Physical mapping of 5S and 45S rDNA loci in pufferfishes (Tetraodontiformes)

Chromosomal features, location and variation of the major and minor rDNA genes cluster were studied in three pufferfish species: Sphoeroides greeleyi and Sphoeroides testudineus (Tetraodontidae) and Cyclichthys spinosus (Diodontidae). The location of the major rDNA was revealed with an 18S probe in two loci for all species. The minor rDNA loci (5S rDNA) was found in one chromosome pair in tetraodontid fishes and four sites located on two distinct chromosomal pairs in C. spinosus. A syntenical organization was not observed among the ribosomal genes. Signal homogeneity for GC/AT-DNA specific fluorochromes was observed in diodontid fish except in the NORs regions, which were CMA₃-positive. Giemsa karyotypes of tetraodontid species presents 2n = 46, having the same diploid value of other Sphoeroides species that have been investigated. On the other hand, the karyotype of C. spinosus, described for the first time, shows 2n = 50 chromosomes (4m + 18sm + 12st + 16a). The foreknowledge of the karyotypic structure of this group and also the physical mapping of certain genes could be very helpful for further DNA sequence analysis.     

Sex chromosome system ZZ/ZW in Apareiodon hasemani Eigenmann, 1916 (Characiformes,Parodontidae) and a derived chromosomal region

Parodontidae fish show few morphological characteristics for the identification of their representatives and chromosomal analyses have provided reliable features for determining the interrelationships in this family. In this study, the chromosomes of Apareiodon hasemani from the São Francisco River basin, Brazil, were analyzed and showed a karyotype with 2n = 54 meta/submetacentric chromosomes, and a ZZ/ZW sex chromosome system. The study revealed active NORs located on pair 11 and additional 18S rDNA sites on pairs 7 and 22. The 5S rDNA locus was found in pair 14. It showed a pericentric inversion regarding the ancestral condition. The satellite DNA pPh2004 was absent in the chromosomes of A. hasemani, a shared condition with most members of Apareiodon. The WAp probe was able to detect the amplification region of the W chromosome, corroborating the common origin of the system within Parodontidae. These chromosomal data corroborate an origin for the ZW system of Parodontidae and aid in the understanding of the differentiation of sex chromosome systems in Neotropical fishes.     

Reconstruction of the Doradinae (Siluriformes-Doradidae) ancestral diploid number and NOR pattern reveals new insights about the karyotypic diversification of the Neotropical thorny catfishes

Doradinae (Siluriformes: Doradidae) is the most species-rich subfamily among thorny catfishes, encompassing over 77 valid species, found mainly in Amazon and Platina hydrographic basins. Here, we analyzed seven Doradinae species using combined methods (e.g., cytogenetic tools and Mesquite ancestral reconstruction software) in order to scrutinize the processes that mediated the karyotype diversification in this subfamily. Our ancestral reconstruction recovered that 2n=58 chromosomes and simple nucleolar organizer regions (NOR) are ancestral features only for Wertheimerinae and the most clades of Doradinae. Some exceptions were found in Trachydoras paraguayensis (2n=56), Trachydoras steindachneri (2n=60), Ossancora punctata (2n=66) and Platydoras hancockii whose karyotypes showed a multiple NOR system. The large thorny catfishes, such as Pterodoras granulosus, Oxydoras niger and Centrodoras brachiatus share several karyotype features, with subtle variations only regarding their heterochromatin distribution. On the other hand, a remarkable karyotypic variability has been reported in the fimbriate barbells thorny catfishes. These two contrasting karyoevolution trajectories emerged from a complex interaction between chromosome rearrangements (e.g., inversions and Robertsonian translocations) and mechanisms of heterochromatin dispersion. Moreover, we believe that biological features, such as microhabitats preferences, populational size, low vagility and migratory behavior played a key role during the origin and maintenance of chromosome diversity in Doradinae subfamily.     

一株降解N-酰基高丝氨酸内酯酵母菌菌株的分离鉴定及其降解特性

利用N酰基高丝氨酸内酯(N-acyl-homoserine lactone,简称AHL)为唯一碳源和能源,筛选得到一株能够降解AHL的菌株R1。常规鉴定和18S rDNA序列分析表明,菌株R1属于红冬孢酵母菌(Rhodosporidium toruloides),定名为R.toruloidesR1。结果显示R.toruloidesR1能利用所测试的3种AHL作为唯一碳源和能源生长,具有降解AHL的能力,其对AHL依赖型胡萝卜欧文氏软腐病菌(Erwinia carotovora subsp.carotovora)的致病有一定的抑制作用。     

棉铃虫18S核糖体RNA基因的序列分析及其分子系统学

克隆并分析了鳞翅目棉铃虫Helicoverpa armigera (Hübner)18S核糖体RNA基因(18S rDNA)的全序列,将该序列与鞘翅目、膜翅目、同翅目、双翅目、捻翅目和弹尾目各一种昆虫的同源保守区进行了比较。序列分析结果显示：鳞翅目和双翅目昆虫在18S rDNA结构上彼此较为相似,捻翅目昆虫的18S rDNA分子结构表现出与其它目昆虫有较大的差异,但相对与弹尾目昆虫的18S rDNA较为接近。该结果支持了有关捻翅目属于一个独立的目级分类阶元的论点。     

1株海洋真菌ZH2.1的鉴定及生物学特性初步研究

从南海海域白姑鱼消化道分离到1株海洋真菌ZH2.1,对其进行了系统鉴定和生物学研究。采用传统的形态学鉴定方法,并结合18S rDNA序列分析确定其归属。18S rDNA序列分析表明其与孔状短小茎点霉Phoma exigua var.exigua在进化位置上最为接近,其18S rDNA在GenBank的登录号为FJ450059。结合形态学观察结果,可认为菌株ZH2.1为茎点霉属真菌。对该菌株的部分生物学特性研究表明,ZH2.1为兼性海洋真菌,最适生长盐度为3%。此外,在微氧条件下也可生长。     

甘草内生真菌的分离及鉴定

植物体内普遍存在着内生菌,为了探寻传统药材甘草的质量与其内生菌之间可能存在的关系。对采自新疆的甘草根部进行了内生真菌的分离及鉴定。选用CYM真菌培养基,对其内生真菌进行分离及纯化,选择其中最占优势的两种菌落分别进行了菌落形态观察及菌丝形态(棉兰染色)观察,以及基因组DNA的18S rDNA和ITS rDNA序列鉴定和系统进化分析。本研究结果表明从甘草根部主要分离到两种内生真菌,分别属于Fusarium镰刀菌属和Gibberella赤霉菌属。     

产紫杉醇内生真菌EFY-21的鉴定与生物学特性研究

利用PCR技术克隆了产紫杉醇内生真菌EFY-21的18S rDNA序列,通过同源性分析,初步确定该菌与拟盘多毛孢属(Pestalotiopsis)有较高的同源性,相似性为99%。为了进一步了解EFY-21的有关生物学特性,分别选用PDA、PSA、查氏、玉米粉琼胶、牛肉膏蛋白胨5种培养基,按照常规方法培养,用十字法测量菌落直径;同时选用查氏培养基为基本培养基,分别观察不同碳源葡萄糖、甘露醇、麦芽糖、果糖、可溶性淀粉,不同氮源KNO3、Ca(N03)2、(NH4)2SO4、NH4N03、(NH4)2HPO4、蛋白胨、尿素,不同培养温度10,15,20,25,28,30,37℃,不同pH值4,5,6,7,8,9对内生真菌菌丝的影响。试验结果表明:EFY-21在PDA培养基上生长最快,生长状况最好;供试的碳氮源中,对EFY-21菌丝生长影响的大小顺序为葡萄糖甘露醇果糖麦芽糖可溶性淀粉;蛋白胨KNO3Ca(N03)2NH4N03(NH4)2HPO4(NH4)2SO4尿素;最适培养温度为25～30℃;最适pH为5～7。     

Cytogenetic analysis of three species of the genus <Emphasis Type="Italic">Haemulon</Emphasis> (Teleostei: Haemulinae) from Margarita Island,Venezuela

This paper describes the karyotype analysis of Haemulon aurolineatum, Haemulon bonariensis and Haemulon plumierii, by Giemsa staining, C-banding, Ag-staining and fluorescent in situ hybridization (FISH), to locate the 18S and 5S rRNA genes. Diploid modal count in the three species was 2n = 48 acrocentric elements. Except for pair 24, which exhibited an unmistakable secondary constriction in all three species, it was not possible to classify them as homologous to each other because differences in chromosome size were too slight between adjacent pairs within a size-graded series. Ag-NOR clusters were located in pair 24 in the three species with signal located on the secondary constriction of these chromosomes. C-banding demonstrated that the three species share the same distribution pattern of the constitutive heterochromatin with centromeric heterochromatic blocks in the 23 chromosome pairs and a pericentromeric block in pair 24 which is coincident with the NORs. FISH experiments showed that 18S rDNA sequences were located coincident with the Ag-NOR site in the three species; however, differences in both the number and chromosome distribution of 5S-rDNA cluster were detected among them. Our data suggest that chromosome evolution of Haemulon has been preserved from major changes in the karyotypic macrostructure, whereas microstructural changes have occurred.     

Microbial diversity and community structure in Fynbos soil

The Fynbos biome in South Africa is renowned for its high plant diversity and the conservation of this area is particularly important for the region. This is especially true in the case of endangered vegetation types on the lowlands such as Sand Fynbos, of which only small fragments remain. The question is thus whether the diversity of the above‐ground flora is mirrored in the below‐ground microbial communities. In order to determine the relationship of the above‐ and below‐ground communities, the soil community composition of both fungal and bacterial groups in Sand Fynbos was characterized over space and time. A molecular approach was used based on the isolation of total soil genomic DNA and automated ribosomal intergenic spacer analysis of bacterial and fungal communities. Soil from four different sites was compared to resolve the microbial diversity of eubacterial and fungal groups on a local (alpha diversity) scale as well as a landscape scale (beta diversity). The community structures from different sites were compared and found to exhibit strong spatial patterns which remained stable over time. The plant community data were compared with the fungal and the bacterial communities. We concluded that the microbial communities in the Sand Fynbos are highly diverse and closely linked to the above‐ground floral communities.     

Patterns of ribosomal gene variation in elite commercial chicken pure line populations

The nucleolus organizer region (NOR) encodes the tandemly repeated 18S, 5.8S and 28S ribosomal (r) RNA genes. The NORs of broiler and layer commercial chicken pure lines were studied to establish the type and extent of genetic variation at this important locus. The parameters studied were gene copy number, repeat size, and diversity of NOR-types. The populations were organized into three groups for analysis including brown-egg broiler (13 lines), brown-egg layer (six lines), and white-egg layer (eight lines). The ribosomal gene copy number average of the white-egg layer populations was significantly lower (329 genes) than that of the brown-egg layers (372 genes); the brown-egg broiler ribosomal gene average was intermediate (350 genes). The white-egg layer populations exhibited a ribosomal repeat unit average size of 36 kb, significantly different from the brown-egg layer and brown-egg broiler average repeat unit size of 32.5 and 33.9 kb, respectively. NOR array size was similar among the three groups (6 mb). The brown-egg broiler populations exhibited polymorphic NOR patterns, intra- and interline, whereas the white-egg layer populations were essentially monomorphic for NOR-type; brown-egg layers exhibited an intermediate level of NOR diversity. Some NOR array characteristics may be a function of breed origin as brown-egg commercial populations, both broilers and layers, have similar breed origins and exhibited similarities for predominant repeat unit size as compared with white-egg layer populations. However, the finding that brown-egg broiler lines typically exhibit a greater number of segregating NOR-types than brown-egg layer lines suggests that the selection schemes of broiler vs. layer pure line populations may also have influenced the degree of variation at this gene complex.     

土地利用变化对土壤真菌群落结构的影响

应用PCR-RFLP和测序分析对川西亚高山米亚罗林区不同土地利用类型的土壤真菌18S rDNA基因进行了多样性和系统发育研究,探讨了土地利用变化对土壤真菌群落结构的影响。在20a龄云杉(Picea likiandensis var balfourianan)人工林和菜地两种类型土壤中,共得到238个阳性克隆,限制性内切酶MspI和RsaI进行RFLP分析后得到56个不同的分类操作单元(OTUs),其中20a龄云杉人工林样地获得137个阳性克隆和37个OTUs,而菜地样地获得101个阳性克隆和19个OTUs。在两类样地中具有不同的优势种群,其中20a龄云杉人工林样地有1个明显优势种群,占总克隆数的20.4%;菜地样地有2个明显优势种群,分别占总克隆数的25.7%和21.8%。对14个克隆进行了序列测定,序列的相似性在86%~99%之间,与GenBank数据库中的序列进行比对,与已知序列的相似性在92%~100%之间。系统发育分析表明,所有的18SrDNA基因被分为3个主要的簇,其中20a龄云杉人工林样地的克隆都聚集在第一和第三簇中,而菜地样地的克隆都聚集在第二簇中。结果说明,两类土壤中具有较为丰富的真菌多样性,而土地利用变化引起了土壤真菌群落结构的明显变化。     

18-26S rDNA在4种重楼属植物中的定位

为探讨rDNA在重楼属Paris L.中的分布规律,利用荧光原位杂交（FISH）对4种重楼属植物的18-26S rDNA进行了定位。所有植物均为二倍体,基因组由A、B、C、D和E5条染色体构成。（1）滇重楼P.polyphylla var.yunnanensis:2n=10=6m+4t,C和D染色体的短臂上各有1个18-26S rDNA位点;（2）长柱重楼P.forrestii:2n=10=6m+4t,B染色体的长臂、C和D染色体的短臂上各有1个位点;（3）五指莲P.axialis:2n=10=6m（2sat）+4t（2sat）+1-2B,C和D染色体的短臂上各有1个位点;在有1个B染色体的细胞中,B染色体没有信号点,而有2个B染色体的细胞中,只有1个B染色体上有信号点,表明B染色体上有基因存在且其分裂不均等;（4）大理重楼P.daliensis:2n=10=4m+2sm+2st+2t,C染色体的短臂上有1个位点。18-26S rDNA位点不仅出现在染色体的次缢痕上,也出现在非次缢痕位点。另外,4个种中C染色体短臂末端均有18-26S rDNA。     

栽培玉米亚种5 S rDNA NTS序列分析及FISH定位方法比较

选取我国7个栽培玉米亚种材料,进行5 S rDNA的非转录间隔区（nontranscribed intergenic spacer,NTS）的序列分析,比较7个亚种材料NTS序列差异并进行聚类分析,探讨其亲缘关系。研究结果表明：7个材料的NTS区GC平均含量为45．67％,核苷酸位点变异位点个数1～15,转换／颠换率为0．83～2．0,特用玉米材料均存在不同程度的缺失;7个材料主要聚为两大类,第一类群中包括甜质、马齿、硬粒、爆裂和蜡质5个亚种材料,第二类群中包括粉质和甜粉2个亚种材料。同时利用荧光原位杂交技术（fluorescence in situ hybridization,FISH）对5 S rDNA进行定位,探针标记分别采用荧光素标记和生物素标记。结果表明：生物素标记检测系统灵敏度高、杂交信号强,更适合于5 S rDNA重复序列的定位检测。     

Genetic diversity of small eukaryotes from the coastal waters of Nansha Islands in China

Population structures and genetic diversity of the small eukaryotic plankton from the coastal waters of the Nansha Islands in China were investigated. Two genes libraries using 18S rDNA of the marine small eukaryotes were constituted, and 323 clones were identified within alveolates (more than 43%), acanthareas, viridiplantaes, and stramenopiles. Many novel clones were detected in the two libraries, including two groups of alveolates and two clades related to both acanthareas and polycystineas. Several sequences unrelated to any other known eukaryotes may represent early branches in the phylogenetic tree. Our results reveal that there is a high diversity and abundance of small eukaryotes in the marine regions of China.     

Phylogenetic analysis of protozoa in the rumen contents of cow based on the 18S rDNA sequences

AIMS: To examine the diversity of protozoa in the rumen contents of cow. METHODS AND RESULTS: Protozoa that inhabit the rumen were detected by PCR using protozoan-specific primers. Libraries of protozoan rDNA sequences were constructed from rumen fluid, solid tissues and epithelium. Twenty-three clones isolated from rumen fluid fell into two genera identified as Entodinium (69.6% of clones) and Epidinium (31.4% of clones). Of the clones isolated from rumen fluid, a moderate number were unidentifiable (30.4%). CONCLUSIONS: The predominant protozoan genus identified in the whole rumen belonged to the Entodinium group (81.1%). Protozoa were not detected in the rumen epithelium. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings suggest that rumen fluid and solid tissues contain different protozoan populations that may play specific roles in rumen function. Quantitative PCR techniques and a more specific set of phylogenetic probes that distinguish between protozoan species are needed to determine the significance of newly identified groups and to determine the distribution of identified protozoan clusters in rumen microbial communities.     

Identification and phylogeny of eukaryotic 18S rDNA phylotypes detected in chlorinated finished drinking water samples from three Parisian surface water treatment plants

Aims:  We performed a preliminary assessment of the eukaryotic 18S rDNA diversity present in finished drinking water samples from three different surface water treatment plants supplying water to the city of Paris (France).
Methods and Results:  A molecular analysis was performed on a sample from each site based on sequencing of PCR amplified and cloned 18S ribosomal RNA genes. Overall, the 18S rDNA sequences combined from all samples could be affiliated to the Amoebozoa (20·8% of the phylotypes), Ciliophora (25%), Metazoa (33·3%), Fungi (8·3%), Cercozoa (4·2%) and unclassified eukaryotes (8·3%) groups.
Conclusions:  The 18S rDNA sequences affiliated to the Amoebozoa, Ciliophora and Metazoa lineages were found to be the most abundant phylotypes observed in the drinking water samples. Phylotypes found to be present in two, or all three, samples (41·7% of the total) may represent groups with members adapted to drinking water treatment plant (DWTP) ecosystem conditions.
Significance and Impact of the Study:  This study shows that finished drinking water can contain 18S rDNA sequences representing a variety of eukaryotic taxa. Further research is needed to better characterize the eukaryotic biodiversity of DWTPs and the effects of the finished drinking water diversity on the downstream water distribution network.     

汉江硅藻水华优势种的形态及18S rDNA序列分析

我国汉江下游在1992、1998、2000、2003年初春曾先后多次发生水华。水华发生期间,硅藻大量增殖,形成肉眼可见的褐色颗粒悬浮于江水中,使江水呈现黄褐色,在以往的研究中认为汉江硅藻水华优势种为小环藻^[1,2],但并未从分类上展开研究。传统上硅藻种类的鉴定主要是以硅藻壳的形态及壳表面上的花纹为依据。目前国内外学者利用18S rDNA基因分析对水华赤潮原因种的鉴定方面也作了许多相关的研究。陈丽芬等利用18SrDNA序列分析确定香港海域一株棕囊藻为球形棕囊藻^[3]。