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1.
太湖似刺鳊鮈染色体组型分析及细胞核DNA含量 总被引：3，自引：0，他引：3
采用PHA、秋水仙素碱腹腔或背部肌肉注射,活体培养法,用空气干燥法制片,运用Micromeasure version 3.3染色体分析软件和Photoshop 7.0对似刺鳊鮈染色体数目和核型进行了分析;同时,取似刺鳊鮈血细胞、尾鳍、鳃、肌肉、性腺和肝脏为材料,以鸡血细胞为DNA标准（2.50pg/2c）,使用EPICS-XL型流式细胞仪测定了似刺鳊鮈细胞核DNA含量。结果显示：染色体众数2n=50,染色体大小的绝对值为0.86～2.32µm,平均长度为1.63µm,未观察到次缢痕及异配型染色体,亦未发现有随体,核型公式为18m+20sm+8st+4t,NF=88。6个组织细胞核DNA含量分别为：尾鳍3.779 pg/2c,鳃4.007 pg/2c,肌肉3.819 pg/2c,性腺(卵巢4.242 pg/2c、精巢1.842 pg/2c),肝脏3.905 pg/2c。尾鳍、鳃、肌肉和肝脏的细胞核DNA含量不存在显著差异（p＞0.05）,但都极显著高于血细胞DNA含量（p＜0.01）;卵巢细胞核DNA含量约为精巢的2倍。 相似文献
2.
采用PHA、秋水仙素碱腹腔或背部肌肉注射,活体培养法,用空气干燥法制片,运用Micromeasure version 3.3染色体分析软件和Photoshop 7.0对似刺鳊(鱼句)染色体数目和核型进行了分析;同时,取似刺鳊鱼句血细胞、尾鳍、鳃、肌肉、性腺和肝脏为材料,以鸡血细胞为DNA标准(2.50 pg/2c),使用EPICS-XL型流式细胞仪测定了似刺鳊(鱼句)细胞核DNA含量.结果显示:染色体众数2n=50,染色体大小的绝对值为0.86～2.32 μm,平均长度为1.63 μm,未观察到次缢痕及异配型染色体,亦未发现有随体,核型公式为18m+20sm+8st+4t,NF=88.6个组织细胞核DNA含量分别为:尾鳍3.779 pg/2c,鳃4.007 pg/2c,肌肉3.819 pg/2c,性腺(卵巢4.242 pg/2c、精巢1.842 pg/2c),肝脏3.905 pg/2c.尾鳍、鳃、肌肉和肝脏的细胞核DNA含量不存在显著差异(P＞0.05),但都极显著高于血细胞DNA含量(P<0.01);卵巢细胞核DNA含量约为精巢的2倍. 相似文献
3.
宽口裂腹鱼为塔里木河水系的特有物种。为建立其尾鳍细胞系，笔者采用组织块法和胰酶消化法相结合，分别用含胎牛血清的DME/F12培养基体外培养尾鳍组织。初步建立宽口裂腹鱼尾鳍细胞系，细胞传代培养至45代。细胞呈悬浮生长，最适培养液为DME/F12,最适胎牛血清质量分数为20%,最适温度为25℃。第10代细胞的群体倍增时间为24.94 h,细胞呈“S”型生长。第6代细胞液氮冷冻保存180 d后复苏，经台盼蓝染色计数，(88.72±0.99)%的宽口裂腹鱼尾鳍细胞系具有活性，复苏后增殖速度快，可正常传代。细菌、真菌、支原体的鉴定未发现污染。第10代细胞线粒体16S rRNA基因测序结果与GenBank中基因序列做一致性对比，宽口裂腹鱼尾鳍细胞系与NC036933.1的一致率达99.91%,从分子水平证明，宽口裂腹鱼尾鳍细胞系来自宽口裂腹鱼。NaCl质量分数为8‰时，宽口裂腹鱼尾鳍细胞系的相对增殖率最高；NaHCO3质量浓度为7 g/L时，宽口裂腹鱼尾鳍细胞系的相对增殖率最高；宽口裂腹鱼尾鳍细胞系的相对增殖率随盐碱度增加呈先升后降趋势。 相似文献
4.
《淡水渔业》2021,51(5)
为了探究吉富罗非鱼(Oreochromis niloticus,GIFT)(♀)与翘嘴鳜(Siniperca chuatsi)(♂)远缘杂交的可行性,对吉富罗非鱼(♀)×翘嘴鳜(♂)子代的胚胎发育、细胞遗传和分子遗传特征进行了初步研究。杂交子代的胚胎存活率仅为(0.34±0.13)%,通过流式细胞技术对正常存活个体进行了DNA相对含量分析,结果显示杂交子代的DNA含量和吉富罗非鱼的DNA含量无显著性差异。染色体数目和核型分析结果显示,杂交子代的染色体数目是2n=44,核型公式是6sm+24st+14t,染色体臂数(NF)=50。杂交子代的染色体数目与其母本相同,核型与其母本相似,二者与父本明显不同。细胞遗传和分子遗传特征的结果显示,吉富罗非鱼(♀)×翘嘴鳜(♂)子代属于雌核发育个体。 相似文献
5.
软骨鱼类染色体研究进展 总被引：2，自引：0，他引：2
在各类脊椎动物中，软骨鱼类是开展染色体研究最少的类群。迄今为止，文献共报道了70种软骨鱼的核型，仅占现生838种软骨鱼类的8．35％，包括31种鲨类，37种鳐类和2种全头类。已报道的软骨鱼类二倍体染色体数为52～104（巴西双鳍电鳐二倍体染色体数为28例外），常存在微染色体。在不同的分类阶元，相对原始的软骨鱼类具有较多的染色体数，端部和亚端部着丝粒染色体较多，多有微染色体。软骨鱼类染色体进化趋势表现为个体染色体数目的减少。中部和亚中部着丝粒染色体数目的增多，并伴随微染色体的消失。软骨鱼类是核DNA含量较大的脊椎动物。已报道144种软骨鱼类的核DNA含量，其变化较大，为3．0～34．1pg／N，全头类的核DNA含量最小，角鲨目和扁鲨目的核DNA含量最大。近年来，在软骨鱼类染色体的C带、核仁组织区（NOR）染色、限制性内切酶显带、特异DNA探针如端粒序列和短散布重复序列（SINE序列）的荧光原位杂交等方面取得了较大进展。现代分子生物学技术也被用于一些软骨鱼类的基因组研究，在重复DNA序列、随体DNA家族和微卫星位点等方面均取得一些成果。 相似文献
6.
用经紫外线灭活的团头鲂精子激活橙黄色锦鲤的卵子,在4℃的水温下冷休克处理卵子30 min使其染色体加倍,获得了孵化率为30.6％、存活率为24.2％的雌核发育锦鲤,表明该雌核发育锦鲤研制方法具有较高的效率.利用外周血细胞培养方法制备了雌核发育锦鲤染色体并利用核型分析方法对其核型进行了分析,发现雌核发育锦鲤染色体数目为100,核型公式为22 m+34 sm+22 st+22 t;表明雌核发育锦鲤为二倍体,其核型与锦鲤母本一致.利用石蜡切片方法,随机选取10尾1龄雌核发育锦鲤性腺进行检测,发现其性腺全部为卵巢;并且在繁殖季节随机检测100尾2龄雌核发育锦鲤,没有l尾能挤出精液,表明雌核发育锦鲤全部为雌性,为证明雌性锦鲤的性别决定类型为XX提供了重要证据.此外,还对雌核发育锦鲤与母本的的SOX基因遗传关系、外形、体色等进行了比较,结果表明雌核发育锦鲤与母本有相同的SOX基因扩增片段,为证明其为雌核发育锦鲤而非杂交后代提供了重要证据;同时,雌核发育锦鲤保留了母本体态优美的特点,且具有色泽更鲜艳等优点,表明该雌核发育方法产生了遗传改良效果.雌核发育锦鲤的获得为该观赏鱼的提纯复壮、遗传改良以及性别决定和性别控制研究奠定了基础. 相似文献
7.
中国胭脂鱼的细胞遗传学分析 总被引：1，自引：0，他引：1
采用植物血凝素、秋水仙素活体注射和肾细胞直接制片法分析了中国胭脂鱼(Myxiocyprinus asiaticus)的染色体核型和Ag-NORs显带;并以中国胭脂鱼血细胞为样本,鸡血细胞DNA含量为标准(2.50 pg),用流式细胞仪测定了中国胭脂鱼的 DNA 含量.结果表明:(1)中国胭脂鱼染色体数目2n=100,核型公式为10 m+4 sm+12 st+74 t, NF=114,未观察到次缢痕及随体,亦未发现有性染色体.(2)中国胭脂鱼染色体具有1对Ag-NORs,位于第1对染色体短臂上,间期核中Ag-NORs的数目为1~2个,未见Ag-NORs的联合现象.(3)PI和DAPI作为荧光染料,得出中国胭脂鱼DNA含量分别为5.50±0.24 pg和6.16±0.03 pg.(4)中国胭脂鱼为四倍体,在特定的分类阶元中属于较原始的鱼类. 相似文献
8.
俄罗斯鲟的细胞遗传学分析 总被引：1，自引：0，他引：1
采用体内注射PHA和秋水仙素,肾细胞短期培养,常规空气干燥法制备俄罗斯鲟(Acipenser gueldenstaedtiBrandt)的染色体,对其肾细胞染色体数目统计分析表明,俄罗斯鲟染色体组由236±2条染色体组成,其中具有明显着丝点的染色体为120条,其余为小染色体(点状染色体);核型公式为80m 16sm 24st,t 116mc,NF=332;采用德国PartecpasⅢ型流式细胞分析仪,以鸡红细胞为标准DNA(含量为2.3pg.N-1),测定俄罗斯鲟体细胞DNA含量为12.24pg.N-1,与染色体分析结果比较倍性一致。综合以上两项结果可得出,俄罗斯鲟为八倍体鱼。 相似文献
9.
对传代296次的牙鲆（Paralichthys olivaceus）鳃细胞系（FG）进行染色体制备和组型分析，并与牙鲆活体鳃细胞组型进行比较。牙鲆活体鳃细胞染色体数目为48，且均为端部着丝粒染色体，染色体组型公式为2n=48，48t，NF=48。而鳃细胞系的染色体数目出现较大变化，有两个分布区问，一个区间是53～79；另一个是119～139。其模式染色体数为68，在模式细胞分裂相中，有30对端部着丝粒染色体和4对中部着丝粒染色体；染色体条数在119以上的分裂相中，除了有很多中部着丝粒染色体以外，小染色体也以很高的频率出现。通过上述研究分析表明，牙鲆鳃细胞系经多次传代后，其染色体与活体鳃细胞染色体相比存在着明显的差别，已经发生了变异，不适合用于牙鲆染色体核型、组型等方面的研究。[中国水产科学，2008，15（3）：483-487] 相似文献
10.
小体鲟染色体核型及倍性的细胞遗传学研究 总被引：1，自引：0，他引：1
采用体内注射PHA和秋水仙素,肾细胞短期培养,常规空气干燥法制备小体鲟(Acipenser dabryanus Dumerii)的染色体,对肾细胞染色体数目统计分析表明,小体鲟染色体组是由116±4条染色体组成,核型公式为40m+34sm+26st+16t±mc,NF 190±;采用德国Partec pasⅢ型流式细胞分析仪,以鸡红细胞为标准DNA(含量为2.3pg/N),测定小体鲟体细胞DNA含量为6.06 pg/N,与染色体分析结果比较倍性一致.综合以上两项结果可以得出,小体鲟为四倍体鲟鱼. 相似文献
11.
Y Wang W Zeng Q Wang Y Li S M Bergmann S Zheng Y Ren C Liu O Chang P Lee 《Journal of fish diseases》2018,41(2):357-364
A novel permanently growing brain cell line from koi (Cyprinus carpio L.) (KB cell line) was established, and its suitability for detection of koi herpesvirus (KHV) was demonstrated in this study. The KB cell line was optimally maintained at 27°C in Leibovitz's L‐15 medium supplemented with 10% foetal bovine serum (FBS). It was subcultured more than 100 times, and chromosome analysis revealed that 51.54% of KB cells at passage 80 maintained the abnormal diploid chromosome number 2n = 96 while the modal chromosome number was 2n = 100. The cell line was cryopreserved in liquid nitrogen at ?196°C and was recovered from storage after 1 year with good cell viability and vitality. The results of virus isolation demonstrated that KB cells were susceptible to KHV, which was shown by the presence of an obvious cytopathic effect and abundant virus particles. The viral titres of KHV in KB reached 105.73TCID50/0.1 ml within 7 days. Immunofluorescence and Western blot assays confirmed that KB replicated KHV. The newly established KB cell line will serve as a useful tool to elucidate KHV disease (KHVD) pathogenesis. 相似文献
12.
Yunbo Wei Tingjun Fan Guojian Jiang Ai Sun Xiaohui Xu & Jing Wang 《Aquaculture Research》2009,40(13):1523-1531
To lay a solid foundation of in vitro investigations of fish viral diseases, cytotechnology and cytotoxicology, a novel fin cell line from brown-marbled grouper, Epinephelus fuscoguttatus , was established and its viral susceptibility was evaluated. The fin tissues, digested with hyaluronidase and collagenase II, were used to initiate primary culture at 24 °C by using 20% foetal bovine serum-Dulbecco's modified Eagle medium/F12 medium, which was further supplemented with carboxymethyl–chitooligosaccharide, basic fibroblast growth factor and insulin-like growth factor-I. The fibroblastic fin cells grew at a steady rate during subsequent subculture and had a population doubling time of 50.6 h at passage 60. The modal diploid chromosome number was 48. A brown-marbled grouper fin cell line (bmGF-1) has been established and subcultured to passage 75 by now. Viral susceptibilities revealed that typical cytopathic effects of bmGF-1 cells emerged after being infected by turbot reddish-body iridovirus (TRBIV) or lymphocystis disease virus (LCDV). However, a large number of TRBIV and LCDV particles were also found in infected bmGF-1 cells. All these indicate that the bmGF-1 cell line has good susceptibility to TRBIV and LCDV, which may serve as a valuable tool for studies of cell–virus interactions and have potential applications in fish virus propagation and vaccine development. 相似文献
13.
A fibroblast cell line (BSF) derived from a caudal fin explant of black sea bream (Mylio macrocephalus) was developed. The optimum fetal bovine serum (FBS) concentration for fibroblast cell line growth was found to be 15–20% v/v FBS and the optimum temperature range for growth was found to be 26–30 °C. The fibroblast cells displayed a diverse distribution in chromosome number with two modal chromosome numbers of 48 and 54. Upon acute heat shock (+8 °C) the cells displayed a 4.1 fold increase in hsp70 and this elevation was not prolonged as hsp70 returned to near basal levels following a 6 h recovery period. The effect of the hsp70 inducer L-azetidine- 2-carboxylic acid was tested and it was found that at a concentration of 10 mM this inducer caused a 2.3 fold increase in hsp70 levels. The sensitivity of the fibroblast cell line to heavy metal exposure was tested by treatment with Cu2+ and it was found that hsp70 was significantly elevated in the presence of micromolar concentrations of Cu2+. The data from this study demonstrates that the established black sea bream fibroblast cell line could serve as a useful in vitro model for stress protein studies. 相似文献
14.
15.
四带小鲃（Puntius tetrazona）是一种具有较高经济价值的小型热带观赏鱼。我们对野生型和长鳍突变型四带小鲃的体长、尾鳍长、尾鳍分节数目和分节长度共4个指标进行了统计分析。结果显示,6月龄野生型（n = 30）与突变型（n = 30）成鱼体长没有显著差异（P = 0.904 2）,但两者在尾鳍长、尾鳍分节数目和分节长度3个指标存在显著差异（P < 0.000 1）。同时,我们对不同体长野生型（n = 80）和突变型（n = 80）尾鳍长度、分节数目和分节长度进行了线性回归分析,结果显示,相对于野生型,突变型尾鳍长度增加速率更快（野生型：k = 0.28,突变型：k = 0.62,P < 0.000 1）,尾鳍分节数目增加频率基本一致（野生型：k = 0.79,突变型：k = 0.82,P = 0.006）,尾鳍分节长度更长（野生型：k = 0.006 8,突变型：k = 0.000 4,P < 0.000 1）。野生型和突变型成鱼尾鳍组织的转录组分析结果显示,转录组从头组装共获得56 271个unigene,野生型组（n=3）和突变型组（n=3）之间共有1 304个差异表达基因,其中971个基因上调,333个基因下调。差异表达基因 KEGG通路显著富集到细胞周期和DNA复制等与细胞增殖相关的信号通路,其中细胞周期蛋白（cyclin A,cyclin B和cyclin E）和细胞周期依赖性激酶（cdk1）等基因显著上调。最后,对ckap2l、ki-67、cdc20、prc1、urgcp和cdks共6个基因的表达水平进行了qRT-PCR分析,其结果与转录组分析基本一致。以上结果表明,长鳍四带小鲃突变基因引起细胞周期相关基因表达量上升,造成尾鳍分节片段的细胞增殖速率加快,尾鳍分节长度增加,呈现为长尾鳍表型。本研究结果为鉴定突变基因和揭示长尾鳍形成的分子机制奠定基础。 相似文献
16.
A continuous cell line [goldfish tail fin (GFTF)] derived from a goldfish tail fin, Carassius auratus, was established and characterized. GFTF cells predominantly consist of fibroblast-like cells that were maintained and subcultured more than 50 times over a period of 15 months. Cells grew at temperatures between 15 and 37°C, with an optimum temperature of 25°C. The growth rate of GFTF cells increased proportionally with the foetal bovine serum (FBS) concentration (5-20%), with optimum growth at 20% FBS. The chromosome numbers were 88-112, with a modal peak of 104 chromosomes. Five known fish viruses were tested to determine susceptibility. Results demonstrated that GFTF is susceptible to snakehead rhabdovirus, spring viraemia of carp virus and channel catfish virus (CCV). In addition, GFTF demonstrated a higher sensitivity to, and increased viral production of, CCV than that observed in the control cell line, channel catfish ovary cells. This suggests that GFTF cells would be useful as a diagnostic tool for viral diseases in this fish species, as well as for the isolation and study of goldfish viruses in the future. Furthermore, these cells were transfected with pEGFP-N1 vector DNA and some fluorescent signals were observed, suggesting that GFTF cells could be a useful tool for transgenic and genetic manipulation studies. 相似文献
17.
Valentin Eckart Takuya Yamaguchi Kati Franzke Sven M. Bergmann Pierre Boudinot Edwige Quillet Motokazu Kawanobe Neila Alvarez de Haro Uwe Fischer 《Journal of fish diseases》2019,42(2):181-187
The production of piscine viruses, in particular of koi herpesvirus (KHV, CyHV‐3) and infectious salmon anaemia virus (ISAV), is still challenging due to the limited susceptibility of available cell lines to these viruses. A number of cell lines from different fish species were compared to standard diagnostic cell lines for KHV and ISAV regarding their capability to exhibit a cytopathic effect (CPE) and to accumulate virus. Two cell lines, so far undescribed, appeared to be useful for diagnostic purposes. Fr994, a cell line derived from ovaries of rainbow trout (Oncorhynchus mykiss), produced constantly high ISA virus (ISAV) titres and developed a pronounced CPE even at high cell passage numbers, while standard cell lines are reported to gradually loose these properties upon propagation. Another cell line isolated from the head kidney of common carp (Cyprinus carpio), KoK, showed a KHV induced CPE earlier than the standard cell line used for diagnostics. A third cell line, named Fin‐4, established from the fin epithelium of rainbow trout did not promote efficient replication of tested viruses, but showed antigen sampling properties and might be useful as an in vitro model for virus uptake or phagocytosis. 相似文献
18.
锦鲤疱疹病毒GZ1301株的分离与鉴定 总被引：3，自引：2，他引：1
2013年4月,广东省一锦鲤养殖场暴发不明病因疾病,濒死锦鲤在塘边游动缓慢直至死亡,死亡率高达100%。现场采样发现,发病锦鲤体长25 cm,眼球凹陷,胸鳍及腹鳍出现出血斑点,解剖发现内脏器官包括肝、脾、肾肿大。细菌分离结果显示,内脏器官肝脏和肾脏中未分离到细菌。提取自然发病鱼的肝、脾、肾、鳃组织DNA作为模板,采用世界动物卫生组织(OIE)推荐的锦鲤疱疹病毒(KHV)检测引物进行PCR扩增,均能扩增出预期大小的特异性产物。NCBI的Blast搜索结果显示,扩增序列与KHV胸苷激酶(thymidine kinase,TK)基因核苷酸序列同源性为99%。病鱼内脏组织研磨过滤除菌后,腹腔注射20尾锦鲤,可复制出与自然发病相似的症状,并于7 d内全部死亡。取病鱼的鳃和肾脏研磨过滤除菌后进行细胞感染实验,结果显示,组织滤液感染CCB细胞后,盲传5代可以观察到典型的细胞病变效应(CPE)。将出现典型CPE的CCB细胞进行超薄切片制备和电镜观察,电镜下病毒呈对称20面体,直径约100 nm。将出现典型CPE的细胞进行间接免疫荧光实验,可以观察到特异性荧光。根据TK基因全长序列建立系统进化树,证实该毒株为KHV亚洲型毒株,暂命名为KHV-GZ1301株。研究结果可为KHV起源进化、分类以及疾病防控提供重要材料。 相似文献
19.
Tor tor is an important game and food fish of India with a distribution throughout Asia from the trans-Himalayan region to the Mekong River basin to Malaysia, Pakistan, Bangladesh and Indonesia. A new cell line named TTCF was developed from the caudal fin of T. tor for the first time. The cell line was optimally maintained at 28°C in Leibovitz-15 (L-15) medium supplemented with 20% fetal bovine serum (FBS). The propagation of TTCF cells showed a high plating efficiency of 63.00%. The cytogenetic analysis revealed a diploid count of 100 chromosomes at passage 15, 30, 45 and 60 passages. The viability of the TTCF cell line was found to be 72% after 6 months of cryopreservation in liquid nitrogen (-196°C). The origin of the cell lines was confirmed by the amplification of 578- and 655-bp sequences of 16S rRNA and cytochrome oxidase subunit I (COI) genes of mitochondrial DNA (mtDNA) respectively. TTCF cells were successfully transfected with green fluorescent protein (GFP) reporter plasmids. Further, immunocytochemistry studies confirm its fibroblastic morphology of cells. Genotoxicity assessment of H?O? in TTCF cell line revealed the utility of TTCF cell line as in vitro model for aquatic toxicological studies. 相似文献
20.
采用体内注射PHA和秋水仙素,肾细胞短期培养,常规空气干燥法制备蓝色鳞鲤染色体,对100个中期分裂相记数统计,确定蓝色鳞鲤（Cyprinus carpio blue var）的染色体数为2n=100。测得核型参数按Levan等的染色体划分标准得出：蓝色鳞鲤有15对中部着丝点染色体（m）;13对亚中部着丝点染色体（sin）;22对端部和亚端部着丝点染色体（st,t）,其染色体总臂数（NF）为156,蓝色鳞鲤核型公式为2n=30m＋26sm＋44st,t。采用流式细胞分析仪测定了蓝色鳞鲤的DNA含量,与鸡血细胞标准对照相比为1．73±0．10,以鸡红细胞DNA含量2．3Pg．N^-1计,则蓝色鳞鲤的体细胞DNA含量为3．99Pg．N^-1。 相似文献