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1.
基于COⅠ序列绒螯蟹属DNA条形码和遗传多样性研究   总被引:1,自引:0,他引:1  
对绒螯蟹属的中华绒螯蟹如东和七里海群体、日本绒螯蟹、合浦绒螯蟹及狭颚绒螯蟹共80条线粒体COⅠ片段进行扩增和测序,并与GenBank中绒螯蟹属的台湾绒螯蟹2条和近方蟹属的绒毛近方蟹19条COⅠ基因序列进行联配分析。结果显示,101条序列包含44种单倍型,序列组成表现明显的碱基偏倚性。中华绒螯蟹如东、七里海群体与日本绒螯蟹间的遗传距离分别为1.210%和1.078%,明显低于COⅠ基因DNA条形码鉴别种的遗传距离为2%的阈值,表明中华绒螯蟹和日本绒螯蟹为同一物种;而合浦绒螯蟹与中华绒螯蟹如东和七里海群体及与日本绒螯蟹的遗传距离分别为4.823%、5.101%以及5.011%,明显大于2%的鉴别阈值,说明合浦绒螯蟹为独立的种。以绒毛近方蟹为外群,基于群体内及群体间的遗传距离构建的邻接树显示,中华绒螯蟹与日本绒螯蟹聚在一起,合浦绒螯蟹则聚成单系。本文测序的5个群体除狭颚绒螯蟹外,其余均具有遗传多样性,单倍型多样性为0.593±0.144~0.779±0.068,核苷酸多样性为0.00156~0.01336;此外,中华绒螯蟹如东群体与日本绒螯蟹、合浦绒螯蟹和中华绒螯蟹七里海群体分别共享单倍型H1、H2和H3,说明这些蟹类可能有种质资源混杂或是遗传污染的现象。  相似文献   

2.
为研究细胞色素CYP2基因在中华绒螯蟹蜕皮及生长发育中的作用,本试验通过逆转录聚合酶链式RT-PCR反应以及cDNA末端快速扩增RACE技术获得了中华绒螯蟹CYP2基因cDNA全长。用实时荧光定量PCR技术,分析该基因在中华绒螯蟹蜕皮过程中各组织的相对表达量。试验结果显示,中华绒螯蟹CYP2的cDNA全长1772bp,编码491个氨基酸序列,包括一个84bp的5′端非编码区,一个212bp的3′端非编码区、一个1475bp的开放阅读框,经BLAST比对,该核苷酸序列与岸蟹、三疣梭子蟹的相似性分别为66%、64%。实时荧光定量PCR结果显示,中华绒螯蟹蜕皮前,CYP2基因在鳃中的表达量相对最高;在肝胰脏、肌肉中表达量略低于鳃;在Y器官、眼柄、胸神经节、脑神经节、肠中少量表达;在心和胃中表达量最低。中华绒螯蟹在不同的蜕皮时期中,通过荧光定量试验得出,肝胰脏中CYP2基因表达量在蜕皮前期和蜕皮期最高;眼柄中CYP2基因表达量从蜕皮期到蜕皮后期有上升趋势;鳃中CYP2基因表达量在蜕皮前期最高;Y器官、脑神经节和肠中CYP2基因表达量在蜕皮期最高;肌肉中CYP2基因表达量在蜕皮前期最高;胸神经节和胃中CYP2基因表达量在蜕皮后期最高;Y器官中CYP2基因在蜕皮前期表达量高于蜕皮后期和蜕皮期。以上试验结果表明,CYP2对中华绒螯蟹蜕皮生长中的调控可能起到重要作用。  相似文献   

3.
中华绒螯蟹辽河种群野生、养殖及"性早熟"个体RAPD分析   总被引:6,自引:0,他引:6  
利用RAPD技术对中华绒螯蟹辽河种群野生、养殖及"性早熟"个体进行比较分析。从S系列的三组引物中筛选出可重复性好的、谱带清晰且扩增结果好的引物,分析了中华绒螯蟹的RAPD基因座位组成,并且检测了S系列随机引物作为中华绒螯蟹遗传变异指标研究中的可能性。从基因水平了解三者之间的差异性。  相似文献   

4.
采用聚丙烯酰胺凝胶电泳分析比较了分布于我国大陆沿海 6个主要水系的两种绒螯蟹 (中华绒螯蟹、日本绒螯蟹 )群体间的生化遗传差异。结果发现 ,(1)辽河、黄河、长江和瓯江中华绒螯蟹群体间的生化遗传差异不显著 ,四个群体间的遗传距离D =0 .0 0 0 6~ 0 .0 0 53,属种内群体间差异。 (2 )珠江和南流江日本绒螯蟹群体间的生化遗传差异也不显著 ,两个群体间遗传距离D=0 .0 0 0 7,尚未达到亚种分化水平。 (3)辽河、黄河、长江和瓯江中华绒螯蟹与珠江和南流江日本绒螯蟹在SOD同工酶表型上有明显差异 ,SOD - 2位点仅在辽河、黄河、长江和瓯江中华绒螯蟹中表达 ,而SOD - 4位点仅在珠江和南流江日本绒螯蟹中表达 ,这些可作为中华绒螯蟹与日本绒螯蟹群体区分的生化遗传标志。辽河、黄河、长江和瓯江中华绒螯蟹群体同珠江和南流江日本绒螯蟹群体间的遗传距离较大 ,D =0 .0 72 1~ 0 .0 74 7。 (4 )聚类分析表明 ,在我国大陆沿海水系分布的绒螯蟹可分为两大类群 ,辽河、黄河、长江和瓯江水系的绒螯蟹属北方类群 ,即中华绒螯蟹 ;珠江和南流江的绒螯蟹属南方类群 ,即日本绒螯蟹。  相似文献   

5.
图们江水系绒螯蟹的形态差异与遗传混杂   总被引:2,自引:1,他引:1  
为了研究图们江水系中华绒螯蟹的种群现状,以图们江水系绒螯蟹为研究材料,将我国黄河和辽河水系的中华绒螯蟹(Eriocheir sinensis),以及日本本土的日本绒螯蟹(Eriocheir japonicus)作为参照对象,应用三种形态多元分析方法与STRUCTURE聚类分析方法,对它们的32个外部形态性状进行分析。判别分析显示,图们江群体的判别准确率最低(83.30%);主成分分析显示图们江群体的12个差异最大的表型性状均位于中华绒螯蟹与日本绒螯蟹之间,呈现为中间类型;传统聚类显示图们江水系绒螯蟹与日本绒螯蟹的形态差异最小;STRUCTURE聚类显示,图们江有60%的个体聚入日本绒螯蟹群体,而只有10%的个体聚入中华绒螯蟹群体,其余30%个体为中华绒螯蟹与日本绒螯蟹的中间类型。形态研究结果表明,图们江水系绒螯蟹为中华绒螯蟹与日本绒螯蟹分布的重叠区与混杂区,但其形态偏向日本绒螯蟹。  相似文献   

6.
中华绒螯蟹RXR基因全长cDNA克隆及表达分析   总被引:4,自引:2,他引:2       下载免费PDF全文
维甲类X受体(retinoid X receptor,RXR),属于核受体超家族(nuclear receptor super family)成员,是一种重要的调控因子,对甲壳动物生长发育和繁殖具有十分重要的作用。本研究根据陆地蟹 RXR基因保守区序列设计上下游引物,采用逆转录聚合酶链反应(RT-PCR)以及cDNA末端快速扩增(RACE)技术克隆中华绒螯蟹(Eriocheir sinensis)RXR基因并进行各组织间的表达分析。序列分析表明中华绒螯蟹RXR cDNA全长1517bp,编码433个氨基酸。经BLASTn和BLASTx分析表明,中华绒螯蟹RXR氨基酸序列与陆地蟹RXR基因的氨基酸序列相似性最高,为96%。系统进化树分析显示,中华绒螯蟹RXR的氨基酸序列与陆地蟹RXR聚为一支。荧光定量PCR结果显示,RXR在所有检测的组织中均有表达,且在中华绒螯蟹Y器官中表达量最高,肝胰腺、鳃和肌肉中有少量表达,心脏、胃、肠、胸神经节和脑神经节中微量表达。在中华绒螯蟹不同蜕皮时期中,Y器官中RXR表达量在D期最高,与AB期、C期呈显著性差异(P<0.05);肌肉中RXR在AB期表达量最高,与其他蜕皮时期呈显著性差异(P<0.05);肝胰腺和鳃中RXR在AB期表达量最高,E期表达量最低,但各蜕皮时期表达量之间差异不显著。  相似文献   

7.
盐度对中华绒螯蟹性早熟生理机制的影响   总被引:8,自引:1,他引:7  
于2004年6月至11月在崇明岛中华绒螯蟹养殖基地进行1秋龄中华绒螯蟹(Eriocheir sinensis)的养殖试验,通过荧光免疫二步抗体检测法和原子吸收等方法,测定了1秋龄蟹血淋巴钙离子和雌二醇水平,研究了不同盐度水体中1秋龄中华绒螯蟹性早熟情况及盐度促进中华绒螯蟹性早熟的生理机制。结果表明:(1)随着水体盐度的升高,1秋龄中华绒螯蟹性早熟率上升,成活率则呈下降趋势;(2)水体盐度升高导致1秋龄未发育雌蟹血淋巴中Ca2 含量上升(P<0.05),但对1秋龄雄蟹作用不显著(P>0.05);(3)池塘养殖90 d之后,高盐度水体中1秋龄未发育雌蟹血淋巴中雌二醇水平显著高于低盐度水体1秋龄未成熟雌蟹(P<0.05);(4)1秋龄性早熟雌蟹血淋巴中雌二醇含量高达(1294.00±595.87)pg/mL,极显著地高于同期的1秋龄未成熟雌蟹(P<0.01)。研究表明:水体盐度升高引起的1龄雌蟹血淋巴中雌二醇水平和Ca2 含量上升是导致1秋龄雌性中华绒螯蟹性早熟的内在生理原因。  相似文献   

8.
李思发 《水产学报》2002,26(6):493-497
用包括Z2和Opp17两个10碱基随机引物在内的10个引物,对来自荷兰斯科克莱(Skodely)、美国加州圣何塞(San Jose)的中华绒螯蟹群体与中国长江水系中华绒螯蟹群体进行RAPD遗传比较分析。结果发现:(1)中华绒螯蟹群体特有的Z2引物扩增的700bp标记带(Z2^700bp),在荷兰与美国2个中华绒螯蟹群体中同时出现,而不出现日本绒螯蟹南流江种群中特有的880bp标记带(Z2^880bp),表明欧洲、美国中华绒螯蟹与中国中华绒螯蟹为同种Eriocheir sinensis,而非日本绒螯蟹Eriocheir japonicus;(2)Opp17引物扩增的947bp片段在中国长江、荷兰及美国3个中华绒螯蟹群体内的出现频率均达100%。结合Z2引物扩增结果,欧洲与美国中华绒螯蟹群体极可能是从中国长江水系中华绒螯蟹引入繁衍的。  相似文献   

9.
蜕皮抑制激素(molt—inhibiting hormone,MIH)属甲壳动物高血糖激素(crustacean hyperglycemic hormone,CHH)家族。MIH抑制Y-器官蜕皮激素的合成。以中华绒螯蟹(Eriocheir japonica sinensis)眼柄总RNA为模板,根据中华绒螯蟹蜕皮抑制激素1(Eriocheir japonica sinensis molt—inhibiting hormone-1,Ers—MIH1)基因序列设计引物,用RT—PCR的方法获得了Ers—MIH1基因成熟肽的cDNA片段。序列分析表明,Ers—MIH1基因成熟肽编码区含有228bp,编码75个氨基酸残基,与已发表的序列一致。将该cDNA片段插入到中间载体pMD18-T中,酶切后再将该片段插入到pGEX-4T-1表达载体中,构建成表达质粒pGEX-4T—MIH1。在BL21细胞中经IPTG诱导表达,得到GST—MIH1的融合蛋白。SDS—PAGE分析表明,经0.1mmol/L IPTG诱导4h,发现大量GST—MIH1融合蛋白表达,在分子量(Mr)为34kD处有1条特异的蛋白质条带。中华绒螯蟹蜕皮抑制激素1融合蛋白的成功表达为进一步深入研究MIH在中华绒螯蟹蜕皮过程中的作用机制奠定了基础。  相似文献   

10.
候利波  陆银月  任秋霖  孔祥会  朱雷  顾伟  孟庆国 《水产学报》2023,47(4):049103-049103
为了探究低氧胁迫对中华绒螯蟹血淋巴细胞影响的分子机制,实验通过使用Nova Seq 6 000测序技术,检测了低氧组(1.5±0.5) mg/L和对照组(6.0±0.5) mg/L分别处理24 h后的中华绒螯蟹血淋巴细胞的转录组数据变化。首先,对测序得到的原始数据进行拼接、注释以及筛选,分析获得的128 614个转录本和128 614条基因,平均长度为2 634 bp (N50),Q30> 92%。其次,以1.2倍为阈值,共筛选出1 687个差异表达基因,其中上调基因899个,下调基因788个。GO和KEGG分析发现,低氧胁迫对中华绒螯蟹血细胞的三羧酸循环、糖酵解途径、ECM-受体相互作用、间隙连接、细胞凋亡、酚氧化酶系统以及其他免疫相关基因等产生了显著影响。最后,随机挑选转录组数据中的10个基因进行实时荧光定量PCR(qRT-PCR)验证,结果显示与转录组数据分析相一致,其中包括6个上调的基因:整合素1、铜/锌超氧化物歧化酶、磷酸肌醇3激酶、磷酸甘油酸突变酶2、PDGF/VEGF相关因子1和relish;4个下调的基因:无脊椎连接蛋白7、热休克蛋白90、线粒体ATP合成酶α和天...  相似文献   

11.
The nutritional values of edible gonad, hepatopancreas and muscle tissues from normal and precocious adult male Chinese mitten crabs (Eriocheir sinensis) were investigated herein. The results showed that the average weight and meat yield (MY) of normal crabs were significantly higher than those of precocious crabs, while the hepatopancreas indexes (HSI) and gonadosomatic indexes (GSI) had no significant differences. Precocious and normal crabs had greater protein and lipid contents, respectively, and the differences were mainly reflected in gonads and hepatopancreas. While assessing the amino acid profiles, both precocious and normal crabs could be high‐quality protein sources, particularly the precocious crabs which showed higher levels of essential amino acids. In fatty acid profiling experiments, precocious crabs revealed higher EPA, ∑n‐3 polyunsaturated fatty acids (PUFA) and ∑n‐3/∑n‐6 ratios than normal crabs; however, DHA and ∑n‐6 PUFA contents were considerably lower. Additionally, contents of nutritious minerals K and Ca in precocious crabs were significantly higher than those in normal crabs, while Zn and Fe were significantly lower. In short, our data indicated that precocious Chinese mitten crabs are also a nutritious food, and even in some respects their nutritional value is higher than normal crabs.  相似文献   

12.
13.
刘红 《水产学报》2006,30(5):577-585
该论文首次报道了经由促雄性腺提取物的注射而在蟹类中引起的性逆转现象。此前关于蟹类促雄性腺活性研究的报道极少,而且蟹类的雄性化均是由促雄性腺的移植所产生。本实验中将锯缘青蟹和中华绒螯蟹的促雄性腺提取物分别注射到刚刚完成性别分化的中华绒螯蟹雌性幼蟹体内,此时幼蟹处于4至5期,壳宽为5~8 mm。注射之后,幼蟹经过大约1~2次蜕皮,此时在注射锯缘青蟹以及中华绒螯蟹的促雄性腺提取物的两组实验幼蟹中均能观察到雄性化现象,而注射生理盐水的对照组实验幼蟹中未能观察到相同现象。由此本实验可以证明促雄性腺确实是蟹类的一种雄性激素,注射促雄性腺提取物能引起雌性幼蟹发生性逆转;同时根据锯缘青蟹和中华绒螯蟹的促雄性腺提取物均能引起中华绒螯蟹雌性幼蟹发生性逆转的现象推测,锯缘青蟹和中华绒螯蟹两种间可能存在促雄性腺的交叉活性;不仅如此,性逆转还能在极低的注射剂量下获得,相当于中华绒螯蟹0.14促雄性腺当量和锯缘青蟹0.06促雄性腺当量。  相似文献   

14.
Although wild Yangtze Eriocheir sinensis juveniles (coin‐sized seed) have better culture performance than the pond‐reared juveniles during the adult crab culture stage, the culture performance of wild‐caught megalopae (WC) remains unknown. Therefore, this study was designed to compare the growth, survival rate, crab yield, feed conversion rate (FCR), precocity, final weight and size distribution between WC and hatchery‐produced megalopae (HP) during the juvenile culture stage. Wild‐caught megalopae had significantly higher body weight, weight gain rate (WGR) and specific growth rate (SGR) during 0–4 weeks but lower WGR and SGR than HP during 4–8 weeks (< .05). Although WC had a significantly lower survival rate and yield of normal juvenile crabs than HP, total yields of normal juveniles and precocious crabs were similar. Wild‐caught megalopae had a significantly higher precocious rate and yield of precocious crabs than HP (< .05). Regardless of sex, no significant differences were found in the gonadosomatic index (GSI), hepatosomatic index (HSI) and final weight of normal juveniles and precocious crabs between WC and HP. As for the size distribution of normal crabs, WC had a higher percentage of small female juveniles (<3 g) but a lower proportion of middle size ranges (3–5.99 g and 6–8.99 g) than HP for females (< .05), and there was no significant difference for males. In conclusion, wild megalopae of the Yangtze E. sinensis had worse culture performance than the hatchery‐produced seeds; therefore, wild megalopae would not be recommended for commercially juvenile culture of E. sinensis.  相似文献   

15.
Precocious puberty is one of the major constraints to the further development of Chinese mitten crab (Eriocheir sinensis) farming industry. Although dietary phospholipids (PL) and highly unsaturated fatty acids (HUFA) supplementation have been shown to enhance the growth of larval E. sinensis in other studies, it is still unknown whether this also leads to a higher precocity rate for juvenile E. sinensis. This study was conducted to investigate the effects of dietary PL and HUFA on precocity, survival, growth and hepatic lipid composition of juvenile E. sinensis. Two diets were formulated with PL [3.95% dry weight (DW)] and HUFA (0.98% DW) supplementation (diet A) and without PL and HUFA supplementation (diet B) and fed to juvenile E. sinensis. Although dietary PL and HUFA levels did not significantly affect the survival and growth performance of juvenile E. sinensis, compared with crabs fed diet A, a higher precocity rate was found among juvenile E. sinensis fed diet B (P=0.051). A higher total lipid content, but significantly lower levels of HUFA and PL (P<0.05) were found in the hepatopancreas of crabs fed diet B than in those fed diet A. Meanwhile, the precocious females had significantly lower hepatosomatic index, arachidonic acid (20:4n‐6), eicosapentaenoic acid (20:5n‐3) and docosahexaenoic acid (22:6n‐3) contents in their hepatopancreas when compared with that of the normal immature juveniles (P<0.05). The results suggest that the occurrence of precocious puberty among farmed juvenile E. sinensis could be reduced by the inclusion of appropriate level of dietary PL and HUFA.  相似文献   

16.
The replacement of fish oil (FO) in Eriocheir sinensis can significantly reduce the cost of E. sinensis cultivation, while several studies have indicated that replacing FO with soybean oil (SO) could significantly reduce the resistance of E. sinensis to disease. However, the molecular mechanisms underlying these effects remain poorly understood. In this study, crabs were fed two diets containing FO or SO, following which a label‐free quantification proteomic analysis was employed. And the activity of enzymes involved in the nonspecific immune response was also measured. Growth performance was undifferentiated between the crabs fed with FO and SO. A total of 519 proteins were identified, and 70 proteins were significantly altered between the crabs fed the two different diets. Five proteins related to the immune response were identified to be differently expressed. C‐type lectin, haemocyanin subunit 6 and cryptocyanin were significantly downregulated, while fatty acid‐binding protein and catalase were highly expressed in the crabs fed SO. The activities of acid phosphatase, alkaline phosphatase, superoxide dismutase and phenoloxidase were all significantly changed in crab fed with different diets. These findings will provide novel insight into the molecular mechanism regarding the replacement of FO on the immune response of E. sinensis and provide evidences for the relationship between nutrition and immunity in E. sinensis.  相似文献   

17.
陈再忠 《水产学报》2005,29(5):630-634
采用磷酸苯二钠法在520nm波长测定了未成熟、早熟和正常成熟中华绒螯蟹肝胰腺与性腺中的酸性磷酸酶(ACP)和碱性磷酸酶(ALP)。结果表明:(1)各组蟹肝胰腺ACP和ALP活性都显著高于性腺;(2)早熟蟹肝胰腺中ACP活性都显著高于未成熟蟹而低于正常成熟蟹,在各发育阶段没有性别差异,但是,早熟雌蟹和正常成熟雌蟹卵巢中ACP活性都较早熟雄蟹和正常雄蟹精巢低,早熟与正常成熟个体没有差异;(3)早熟雌蟹肝胰腺ALP活性显著高于未成熟和正常成熟雌蟹,而与三组雄蟹处于同一水平,但是,早熟和正常成熟雌雄蟹的性腺中ALP活性没有显著差异。由此得出结论:(1)性腺发育所需营养物质的动员或合成主要与肝胰腺ACP有关;(2)性早熟的发生与肝胰腺中ACP和ALP活性的显著升高有关。  相似文献   

18.
MicroRNAs (miRNAs) represent a class of endogenous small non‐coding RNAs with 18–25 nucleotides that are involved in diverse biological processes. In the present study, differentially expressed miRNAs in four different metamorphic stages of Eriocheir sinensis were comprehensively detected and validated from high‐throughput sequencing miRNA data sets. We identified and characterized 597 known miRNAs and 901 novel miRNAs from the four small RNA libraries. We also demonstrated the evolutionary conservation of miRNAs from Chinese mitten crab with those from other species. Analyses of miRNA expression profiles during brachyurization development of the Chinese mitten crab revealed that 86 mature miRNAs were dysregulated, 47 of which were significantly differentially expressed. The results of real‐time quantitative PCR analyses for a few selected miRNAs were similar to those of deep sequencing analyses. Intriguingly, the results of Gene Ontology annotation and KEGG pathway analyses indicated that miRNA gene clusters and families were consistently dysregulated at four different metamorphic stages, although they were enriched at various levels. Functional enrichment analysis revealed the versatile physiological roles (over six signal pathways) of these miRNAs. The results suggest that these miRNAs might play potential key roles in the regulation of gene expression in the brachyurization development of the Chinese mitten crab. Particularly, miR‐2a‐3p, miR‐305‐5p, miR‐263a‐5p and miR‐7‐5p may play major regulatory roles in the metamorphosis of E. sinensis.  相似文献   

19.
转录组测序研究三角帆蚌珍珠颜色相关基因   总被引:3,自引:2,他引:1  
为筛选影响三角帆蚌珍珠颜色的候选基因,采用转录组测序平台Illumina Hi Seq TM2500对蚌壳珍珠层颜色为紫色和白色的三角帆蚌中央膜组织进行高通量测序。所得序列经质控、组装后比对到NR、Swiss-Prot、COG、KOG、KEGG、GO和Pfam数据库中注释,并进行差异基因聚类分析。结果显示,获得干净数据共72 800 581 bp,拼接获得了89 529个Unigene,差异表达基因2644个,其中上调表达基因1323个,下调表达基因1321个。根据GO功能分类可分为分子功能、细胞组分、生物过程等3大类50分支;根据KEGG代谢通路分析可以分为187类。对差异表达基因分析发现,部分基因与色素合成(眼黄素、黑色素、喋啶色素)及金属离子转运相关,还有6个细胞色素P450,3个细胞色素b561和1个细胞色素b560。研究表明,这些基因可能在珍珠颜色形成中发挥作用。  相似文献   

20.
中华绒螯蟹大颚器激素生物合成与性早熟的关系   总被引:15,自引:1,他引:14       下载免费PDF全文
赵维信 《水产学报》2003,27(4):289-294
运用放射化学方法跟踪测定中华绒螯蟹早熟个体和正常发育个体的大颚器合成和分泌的激素-甲基法尼酯(methyl farnesoate,MF),发现早熟雌、雄蟹当年9月的MF合成速率约为正常蟹的2倍,早熟蟹的MF在当年秋季(10月或11月)均达到峰值,之后:MF合成速率显著降低,而成熟系数继续升高。正常发育的雌、雄幼蟹,当年至翌年4月的MF合成速率始终维持较低水平,翌年6月开始,MF合成速率迅速增大,至翌年10月的MF合成速率已超过早熟蟹的峰值,11月的MF合成速率明显降低,性腺发育至Ⅳ期末。研究结果表明:早熟蟹大颚器的提早发育,大量合成和分泌促性腺激素-MF,刺激卵巢或精巢发育成熟是导致性早熟的内分泌因素。  相似文献   

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