Re-examination of the phylogeny of Rhacophoridae (Anura) based on mitochondrial and nuclear DNA

The phylogenetic relationships among rhacophorid frogs are under dispute. We use partial sequences of three mitochondrial (12S rRNA, 16S rRNA, and cytochrome b) and three nuclear protein-coding (Rag-1 rhodopsin exon 1, and tyrosinase exon 1) genes from 57

几类“无线粒体”原生动物进化地位的探讨* ——来自DNA 拓扑异构酶Ⅱ的系统分析证据

以贾第虫、毛滴虫、内变形虫和微孢子虫等为代表的几类寄生原生动物曾因为“无线粒 体”, 再加上其他一些似乎介于原核和真核细胞之间的原始特点和分子系统树中它们往往处在真 核生物的最基部等证据, 而被有些人认为是目前已知的一类最原始的真核生物类群——Archezoa, 其进化地位应该是处在真核细胞通过“内共生”产生线粒体之前的极早阶段. 这一观点一度被部 分学者认为对探讨真核细胞(生物)的起源、进化极为重要, 是进化生物学上的重要突破. 然而, 近 年来不断有研究对此提出质疑. 我们首先扩增、测序并鉴定了蓝氏贾第虫、阴道毛滴虫和痢疾内 变形虫的DNA 拓扑异构酶Ⅱ序列, 再结合GenBank 数据库中已有的脑炎微孢子虫和其他一系列 处在不同进化地位的真核生物的相应序列数据, 用多种方法构建出分子系统树, 对这些“无线粒 体”原生动物的进化地位进行了探讨. 结果表明, 由于DNA 拓扑异构酶Ⅱ的特点和可以克服“长 枝吸引”等以往分子系统分析中的不足, 所构建的系统树不仅能有效地反映出已普遍接受的真核 生物各主要类群的系统关系, 而且显示出这些“无线粒体”原生动物不同于以前系统树所推测的 进化地位: 它们并非是最早分支出来的真核生物, 而是在具有线粒体的生物如动基体类或菌虫类 等之后才分化的、分别属于不同进化地位的类群. 结合近来在它们中发现了类似线粒体细胞器的 证据, 我们认为这些“无线粒体”的原生动物虽然其中有些种类(如以贾第虫为代表的双滴虫类)进 化地位很低等, 但总体上并非过去所认为的那么极端原始, 它们应该是线粒体产生之后因适应长 期的无氧条件下的内寄生生活方式才分别分化出来的多源发生的不同生物类群.

滇池铜绿微囊藻(M．aeruginosa zosa Kutz)的分离培养与总DNA提取的改进

利用稀释法结台平板分离培养法将采自滇池水华中的铜绿微囊藻进行了成功的 分离纯化对比纯化培养前后，发现纯化后该藻的生长速度明显加快，密度大为提高，约为纯化前 的5倍用乙醇乙醚混合液剐该藻作预处理后，破坏了其胶质被．再经常规方法提取总DNA，其得 率提高3 4倍

DNA sequence analysis of the triose phosphate isomerase gene from isolates of Giardia lamblia

Objective To confirm the genetic relation between Giardia lamblia (G. lamblia) isolates from different geographic regions of China and other countries. Methods Genomic DNA were extracted from the trophozoites or cysts of Giardia lamblia. The triose phosphate isomerase (tim) gene was amplified using polymerase chain reaction (PCR) technique. PCR products were digested with endonuclease and sequenced. The data of sequencing were analyzed with the DNAstar software and compared with that of the isolates acquired from GenBank. Results Of nine isolates of Giardia lamblia from China (C1, C2, CH2 and CH3), Cambodia (CAM), Australia (A1 and A2) and America (BP and CDC), respectively, 3 (A1, A2 and CAM) fit into Group 1 (WB), 2 (CH2 and CH3) into Group 2, and 4 (C1, C2, BP and CDC) into Group 3 (GS). The results confirmed the genetic relatedness of G. lamblia isolates from all over the world. Conclusion Genotyping isolates of G. Lamblia provides important information for establishing the phylogenetic relationship or for the epidemiological evaluation of the spreading of this organism.

Phylogenetic positions of several amitochondriate protozoa - Evidence from phylogenetic analysis of DNA topoisomerase II

Several groups of parasitic protozoa, as represented by Giardia, Trichomonas, Entamoeba and Microsporida, were once widely considered to be the most primitive extant eukaryotic group - Archezoa. The main evidence for this is their 'lacking mitochondria' and possessing some other primitive features between prokaryotes and eukaryotes, and being basal to all eukaryotes with mitochondria in phylogenies inferred from many molecules. Some authors even proposed that these organisms diverged before the endosymbiotic origin of mitochondria within eukaryotes. This view was once considered to be very significant to the study of origin and evolution of eukaryotic cells (eukaryotes). However, in recent years this has been challenged by accumulating evidence from new studies. Here the sequences of DNA topoisomerase 11 in G lamblia, T vaginalis and E histolytica were identified first by PCR and sequencing, then combining with the sequence data of the microsporidia Encephalitozoon cunicul and other eukaryotic groups of different evolutionary positions from GenBank, phylogenetic trees were constructed by various methods to investigate the evolutionary positions of these amitochondriate protozoa. Our results showed that since the characteristics of DNA topoisomerase 11 make it avoid the defect of 'long-branch attraction' appearing in the previous phylogenetic analyses, our trees can not only reflect effectively the relationship of different major eukaryotic groups, which is widely accepted, but also reveal phylogenetic positions for these amitochondriate protozoa, which is different from the previous phylogenetic trees. They are not the earliest-branching eukaryotes, but diverged after some mitochondriate organisms such as kinetoplastids and mycetozoan; they are not a united group but occupy different phylogenetic positions. Combining with the recent cytological findings of mitochondria-like organelles in them, we think that though some of them (e.g. diplomonads, as represented by Giardia) may occupy a very low evolutionary position, generally these organisms are not as extremely primitive as was thought before; they should be polyphyletic groups diverging after the endosymbiotic origin of mitochondrion to adapt themselves to anaerobic parasitic life.

Identification, characteristic and phylogenetic analysis of type II DNA topoisomerase gene in Giardia lamblia

The genes encoding type II DNA topoisomerases were investigated in Giardia lamblia genome, and a type IIA gene, GlTop 2 was identified. It is a single copy gene with a 4476 by long ORF without intron. The deduced amino acid sequence shows strong homology to eukaryotic DNA Top 2. However, some distortions were found, such as six insertions in the ATPase domain and the central domain, a similar to 100 as longer central domain; a similar to 200 as shorter C-terminal domain containing rich charged residues. These features revealed by comparing with Top 2 of the host, human, might be helpful in exploiting drug selectivity for antigiardial therapy. Phylogenetic analysis of eukaryotic enzymes showed that kinetoplastids, plants, fungi, and animals were monophyletic groups, and the animal and fungi lineages shared a more recent common ancestor than either did with the plant lineage; microsporidia grouped with fungi. However, unlike many previous phylogenetic analyses, the "amitochondriate" G. lamblia was not the earliest branch but diverged after mitochondriate kinetoplastids in our trees. Both the finding of typical eukaryotic type IIA topoisomerase and the phylogenetic analysis suggest G. lamblia is not possibly as primitive as was regarded before and might diverge after the acquisition of mitochondria. This is consistent with the recent discovery of mitochondrial remnant organelles in G. lamblia.

Mitochondrial DNA sequence diversity and origin of Chinese domestic yak

In order to clarify the origin and genetic diversity of yak in China, we analysed mitochondrial DNA (mtDNA) control region sequences (similar to 891 bp) in 52 individuals from four domestic yak (Poephagus grunniens) breeds, as well as from a hybrid betwee

Mitochondrial DNA mutation m.3635G > A may be associated with Leber hereditary optic neuropathy in Chinese

Leber hereditary optic neuropathy (LHON) was the first disease to be linked to the presence of a mitochondrial DNA (mtDNA) mutation. Nowadays over 95% of LHON cases are known to be caused by one of three primary mutations (m.11778G>A, m.14484T>C, and m.34

Genetic diversity of Chinese domestic goat based on the mitochondrial DNA sequence variation

The aim of this study was to characterize the genetic diversity of domestic goat in China. For this purpose, we determined the sequence of the mitochondrial DNA (mtDNA) control region in 72 individuals of the Yangtze River delta white goat, and reanalysed

卷象科昆虫无损形态的DNA提取方法

提取DNA是所有分子生物学实验的第一步,但在昆虫分子生物学研究中,常规提取总DNA的方法不能保留昆虫所有的形态特征.显然,这不适用于对于体形较小的珍稀标本.以鞘翅目卷象科(Coleoptera: Attelabidae)昆虫为实验材料,提供了一种新的取材方法,取下整个腹部,消化其肌肉组织后收回骨化腹板.该方法可在提取昆虫总DNA的同时保存全部外形特征及其生殖器.

DNA 拓扑异构酶Ⅱ的结构和作用原理

1976年美国国立卫生研究所的 Gellert 等从大肠杆菌中提取到解旋酶 (gyrase, 也叫细菌DNA 拓扑异构酶Ⅱ ,以下简称 TopoⅡ )以来, 对 TopoⅡ的作用机制一直存在争议. Tse (1980)报道, Topo Ⅱ能同时打断 DNA 的两条链,TopoⅡ的两个亚基键分别与断开的 DNA 的两个5'末端连接, 并互相错开4个碱基对，以便让另一股 DNA 双链通过。