虾夷扇贝养殖群体及其子代的遗传多样性分析     

韩斐斐  张继彪  李莉  张国范  闫喜武 《海洋科学》2012,36(9):1-8

虾夷扇贝(Patinopecten (Mizuhopecten) yessoensis)于1982年从日本引进,在中国已经成为一个很重要的产业.本研究利用微卫星标记,对獐子岛海域2个养殖群体虾夷扇贝(大耗岛、褡裢岛),1个日本野生群体遗传多样性进行了评估,并对2个养殖群体的群体内繁育和群体间杂交后代进行了遗传和生长检测.结果表明:养殖群体与日本野生群体相比,其遗传多样性没有显著降低,养殖群体遗传多样性较高,养殖状况暂时良好.TL群体和DH群体遗传距离很近, DH、TL群体和RB群体的遗传距离远大于 TL群体和 DH群体间的遗传距离.说明2个养殖虾夷扇贝群体遗传分化较小.分别对4个子代群体的壳长、壳高在幼苗培育第1、5、10、15、20、100、190天进行测量,结果表明生长性状差异不显著.4个子代群体的遗传多样性差异也不显著,说明养殖群体的遗传背景不清楚,种质资源混交较严重.本研究结果反映了我国虾夷扇贝种质资源评估状况,并为虾夷扇贝的健康养殖提供基础数据.  相似文献   

Distribution of Legionella pneumophila in Thermal Pools          下载免费PDF全文

Maciej Walczak  Aleksandra Burkowska‐But  Maria Swiontek Brzezinska  Arkadiusz Krawiec 《洁净——土壤、空气、水》2016,44(5):532-537

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Optimization of fluorescence in situ hybridization （FISH） for the identification of two polar coccoid green algae species     

高小艳  李运广  李会荣  陈雯莉  罗玮 《极地研究(英文版)》2010,21(2):167-179

Standard FISH protocols using fluorochrome-labeled oligonucteotide probes have been successfully applied for in situ detection.However,optimized protocols of FISH for specific eukaryotes in marine environments are often not developed.This study optimized the conditions of fluorescence in situ hybridization (FISH) by using two polar isolated microalgae.The modified conditions were as follows: (1) 10 mg·mL-1 lysozyme solution pretreatment at 37℃ for 30 min;(2) the hybridization buffer including 20% formamide;(3) the hybridization condition was 47℃ for 6 h. The cells enumerated by FISH were compared with those enumerated by flow eytometry (FCM) and DAPI to confirm the cell loss and hybridization efficiency.The optimized protocol was also successfully applied to Arctic Ocean samples,which were found to be dominated by Micromonas sp.The modified protocol showed a high relative efficiency and could be successfully applied for the detection of specific microbial eukaryotes in environmental samples.  相似文献   

文昌鱼胚胎整装荧光原位杂交技术的建立     

喻丹  张培军 《海洋科学》2005,29(6):1-3

文昌鱼(Branchiostoma belcheri tsingtauense)是进化发育生物学研究的重要模式生物,文昌鱼胚胎整装荧光原位杂交(WFISH,whole-mounted fluorescent in situ hybridization)技术将有助于鉴定文昌鱼胚胎发育过程中具有重要调控作用的功能基因。报告了文昌鱼胚胎整装荧光原位杂交技术,用以快速灵敏分析特定基因在文昌鱼胚胎发育过程中的时空表达图式。用体外转录合成的地高辛标记文昌鱼FGFR基因的反义RNA探针,检测到FGFR在文昌鱼原肠胚中表达于发生内陷的中内胚层细胞中,而预定发育为外胚层的细胞不表达FGFR。  相似文献   

核酸探针原位杂交检测白斑综合症病毒的组织特异性   总被引：2，自引：0，他引：2  

吕玲  何建国  邓敏  翁少萍  左涛 《热带海洋学报》2000,19(4):86-91

从白斑综合症病毒 (whitespotsyndromevirus ,简称WSSV)部分基因组文库中得到核酸探针 ,采用原位杂交检测方法检测对虾甲壳下表皮、胃、后肠、鳃、触角腺的上皮细胞、淋巴器官的基质细胞、肌细胞、心肌细胞及结缔组织细胞 ,原位杂交结果均为阳性。对虾甲壳下表皮的上皮细胞、胃的上皮细胞对病毒较其它部位敏感 ,感染程度高 ;中肠上皮细胞、肝胰腺上皮细胞、淋巴器官内皮细胞未发现感染病毒 ,原位杂交用于WSSV检测 ,比组织切片HE染色更准确、更敏感。在同一组织的同一部位 ,原位杂交所得到的阳性细胞个数比HE多。探针大小不同 ,敏感性也有变化。短的探针 41 3bp敏感性较高 ,长的探针敏感性降低。  相似文献   

刺参(Apostichopus japonicus)高温胁迫消减cDNA文库的构建与分析   总被引：1，自引：1，他引：0       下载免费PDF全文

吉成龙  孙国华  杨建敏  宋志乐  刘相全  王卫军 《海洋与湖沼》2011,42(1):60-66

采用抑制性消减杂交技术(SSH)研究刺参高温胁迫下差异表达的基因.分别以高温实验组为检测组(tester)、常温对照组为驱动组(driver),进行正向抑制性消减杂交;以常温组为tester、高温组为driver,进行反向消减杂交,成功构建了刺参正反双向差异消减文库.从两个文库随机挑选384个白斑克隆进行斑点杂交进一步...  相似文献   

华北克拉通孔兹岩带古元古代凉城石榴石花岗岩成因机制及其岩石学意义     

王洛娟  郭敬辉  彭澎 《岩石学报》2021,37(2):375-390

在华北克拉通孔兹岩带东段凉城地区分布有大规模古元古代石榴石花岗岩。凉城石榴石花岗岩是产于麻粒岩相变沉积岩中的原地-半原地花岗岩,伴生有徐武家岩体为代表的辉长苏长岩小侵入体群。凉城石榴石花岗岩富含石榴子石(5%~15%,局部可达25%),常见紫苏辉石,推测形成于高温(>850~900℃)、高压(8~10kbar)条件下,不同于典型的S型花岗岩。地球化学特征上,石榴石花岗岩具有强过铝的地球化学属性(A/CNK=1.1~1.6),显示低SiO₂(55%~75%)和富MgO+FeO^T(5%~14%)的特征,已经偏离了正常S型花岗岩的成分范围。凉城石榴石花岗岩低硅富镁铁的成分特征很可能是由残留体(石榴石岩)带入和辉长苏长岩物质添加造成,残留体和辉长苏长岩的物质贡献比例约占20%~40%。年代学研究表明凉城石榴石花岗岩形成时代为1.93~1.92Ga,与辉长苏长岩形成时代(1.93Ga)和变质沉积岩记录的超高温变质作用时代(1.92Ga)一致,指示凉城石榴石花岗岩是幔源基性岩浆侵入麻粒岩相变质沉积岩中引起变沉积岩在高温-超高温条件下大规模部分熔融的产物。大规模出露的高温石榴石花岗岩及多点分布的超高温麻粒岩共同反映了集宁-凉城区域上超高温的特点。  相似文献   

Detection of Prorocentrum donghaiense using sandwich hybridization integrated with nuclease protection assay   总被引：1，自引：0，他引：1  

CHEN Jie  ZHEN Yu  MI Tiezhu  YU Zhigang 《海洋学报(英文版)》2009,28(2):121-126

Prorocentrum donghaiense is an important harmful algae bloom (HAB) causing creature in China’s seas, and the conventional visual detection can not cope with long-term monitoring and high-throughput sampling projects. An assay for P. donghaiense with sandwich hybridization integrated with nuclease protection assay (NPA-SH) was established. Tests with mixed samples and spiked field ones confirmed its good specificity and sensitivity. The cell number of P. donghaiense correlated well with the optical density, ...  相似文献   

Identification of Provocentrum minimum and Takayama pulchella by fluorescence in situ hybridization through epifluorescence microscopy and flow cytometry     

HOU Jianjun  LAI Hongyan  HUANG Bangqin  CHEN Jixin 《海洋学报(英文版)》2009,28(2)

Partial rDNA sequences of Prorocentrum minimum and Takayama pulchella were amplified,cloned and sequenced,and these sequence data were deposited in the GenBank.Eight oligonucleotide probes(DNA probes)were designed based on the sequence analysis.The probes were employed to detect and identify P.minimum and T. pulchella in unialgal and mixed algal samples with a fuorescence in situ hybridization method using flow cytometry.Epifluorescence micrographs showed that these specific probes labeled with fluorescein isothiocyanate entered the algal cells and bound to target sequences,and the fluorescence signal resulting from whole-cell hybridization varied from probe to probe.These DNA probes and the hybridization protocol we developed were specific and effective for P.minimum and T. pulchella,without any specific binding to other algal species.The hyrbridization efficiency of difierent probes specific to P.minimum was in the order:PMl8S02＞PM28S02＞PM28S01＞PM18S01,and that of the probes specific to T. pulchella was TP18S02＞TP28S01＞TP28S02＞TP18S01.The djfferent hybridization efficiency of the DNA probes could also be shown in the fuorescent signals between the labeled and unlabeled cells demonstrated using flow cytometry.The DNA probes PM18S02,PM28S02,TPl8S02 and TP28S01,and the protocol,were also useful for the detection of algae in natural samples.  相似文献   

Development of taxonomic rRNA-targeted probes of two harmful algae: Prorocentrum minimum and Karenia mikimotoi by fluorescence in situ hybridization     

CHEN Guofu  WANG Quanfu  ZHANG Chunyun  ZHANG Baoyu  WANG Guangce  LU Douding  XU Zhong  YAN Peishen 《海洋学报(英文版)》2013,32(2):66-75

Harmful algal blooms recently have been under the spotlight throughout the world, because of their negative impact on the marine environment, aquaculture, fisheries as well as public health. The development of methods for rapid and precise identification and quantification of causative species is essential for the warning and monitoring of blooms, among which the techniques based on taxonomic probes are the most favored. In this study, two harmful algae, i.e., Prorocentrum minimum and Karenia mikimotoi were taken into consideration. The partial large subunit rDNA (D1-D2) of both species were firstly PCR-amplified, cloned and sequenced. The obtained sequences were then introduced to carry out alignment analysis for gene specific regions. Three respective candidate probes for each species were designed and used to screen the optimal probe by performing fluorescence in situ hybridization (FISH) tests. The results showed that the probes Pmin0443 and Kmik0602 displayed the best hybridization for P. minimum and K. mikimotoi, respectively. Both the specific (taxonomic) (Pmin0443 and Kmik0602) and the control probes (UniC0512 and UniR0499) were used for cross-reactivity tests with other microalgae in our laboratory. The probes Pmin0443 and Kmik0602 are specific and could be served as taxonomic probes introduced into the techniques targeting rRNA, such as FISH, sandwich hybridization, and DNA-microarray assay of P. minimum and K. mikimotoi in the future. Finally, FISH analyses with both probes were performed on the simulated field samples. The probes could hybridize exclusively with the target cells well, and no significant difference (p >0.05) was observed in the cell densities of the samples determined by FISH and light microscopy (LM). All suggest that the probes are specific and could be introduced into FISH for the monitoring of both harmful algae.  相似文献