Molecular markers for identifying a new selected variety of Pacific white shrimp Litopenaeus vannamei     

于洋  张晓军  刘敬文  李富花  黄皓  李义军  刘小林  相建海 《中国海洋湖沼学报》2015,33(1):1-10

Selective breeding of the Pacific white shrimp Litopenaeus vannamei during the last decade has produced new varieties exhibiting high growth rates and disease resistance.However,the identification of new varieties of shrimps from their phenotypic characters is difficult.This study introduces a new approach for identifying varieties of shrimps using molecular markers of microsatellites and mitochondrial control region sequences.The method was employed to identify a new selected variety,Kehai No.1(KH-1),from three representative stocks(control group):Zhengda;Tongwei;and a stock collected from Fujian Province,which is now cultured in mainland China.By pooled genotyping of KH-1 and the control group,five microsatellites showing differences between KH-1 and the control group were screened out.Individual genotyping data confirmed the results from pooled genotyping.The genotyping data for the five microsatellites were applied to the assignment analysis of the KH-1 group and the control group using the partial Bayesian assignment method in GENECLASS2.By sequencing the mitochondrial control regions of individuals from the KH-1 and control group,four haplotypes were observed in the KH-1 group,whereas14 haplotypes were obtained in the control group.By combining the microsatellite assignment analysis with mitochondrial control region analysis,the average accuracy of identification of individuals in the KH-1group and control group reached 89%.The five selected microsatellite loci and mitochondrial control region sequences were highly polymorphic and could be used to distinguish new selected varieties of L.vannamei from other populations cultured in China.  相似文献   

凡纳滨对虾C-型凝集素LvLec2 对不同刺激的免疫应答     

罗展  张继泉  李富花  柳承璋  相建海 《海洋科学》2010,34(11):103-110

克隆获得了与中国明对虾(Fenneropenaeus chinensis)C-type lectin 3同源的C-型凝集素LvLec2基因序列,并通过Real-time PCR研究了其在脂多糖(lipopolysaccharide,简称LPS)、灭活溶壁微球菌(Micrococcus lysodeikticus)和白斑综合症病毒(white spot syndrome virus,简称WSSV)刺激后的转录表达变化。结果表明,LvLec2编码157个氨基酸,含有1个糖识别结构域(carbohydrate recognition domain,简称CRD),其CRD结构域中具有决定糖结合特异性的基序"EPS"(Glu118-Pro119-Ser120)序列。系统进化树分析表明LvLec2与已发表的凡纳滨对虾(Litopenaeus vannamei)C-型凝集素亲缘较远。LPS、灭活溶壁微球菌和WSSV刺激后,LvLec2基因在肝胰脏中的转录表达有明显的变化但表达模式不同。LvLec2可能作为模式识别受体参与了对虾对病原的识别或防御。  相似文献   

BAC end sequencing of Pacific white shrimp <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>: a glimpse into the genome of Penaeid shrimp     

赵翠  张晓军  柳承璋  郇聘  李富花  相建海  黄超 《中国海洋湖沼学报》2012,30(3):456-470

Little is known about the genome of Pacific white shrimp (Litopenaeus vannamei). To address this, we conducted BAC (bacterial artificial chromosome) end sequencing of L. vannamei. We selected and sequenced 7 812 BAC clones from the BAC library LvHE from the two ends of the inserts by Sanger sequencing. After trimming and quality filtering, 11 279 BAC end sequences (BESs) including 4 609 paired- ends BESs were obtained. The total length of the BESs was 4 340 753 bp, representing 0.18% of the L. vannamei haploid genome. The lengths of the BESs ranged from 100 bp to 660 bp with an average length of 385 bp. Analysis of the BESs indicated that the L. vannamei genome is AT-rich and that the primary repeats patterns were simple sequence repeats (SSRs) and low complexity sequences. Dinucleotide and hexanucleotide repeats were the most common SSR types in the BESs. The most abundant transposable element was gypsy, which may contribute to the generation of the large genome size of L. vannamei. We successfully annotated 4 519 BESs by BLAST searching, including genes involved in immunity and sex determination. Our results provide an important resource for functional gene studies, map construction and integration, and complete genome assembly for this species.  相似文献   

Rapid, sensitive detection of Vibrio anguillarum using loop-mediated isothermal amplification     

高宏伟  李富花  张晓军  王兵  相建海 《中国海洋湖沼学报》2010,28(1):62-66

Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming. The rapid detection and identification of V. anguillarum, and other pathogens that infect marine organisms, is crucial to effective disease management. In this study, we developed a loop-mediated amplification (LAMP) assay to detect V. anguillarum in an hour in a single tube without the need for thermal cycling. Conserved regions of the metalloproteinase (empA) gene of V. anguillarum served as the targets for primer design. A fragment of the empA gene was amplified at 65°C in the presence of the primer mixture and Bst DNA polymerase. In the optimized LAMP assay, 6.7 pg of V. anguillarum DNA could be detected. Six strains of V. anguillarum and 17 strains of non-V. anguillarum bacteria were used in this study to evaluate the species specificity of the primers. The six V. anguillarum strains gave a positive result in the LAMP assay. This method was also validated in V. anguillarum-infected fish. This LAMP method is more sensitive than PCR in the detection of V. anguillarum and shows good species specificity. The LAMP assay is therefore an effective method for the quick detection of V. anguillarum both in the laboratory and in the field.  相似文献   

Construction of bacterial artificial chromosome libraries for Zhikong Scallop Chlamys farreri     

张洋  张晓军  张洪斌  李富花  相建海 《中国海洋湖沼学报》2008,26(2):215-218

Two Large-insert genomic bacterial artificial chromosome （BAC） libraries of Zhikong scallop Chlamys farreri were constructed to promote our genetic and genomic research. High-quality megabase-sized DNA was isolated from the adductor muscle of the scallop and partially digested by BamH I and Mbo I, respectively. The BamH I library consisted of 53 760 clones while the Mbo I library consisted of 7 680clones. Approximately 96 % of the clones in BamH I library contained nuclear DNA inserts in average size of 100 kb, providing a coverage of 5.3 haploid genome equivalents. Similarly, the Mbo I library with an average insert of 145 kb and no insert-empty clones, thus providing a genome coverage of 1.1 haploid genome equivalents.  相似文献   

Chromosome Behavior of Heat Shock Induced Triploid in Fenneropenaeus Chinensis     

张晓军  李富花  相建海 《中国海洋湖沼学报》2003,21(3):222-228

INTRODUCTIONTriploidanimalsareusefulbecausetheyaresterile ,characterizedbypositivegrowthinthere productiveseason ,usefulfortesting ,forintroductionofnon nativespecies ,andforprotectionofde velopedstrains.Theinductionoftriploidyhadbeenreportedinmanyaquaculturespecies .Triploidshavebeensuccessfullyinducedinshrimpusingtemperatureshockorchemicalshock (CB ,6 DMAP)in 4species :Sicyoniaingentis (Xiangetal.,1 991 ) ,Fenneropenaeuschinensis (Xiangetal.,1 992 ;Daietal.,1 993;Baoetal.,1 993;Li…  相似文献   

Heat-shock protein 70 expression in shrimp Fenneropenaeus chinensis during thermal and immune-challenged stress   总被引：4，自引：0，他引：4  

郭振宇焦传珍  相建海 《中国海洋湖沼学报》2004,22(4):386-391

Using western immunoblotting, we obtained heat-shock protein 70 (HSP70) induction data and distribution in different tissues from shrimp Fenneropenaeus chinensis during thermal and immune-challenged stresses. This is probably the first report of the effects of various stressors on the expression of HSP70 in shrimp.HSP70 was prominently induced in hepatopancreas and gills, but not in muscle, eyestalk and hemolymph, when the shrimp were exposed to heat shock and Vibrio anguillavium-challenged stresses. Cold shock and WSSV treatment had no significant effects on the levels of HSP70 expression in all tissues examined. HSP70 induction was greatest after 2 h exposure to heat shock stress, which was elevated after acute heat shock exposure of 10℃ above ambient temperature.  相似文献   

CHARACTERIZATION OF LACTATE DEHYDROGENASE ISOZYME PATTERN AND MORPHOLOGY OF THREE MARINE FISH CELL LINES     

郭华荣  张士璀  李红岩  童裳亮  相建海 《中国海洋湖沼学报》2002,20(3):226-231

Three continuous marine fish cell lines of FG (i. e., Hounder Gill) from flounder (Paralichthys olivaceus) gill, SPH (i. e. , Sea Perch Heart) from sea perch (Lateolabrax japonicus) heart and RSBF (i.e., Red Sea Bream Fin) from red sea bream (Pagrosomus major) fin, were characterized by lactate dehydrogenase (LDH) isozyme and morphological analysis. The LDH isozyme patterns of these three cell lines and their corresponding tissues of origin were investigated and compared. The results showed: (1) No difference was found in the LDH isozyme patterns of FG and flounder gill tissue. However, the LDH isozyme patterns of SPH and RSBF were significantly different from their corresponding tissues of origin; (2) LDH isozyme patterns of FG, SPH and RSBF were markedly different from each other and could serve as genetic markers for species identification and detection of cross contamination. Morphological change analysis of these three cell lines in comparison to their original tissues indicated that FG cells still appeared epithelioid without morphological transformation. However, morphological changes were found in SPH and RSBF compared to their original tissues. Therefore, the cellular morphology was still plastic in the relatively stable culture conditions, and it was possible that change of LDH patterns wasrelated to morphological changes of fish cells in vitro.  相似文献   

刀额新对虾染色体核型及细胞核DNA含量   总被引：9，自引：0，他引：9  

张晓军  周岭华  相建海 《海洋与湖沼》2002,33(3):225-231

以刀额新对虾胚胎、无节幼体、卵巢、精巢等为材料 ,空气干燥法制备染色体 ,并初步进行核型分析。结果表明 ,刀额新对虾的染色体数目为 2n=78,n =39,核型为 2n=78=40M 1 0SM 1 4ST 1 4T。以刀额新对虾血淋巴、卵巢、肌肉、鳃为材料 ,以鸡血细胞为DNA标准 (2 50pg/ 2c) ,使用PartecCCAⅡ型流式细胞仪测定了刀额新对虾细胞的基因组DNA含量 ,其值为鸡血细胞的 1 75倍 ,绝对含量为 4 375pg/ 2c。  相似文献   

专家系统开发工具(TUILI语言)与海洋生物学研究          下载免费PDF全文

黄勃  刘瑞玉  崔玉珩  相建海 《海洋科学》1993,17(5):15-16

专家系统(Expert System简称ES)是在人工智能(Artifical Intelligence简称AI)的研究过程中产生的。不管人们如何看待AI,但都很现实地承认:解决某一领域内的具体问题,ES是不可低估的手段,有时甚至可超过同行专家。EH.Shorfliffe等人完成的用于诊断和治疗感染性疾病MYCIM系统收到很好效果;斯坦福(Startford)国际研究所(SDI)研制的矿藏勘测专家系统PROSPECTOR,在华盛顿州找到了专家未曾找到的一处钼矿。  相似文献