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1.
罗氏沼虾繁殖行为的再描述   总被引:1,自引:0,他引:1       下载免费PDF全文
王春  成永旭 《动物学杂志》2009,44(4):102-110
2007年6~9月对采自孟加拉国库尔纳专区羌纳县布苏尔河原产地罗氏沼虾(Macrobrachium rosenbergii)的繁殖行为进行了观察研究。配合摄像机录像技术,重点观察并记录了野生罗氏沼虾生殖过程中的主要行为特征。本文所涉及的繁殖过程特指在人工操控条件下,自两性遭遇、识别至雌虾抱卵为止这段时间。选择体重在80.0g以上的雄虾、40.0g以上的雌虾分别在当地河堤外侧的池沼中驯化适应15d,然后,在室内水泥池中观察其繁殖行为。罗氏沼虾在繁殖过程中呈现的典型行为主要有:两性遭遇及识别,雄虾的占位,雌虾的蜕壳,雄虾俘获雌虾及其交媾前的守卫、交媾及精荚传递,雄虾交媾后的守卫及释放雌虾,雌虾产卵及抱卵。其中,雄虾的配偶守卫行为是有效地保证两性生殖成功的交配策略之一。  相似文献   

2.
不同形态型雄性罗氏沼虾的交配能力   总被引:1,自引:1,他引:0  
王春  成永旭 《应用生态学报》2010,21(4):1043-1048
从行为生态学角度,对采自孟加拉国库尔纳专区羌纳县布苏尔河罗氏沼虾原产地池塘种群中3种不同形态型雄虾的繁殖行为及交配能力进行了初步研究.结果表明:社群中蓝螯型(BC)、橙螯型(OC)和小型(SM)雄性罗氏沼虾的交配行为模式没有明显的区别,但BC、OC和SM雄虾在交媾前守卫、交媾及精荚传递和交媾后守卫3个典型行为中投入的时间存在明显差异.BC雄虾交媾前守卫、交媾及精荚传递用时明显比OC、SM雄虾短,而交媾后守卫用时明显比OC、SM雄虾长.SM雄虾在交媾前守卫和精荚传递上投入的时间最长,而交媾后守卫用时最短.在雌虾体质量为雄虾体质量的80%,且♀∶♂为8∶1性比条件下,BC、OC和SM雄虾在120h内平均交媾次数分别为4.6、2.2、1.3,BC雄虾具有较强的连续交媾能力.罗氏沼虾的交配能力与其体质量相关,雄、雌虾的体质量相匹配才能保证成功交媾.否则会出现"假交"现象.  相似文献   

3.
罗氏沼虾受精机制的细胞学研究   总被引:2,自引:0,他引:2  
王玉凤  堵南山  赖伟 《动物学报》1998,44(2):200-202
运用细胞学方法对罗氏沼虾的受精机理进行了初步研究,从卵巢中取邮的成熟卵,表面略有皱褶,无受精孔,不同于一般的节肢动物。成熟精子基部侧边分布许多成束的管状结构,其末端呈连续泡状。精卵接触时,这些泡破裂,膜与卵表膜融合,泡内的糖复合物对精卵识别,粘附可能起着重要和,从而使精子以基部侧边附着卵子表面,。  相似文献   

4.
为了发掘罗氏沼虾(Macrobrachium rosenbergii)免疫相关基因, 研究Rab蛋白(Ras-related proteins inbrain)在罗氏沼虾免疫应答中发挥的作用, 研究采用RACE-PCR技术克隆了罗氏沼虾Rab11基因全长cDNA序列, 记为MrRab11。全长1381 bp, 包括226 bp的5'UTR, 511 bp的3'UTR和645 bp的开放阅读框, 编码214个氨基酸, 含有一个Rab结构域。氨基酸序列比对显示, 罗氏沼虾与冈比亚按蚊(Anopheles gambiae)、蚤状溞(Daphniapulex)、叶蝉(Homalodisca vitripennis) Rab11一致性分别为82%、83%和82%。软件预测, MrRab11编码的蛋白分子量约为23.75 kD, 等电点约为5.34。实时荧光定量表达分析表明, MrRab11基因在罗氏沼虾各组织中都有表达, 肝胰腺中的表达量最高, 其次是肌肉和肠道。在阴沟肠杆菌(Enterobacter cloacae)感染12h后, 罗氏沼虾肝胰腺中MrRab11的表达量上升, 显著高于对照组(P0.05), 推测这是MrRab11对阴沟肠杆菌的应激表达,MrRab11在肝胰腺中参与了罗氏沼虾免疫应答过程。  相似文献   

5.
为探求罗氏沼虾精氨酸激酶(Macrobrechium rosenbergii arginine kinase,MrAK)基因特征和与病毒感染的相关性,研究通过AK基因mRNA全长克隆,并采用QPCR检测病毒感染前后不同时间点罗氏沼虾幼体AK基因的转录差异。最终克隆得到的AK序列全长为1740 bp(GenBank:KT970484),其开放阅读框包含1068个碱基,编码356个氨基酸;同源性分析显示MrAK基因编码区蛋白与多齿新米虾、南极磷虾、鲑鱼海虱和安氏伪镖水蚤的同源性分别为97%、82%、83%和79%;系统进化树分析表明,该基因与多齿新米虾的AK聚在同一分支簇,而蟹、对虾和螯虾聚在另一分支;氨基酸结构分析表明,其存在一个可能的ATP:胍基磷酸转移酶的活性位点(Cys286-Pro287-Thr288-Asn289-Leu290-Gly291-Thr292);病毒感染罗氏沼虾幼体后,其AK基因表达在第9h开始出现显著性上调,在12h时达到峰值,随后开始降低。以上研究结果表明,AK基因在无脊椎甲壳动物中存在多样性,其蛋白结构存在保守性,并且其在罗氏沼虾病毒感染过程中起到潜在的能量供给调节作用。  相似文献   

6.
对刚孵化后的罗氏沼虾(Macrobrachium rosenbergii)亲虾用高低两个浓度的壬基酚(nonylphenol,NP,100μg/L和0·01μg/L)和雌二醇(estradiol,E2,1μg/L和0·01μg/L)进行浸泡处理,分别于3d和5d对罗氏沼虾肝胰腺和卵巢中卵黄蛋白原(vitellogenin,VTG)基因表达变化进行半定量分析。结果显示,NP和E2能够提高罗氏沼虾肝胰腺和卵巢中卵黄蛋白原VTG基因的表达。100μg/LNP对罗氏沼虾表现出雌激素效应,0·01μg/LNP作用效果不明显;而E2则在1μg/L和0·01μg/L两个浓度下均对罗氏沼虾有较强的雌激素效应。在100μg/LNP作用下,卵巢VTG表达量3d和5d均保持较高水平,无明显下降,其他实验组均是3dVTG基因表达量升高,5d后表达量较3d表达量有所降低。结果表明,与其他动物一样,NP对罗氏沼虾具有内分泌干扰作用。  相似文献   

7.
对虾白斑综合征病毒(white spot syndrome virus,WSSV)是一种能够感染虾类并且造成其大面积死亡的环状双链DNA病毒。WSSV有多种分离株,其毒力有所差异。从克氏原螯虾(Procambarus clarkii)中分离得到1株WSSV新分离株WSSV-CN-Pc,其毒力尚不清楚。本研究采用肌肉注射和经口注射的方法,以WSSVTW型作为阳性对照,分别对克氏原螯虾(P.clarkii)和罗氏沼虾(Macrobrachium rosenbergii)进行活体实验。实验结果显示:肌肉注射WSSV-CN-Pc和WSSV-TW的克氏原螯虾均在第6天出现100%的死亡;罗氏沼虾在肌肉注射WSSV-TW后未出现死亡,但在注射WSSV-CN-Pc后的第9天死亡率达100%。经口注射WSSV-CN-Pc和WSSV-TW的克氏原螯虾均在第16天出现100%的死亡;罗氏沼虾经口注射WSSV-CN-Pc后的第19天死亡率为100%,但注射WSSV-TW的实验组并未出现死亡。结果表明,对于克氏原螯虾,WSSV-CN-Pc具有和WSSV-TW相似的毒力,而对罗氏沼虾存在明显的毒力差异。提示克氏原螯虾是WSSV传播途径中的重要因素。  相似文献   

8.
罗氏沼虾白尾病(White tail disease,WTD)是流行于罗氏沼虾苗种阶段的重要疾病,该病病原被确定为罗氏沼虾野田村病毒(Macrobrachium rosenbergii Nodavirus,MrNV),为了建立罗氏沼虾野田村病毒中国分离株(MrNV-chin)的便捷、灵敏、特异的分子诊断方法,本研究根据罗氏沼虾野田村病毒RNA2基因保守序列的6个区域设计了4条特异性引物和1条检测探针,将环介导等温扩增技术与侧向流层析试纸(Lateral flow dipstick,LFD)相结合,建立了MrNV的RT-LAMP-LFD检测方法。结果表明,该方法的灵敏度是常规琼脂糖凝胶电泳的10倍;从核酸提取到检测结果判定仅需45min;反应特异性强,对其他虾类病毒WSSV、IHHNV、IMNV、TSV和YHV扩增结果均为阴性;操作简单,不需要复杂的仪器,在61℃水浴反应即可;利用LFD试纸显示结果,肉眼可直接观察判定。作为一种快速灵敏实用的分子诊断技术,该方法有利于罗氏沼虾白尾病的诊断与预防。  相似文献   

9.
罗氏沼虾(Macrobrachium rosenbergii)和日本沼虾(M.nipponense)经在我国得到广泛的养殖,产生巨大的经济效益.祁连沼虾(M. qilianensis)是自然分布在我国甘肃省的土著虾种,因其外部形态符合沼虾属的特征,而被前人归入沼虾属.为了从分子生物学的角度理解罗氏沼虾、日本沼虾与祁连沼虾的遗传差异,为合理开发和利用沼虾资源提供理论基础,作者对这3种沼虾的线粒体COI基因序列进行研究.从甘肃、浙江等地分别采集这三种沼虾的样本各10尾,共30尾,其中祁连沼虾是野生样本,而罗氏沼虾和日本沼虾都是养殖样本.通过PCR方法扩增线粒体COI基因,并测序.通过比对,获得一致序列649 bp.在30个样本中共检测到169个变异位点,占总变异的26.04%;共检测到7种单倍型.3种沼虾的核苷酸多态性分别为:罗氏沼虾0.411%、日本沼虾0.092%、祁连沼虾0.031%.野生的祁连沼虾遗传多样性远远低于养殖的罗氏沼虾和日本沼虾.三种沼虾单倍型之间的Kimura双参数遗传距离在19.87%~23.84%,三者之间的遗传距离较大,提示三者均为有效种.为进一步确定这三种沼虾在长臂虾科的分类地位, 我们从NCBI数据库中下载了长臂虾科的其它种类的COI序列进行系统发生分析.用NJ法构建的分子系统树显示:日本沼虾和罗氏沼虾与沼虾属的其它种类聚成一枝,而祁连沼虾与同亚科的脊尾白虾(Exopalaemon carinicauda)和长角长臂虾(Palaemon debilis)较沼虾属另10种虾的遗传距离近,即祁连沼虾与白虾属及长臂虾属聚成另一枝.凶此,COI序列的结果不支持祁连沼虾归入沼虾属.但其分类地位应该综合多方面证据重新进行分析确定.  相似文献   

10.
通过对日本沼虾(Macrobrachium nipponense)3个群体线粒体DNA 16S rRNA基因片段进行扩增和测定,得到长度为495bp的片段,其碱基A、T、G和C的平均含量分别为28.6%、36.1%、22.7%和12.5%,AT含量明显高于GC含量。通过对日本沼虾16SrRNA基因片段遗传特征的研究发现其种内变异很小,在3个群体中只有5个位点发生转换。另外,利用其454bp的同源序列,以中国明对虾(Fenneropenaeus chinensis)为外群探讨了沼虾属日本沼虾、罗氏沼虾(M.rosenbergii)等8种沼虾的系统进化关系。用MEGA3.1软件中的NJ法构建的分子进化树,日本沼虾3个群体先聚在一起后与海南沼虾聚在一起;另外,罗氏沼虾与马氏沼虾、短腕沼虾与贪食沼虾亲缘关系较近先聚在一起,然后再与大臂沼虾和等齿沼虾聚在一起,最后才与外群中国明对虾聚在一起。  相似文献   

11.
To determine the molecular basis of transformation defects in Haemophilus influenzae, the fate of genetically marked, (32)P-labeled, heavy deoxyribonucleic acid (DNA) was examined in three mutant strains (rec(1) (-), rec(2) (-), and KB6) and in wild type having (3)H-labeled DNA and a second genetic marker. Transforming cells upon lysis with digitonin followed by low-speed centrifugation are separable into the supernatant fraction, containing mainly the unintegrated donor DNA, and the pellet, containing most of the resident DNA along with integrated donor DNA. Electron micrographs of digitonin-treated cells also indicate that the resident DNA is trapped inside a cellular structure but that cytoplasmic elements such as ribosomes are extensively released. DNA synthesis in digitonin-treated cells is immediately blocked, as is any further integration of donor DNA into the resident genome. Isopycnic and sedimentation analysis of supernatant fluids and pellets revealed that in strains rec(2) (-) and KB6 there is little or no association between donor and resident DNA, and thus there is negligible transfer of donor DNA genetic information. In these strains, the donor DNA is not broken into pieces of lower molecular weight as it is in strain rec(1) (-) and in the wild type, both of which show association between donor and recipient DNA. In strain rec(1) (-), although some donor DNA atoms become covalently linked to resident DNA, the incorporated material does not have the donor DNA transforming activity.  相似文献   

12.
A new method has been developed for determination of DNA synthesis during cell proliferation. The method is based on the metabolism of [U-(13)C(6)]glucose to deoxyribose (DR) and then incorporation of [U-(13)C(5)]DR into newly synthesized DNA. Extracted cellular DNA is subjected to HCl hydrolysis (2 h at 100 degrees C), which converts DR into levulinic acid. The (13)C enrichment in DR is determined in the trimethylsilyl derivative of levulinate using gas chromatography-mass spectrometry. The method is rapid and sensitive. It can precisely determine (13)C enrichment below 1 at.% excess in as little as 4 ng DNA. We have used this method to determine the rate of cell proliferation in vitro and the level of DR in a given amount of DNA. The current approach has significant advantages over previously described methods and overcomes several difficulties related to the determination of DNA synthesis both in vivo and in vitro.  相似文献   

13.
9-(2-Phosphonylmethoxyethyl)adenine (PMEA) is a new antiviral compound with activity against herpes simplex virus (HSV) and retroviruses including human immunodeficiency virus. Although it has been suggested that the anti-HSV action of PMEA is through inhibition of the viral DNA polymerase via the diphosphorylated metabolite of PMEA (PMEApp), no conclusive evidence for this has been presented. We report that in cross-resistance studies, a PMEA-resistant HSV variant (PMEAr-1) was resistant to phosphonoformic acid, a compound which directly inhibits the HSV DNA polymerase. In addition, phosphonoformic acid-resistant HSV variants with defined drug resistance mutations within the HSV DNA polymerase gene were resistant to PMEA. Furthermore, the HSV DNA polymerase purified from PMEAr-1 was resistant to PMEApp in comparison with the enzyme from the parental virus. Moreover, PMEA inhibited HSV DNA synthesis in cell culture. These results provide strong evidence that HSV DNA polymerase is the major target for the anti-viral action of PMEA. Further studies showed that HSV DNA polymerase incorporated PMEApp into DNA in vitro, while the HSV polymerase-associated 3'-5' exonuclease was able to remove the incorporated PMEA. Thus, the inhibition of HSV DNA polymerase by PMEApp appears to involve chain termination after its incorporation into DNA.  相似文献   

14.
M Kann  A Bischof    W H Gerlich 《Journal of virology》1997,71(2):1310-1316
Hepadnaviruses contain a DNA genome, but they replicate via an RNA intermediate, synthesized by the cellular RNA polymerase II in the nucleus of the infected cell. Thus, nuclear transport of the viral DNA is required in the viral life cycle. Protein-free DNA is only poorly imported into the nucleus, so one or more of the viral proteins must be involved in the transport of the viral genome. In order to identify these viral proteins, we purified woodchuck hepadnavirus (WHV) core particles from infected woodchuck liver, isolated WHV DNA, and extracted the covalent complex of viral polymerase from the particles using urea. Intact core particles, the polymerase-DNA complex, or protein-free WHV DNA from core particles was added to digitonin-permeabilized HuH-7 cells, in which the cytosol was substituted by rabbit reticulocyte lysate (RRL) and an ATP-generating system. The distribution of the viral genome was analyzed by semiquantitative PCR or by hybridization in total nuclei, RRL, nuclear membranes, and nucleoplasm. The polymerase-DNA complex was efficiently transported into the nucleus, as indicated by the resistance of the nucleus-associated DNA to a short-term treatment with DNase I of the intact nuclei. The DNA within core particles stayed mainly in the cytosol and remained protected against DNase I. A minor part of the encapsidated DNA was bound to nuclei. It was protected against DNase I but became accessible after disruption of the nuclei. Deproteinized viral DNA completely remained in the cytosol. These data show that the viral polymerase is probably sufficient for mediating the transport of a hepadnavirus genome into the nucleus and that the viral core particles may release the genome at the nuclear membrane.  相似文献   

15.
Agents discriminating between DNA polymerase alpha and DNA polymerases of class delta (polymerase delta or epsilon) were used to characterize steps in the synthesis of the lagging DNA strand of simian virus 40 during DNA replication in isolated nuclei. The synthesis of lagging-strand intermediates below 40 nucleotides, termed DNA primers (T. Nethanel, S. Reisfeld, G. Dinter-Gottlieb, and G. Kaufmann, J. Virol. 62:2867-2873, 1988), was selectively inhibited by butylphenyl dGTP or by neutralizing DNA polymerase alpha monoclonal antibodies. The synthesis of longer lagging chains of up to 250 nucleotides (Okazaki pieces) was affected to a lesser extent, possibly indirectly, by these agents. Aphidicolin, which inhibits both alpha- and delta-class enzymes, elicited the opposite pattern: DNA primers accumulated in its presence and were not converted into Okazaki pieces. These and previous data suggest that DNA polymerase alpha primase synthesizes DNA primers, whereas another DNA polymerase, presumably DNA polymerase delta or epsilon, mediates the conversion of DNA primers into Okazaki pieces.  相似文献   

16.
H Guo  C Xu  T Zhou  TM Block  JT Guo 《PloS one》2012,7(8):e43270
Synthesis of the covalently closed circular (ccc) DNA is a critical, but not well-understood step in the life cycle of hepadnaviruses. Our previous studies favor a model that removal of genome-linked viral DNA polymerase occurs in the cytoplasm and the resulting deproteinized relaxed circular DNA (DP-rcDNA) is subsequently transported into the nucleus and converted into cccDNA. In support of this model, our current study showed that deproteinization of viral double-stranded linear (dsl) DNA also took place in the cytoplasm. Furthermore, we demonstrated that Ku80, a component of non-homologous end joining DNA repair pathway, was essential for synthesis of cccDNA from dslDNA, but not rcDNA. In an attempt to identify additional host factors regulating cccDNA biosynthesis, we found that the DP-rcDNA was produced in all tested cell lines that supported DHBV DNA replication, but cccDNA was only synthesized in the cell lines that accumulated high levels of DP-rcDNA, except for NCI-H322M and MDBK cells, which failed to synthesize cccDNA despite of the existence of nuclear DP-rcDNA. The results thus imply that while removal of the genome-linked viral DNA polymerase is most likely catalyzed by viral or ubiquitous host function(s), nuclear factors required for the conversion of DP-rcDNA into cccDNA and/or its maintenance are deficient in the above two cell lines, which could be useful tools for identification of the elusive host factors essential for cccDNA biosynthesis or maintenance.  相似文献   

17.
Friend murine leukemia virus (F-MuLV) is a replication-competent, ecotropic, NB-tropic retrovirus which produces a rapidly fatal erythroleukemia in susceptible strains of mice. We previously molecularly cloned the entire F-MuLV genome. Transfection of this cloned DNA into NIH 3T3 mouse fibroblasts produces a virus with the same leukemia-inducing characteristics as F-MuLV. To identify which portion of the F-MuLV genome is responsible for causing leukemia, we made recombinant viruses between subgenomic fragments of F-MuLV DNA and another retrovirus--Amphotroph clone 4070. Amphotroph clone 4070 is a replication-competent, amphotrophic, N-tropic virus which does not produce any detectable malignancy in mice. A 2.4-kilobase-pair fragment of F-MuLV DNA was isolated. This DNA fragment encompassed approximately 700 base pairs from the 3' end of the F-MuLV pol gene and 1.7 kilobase pairs of the env gene including all of gp70 and the N-terminal four-fifths of p15E. A molecularly cloned fragment of Amphotroph DNA was ligated to the 2.4-kilobase-pair F-MuLV DNA, and an 8.3-kilobase-pair hybrid F-MuLV-Amphotroph DNA was subcloned into a new plasmid (p5a25-H). Transfection of p5a25-H DNA into fibroblasts resulted in the production of a replication-competent, ecotropic, N-tropic retrovirus--5a25-H virus. Inoculation of this virus into newborn NIH Swiss mice caused leukemia within 4 to 6 months. The disease caused by 5a25-H was pathologically and histologically indistinguishable from the disease caused by F-MuLV. We conclude that the F-MuLV sequences needed to cause disease are contained in these 2.4 kilobase pairs of DNA.  相似文献   

18.
19.
A method has been devised for directly detecting and monitoring genetically engineered microorganisms (GEMs) by using in vitro amplification of the target DNAs by a polymerase chain reaction and then hybridizing the DNAs with a specific oligonucleotide or DNA probe. A cloned 0.3-kilobase napier grass (Pennisetum purpureum) genomic DNA that did not hybridize to DNAs isolated from various microorganisms, soil sediments, and aquatic environments was inserted into a derivative of a 2,4-dichlorophenoxyacetic acid-degradative plasmid, pRC10, and transferred into Escherichia coli. This genetically altered microorganism, seeded into filter-sterilized lake and sewage water samples (10(4)/ml), was detected by a plate count method in decreasing numbers for 6 and 10 days of sample incubation, respectively. The new method detected the amplified unique marker (0.3-kilobase DNA) of the GEM even after 10 to 14 days of incubation. This method is highly sensitive (it requires only picogram amounts of DNA) and has an advantage over the plate count technique, which can detect only culturable microorganisms. The method may be useful for monitoring GEMs in complex environments, where discrimination between GEMs and indigenous microorganisms is either difficult or requires time-consuming tests.  相似文献   

20.
In this report we propose a model in which after the herpes simplex virus (HSV) capsid docks at the nuclear pore, the tegument protein attached to the capsid must be cleaved by a serine or a cysteine protease in order for the DNA to be released into the nucleus. In support of the model are the following results. (i) Exposure of cells at the time of or before infection to l-(tosylamido-2-phenyl) ethyl chloromethyl ketone (TPCK), a serine-cysteine protease inhibitor, prevents the release of viral DNA or expression of viral genes. TPCK does not block viral gene expression after entry of viral DNA into the nucleus. (ii) The tegument protein VP1-2, the product of the U(L)36 gene, is cleaved shortly after the entry of the HSV 1 (HSV-1) virion into the cell. (iii) The proteolytic cleavage of VP1-2 does not occur in cells that are infected with HSV-1 under conditions that prevent the release of the viral DNA into the nucleus. (iv) The proteolytic cleavage of VP1-2 occurs only after the capsid is attached to the nuclear pore. Thus, TPCK prevented the release of HSV-1 DNA into the nucleus when added to medium 1 hour after infection with tsB7 at 39.5 degrees C followed by a shift down to the permissive temperature. The ts lesion maps in the U(L)36 gene. At the nonpermissive temperature, the capsids accumulate at the nuclear pore but the DNA is not released into the nucleus.  相似文献   

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