斑鳢、乌鳢及其杂种细胞核DNA流式含量分析

以斑鳢(Channa maculata)、乌鳢(C.argus)及其正交杂种斑乌鳢(斑鳢♀×乌鳢♂)和反交杂种乌斑鳢(乌鳢♀×斑鳢♂)的红细胞为材料,以鸡(Gallus gallus)血细胞为DNA标准(2.5 pg/2c,2c指2倍体),采用流式细胞仪测定了这4种鱼的细胞核DNA含量。斑鳢、乌鳢、斑乌鳢及乌斑鳢这4种鱼血细胞DNA的绝对含量分别为(1.488±0.035)pg/2c、(1.489±0.034)pg/2c、(1.522±0.077)pg/2c和(1.520±0.033)pg/2c。斑鳢和乌鳢的细胞核DNA含量差异不显著(P0.05),斑鳢和乌鳢与两种杂交鳢的DNA含量差异显著(P0.05),两种杂交鳢之间的细胞核DNA含量差异不显著(P0.05)。杂交鳢细胞核DNA含量显著高于亲本,可以作为杂种鉴定的依据。     

泉水鱼的细胞遗传学分析

以泉水鱼(Pseudogyrinocheilus prochilus)的肾为材料,采用体内注射植物血球凝集素(PHA)和秋水仙素、肾组织细胞短期培养、常规空气干燥法制备泉水鱼染色体标本,对其核型进行分析。以泉水鱼外周血细胞为样本,鸡(Gallus gallus)血细胞DNA含量(2.50 pg/2c,2c指二倍体)为标准,用流式细胞仪测定了泉水鱼的DNA含量。结果表明:(1)泉水鱼的染色体数量为2n=50,核型公式为12m+14sm+14st+10t,总臂数NF=76,未发现性别相关的异型染色体;(2)泉水鱼的DNA含量为鸡血对照的(1.05±0.04)倍,其绝对DNA含量为(2.62±0.10)pg/2c。泉水鱼的染色体数目和DNA含量显示出二倍体的特征。     

宽体沙鳅的染色体核型与DNA含量分析

为了解宽体沙鳅(Sinibotia reevesae)的种质特征,以野生宽体沙鳅为材料,采用腹腔注射植物血球凝集素(PHA)和秋水仙素,肾组织细胞短期培养、常规空气干燥法制备染色体标本,并对其核型进行分析。以鸡(Gallus gallus)血细胞DNA含量(2.50 pg/2c,2c指二倍体)为标准,用流式细胞仪测定宽体沙鳅外周血细胞的DNA含量。结果表明:(1)宽体沙鳅的染色体数目为2n=96,核型组成公式为2n=36m+14sm+20st+26t,染色体总臂数NF=146;未发现与性别相关的异型染色体。(2)宽体沙鳅的DNA含量为(2.60±0.36)pg/2c。通过与其他26种鳅科鱼类核型进行比较,发现宽体沙鳅属于鳅科鱼类中的特化类群,其染色体核型经历了罗伯逊易位和染色体多倍化等过程。本研究结果可为宽体沙鳅种质资源保护和细胞遗传学研究提供基础资料。     

三个鲫品系DNA含量的比较研究

采用流式细胞术 (FCM)对红鲫、彭泽鲫、异育银鲫进行红血球DNA含量的检测分析比较 ,以鉴定它们的倍性。结果显示 ,红鲫红血球的DNA含量是 3 0pg ,彭泽鲫是 4 7pg ,异育银鲫是 4 8pg。显而易见 ,彭泽鲫的DNA含量是二倍体红鲫的 1 57倍 ,异育银鲫的DNA含量是红鲫的 1 6倍。采用肾细胞直接制作染色体的方法进行红鲫、彭泽鲫、异育银鲫的染色体倍性鉴定 ,结果红鲫的染色体数目是 10 0 ,为二倍体 (2n =10 0 ) ,彭泽鲫的染色体数目是 162 ,为三倍体 (3n =162 ) ,异育银鲫的染色体数目是 156— 162 ,为三倍体 (3n =156— 162 )。研究证明 :不同品系鲫的DNA含量高低与染色体的倍性有显著的正相关性     

黄喉拟水龟转铁蛋白基因的克隆以及表达特征分析

转铁蛋白在铁的新陈代谢中起着重要的作用,参与细菌感染后的免疫反应。研究克隆了黄喉拟水龟Tf基因组DNA全长序列,并对Tf基因的序列特征进行了分析。序列分析表明,黄喉拟水龟Tf是由两个相似结构域构成的单一肽链,每个结构域包含两个亚基,它们相互作用形成一个深的、亲水的铁结合位点。其基因组DNA由17个外显子和16个内含子组成,与其他脊椎动物Tf基因结构相似,显示了黄喉拟水龟Tf基因在结构上的保守性。同源性分析表明,黄喉拟水龟的Tf基因与鸟类、爬行类的Tf基因同源性最高,约为75%—97%;与哺乳类Tf基因的同源性约为65%—75%;与爪蟾等两栖类的Tf基因也有一定的同源性。荧光定量PCR结果显示,黄喉拟水龟人工感染粘质沙雷氏菌后,Tf基因在其肝脏、脾脏、肾脏及心脏各组织中的表达量均有上调的趋势,这与其他动物经病原刺激后的表达特征具有相似性。       

十四种淡水鱼的DNA含量

本实验采用血涂片,Feulgen染色,显微荧光光度法对分属4目7科的14种淡水鱼的红细胞核DNA含量进行了初步测定。在这些鱼类中,二倍体染色体数目和DNA含量存在着明显的相关性:2n=50左右的类型的DNA含量大多在2.0—2.9微微克(pg);而2n=100的鲤鱼和泥鳅则具有相对高的DNA含量,分别为3.5和4.6pg。分类地位较高的黄鳝和乌鳢具有较低的DNA含量,分别为1.6和1.3pg。鲤科鱼类中,染色体数目2n=50和2n=48的两种类型并不存在一致性的显著差异,这可能反映了这两类核型在进化上的近缘程度。泥鳅的DNA含量大约是大鳞付泥鳅的DNA含量(2.2pg)的2倍,证明了泥鳅是一种四倍体类型的鱼。     

黄喉拟水龟对冬眠期间急性高温胁迫的氧化应激响应

全球气候变暖加剧,暖冬现象出现的频率及强度不断增加,外温动物冬眠期间将面临更加频繁的高温胁迫。但目前,高温胁迫对爬行动物冬眠期间氧化应激以及抗氧化防御系统影响的研究还较为缺乏。本研究以黄喉拟水龟(Mauremys mutica)南北两个种群的一龄幼龟为研究对象,探究了冬眠期间(6℃)幼龟受高温胁迫(25℃)后的脂质过氧化损伤标志物丙二醛(MDA)含量,以及抗氧化防御和修复系统的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)的活性变化。结果显示,南种群在经历高温胁迫后肝脏MDA含量高于北种群,且放回6℃冬眠条件恢复24 h后仍在升高;北种群肌肉MDA含量受高温胁迫后上升,但在冬眠温度下恢复24 h后即降至对照水平。两种群SOD活力受高温刺激影响不显著。同时,黄喉拟水龟南种群肝脏中CAT的活力在高温刺激2 h后达到最高,并且各处理组间南种群CAT活力均高于北种群;北种群幼龟肝脏GSH-Px活力在高温刺激12 h达到最高。综上,本研究表明,高温胁迫对冬眠期间黄喉拟水龟机体造成一定的氧化损伤,南方种群可能更易受到冬眠期间温度波动的影响,但南方种群机体具有更强的抗氧化酶活力以应对高温胁迫的威胁。     

长江江豚基因组大小测定

本研究采用流式细胞术,以公鸡(Gallusdomesticus)红血细胞DNA含量为标准,测定了23头长江江豚(Neophocaenaphocaenoidesasiaeorientali)的基因组大小(或称C值)。实验过程中采用了保存在3中不同条件下的长江江豚的全血样品,用3种不同的方法提取白细胞。为了获得本实验所用的公鸡红血细胞DNA含量的准确值,首先以人(Homosapiens)的C值为标准,对其进行了校正。然后其C值(2C=2.35pg)用于长江江豚的基因组大小测定。结果发现:长江江豚的单倍体DNA含量为3.27pg/C,由此得出其基因组大小为3.17×109bp;雌性和雄性的C值分别为3.25pg和3.29pg,野外长江江豚和豢养长江江豚的C值分别为3.30pg和3.05pg。对雌雄个体之间以及不同生长条件下长江江豚的C值应用独立样本t检验分析,发现:1)不同性别的长江江豚基因组大小之间没有明显的差异;2)豢养条件下的长江江豚和野生长江江豚之间的基因组大小有明显差异,豢养条件下的长江江豚的基因组明显小于野生条件下的长江江豚。据此推测必要环境因子的变化可能会对长江江豚的基因组DNA含量造成影响。     

黄喉拟水龟和乌龟盾片的变异

龟类盾片的数目、形状和排列方式是鉴别属种的重要特征之一。我们在进行皖南龟类区系调查时,采到了盾片的数目和形状与记载的黄喉拟水龟(Mauremysmutica)和乌龟(Chinemysreevsii)差异较大的标本,经研究认为系个体变异。今简单报道如下:一、黄喉拟水龟Mauremysmutica关于黄喉拟水龟的属名,国内外有二种意见:田婉淑、江耀明(1986)和胡淑琴等(1962)采用Clemmysmutica;赵尔宓(1986)采用Mauremysmutica。McDowell(1964)将黄喉水龟(Clemmysmutica)订正为黄喉拟水龟(Mauremysmutica),并有详细的叙述。我们检查了此龟的骨骼,同意McDowell和赵尔…     

Evidence that 19-hydroxyandrostenedione is secreted by the adrenal cortex and is under the control of ACTH and the renin-angiotensin system in man

Plasma 19-hydroxyandrostenedione (19-OH-A-dione) concentrations in man were evaluated using a specific and sensitive radioimmunoassay. Plasma 19-OH-A-dione concentrations (mean ± SE) in normal subjects are 151 ± 14 pg/ml (n=13) in males and 141 ± 9 pg/ml (n=14) in females. Plasma 19-OH-A-dione (mean ± SE) rises significantly during ACTH stimulation (116 ± 25 pg/ml vs 288 ± 38 pg/ml; P<0.01; n=5), declines significantly during dexamethasone suppression (180 ± 30 pg/ml vs 36 ± 14 pg/ml; P<0.01; n=4) and rises significantly during angiotensin II infusion (89 ± 10 pg/ml vs 159 ± 27 pg/ml; P<0.05; n=5). Plasma 19-OH-A-dione in the adrenal vein is much higher than that in the inferior vena cava (2076–3076 pg/ml vs 115–184 pg/ml; n=2). These results demonstrate that 19-OH-A-dione is directly secreted by the adrenal cortex and is under the control of ACTH and the renin-angiotensin system.     

Temperature dependence-independence of antifreeze turnover in Eurosta solidaginis (Fitch)

Temperature effects on antifreeze metabolism were investigated in two populations (northern and southern) of the golden rod gallfly, Eurosta solidaginis. Sorbitol production was temperature dependent and was triggered by short-term exposure to < +10°C. The maximal rate sorbitol synthesis occurred at 0°C. For both populations, sorbitol was rapidly catabolized during warm acclimation at +20°C. During the first 12 h of warm acclimation, sorbitol levels decreased by 36% (19.7 ± 0.6) to 12.6 ± 1.2 μg/mg) and by 83% (to 3.3 ± 1.7 μg/mg) after 48 h in the northern population. The southern population decreased sorbitol levels 64% (11.8 ± 0.69 to 4.2 ± 0.62) after 48 h. The southern population resynthesized more sorbitol than did the northern population upon re-exposure to 0°C. Glycerol levels increased linearly during the experimental period independent of temperature.     

Cytogeography of Cenchrus ciliaris (Poaceae) in Tunisia

Belonging to the genus Cenchrus with 16–22 species, Cenchrus ciliaris L. (syn. Pennisetum ciliare (L.) Link, buffelgrass) is a perennial, common in warmer regions of both hemispheres, growing as a C4 grass in a wide range of habitats. In the present study we determined chromosome number and nuclear DNA content (2C DNA) for 28 natural populations collected from northern to southern Tunisia. Three ploidy levels were found: one tetraploid population (2n?=?4x?=?36), three pentaploid (2n?=?5x?=?45), and 24 hexaploid populations (2n?=?6x?=?54). The hexaploid chromosome number has already been reported for Tunisian populations of C. ciliaris but tetraploid and pentaploid (2n?=?45) are new for this area. The tetraploid population was found in the semi-arid north; pentaploids were mostly on the northern side of the arid region, while the hexaploids were located mainly in the arid southern Tunisian and Saharan region. 2C DNA values, assessed using flow cytometry, correlated with chromosome counts. Nuclear DNA content ranged from 2C?=?3.03 to 4.61 pg, revealing three ploidy levels corresponding to 4x, 5x, 6x, and mean 2C DNA amounts were of 3.03, 3.7 and 4.48 pg, respectively. Each cytotype produced viable pollen. Flow cytometric seed screening neither proved nor disproved apomixis. The most frequent hexaploid populations seem best adapted to arid conditions in southern Tunisia. The monoploid value, 1Cx, was constant. The existence of pentaploid cytotype suggests hybridization ability between tetraploids and hexaploids. It appears that polyploidization is the major evolutionary mechanism in the speciation of C. ciliaris.     

Genome evolution in the genus Sorghum (Poaceae)

BACKGROUND AND AIMS: The roles of variation in DNA content in plant evolution and adaptation remain a major biological enigma. Chromosome number and 2C DNA content were determined for 21 of the 25 species of the genus Sorghum and analysed from a phylogenetic perspective. METHODS: DNA content was determined by flow cytometry. A Sorghum phylogeny was constructed based on combined nuclear ITS and chloroplast ndhF DNA sequences. KEY RESULTS: Chromosome counts (2n = 10, 20, 30, 40) were, with few exceptions, concordant with published numbers. New chromosome numbers were obtained for S. amplum (2n = 30) and S. leiocladum (2n = 10). 2C DNA content varies 8.1-fold (1.27-10.30 pg) among the 21 Sorghum species. 2C DNA content varies 3.6-fold from 1.27 pg to 4.60 pg among the 2n = 10 species and 5.8-fold (1.52-8.79 pg) among the 2n = 20 species. The x = 5 genome size varies over an 8.8-fold range from 0.26 pg to 2.30 pg. The mean 2C DNA content of perennial species (6.20 pg) is significantly greater than the mean (2.92 pg) of the annuals. Among the 21 species studied, the mean x = 5 genome size of annuals (1.15 pg) and of perennials (1.29 pg) is not significantly different. Statistical analysis of Australian species showed: (a) mean 2C DNA content of annual (2.89 pg) and perennial (7.73 pg) species is significantly different; (b) mean x = 5 genome size of perennials (1.66 pg) is significantly greater than that of the annuals (1.09 pg); (c) the mean maximum latitude at which perennial species grow (-25.4 degrees) is significantly greater than the mean maximum latitude (-17.6) at which annual species grow. CONCLUSIONS: The DNA sequence phylogeny splits Sorghum into two lineages, one comprising the 2n = 10 species with large genomes and their polyploid relatives, and the other with the 2n = 20, 40 species with relatively small genomes. An apparent phylogenetic reduction in genome size has occurred in the 2n = 10 lineage. Genome size evolution in the genus Sorghum apparently did not involve a 'one way ticket to genomic obesity' as has been proposed for the grasses.     

Geranium kikianum sp. nov. (Geraniaceae) from the southern Peloponnese,Greece

Geranium kikianum Kit Tan & G. Vold sp. nov. (Geraniaceae) is illustrated and described as a new species endemic to Greece. It occurs beside streams and other wet places in open Pinus nigra forest at the foothills of Mt Taigetos in the southern Peloponnese. It is closely related to G. macrorrhizum, a species more widely distributed in southern and central Europe, ranging from the southern Alps to the Balkan Peninsula. It differs from the latter by its smaller and narrower deflexed petal limb, slender claw and occurrence in wet habitats. The genome sizes of G. kikianum and of Greek populations of G. macrorrhizum are presented here for the first time. The nuclear DNA content (2C values) of 2.84 pg for G. kikianum and 2.83 and 2.87 pg for two G. macrorrhizum populations from Greece probably corresponds to a 2n=92 cytotype while the smaller genome size of 1.45 pg in a population of G. macrorrhizum from Mt Smolikas, Greece may correspond to a 2n=46 cytotype.     

Genome size variation in the common frog Rana temporaria

Genome size variation in the common frog (Rana temporaria) was investigated with flow cytometry in three latitudinally separated populations in Sweden to see whether it could provide a useful tool for sex-identification in this species. Depending on the sex and population, per cell DNA content (2C value) varied from 8.823 to 11.266 pg with a mean (+/- SE) 2C value of 9.961+/-0.083 pg. Analysis of variance revealed significant differences in genome size among populations and between sexes. Females had ca 3% larger genomes (x=10.133+/-0.068 pg) than males (x=9.832+/-0.068 pg) in all of the populations (sex x population interaction: P>0.10). Individuals from the southern-most population had significantly (x=9.330+/-0.081 pg) smaller genomes than those from the more northern populations (x=10.032+/-0.085 and x=10.584+/-0.085 pg, respectively). These results are in line with the interpretation that males in the common frog are the heterogametic sex, and that there exists large (up to 12%) geographic variation in genome size in this species. However, the sex differences in the genome size are too small to be useful in individual sex identification.     

Mosaic individuals found in genetically manipulated northern pike (Esox lucius) using flow cytometry

The purpose of this study was to investigate abnormal larvae produced in the experimental induction of gynogenetic northern pike ( Esox lucius L.), using a flow cytometry method. We reported for the first time the mosaic individuals found in genetically manipulated northern pike. Two types of abnormal larvae were obtained after eggs were fertilized with UV-irradiated sperm and then exposed to heat shock treatment. One type had malformations frequently associated with haploid syndrome and were proven to be haploids. The other type had a similar body size and appearance as diploids but with curved body and a swirling swimming pattern. Individuals of the second type were proven to be mosaics containing both 1n (59.5 ± 14.7% mean ± SD) and 2n (29.7 ± 13.3%) cell populations. The ratio of 1n:2n cell populations ranged from 80.6:10.6 to 38.5:44.7. There was no significant difference in the relative nuclear DNA content and cell size between 1n and 2n cell populations of the mosaic individuals as compared to haploid cells and diploid cells, respectively. The nuclear DNA content of the northern pike was estimated to be 2.07 ± 0.06 pg ( n  = 10) given that rainbow trout has a red blood cell nucleus DNA content of 4.66 pg.     

Microsatellite Markers Reveal Strong Genetic Structure in the Endemic Chilean Dolphin

Understanding genetic differentiation and speciation processes in marine species with high dispersal capabilities is challenging. The Chilean dolphin, Cephalorhynchus eutropia, is the only endemic cetacean of Chile and is found in two different coastal habitats: a northern habitat with exposed coastlines, bays and estuaries from Valparaíso (33°02′S) to Chiloé (42°00′S), and a southern habitat with highly fragmented inshore coastline, channels and fjords between Chiloé and Navarino Island (55°14′S). With the aim of evaluating the potential existence of conservation units for this species, we analyzed the genetic diversity and population structure of the Chilean dolphin along its entire range. We genotyped 21 dinucleotide microsatellites for 53 skin samples collected between 1998 and 2012 (swab: n = 8, biopsy: n = 38, entanglement n = 7). Bayesian clustering and spatial model analyses identified two genetically distinct populations corresponding to the northern and southern habitats. Genetic diversity levels were similar in the two populations (He: 0.42 v/s 0.45 for southern and northern populations, respectively), while effective size population was higher in the southern area (Ne: 101 v/s 39). Genetic differentiation between these two populations was high and significant (F_ST = 0.15 and R_ST = 0.19), indicating little or no current gene flow. Because of the absence of evident geographical barriers between the northern and southern populations, we propose that genetic differentiation may reflect ecological adaptation to the different habitat conditions and resource uses. Therefore, the two genetic populations of this endemic and Near Threatened species should be considered as different conservation units with independent management strategies.     

Quantitative analysis of DNA in sea urchin eggs and subcellular distribution of DNA in the eggs

An improved procedure for analyzing DNA in animal eggs which contain a great deal of other chromogenic materials has been developed. As a result the following DNA contents were estimated: 3.59 ± 0.39 pg DNA/egg for H. pulcherrimus, 5.06 ± 0.30 pg DNA/egg for Ps. depressus, and 3.78 ± 0.33 pg DNA/egg for A. crassispina. Egg nuclei contain the haploid level of DNA. Most of the cytoplasmic DNA was found to be associated with the mitochondria (more than 90%) and only a small portion was detected in the yolk granule fraction.