝亦中空。皮层细胞叉状排列或稍背斜形;生枝枝丛（ampullae）只有一条主枝或1分枝;果胞枝由1～3个细胞组成,有的果胞亦产生由1～4个细胞组成的侧枝。其特征与蜈蚣藻属（GrateloupiaC．Ag）不同。建议将其从蜈蚣藻属中移出另成立一个新属管形藻属SinotubimorphaW．X．LietZ．F．Dinggen,nov．该属模式种为管形藻S,porracea（Mert．）W．X．LietZ．F．Dingcomb．nov,采于青岛1984年。     

不同前处理方法对微拟球藻脂肪酸组成的影响

采用直接甲酯化、乙醚抽提、正己烷提取等3种方法对微拟球藻进行前处理,用索氏法提取油脂,经气相色谱分析,对比前处理方法对微拟球藻油脂提取率及脂肪酸组分的影响。结果表明:乙醚提取藻粉油脂的提取率(微藻的油脂质量分数)为12.70%,正己烷提取藻粉油脂的提取率为11.00%,乙醚提取油脂的提取率大于正己烷的提取率;直接甲酯化后测得微拟球藻脂肪酸中多不饱和脂肪酸(Polyunsaturated Fatty Acids,PUFAs)质量分数为20.04%,多不饱和脂肪酸中二十碳五烯酸(Eicosapentaenoic Acid,EPA)含量最丰富,占总脂肪酸13.0%;经过乙醚和正己烷提取后的油脂,脂肪酸组成更加简单,PUFAs含量明显增加,乙醚提取油脂EPA含量是直接甲脂化法的2.3倍,正己烷提取所得油脂EPA含量是直接甲脂化法的2.08倍。