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1.
转录组测序可在各种环境条件下对物种进行高通量测序,通过基因结构分析和功能注释,探讨特定条件下相关基因的功能和表达情况。该研究分别获取低盐胁迫组(盐度0.2)和自然海水组(盐度31)脊尾白虾(Exopalaemon carinicauda)样品,通过Illumina平台进行转录组测序,获得13.92 Gb高质量测序数据,组装得到111 618条转录本和72 734条Unigenes,其中有22 879条Unigenes得到注释,比对到Nr数据库的Unigenes为21 931条;筛选出1 492条脊尾白虾低盐胁迫差异显著表达基因,包括829条上调基因和663条下调基因,其中有810条差异表达基因得到注释。通过差异表达基因GO功能注释和富集分析及KEGG通路富集分析,锁定大量脊尾白虾在自然海水和淡水环境下的差异表达基因,为深入探讨脊尾白虾在低盐胁迫条件下的生理保护机制提供了技术支撑。  相似文献   

2.
为了探究低氧胁迫下中华圆田螺(Cipangopaludina cathayensis)肝脏组织基因的差异表达,本研究通过高通量测序技术,分析中华园田螺低氧胁迫组(2.5 mg/L)和常氧组(6.9 mg/L)某些基因的差异表达,并对差异基因进行生物信息学分析,进一步采用实时荧光定量PCR (RT-qPCR)对关键差异表达基因进行验证。结果显示,测序共获得232 379条基因(unigenes),与对照组比较,低氧胁迫组筛到176个差异基因,包含64个上调基因和112个下调基因。GO功能注释分析显示,差异基因主要富集在生物学过程中的几丁质代谢过程和含氨基葡萄糖的复合代谢过程,细胞组分中的胶原三聚体成分,分子功能中的几丁质结合功能和糖衍生物结合功能。KEGG通路富集分析显示,差异基因主要集中于环境信息处理、遗传信息处理、代谢和生物系统这4大类通路。6个关键差异基因的RT-qPCR结果显示,热休克蛋白70B2和热休克蛋白β-6基因表达量上调,几丁质酶蛋白4、α-1胶原蛋白(XIV)、α-4胶原蛋白(XIV)、5-磷酸酶蛋白基因表达量下调,证实了转录组测序结果的可靠性。本研究发现,低氧胁迫激活了中华圆田螺适应缺氧的生理活动,并获得了低氧胁迫下中华圆田螺肝脏组织中相关功能基因的表达信息,为深入研究中华圆田螺响应低氧胁迫的调控机制提供了基础数据和理论依据。  相似文献   

3.
长臂虾科线粒体基因组特征分析及分子标记探讨   总被引：1，自引：0，他引：1  
通过常规PCR和长PCR扩增获得脊尾白虾的线粒体DNA,采用鸟枪法测序、序列组装获得脊尾白虾的线粒体基因组全序列.结合罗氏沼虾和沼虾的线粒体基因组,分析了长臂虾线粒体基因组的基本特征、基因排列、蛋白质编码基因和基因变异程度等.与泛甲壳动物线粒体基因组的原始排列相比,2个沼虾的线粒体基因组的基因排列完全一致;而脊尾白虾线粒体基因组却存在2个tRNA基因的易位.长臂虾科线粒体基因组主编码基因的变异特征分析揭示了coxl基因多态位点的比例最低,仅为25.5%;而3个基因(nad2、nad4和nad5)不仅多态位点的比例较高,而且基因的长度较长,从而拥有最丰富的多态位点.因此,nad2、nad4和nad5基因可以作为分子标记,用于分析长臂虾不同群体之间的遗传多样性,为长臂虾生物多样性的保护及合理利用其生物资源提供更多保障.  相似文献   

4.
脊尾白虾线粒体基因组应答饥饿的甲基化特征分析   总被引：1，自引：1，他引：0  
为研究脊尾白虾应答饥饿的线粒体基因组表观遗传学特征,本研究在筛选适宜脊尾白虾线粒体基因组甲基化位点分析的BSP引物基础上,对不同饥饿期该虾线粒体基因组的甲基化特征进行了分析。共设计覆盖脊尾白虾线粒体全基因组序列的BSP引物51对,经验证其中10对引物可用于脊尾白虾线粒体基因组的甲基化特征分析,选用其中具有稳定扩增产物的5对引物分别对饥饿10、20和30 d的脊尾白虾线粒体基因组的甲基化特征进行了分析,结果表明,脊尾白虾耐受饥饿的平均死亡时间为19.8 d,最长耐受时间为48 d,在不同饥饿阶段线粒体基因组均可发生甲基化现象,且在第10天时甲基化比例最高,而第20和30天时甲基化比例反而降低;甲基化主要发生在ND基因位点上,据此推测这种甲基化可能用来调节能量代谢相关的电子传递链过程。本研究首次揭示了脊尾白虾线粒体基因组存在甲基化现象,为展开虾类线粒体基因组表观遗传学研究奠定了理论基础。  相似文献   

5.
以脊尾白虾(Exopalaemon carinicauda)为试验对象,探讨了蒽对脊尾白虾存活、免疫和能量分配的影响。将同一批亲虾繁殖的幼虾放在试验室暂养7 d,挑选健康活泼和规格相似的幼虾作为试验对象,推算蒽对脊尾白虾的半致死浓度和安全浓度。试验设置8个浓度梯度,每个处理设3个重复。在试验过程中,每个水族箱加入5只白虾,经试验计算蒽对脊尾白虾24、48、72和96 h半致死浓度分别为21.53、16.31、11.76和9.06 mg/L,安全浓度为2.81 mg/L。在安全浓度下胁迫192 h,测定血清酚氧化酶和超氧化物歧化酶的活性,发现蒽对脊尾白虾免疫有着一定程度的抑制作用。随后在安全浓度下试验21 d,设两个处理,每个处理设3个重复,每个重复投放10只白虾,待试验结束后,选取白虾的肌肉部分进行能量分配测定,计算出蛋白含量、脂肪含量、糖元含量和肌肉的总能量储备以及对应能量分配的变化,结果发现对照组与处理组总能量储备相差240.67 mJ·mg^-1(湿重),能量消耗相差252 mJ·mg^-1(湿重),细胞能量分配相差11.33 mJ·mg^-1·h^-1(湿重)。以上结果表明,蒽明显影响脊尾白虾的存活、免疫和代谢。  相似文献   

6.
为研究低氧胁迫对大口黑鲈"优鲈3号"呼吸代谢和细胞凋亡的影响,在溶解氧(DO)为(0.70±0.05) mg/L的条件下,测定了试验鱼肝脏及脑组织hif-1α、glut-1和caspases基因的表达。结果显示:与DO为(7.50±0.50)mg/L的常氧对照组相比,低氧胁迫能引起大口黑鲈"优鲈3号"肝脏及脑组织hif-1α、glut-1表达水平的显著上调,同时也可诱导caspase 3和caspase 9基因的表达,致使细胞发生凋亡;复氧24 h后,hif-1α、glut-1基因在试验鱼肝脏及脑组织中的表达量均恢复至常氧水平,caspases基因在脑组织中的表达量极显著高于对照组(P0.01),caspase 3在肝组织中的表达量显著高于对照组(P0.05),caspase 9则恢复至常氧水平。试验结果初步表明,hif-1α和glut-1是大口黑鲈"优鲈3号"低氧信号传导途径中的重要组成部分,低氧胁迫可能会激活线粒体凋亡通路,从而可能会诱导细胞凋亡的发生。  相似文献   

7.
为研究不同发育阶段脊尾白虾线粒体基因组表观遗传学特征,本研究在筛选硫化测序引物的基础上,对不同发育期的脊尾白虾线粒体基因组甲基化特征进行了分析。经引物筛选验证,选取了4对引物在脊尾白虾的受精卵、幼体期、仔虾期和成虾期共4个发育期进行甲基化特征的分析,结果表明,除ND5基因3′端序列(21#引物扩增区)无甲基化现象外,COX3基因的起始区序列(13#引物扩增区)、COX3基因的3′端序列(16#引物扩增区)和ND5基因5′端序列(19#引物扩增区)均存在不同程度的甲基化现象。其中,COX3基因的起始区序列仅在受精卵孵化期和幼体期存在甲基化现象,COX3基因3′端序列在除受精卵外的其他3个时期均存在甲基化现象,且以上甲基化率随个体发育阶段的提高呈下降趋势;ND5基因5′端序列在4个不同发育期均存在甲基化现象,但甲基化率无显著差异。甲基化主要发生在COX3基因和ND5基因位点上,推测脊尾白虾在生长发育过程中通过调节线粒体基因组甲基化来调节机体能量代谢过程。本研究首次从整个发育周期角度探讨了线粒体基因组甲基化特征,以期为进一步揭示甲壳类生长发育相关遗传机制提供帮助。  相似文献   

8.
为探究长期盐度胁迫条件下红螯光壳螯虾肠道基因表达和代谢通路的变化。实验采用Illumina Hiseq 2 500平台对0、5、10和15这4个盐度下养殖5周的红螯光壳螯虾的肠道组织进行了高通量测序。研究共获76 534个unigenes,通过与NR、NT、KO、Swissprot、PFAM、GO和KOG数据库进行比对,成功注释37 378个unigenes。通过引入FPKM(Fragments Per Kilo bases per Million fragments)来估算基因表达水平,基于NR和Pfam两个数据库的蛋白注释结果,448 362个unigenes在GO数据库得到注释。根据KO功能注释与Pathway的联系,9 483个unigenes在KEGG数据库被注释分类到34条通路途径。通过DEGseq进行基因表达差异分析,以盐度0为对照组,显著差异基因筛选条件设置为Q value<0.05且差异倍数|Fold Change|>2,盐度5、10和15组分别获得差异基因2 733、91和236个,其中盐度5组共有2 068个基因显著上调,665个基因显著下调。将所有差...  相似文献   

9.
为探讨盐碱水环境对脊尾白虾(Exopalaemon carinicauda)基因组DNA甲基化的影响,本研究利用MethylRAD-Seq技术探究了长期盐碱水养殖组(SAS)和正常海水养殖对照组(SW)脊尾白虾鳃组织中的DNA甲基化水平,并对关键通路和基因进行了差异表达分析。结果显示,脊尾白虾鳃组织基因组中CG和CWG位点(W=A或T)分别检测到2 347 003和416 176处甲基化,甲基化普遍存在于基因组的基因间区和内含子区域,共筛选到8805个(8189个CG-DMSs和616个CWG-DMSs)差异甲基化位点,盐碱水环境下DNA甲基化水平略有增强。通过KEGG富集分析发现,DMS所在差异表达基因显著富集在HIF-1信号通路和剪接体通路,通路中hif-p、hk和sf3b1等关键基因在脊尾白虾盐碱水环境适应中可能发挥着重要作用;对SW和SAS组差异甲基化基因(DMG)进行筛选,得到158个CG-DMGs和94个CWG-DMGs,其中,富集到脂质代谢和囊泡介导的转运通路中的DMG最多;此外,有一些DNA甲基化位点与基因表达呈负相关,表明DNA甲基化与基因调控之间存在复杂的联系,大部分基因组DNA甲基化对基因表达有正调控效应。本研究结果首次分析了在盐碱水环境下脊尾白虾鳃组织的DNA甲基化水平特征,为解析甲壳类盐碱水环境适应机制提供了基础信息。  相似文献   

10.
为研究Cd2+胁迫下脊尾白虾(Exopalaemon carinicauda)甘露糖集合凝集素(MBL)在肝胰腺中表达量的变化特征,本研究利用重金属镉(Cd2+)对脊尾白虾进行96 h急性毒性实验。实验共设置5组Cd2+浓度胁迫(0、0.01、0.0175、0.021和0.0278 mmol/L),分别在Cd2+胁迫后0、3、6、12、24、48、72和96 h共8个时间点取样。实时荧光定量结果显示,高浓度Cd2+(0.0175、0.021和0.0278 mmol/L)胁迫下,脊尾白虾MBL基因的表达量先呈上升趋势,在72 h达到峰值后随后下降,但各时间点表达量均与对照组存在显著性差异(P<0.01);Cd2+浓度为0.01 mmol/L时,脊尾白虾MBL基因的表达量整体呈下降趋势。进一步采用RNA技术干扰该基因表达后,发现Cd2+胁迫后的脊尾白虾个体死亡率显著增加。本研究表明,脊尾白虾MBL在应答Cd2+胁迫过程中可对机体起到一定的保护作用。  相似文献   

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12.
The environmental processes associated with variability in the catch rates of bigeye tuna in the Atlantic Ocean are largely unexplored. This study used generalized additive models (GAMs) fitted to Taiwanese longline fishery data from 1990 to 2009 and investigated the association between environmental variables and catch rates to identify the processes influencing bigeye tuna distribution in the Atlantic Ocean. The present findings reveal that the year (temporal factor), latitude and longitude (spatial factors), and major regular longline target species of albacore catches are significant for the standardization of bigeye tuna catch rates in the Atlantic Ocean. The standardized catch rates and distribution of bigeye tuna were found to be related to environmental and climatic variation. The model selection processes showed that the selected GAMs explained 70% of the cumulative deviance in the entire Atlantic Ocean. Regarding environmental factors, the depth of the 20 degree isotherm (D20) substantially contributed to the explained deviance; other important factors were sea surface temperature (SST) and sea surface height deviation (SSHD). The potential fishing grounds were observed with SSTs of 22–28°C, a D20 shallower than 150 m and negative SSHDs in the Atlantic Ocean. The higher predicted catch rates were increased in the positive northern tropical Atlantic and negative North Atlantic Oscillation events with a higher SST and shallow D20, suggesting that climatic oscillations affect the population abundance and distribution of bigeye tuna.  相似文献   

13.
Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae(GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the ?rst time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in ?ve provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding36.9%(62/168), 37.5%(63/168) and 19.6%(33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia(95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010,but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during2007–2011, III onlyoccurred in 2012. 14 different PFGE types, including 4 new types(N, O,P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9%(141/168) of the total GBS strains. Ia corresponded to 11 PFGE types(A–H, N–P), in which type D predominated(51%). Ib represented 3 genotypes(I, J and Q) and III harbored only 2genotypes(N and F). Type N and Fsynchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years.Although Iastillpredominated, new rareserotypeIII alreadyoccurred in China.  相似文献   

14.
The purpose of this work was to evaluate the suitability of a method based on the presence of two restriction sites (for Hae III and Hindf I) in the mitochondrial NADH dehydrogenase subunit 5 (mt ND5) gene to identify Scomber species. The evaluation was performed on 144 reference and market samples by sequencing of the entire 505-bp fragment of the mt ND5 gene and of a 464-bp fragment of the Kocher fragment of the cytochrome b gene (mt Cytb). Sequence analysis of any of the two fragments allows the identification of each of the four Scomber species, but S. japonicus and S. colias had the same restriction sites at the ND5 amplicon and would not have been differentiated by this analysis. Similarly, loss of the Hae III site in some S. scombrus individuals would have misidentified them as not being Scomber. All the market products were correctly labeled except one acquired in Spain labeled as originating in the Atlantic and containing S. japonicus.  相似文献   

15.
In this experiment, a feeding trial was performed to determine the effects of fructooligosaccharide (FOS) on growth performance, digestive enzyme activity and immune response of Japanese sea bass, Lateolabrax japonicus juveniles (initial weight 38.3 ± 0.5 g), and the fish were examined following feeding with six levels of FOS (0, 0.5, 1, 2, 4 and 6 g/kg) for 28 days. Significant enhancement of weight gain (WG) and specific growth rate (SGR) was found in fish fed 1 g/kg FOS incorporated diets (p < .05), while the feed conversion ratio (FCR) in the 1, 2 g/kg FOS groups reduced significantly compared with the control (p < .05). Besides, the crude lipid in the 4, 6 g/kg FOS groups increased significantly compared with the control (p < .05). On the other hand, the erepsin and lipase activities significantly elevated in intestine of fish fed 2 g/kg FOS (p < .05) and the lysozyme activity in serum of fish fed 2 g/kg FOS were significantly higher than that in the control (p < .05). Moreover, the alkaline phosphatase activities in serum of fish fed 0.5, 1, 2 g/kg FOS were significantly higher than in control (p < .05). Regression analysis showed that the relationships between dietary FOS levels and either SGR, FCR, erepsin or lysozyme activities were best expressed by regression equations, and the optimal inclusion levels are 1.37, 1.80, 3.06, 3.11, 1.93 and 1.80 g/kg for SGR, FCR, erepsin, lipase, lysozyme and total superoxide dismutase activities, respectively. Overall, this study revealed that FOS incorporated diets could beneficial for L. japonicus culture in terms of increasing the growth, digestion and immune activities. Under the present experimental condition, the optimal supplementary level of FOS in the diet of L. japonicus is 1–3 g/kg.  相似文献   

16.
Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL?1 (1.25 ng well?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation.  相似文献   

17.
Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality).  相似文献   

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19.
A 5 × 3 factorial growth trial was conducted to evaluate optimal dietary protein and lipid levels (dietary protein level, DP; dietary lipid level, DL) for juvenile Sillago sihama (S. sihama) (2.0 ± 0.02 g, initial weight). Fish were fed 15 diets containing 5 DPs (350, 400, 450, 500 and 550 g/kg) and 3 DLs (60, 90 and 120 g/kg) for 8 weeks. The interaction between proteins and lipids significantly influenced the feed conversion ratio, condition factor, body composition, antioxidant indices and lipase activity (p < .05). DP 450 g/kg showed the highest average final body weight. DPs 500 and 550 g/kg significantly decreased the protein efficiency ratio (p < .05). DL 120 g/kg showed the highest percentage weight gain. The low feed conversion ratio was found in diets P45L12, P55L9 and P55L12. Diet P45L12 showed high superoxide dismutase activities. DP 450 g/kg showed the lowest average malondialdehyde content. Lipase activity was increased by increasing DP (p < .05) with a fall at DP 550 g/kg. Under the present experimental conditions, the optimal DP for S. sihama was 450 g/kg under the DL 120 g/kg.  相似文献   

20.
Cholecalciferol (Vitamin D3) increased plasma inorganic phosphate concentration in American eels,Anguilla rostrata, in a dose-dependent fashion. This response was more marked in phosphate loaded fish. In control as well as phosphate loaded eels the hyperphosphatemic response to D3 was associated with a sharp reduction in renal phosphate clearance relative to14C-polyethelene glycol (PEG) clearance. Glomerular filtration and urine flow rates were not affected by D3. As renal phosphate clearance, even in phosphate loaded eels, never significantly exceeded that of PEG, it is suggested that D3 reduced the relative clearance rate of phosphate by increasing renal phosphate reabsorption rather than by reducing the tubular secretion of phosphate.  相似文献   

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