黄鳍东方鲀精子的超微结构

在透射电子显微镜及环境扫描电子显微镜下观察黄鳍东方鲀精子的超微结构。结果表明:黄鳍东方鲀精子由头部和尾部两部分组成。头部呈长圆柱形,无顶体,核内有小空隙,空隙中有少量电子致密小颗粒存在;核后端正中有凹陷入核内的植入窝,中心粒复合体位于植入窝内,由近端中心粒、中心粒间体和基体组成;袖套位于核的后端,内含线粒体球和含有电子致密物质的小囊泡。尾部细长,自头部后端的正中部向后伸出,近核端部分很短,核心结构是轴丝,鞭毛起始于袖套腔,远核端有侧鳍;轴丝为典型的"9+2"微管对复合体结构。     

基于纳米尺度的碎屑岩骨架矿物结构研究

对胜利油田胜坨地区碎屑岩储层岩芯样品和岩石薄晶片分别采用场发射扫描电镜、透射电镜进行镜下观察与能谱测试分析.结果表明:碎屑岩中骨架矿物石英和长石中存在纳米尺度的结构特征;石英镜下可见类蚀像凹坑、雏晶单晶、平行脊线、晶体位错、类包裹体晶内核等5种纳米尺度的结构;长石镜下可见晶面平行凹痕、溶蚀针、纳米级薄层断阶、长石高岭石化、晶体位错和超微包裹体等6种纳米尺度的结构;这些结构大多在几百纳米的尺度范围内,有的甚至只有几十纳米;矿物在纳米尺度上表现出的结构特征与其本身的硬度变化、晶体晶格缺陷、晶体生长过程以及成岩作用的显微表现和显微构造应力改造作用有关.对碎屑岩骨架矿物中纳米尺度结构特征的进一步研究有望为岩石显微变形的应力作用机制和方式以及溶解作用等成岩作用模式做出有效分析,进而为油气勘探开发提供新的探索方式.     

Molecular phylogeny of oligotrich genera Omegastrombidium and Novistrombidium (Protozoa, Ciliophora) for the systematical relationships within Family Strombidiidae

The phylogeny of the oligotrich ciliates is currently a hot debate despite the availability of both morphological and molecular data. In the present paper, further small subunit rRNA (SS rRNA) genes were analyzed from the Genera Omegastrombidium and Novistrombidium, as well as from Strombidium, and combined with three new SS rRNA sequences from Strombidium basimorphum, S. sulcatum population QD-1, and Novistrombidium testaceum population GD. The phylogenetic positions of these organisms were inferred using Bayesian inference, Maximum Likelihood, and Maximum Parsimony methods. The main results are: (1) the SS rRNA gene sequence analyses match the recent findings about the molecular evolution of oligotrichs, indicating that the family Strombidiidae is paraphyletic; (2) the Genus Omegastrombidium is separated from the Genus Strombidium, as shown in recent cladistic analyses; (3) morphospecies in Genus Novistrombidium, based on similarity of somatic ciliature, are separated from each other in all topological trees, indicating that this genus could be a paraphyletic group; (4) the molecular data indicate a possibility of paraphyly for the genus Strombidium; and (5) the similarities of the SS rRNA gene of specimens identified as S. sulcatum and S. inclinatum are 99.8%–100%. However, present knowledge on the oligotrichs sensu stricto is still insufficient and further studies based on both molecular and other technologies are required.     

Screening for lipophilic marine toxins and their potential producers in coastal waters of Weihai in autumn, 2020

Lipophilic marine toxins (LMTs) produced by some microalgae in the sea could accumulate in shellfish and pose potential threats to the health of seafood consumers. Phytoplankton and shellfish samples were collected from coastal waters of Weihai in Shandong Peninsula, China in autumn, 2020, and screened for lipophilic marine toxins and their potential producers using liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis and high throughput sequencing of partial DNA (V4 region of the 18S rRNA gene) extracted from phytoplankton. Pectenotoxin-2 (PTX2), trace amounts of azaspiracid (AZA1 or AZA40), and 13-desmethyl spirolide C (13-DesMe-C) were detected in phytoplankton samples, while PTX2 and gymnodimine (GYM) were detected in shellfish samples. The toxin content in shellfish samples was much lower than the regulatory limit or values reported previously. Results suggest that lipophilic marine toxins should have low risk in coastal waters of Weihai in autumn. Based on the data of high throughput sequencing, the OTUs were assigned to 5 identified species of Alexandrium, including A. ostenfeldii capable of producing 13-DesMe-C and GYM. Two OTUs were found closely related to the toxic species in genus Dinophysis, but it is impossible to assign them to any identified species due to the low resolving power of the V4 region for Dinophysis. The OTUs could not be assigned to any identified species in the genus Azadinium, suggesting the existence of unidentified species in this region.

     

Preparation and charaterization of self-assembled nanoparticles based on linolenic-acid modified chitosan

1 IntroductionHydrophobically modified water-soluble polymers,also known as hydrophobized polymers, are thosemodified by hydrophobic groups such as alkyl,arolkyl, and deoxycholic acid. In water, they canself-aggregate due to their intro- and/or intermolecularhydrophobic interaction. By this process, unique hy-drogel nanoparticles with hydrophobic core and hy-drophilic shell can be prepared, which is a new ap-proach to preparing the monodispersed hydrogelnanoparticles (Whitesides et al., 1991; …     

Diversity and community structure of eukaryotic microalgae in surface sediments in the central Bohai Sea,China, based on a metabarcoding approach

Sediment samples were collected at 17 stations in the central Bohai Sea, China, and the diversity and community structure of eukaryotic microalgae were assessed by metabarcoding the V4 region of 18S rDNA. A total of 930 operational taxonomic units (OTUs) were detected for microeukaryotes, including 98 algal OTUs. The algal communities comprised 42 genera belonging to 19 classes of six phyla, and they were dominated by chrysophytes and dinoflagellates. Dinoflagellates were also the most diverse microalgal group. The nano-sized dinoflagellates Biecheleria halophila and Azadinium trinitatum occurred abundantly in the study area; however, they have not been reported previously, as they may be overlooked or misidentified in light microscopy. Many pico-sized chlorophytes were detected in the sediment samples. Sixteen of the detected OTUs were assigned to potentially harmful and/or bloom-forming microalgae, suggesting some potential risks of harmful algal blooms in the central Bohai Sea. The capacity of metabarcoding to detect morphologically cryptic and small species makes this method a sufficiently sensitive means of detection for assessing eukaryotic microalgae in sediments.

     

Studies on the Tolerating Mechanism for Sulfide in Urechis Unicinctus (Echiura:Urechidae)-cytological observation on Urechis Unicinctus in Different Hydrogen Sulfide Environment

This study on the cytological changes of the body wall ,aspiratory intestine and crissal bursa in Urechis unicincuts by light microscope(LM) and transmission electron microscope(TEM) showed that the difference between the body wall and the natural environment rich in H2S was not obvious,that the wall color of the aspiratory intestine in H2S rich environment changed from normal semitransparency to dark brown.that its epithelia were disassembled and the electron density of its cytoplasm matrix was lower;and that in H2S rich environment many basoplhilic granules occurred in the epithelia of the crissal bursa.Granules with single membrane and myelinefingure were found with TEM.     

Histological study on the skin of Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

Histological development of Japanese flounder Paralichthys olivaceus larval skin and ultrastructural difference of skin between reared normal and malpigmented Japanese flounder were studied with light microscopy (LM) and transmission electron microscopy (TEM). The results show that the skin develops slowly before the metamorphosis, while at the onset of metamorphosis, the skin develops quickly and becomes complete in structure till about 50d after being hatched. Ultrastructural observation on the normal and malpigmented skins shows that the iridophore and melanophore are adjacent to each other. Profile and structure of the two kinds of pigmcnt cells are more complete in the skin of normal ocular side than in the skin of pigmented blind side. The ultrastructure of typical chloride cell was observed in the skin of Japanese flounder larvae for the first time.     

Isolation and characterization of pathogenic <Emphasis Type="Italic">Listonella anguillarum</Emphasis> of diseased half-smooth tongue sole (<Emphasis Type="Italic">Cynoglossus semilaevis</Emphasis> Günther)

Studies were conducted to determine the cause of the acute mortality of half-smooth tongue sole Cynoglossus semilaevis Günther juveniles in a fish farm in Jimo, Shandong Province, China, in June 2006. Gross signs of the diseased tongue sole included several petechiae and ecchymoses on the body and fin necrosis and hemorrhagic lesion at the base of the fin. Bacteria were isolated from kidney, liver and hemorrhagic lesions of the diseased tongue sole. Among14 strains, SJ060621 was proved to be highly virulent to juvenile tongue sole with LD₅₀ value of <1.0×10⁵ colony forming units (CFU) mL⁻¹, while the remaining 13 were avirulent. Among the 16 antibiotics tested, SJ060621 was sensitive to gentamicin and nitrofurantoin. It was identified as Listonella anguillarum with conventional plate and tube tests in combination with API 20E analysis. 16S rRNA gene and partial HSP60 gene sequenceing analysis revealed that the strain was highly homologous with L. anguillarum. Examination of the infected musculature by electron microscopy indicated numerous bacteria and lots of macrophages containing phagocytosed bacteria. Histopathological investigations revealed severe necrotic degenerative changes in the infected organs. Indirect immunofluorescence assay (IFA) was employed to detect the location of occurrence of bacteria, and bacteria were found in aggregations in the inflammatory areas in musculature.     

Morphological observation of Epithemia ocellata （Ehr.） Kuetz. （Bacillariophyceae） from China

The species of Epithemia ocellata （Ehr.） Kuetz. from Northeastern China were observed by scanning electron microscopy （SEM）. The studies on silica valve formation in different stages were made. At the early stage, a Y-shaped raphe sternum was found. Areolae were made from virgae and vimines. In immature valves, the canal formation before the raphe fissure is visible. The raphe system, virgae and vimines （areolae） might be the early siliceous elements of the valve during morphogenesis. The formations of virgae and vimines are similar to that of Gephyria media W. Arnott. Rows of silica papillae formed gradually on the face of the valve. After the areolae were covered with silica and form rows of papillae, the outer valve surface could become mature. Details of the internal costae were observed in mature valves.     

Phylogenetic relationships within the genus Aspidisca (Protozoa, Ciliophora, Euplotida) revealed by ITS1-5.8S-ITS2 region sequences

The internal transcribed spacer regions (ITS1 and ITS2) and 5.8S rRNA genes were sequenced in six populations of four Aspidisca species, namely A. leptaspis, A. orthopogon, A. magna and A. aculeata. Phylogenetic trees were constructed by means of Bayesian inference (BI), Maximum Parsimony (MP), Neighbor-Joining (NJ), and Maximum Likelihood (ML) to assess the inter- and intra-species relationships within the genus Aspidisca. All trees show similar topologies with stable supports and indicate that: (1) four well known groups, i.e., Oligotrichia, Stichotrichia, Choreotrichia and Hypotrichia, are distinctly outlined within the class Spirotrichea, and all are monophyletic other than Hypotrichia; (2) members of Aspidisca can be distinguished well, based on the ITS1-5.8S-ITS2 region sequences, and A. leptaspis and A. magna shared a closer relationship than other species; (3) Aspidisca and Euplotes branch early in the subclass Hypotrichia. To compare the phylogenetic relationships based on different genes, SSU rRNA trees were also constructed with nearly the same species inclusion, which revealed different topologies of inter-species, inter-genera and inter-subclasses.     

A transformation model forLaminaria Japonica (Phaeophyta,Laminariales)

A genetic transformation model for the seaweedLaminaria japonica mainly includes the following aspects:

1. 1.

   The method to introduce foreign genes into the kelp,L. japonica

   Biolistic bombardment has been proved to be an effective method to bombard foreign DNA through cell walls into intact cells of both sporophytes and gametophytes. The expression ofcat andlacZ was detected in regenerated sporophytes, which suggests that this method could induce random integration of foreign genes.

   Promoters to drive gene expression

2. 2.

   The CaMV35S promoter was first used by us to induce the expression of GUS gene in brown algae. But results of further studies suggested that CaMV35S could be a tissue-specific promoter. Our use of SV40 promoter resulted in both transient and stable expression oflacZ andcat in sporophytes or gametophytes. No GUS or LacZ background was found in either sporophytes or gametophytes.The regeneration route of transgenic kelp

   The regeneration efficiency of explants is still very low. By using female gametophytes as gene hosts and parthenogenesis as regeneration route, CAT activity and LacZ activity were detected in regenerated sporophytes of parthenogenetic kelp. li]4.|The way to select transgenic kelp

3. 1.

   Results of sensitivity tests showed that kelp was only sensitive to chloramphenicol and hygromycin among many antibiotics. The regenerated sporophytes by parthenogenesis were more sensitive to hygromycin than to chloramphenicol. Resistant kelp was created by transforming female gametophytes with pSV40-CAT and stimulating parthenogenesis followed by selection in medium with lethal concentration of chloramphenicol.

   Safety consideration of transgenic kelp

   L. japonica was originally introduced from Japan. In China it is a cultured population. The possibility of its negative impact on natural populations is very low. 2) The vectors and target genes used for transformation should be restricted in order to avoid any negative impacts on human health and environment. 3) Specially devised containers (3.6 L, made of 200 μm membrane) were used to ensure that the kelp cannot escape or be eaten by marine animals. 4) To avoid the release of spores, it is very necessary to harvest the kelp at suitable age before the sporangium forms.

     

C_(17)-fengycin B,produced by deep-sea-derived Bacillus subtilis,possessing a strong antifungal activity against Fusarium solani

Root rot disease caused by Fusarium solani is the most devastating disease of the tomato and legume crops in China.The metabolites of Bacillus species can inhibit many fungal diseases.In this study,the metabolites of deep-sea-derived bacterium Bacillus subtilis 2 H11 can significantly inhibit the growth of F.solani.The metabolite C_(17)-fengycin B,one of the cyclic lipopeptides,was identified by the combination of silica column chromatography,high-performance liquid chromatography(HPLC),high-energy collision induced dissociation mass spectrometry(HCD-MS) and tandem mass spectrometry(HCD-MS/MS).The results of scanning electron microscopy(SEM) and transmission electron microscopy(TEM) showed that C_(17)-fengycin B could destroy the structure of the hyphae and spores of F.solani.The antifungal activities of C_(17)-fengycin B against F.solani were tested at concentrations ranging from 0.05 mg/mL to 0.20 mg/mL.The results indicated that C_(17)-fengycin B inhibited the growth of F.solani with antifungal index of 89.80%at 0.20 mg/mL,and the antifungal activity of C_(17)-fengycin B was further verified by the pot experiment.In addition,the cytotoxicity experiment showed that C_(17)-fengycin B had good biocompatibility and was a potential candidate for the development of biocontrol pesticide in the future.     

Molecular identification,expression pattern,and in-vitro bioactivity analysis of insulin-like growth factor 2 in olive flounder Paralichthys olivaceus

Insulin-like growth factors (IGFs) are key regulators of development and growth. Here, we characterized the igf2 gene from olive flounder (Paralichthys olivaceus) and determined its temporal and spatial expression. We set up an in-vitro protein expression system in eukaryotic human embryonic kidney (HEK293T) cells and explored its effects on cell proliferation. The flounder igf2 cDNA contained a 648-bp open reading frame (ORF) encoding a protein of 215 amino acids (aa), which spanned the complete signal peptide (47 aa), mature peptide (70 aa), and E domain (98 aa). In adult flounder, igf2 mRNA was detected in all selected tissues. In early development, igf2 mRNA was detected throughout development from unfertilized eggs to hatching-stage embryos. In-situ hybridization analysis indicated that igf2 mRNA was specially expressed in the brain region, floor plate, hypochord, otic vesicle, and pectoral fin during embryogenesis. Western blotting analysis indicated that the soluble recombinant flounder IGF2 protein was successfully produced through eukaryotic expression in HEK293T cells. In addition, the recombinant IGF2 protein significantly promoted the proliferation of human cervical carcinoma (HeLa) and HEK293T cells. These results provide new information about the structural and functional conservation, expression patterns, and biological activity of the igf2 in teleosts.

     

16S rRNA gene phylogenesis of culturable predominant bacteria from diseased Apostichopus japonicus (Holothuroidea, Echinodermata)

Cultured Apostichopus japonicus in China suffers from a kind of skin ulceration disease that has caused severe economic loss in recent years. The disease, pathogens of which are supposed to be bacteria by most researchers, is highly infectious and can often cause all individuals in the same culture pool to die in a very short time. The 16S rRNA gene phylogenesis of the culturable bacteria from the lesions of diseased individuals was conducted to study the biodiversity of the bacterial communities in the lesions and to identify probable pathogen(s) associated with this kind of disease. S. japonica samples were selected from a hatchery located in the eastern part of Qingdao, China. Bacterial universal primers GM5F and DS907R were used to amplify the 16S rRNA gene of bacteria colonies, and touchdown PCR was performed to amplify the target sequences. The results suggest that γ- proteobacteria (Alteromonadales and Vibrionales) of CFB group, many strains of which have been also determined as pathogens in other marine species, are the predominant bacterial genera of the diseased Apostichopus japonicus individuals.     

Observation of female gamete (Carpogonia) germination of Porphyra yezoensis and P. oligospermatangia

Several papers have reported that part or whole leafy thallus seemingly consisting of zygotospores can give rise to both blades and conchocelis in the same culture ofPorphyra. Study on samples of wild and cultivatedPorphyra yezoensis andP. oligospermatangia were conducted to clarify the origination of the young blades in the culture. It is confirmed that single cells on the blade of both species, which normally intermixed with zygotospores, germinated into young blades. TEM and SEM observation has shown that the single cells ofPorphyra yezoensis had typical features of female gamete (carpogonia) but archeospore. Therefore, the female gametes are responsible in developing leafy thalli. This project was sponsored by the NSFC (No. C-0205-05-39770593)     

Daptonema parabreviseta sp. nov. (Xyalidae,Nematoda) from the Jiaozhou Bay of the Yellow Sea,China

A new species of free-living marine nematode of the genus Daptonema Cobb, 1920 is described from the sublittoral sediment of the Jiaozhou Bay, Qingdao. Daptonema parabreviseta sp. nov. is characterized by having narrowed anterior end; small funnel-shaped buccal cavity; short cephalic setae; spicules with bulbiform proximal end and pointed distal end; gubercunalum tubular shape with a blunt dorsal apophysis. The female genital system presents a spacious postvulval sac. The new species closely resembles Daptonema brevisetosum Thanh & Gagarin, 2009. However, it differs from Daptonema brevisetosum by having relatively narrowed head (10 μm vs 20 μm); spicules with bulbiform proximal end and pointed distal end, not bifurcated at distal end. Female genital system with a spacious postvulval sac (vs absent).

     

Effects of temperature and salinity on survival,growth and DNA methylation of juvenile Pacific abalone,Haliotis discus hannai Ino

Temperature and salinity are two of the most potent abiotic factors infl uencing marine mollusks.In this study,we investigated the individual and combined ef fects of temperature and salinity on the survival and growth of juvenile Pacifi c abalone,Haliotis discus hannai Ino,and also examined the DNA methylation alteration that may underpin the phenotypic variation of abalone exposed to dif ferent rearing conditions.The single-factor data showed that the suitable ranges of temperature and salinity were 16–28°C at a constant salinity of 32,and 24–40 at a constant temperature of 20°C,respectively.The two-factor data indicated that both survival and growth were signifi cantly af fected by temperature,salinity and their interaction.The optimal temperature-salinity combination for juveniles was 23–25°C and 30–36.To explore environment-induced DNA methylation alteration,the methylation-sensitive amplifi ed polymorphism(MSAP)technique was used to analyze the genomic methylation profi les of abalone reared in optimal and adverse conditions.Neither temperature nor salinity induced evident changes in the global methylation level,but 67 and 63 dif ferentially methylated loci were identifi ed in temperature and salinity treatments,respectively.The between-group eigen analysis also showed that both temperature and salinity could induce epigenetic dif ferentiation in H.discus hannai Ino.The results of our study provide optimal rearing conditions for juvenile H.discus hannai Ino,and represent the fi rst step toward revealing the epigenetic regulatory mechanism of abalone in response to thermal and salt stresses.     

Two new species of Sellaphora (Bacillariophyta) from a deep oligotrophic plateau lake, Lake Fuxian in subtropical China

We describe two new species of Sellaphora from Lake Fuxian, Yunnan Plateau, China. Based on both light and scanning electron microscopy, these species are described as S. yunnanensis sp. nov. and S. sinensis sp. nov. The primary features of S. yunnanensis are: elliptical to linear-elliptical valves with broadly rounded ends, straight filiform raphe, almost straight central endings and small, slightly expanding central pores, small central area, symmetrical or slightly asymmetrical central nodule. The primary features of S. sinensis are: elliptical valves, obtusely rounded ends, similar raphe and axial area, transapically less expanded central area, larger, elliptical central nodule. We compare these species to those of a similar shape and morphology.     

Primary molecular basis of androgenic gland endocrine sex regulation revealed by transcriptome analysis in Eriocheir sinensis

In crustaceans, the male sexual differentiation and maintenance are specially regulated by androgenic gland (AG). However, little is known about the genes involved in the regulation process. RNA-Seq was performed on AG with ejaculatory duct (AG_ED) and ejaculatory duct (ED) as control in Eriocheir sinensis, one of the most important economic and fishery crabs with typically sex dimorphism. A total of 925 unigenes were identified as differentially expressed genes (DEGs) and the expression of nine genes randomly selected was confirmed by qRT-PCR. 667 unigenes were up-regulated in AG_ED, being supposed to be AG preferential genes. Among them, the full length of insulin-like androgenic gland factor (IAG) cDNA named as Es-IAG was obtained as a logo gene of AG, which together with the genes insulin-like receptor (INR), and single insulin binding domain protein (SIBD), might constitute the sex regulation pathway. Several sex related genes were identified, and their function will have to be investigated. Also, the identification of juvenile hormone epoxide hydrolase 1 (JHEH1), ecdysteroid 22-hydroxylase (DIB) and ecdysone receptor (ECR) preliminarily clarified the molecular regulation mechanism of eyestalk-AG-testis axis, which plays important roles in molting and reproduction. The results will enhance our understanding for the molecular basis of the AG involved in male sex regulation in crabs.