Molecular characterization and expression profiles of four transformer-2 isoforms in the Chinese mitten crab Eriocheir sinensis

The transformer-2(tra-2) gene plays a key role in the regulatory hierarchy of sexual differentiation in somatic tissues and in the germline of Drosophila melanogaster.In this study,sequences and expression profiles of tra-2 in the Chinese mitten crab Eriocheir sinensis were characterized.Four tra-2 isoforms,designated as Estra-2a,Estra-2b,Estra-2c,and Estra-2d,were isolated.They all contained an RNA-recognition motif(RRM) and a linker region,which shared high similarity with other reported tra-2s.Sequence analysis revealed that Estra-2a,Estra-2b and Estra-2c are encoded by the same genomic locus and are generated by alternative splicing of the pre-mRNA.Compared with the other three isoforms,Estra-2d lacks the RS2 domain.Quantitative real-time PCR showed that all four isoforms were highly expressed in the fertilized egg,and in the 2-4 cell and blastula stages compared with larval stages(P0.01),suggesting their maternal origin in early embryonic developmental stages.Notably,Estra-2a was highly expressed in male somatic tissues,while Estra-2c was significantly highly expressed in the ovary.These results suggest that Estra-2c is involved in sexual differentiation of the Chinese mitten crab.Our findings provide basic information for further functional studies of the tra-2 gene/protein in this species.     

EFFECTS OF TEMPERATURE AND Ca^2＋ ON THE LARVAL DEVELOPMENT OF THE DECAPODA CRUSTACEAN ： ERIOCHEIR SINENSIS

With increasing demand in China for the mitten crab larvae, understanding its survival mechanism gets more important. This research focused on the effects of temperature and Ca^2 on the larval growth and development. Eriocheir sinensis larvae were reared in laboratory under 21 different combinations of temperature (15, 20, 25℃) and Ca^2 content (120, 130, 140, 150, 160, 170, 180mg/l) and constant salinity (20) and pH (8), The results suggested that the survival rate increases with temperature and Ca^2 content. These combinations of temperature and Ca2 content maximized survival rate inour study and it may be the optimum water environmental conditions for culturing the larvae. To predict surviving larvae number under different water environmental conditions, 21 dynamic mathematical models were developed. This for the first time observation of the zoeal Ⅵ larvae of the Changjiang River E.sinensis population showed that they occurred under stressed water environmental conditions: temperature of 15℃ and Ca^2 content of 120,130mg/l.     

Effects of fishes on the growth and survival of Chinese mitten crab,Eriocheir sinensis

This experiment on the effects of fishes (Pelteobagrus fulvidraco, Channa argus, Cyprinus carpio) on growth and survival of juvenile Chinese mitten crab,Eriocheir sinensis was conducted in cages placed in Bao'an Lake. T-tests showed that effects of fishes on the carapace width growth were not significant as compared with the control (no fish). But the effects of these fishes on crab survival were negative. The causes of negative effects of fishes on crab survival were simply discussed on the basis of food habits. This research was financially supported by the Department of Agriculture (96-008-02-03), P. R. China, and Chinese Academy of Sciences (Key Project: KZ951-A1-102).     

Morphological and functional studies on the epidermal cells of amphioxus (Branchiostoma belcheri tsingtauense) at different developmental stages

Epidermal cells of amphioxus at different developmental stages were investigated by electron microscopy and colloidal carbon tracing experiments. Amphioxus epidermal cells showed different ultrastructural characteristics at larval and adult stages. The epidermal cells at all larval stages studied (24–96 h) had numerous vesicles containing electron dense materials in their apical cytoplasm. In tracing experiments, carbon particles were found in apical vesicles and interoellular spaces. Under scanning electron microscope, many crater-like protrusions were observed on the surface of the cells. These results indicated that amphioxus larval epidermal cells may be capable of endocytosis. The epidermal cells of 3-month and adult amphioxus were obviously secretory ones characterized by well-developed peripheral filaments, a prominent Golgi apparatus and abundant apical secretory vesicles. This study also showed that adult amphioxus body surface mucus contained lectin that could agglutinate human red blood cells. The authors propose that the epidermal cells of amphioxus larva and adult may contribute to the immune defense of the amimal by different means. Project 3860811 supported by NSFC and study also supported by the Shandong Natural Science Foundation (Grant No. 92D1144).     

Statistical optimization of medium composition and culture condition for the production of recombinant anti-lipopolysaccharide factor of Eriocheir sinensis in Escherichia coli

Anti-lipopolysaccharide factors (ALFs) are important antimicrobial peptides that are isolated from some aquatic species. In a previous study, we isolated ALF genes from Chinese mitten crab, Eriocheir sinensis. In this study, we optimized the production of a recombinant ALF by expressing E. sinensis ALF genes in Escherichia coli maintained in shake-flasks. In particular, we focused on optimization of both the medium composition and the culture condition. Various medium components were analyzed by the Plackett-Burman design, and two significant screened factors, (NH4)2SO4 and KH2PO4, were further optimized via the central composite design (CCD). Based on the CCD analysis, we investigated the induction start-up time, the isopropylthio-D-galactoside (IPTG) concentration, the post-induction time, and the temperature by response surface methodology. We found that the highest level of ALF fusion protein was achieved in the medium containing 1.89 g/L (NH4)2SO4 and 3.18 g/L KH2PO4, with a cell optical density of 0.8 at 600 nm before induction, an IPTG concentration of 0.5 mmol/L, a post-induction temperature of 32.7°C, and a post-induction time of 4 h. Applying the whole optimization strategy using all optimal factors improved the target protein content from 6.1% (without optimization) to 13.2%. We further applied the optimized medium and conditions in high cell density cultivation, and determined that the soluble target protein constituted 10.5% of the total protein. Our identification of the economic medium composition, optimal culture conditions, and details of the fermentation process should facilitate the potential application of ALF for further research.     

Transcriptome analysis of the Chinese grass shrimp Palaemonetes sinensis (Sollaud 1911) and its predicted feeding habit

Journal of Oceanology and Limnology - Chinese grass shrimp Palaemonetes sinensis is an economically important freshwater shrimp in China. However, wild resources of grass shrimp have declined...     

Effects of salinity on embryonic and larval development of Chinese mitten crab Eriocheir sinensis(Decapoda: Brachyura) and salinity-induced physiological changes

To investigate the effects of salinity on early development of Chinese mitten crab(Eriocheir sinensis),and the salinity tolerance mechanism of embryos,different developmental stages of embryos(gastrula,eyespot and pre-hatching stage),and hatched stage I zoea and megalopa,were exposed to a range of salinities(1,5,10,15(control),20,25,30,35 and 40).Hatching,survival and molting were monitored.Effects of 24-hour hypersaline(35)and hyposaline(1)stress on egg diameter,water content,Na~+/K~+-ATPase(NKA)activity,and crustacean hyperglycemic hormone(CHH)gene mRNA expression in embryos and megalopa,are reported.Embryos are more tolerant of low(≤5)than high(≥25)salinities,with optimum ranges for gastrula and pre-hatching stage embryos being 5-20,and for eyespot embryo and stage I zoea,10-20.Most megalopa can molt to the first juvenile instar by day 5 at salinities between 1 and 40,whereas molting of megalopa stages was delayed at 40.Hypersaline conditions resulted in a loss of moisture,reduction of egg volume,and a significant increase in NKA activity and CHH mRNA expression at some developmental stages.Hyposaline conditions did not affect moisture content or egg volume,but resulted in decreased NKA activity and CHH mRNA expression in embryos.For megalopa stages,NKA activity was significantly upregulated following both hypo-and hypersaline stress.Our results suggest high salinity will inhibit development and hatching of E.sinensis embryos,and low salinity will affect the survival of their stage I zoea.Increased NKA activity in embryos and megalopa stages might indicate a hyporegulation response under hypersaline conditions.These findings provide useful information for spawning ground protection of indigenous E.sinensis and enrich the knowledge of embryonic tolerance mechanisms of hyperregulating crustaceans following osmotic stress.     

Effects of florfenicol exposure on growth,development and antioxidant capacity of flounder Paralichthys olivaceus larvae at different developmental stages

Antibiotics are widespread in various environmental media,and may pose a potential threat to aquatic ecosystems and non-target aquatic organisms.Florfenicol(FLO) is one of the most commonly used antibiotics in aquaculture,and extensively used to substitute chloramphenicol with its strong sterilization and low adverse ef fect.In this study,flounder Paralichthys olivaceus,an important economic fish species in seawater was used as an experimental subject.Five exposure concentrations of FLO(including environmentrelated concentrations) were set at 0,0.01,0.1,1,and 10 mg/L.Ef fects of FLO exposure for 168 h on growth and development,motor behavior,antioxidant enzyme activity,malondialdehyde(MDA) content,and thyroid hormone level of P.olivaceus larvae were studied in pre-larvae(1 dpf) and post-larvae(20 dpf).The results show that the short-term FLO exposure could promote the larvae growth to some degrees,but inhibit them as the exposure time prolonged.For pre-larvae,FLO at 0.01 mg/L could stimulate the motor nerve system and increase the swimming ability,but inhibited it at 1 mg/L.With the increasing dosage of FLO,the superoxide dismutase(SOD) and MDA contents were elevated,reaching the maximum in the 1 mg/L FLO group.The pre-larvae were more sensitive than the post-larvae to FLO in the environment,and the growth and immune resistance could be damaged with long exposure.Post-larvae were more tolerant to external pollutants,FLO at 1 mg/L could promote the motor behavior and reduce SOD and MDA contents.Therefore,FLO can be used as an antibiotic at a proper concentration but as a drug to prevent disease in a long-term way.     

Quantitative division of succession and spatial patterns among different stand developmental stages in Changbai Mountains

Broadleaf-Korean pine forests exhibit high species richness and distinctive species composition, which are currently becoming more dominant among natural forests in Changbai Mountains of northeastern China. Understanding the ecological process of restored vegetation is quite important for ecosystem reconstruction. Distinguishing stand development stages and analyzing the dynamic spatial patterns could provide insights into significant community coexistence mechanisms. In the present study, eight permanent study areas were established according to the substituting space for time method in Changbai Mountains of north-eastern China. The optimal division method was used to quantify the successional series into different stand development stages, and the point pattern analysis method(L(r) function) was used to analyse the dynamic changes in spatial patterns and interspecific associations. Our results suggested that:(1) The stand development process was divided into five stages: the first three stages were poplar-birch secondary forests in different stages of recovery, the fourth stage was thespruce-fir mixed forest, and the last stage was the primary broadleaf-Korean pine forest;(2) The spatial pattern showed an aggregated distribution at a small scale and changed to a random distribution as the scale increased in poplar-birch secondary forests, but the spatial pattern appeared to be randomly distributed in spruce-fir mixed forest and broadleafKorean pine forest;(3) The interspecific associations between pioneer species and climax species changed from negative to positive among the different stand stages, and environmental resources were shared among these species. Interspecific differences in shade tolerance among the tree species were key determinants of forest dynamics and structure. Our study is vital to the understanding of the forest development; thus, the spatial change features should receive greater attention when forest management is being planned and restoration strategies are being developed for the Changbai Mountains.     

邹城台钻孔应变更换探头前后数据精度评价

利用相对标定、四分量自检分析、M2潮汐因子相对中误差计算及与同台站TJ-2体应变观测资料进行相关性对比等方法,计算邹城地震台RZB-2分量式钻孔应变2020-01~2021-07更换探头前后仪器的内精度、自检情况及M2波潮汐因子精度,综合分析分量式钻孔应变观测系统的稳定性和观测数据的可靠性。结果表明,更换探头后邹城台RZB-2分量式钻孔应变数据内精度高,两组面应变符合四分量自检方程,相关系数接近1,面应变与体应变的面应变数据的相关性也接近1,仪器观测系统稳定,观测数据可靠。但2021年台站周边新建工厂导致干扰增多,数据潮汐因子精度较差,平均值为0.07左右,建议做好环境保护工作,以提高观测数据质量。     

向家坝水电站蓄水前后库区地震活动特征

对比向家坝库区蓄水前后地震活动特点,结合库尾段地震时序与库水位的变化关系及震中区地层岩性、构造特征等,认为发生在库尾段的地震属于构造型水库诱发地震。地震震中呈北西向密集条带状分布,虽然地表没有明显的控震构造,但不排除在2 km之下存在隐伏断层的可能。该区段位于马边-盐津断裂带的中段,水库蓄水前地震活动十分微弱,蓄水后发生不同于以往的地震活动。     

利用EOF分析方法研究康定MS6.3地震前后重力时空分布异常特征

对康定MS6.3地震前后的流动重力观测数据采用正交经验函数(EOF)分析方法进行处理,分析主要模式与资料时段的MS5.0以上地震震中位置的关系,探讨主要模式代表的物理含义。结果表明,川滇地区7期累积重力观测资料中大于200 μGal的数据是偶然误差的概率大于95%。第二模式规格向量和累积重力变化均具有相似的分区特征,2013~2014年6次MS6.0以上地震和第二模式规格向量正负转换带有超过83.3%的对应率,具备重力长期变化模式。第一模式地震前后规格值序列明显的同震信号、空间相似的4象限分布以及2014年4次MS6.0以上地震和第二模式规格向量正负转换带100%的对应率表明,该模式代表重力短期变化模式。康定MS6.3地震前第一模式包含4象限特点,震中位置规格值不断增大,且地震前后变化从负转为正。     

2017年南丹Ms4.0地震前后重力变化

利用广西及邻区2014~2017年流动重力观测资料,系统分析区域重力场变化及其与2017-07-15广西南丹MS4.0地震的关系,并结合GPS观测数据与地震地质调查成果,探讨区域重力场变化的时空分布特征及机理。结果表明：1）南丹M_S4.0地震前后,震区附近重力异常变化与主干断裂关系密切,反映沿控震断裂在2014~2017年间发生了引起地表重力变化效应的地壳变形和构造活动;2）差分重力图像表明,南丹M_S4.0地震前重力变化为“局部重力异常→四象限重力异常→重力反向变化发震”的过程;3）重力变化与GPS 观测反映的水平运动表明,南丹M_S4.0地震位于重力四象限分布中心部位及面压缩峰值附近,证明重力场和形变场动态变化对中强地震地点预测具有指示意义;4）天峨-南丹-环江一带是重力变化最剧烈地区,亦是水平形变面压缩过渡带,同时位于莫霍面等深线强烈变化地段、M_L3.0地震围空区、沿主断裂存在的低b值异常区。结合地震活动图像、定点前兆观测异常综合分析认为,该地区仍存在发生中强地震的可能。     

Dynamic changes in lakes in the Hoh Xil region before and after the 2011 outburst of Zonag Lake

Zonag, Kusai, Hedin Noel and Yanhu Lakes are independent inland lakes in the Hoh Xil region on the Qinghai-Tibet Plateau. In September2011, Zonag Lake burst after the water level had increased for many years. Floods flowed through Kusai and Hedin Noel Lakes into Yanhu Lake; since then, the four small endorheic catchments merged into one larger catchment. This hydrological process caused the rapid shrinkage of Zonag Lake and continuous expansion of Yanhu Lake. In this study,based on satellite images, meteorological data and field investigations, we examined the dynamic changes in the four lakes and analyzed the influencing factors. The results showed that before 2011, the trends in the four lake areas were similar and displayed several stages. The change in the area of Zonag Lake corresponded well to the change in annual precipitation(AP), but the magnitude of the change was less than that of a non-glacier-fed lake. Although increased precipitation was the dominant factor that caused Zonag Lake to expand, increased glacier melting and permafrost thawing due to climate warming also had significant effects. After the 2011 outburst of Zonag Lake, due to the increasing AP and accelerating glacier melting, the increases in water volume of the three lakes were absorbed by Yanhu Lake, and Yanhu Lake expanded considerably. According to the rapid growth rates in water level and lake area, Yanhu Lake is likely to burst in 1-2 years.     

Identification of suitable reference genes for quantitative gene expression analysis in clam Cyclina sinensis under salinity stress and Vibrio infection

The appropriate reference gene is a prerequisite for accurate normalization of gene expression level, and research on suitable reference genes in clam Cyclina sinensis is scarce. To improve the situation, we selected five commonly used housekeeping genes, including β-actin, Elongation factor 1-α (EF1-α), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 40S ribosomal protein S18 (RPS18), and Tubulin α (TUB-α), then evaluated their expression stability in different adult tissues and under different experimental treatments (salinity stress and Vibrio parahaemolyticus infection). Their expression stability was analyzed by three frequently used programs, geNorm, NormFinder, and BestKeeper. This analysis indicated that multiple genes should be used for normalization, and we concluded that the reference gene combination GAPDH-RPS18-β-actin, should be used for qRT-PCR analysis in different tissues of C. sinensis under normal physiological conditions. For the clams under salinity stress and Vibrio infection, EF1-α-GAPDH-RPS18 was recommended as the gene combination for qRT-PCR normalization. TUB-α was generally poorly ranked by all programs, and should not be used in future studies. This study should provide fundamental support for accurate quantitative gene expression analysis of this species.

     

2012年新源和静6.6级地震前后P波速度结构的演化

基于新疆区域地震台网记录的地震观测报告,采用有限差分层析成像方法反演新疆天山中东段地区的地壳P波速度结构,获得了2012年新源和静6.6级地震前后该区域12 km和24 km深度处半年尺度的P波速度结构的时间演化过程。结果表明,12 km深度处震前震源区及附近区域P波速度差在空间上形成NNE向变化剧烈的高值异常区,异常幅度达0.3 km/s。这种明显的速度差异表明,将半年尺度不同时间段相同深度的P波速度进行差分析,能较好地反映一定区域内地壳介质特性的变化。     

岷县-漳县M6.6地震前后的形变范数变化特征

采用分布在南北地震带上10个洞体应变台的观测资料,基于小波分解和范数计算,提取2013-07-22甘肃岷县-漳县M6.6地震前异常信息。结果表明,除肃南台外,其余9个台均出现震前异常信息,昭通台、小庙台最早出现异常,在2013-06初（约在震前50 d）,距震中最近的宕昌台在7月初出现异常。结合其他学者的研究,对异常信息特征进行讨论认为,这次地震与南北地震带东侧块体自南向北运动有密切关系,而震后的形变传递过程是一个反向过程。同时也证明,用范数计算方法提取震前形变异常是简单、有效和可靠的。     

南水北调前后海河流域陆地水储量变化分析北大核心CSCD

基于GRACE及GRACE-FO重力卫星数据,通过滑动T检验确定海河流域陆地水储量突变的特征时间点,分析其时空演变特征;结合多变量趋势分析和贡献率量化法探究陆地水储量变化成因,讨论南水北调工程的贡献。结果表明,在南水北调前(2004-01~2015-01)后(2015-01~2020-10),海河流域陆地水储量的衰减趋势分别为-17.19 mm/a和-13.49 mm/a,缓解约24%,缓解趋势由南到北逐渐增大;人类活动与气候变化对流域陆地水储量变化的贡献率比为7∶3,人为耗水是海河流域陆地水储量常年处于亏损状态的主要原因;调水量的贡献率和趋势呈现年际增大和升高态势,预示南水北调工程在改善区域陆地水储量方面具有巨大潜力。