TWO DIFFERENT SPECIES OF EUGLENA,E. GENICULATA AND E. MYXOCYLINDRACEA (EUGLENOPHYCEAE), ARE VIRTUALLY GENETICALLY AND MORPHOLOGICALLY IDENTICAL1

We investigated the similarity of a single Euglena myxocylindracea strain, isolated originally by Bold and MacEntee, to several Euglena geniculata strains on both morphological and DNA levels. We found the three DNA stretches, consisting of fragments coding for the parts of cytoplasmic and chloroplast small subunit rRNA, and the internal transcribed spacer (ITS2) of cytoplasmic rDNA, with the combined length of 4332 nucleotides, are identical in E. myxocylindracea and E. geniculata, strain SAG 1224‐4b. Morphological differences between E. myxocylindracea and any E. geniculata strain examined were well within the range of E. geniculata variability as well. The only difference behind the distinction of E. myxocylindracea from E. geniculata is the presence of the second chloroplast in the latter. However, we were able to induce the appearance of the second chloroplast in the cells of E. myxocylindracea and its disappearance in the cells of E. geniculata by changing the composition of the culture media. We therefore conclude that E. myxocylindracea Bold and MacEntee should be regarded as an environmental form of E. geniculata Dujardin. For the first time the morphology of E. geniculata chloroplasts was shown as revealed by confocal laser microscopy.     

Molecular phytogeny of conifers using RFLP analysis of PCR-amplified specific chloroplast genes

We investigated the molecular phylogeny of conifers using restriction endonuclease fragment length polymorphism of six polymerase chain reaction-amplified chloroplast genes — frxC, rbcL, psbA, psbD, trnK, and 16S. We detected 227 total site changes among species, representing 23, 26, 38, 48, 67, and 25 site changes in frxC, psbA, psbD, rbcL, trnK and 16S, respectively. The mean nucleotide substitution was 10.75% (SD 0.573) among species in five families. Forty maximally parsimonious trees were obtained using the Wagner parsimony method, and a 50% majority-rule consensus tree was obtained from them. Data analysis produced similar basic patterns when both the Wagner parsimony and the neighbor-joining methods were applied, and the main lineages were clearly separated. Taxaceae and Cephalotaxaceae species were used as the out-groups when applying Wagner parsimony methods. With the Wagner method, the consistency index was 0.510, the retention index was 0.879, and tree length was 435 steps. Our results indicated that Cupressaceae and Taxodiaceae are closely related families and that Sciadopitys verticillata is the basal lineage of Cupressaceae and Taxodiaceae. The neighbor-joining tree is similar to the 50% majority-rule consensus of the 40 Wagner parsimony trees except for the position of Keteleeria daversifolia, the Picea and Cedrus group, and the divergence within Cupressaceae.     

Molecular genetic characterization of H5N1 influenza virus strains isolated from poultry in the Kurgan region in 2005

In the second half of 2005, a large-scale outbreak of influenza in poultry and wild birds was caused by a highly pathogenic H5N1 influenza virus in Russia. The level of pathogenicity is a polygenic trait, and most individual genes contribute to the influenza A virus pathogenicity in birds, animals, and humans. The full-length nucleotide sequences were determined for H5N1 strains isolated in the Kurgan region (Western Siberia). The structure of viral proteins was analyzed using the deduced amino acid sequences. The receptor-binding site of hemagglutinin (HA) in strains A/chicken/Kurgan/05/2005 and A/duck/Kurgan/08/2005 was typical for avian influenza viruses and contained Glu and Gly at positions 226 and 228, respectively. The structure of the basic amino acid cluster located within the HA cleavage site was identical in all isolates: QGERRRKKR. According to the neuraminidase structure, all H5N1 isolates from the Kurgan region were assigned to the Z genotype. Amino acid residues typical for the avian influenza virus were revealed in 30 out of 32 positions of M1, M2, NP, PA, and PB2, determining the host range specificity. One of the strains contained Lys at position 627 of PB2. Isolates from the Kurgan region were shown to have a remantadine-sensitive genotype. Both strains contained Glu at position 92 of NS1, indicating that the virus is interferon-resistant. Phylogenetic analysis related the Kurgan isolates to subclade 2 of clade 2 of highly pathogenic H5N1 influenza viruses.     

Description of a new species of Phylloscopus warbler from Vietnam and Laos

A new species of Phylloscopus warbler, which we name Phylloscopus calciatilis Limestone Leaf Warbler, is described from central and northern Vietnam and central and northern Laos; it probably also breeds in southernmost China. In morphology, the new species is very similar to Sulphur-breasted Warbler Phylloscopus ricketti , but it is smaller with a proportionately larger bill and rounder wing. Its song and calls are diagnostic. Based on mitochondrial and nuclear DNA, the new species is most closely related to P. ricketti and Yellow-vented Warbler Phylloscopus cantator , and it is inferred to be sister to the latter. The mitochondrial divergences between these three species are at the low end of the variation found in other species of Phylloscopus and Seicercus warblers, but greater than in other taxa generally treated as subspecies. Possible introgressive hybridization between the new species and P. ricketti is discussed, but more data are needed to establish whether it does occur and, if it does, to what extent. The new species appears to have a restricted breeding range in limestone karst environments, where it is locally common and therefore not under any immediate threat. In view of the recognition of the new species, all previous records of P. ricketti sensu lato need to be re-evaluated.     

葡萄Actin基因家族的鉴定及进化和表达分析

采用生物信息学方法从葡萄(Vitis vinifera Linn.)全基因组中鉴定Actin基因家族,并对各基因的染色体定位和结构特征,编码蛋白质的理化性质、亚细胞定位、二级结构、三级结构和系统进化,以及不同组织的基因表达进行研究.结果表明:葡萄Actin基因家族16个基因分布在12条染色体上.16个基因的结构特征及其编码蛋白质的理化性质差异较大.16个基因的长度及其内含子总长度的变化范围较大,编码序列(CDS)和外显子总长度的变化范围较小.除登录号GSVIVG01008254001和GSVIVG01014035001的基因外,其他14个基因的GC含量均低于其CDS的GC含量.除登录号GSVIVG01008254001的基因外,其他15个基因编码的蛋白质的理论相对分子质量为12534.54~82612.33,理论等电点为pI 4.92~pI 9.13.16个基因编码蛋白质的消光系数为14105~73645,脂肪族氨基酸指数为65.54~92.06,其中9个为稳定蛋白,7个为不稳定蛋白.除登录号GSVIVG01014035001的基因外,其他15个基因编码的蛋白质均为亲水性蛋白.登录号GSVIVG01016517001的基因编码的蛋白质定位于细胞质和细胞核,其他15个基因编码的蛋白质定位于细胞质.二级结构和三级结构显示:葡萄Actin基因家族16个基因编码的蛋白质均由α螺旋、无规则卷曲和延伸链构成,且总体以无规则卷曲为主.系统进化分析和不同组织的基因表达分析结果显示:与拟南芥〔Arabidopsis thaliana(Linn.)Heynh.〕相似,葡萄Actin基因家族16个基因编码的蛋白质分为3个亚家族,ClassⅡ亚家族(营养型)包括登录号GSVIVG01003099001和GSVIVG01026580001的基因编码的蛋白质,这2个基因在所有组织中的表达均较高;ClassⅢ亚家族(生殖型)包括登录号GSVIVG01033494001、GSVIVG01024980001和GSVIVG01016550001的基因编码的蛋白质,这3个基因在花粉、雄蕊和花中的表达均较高;ClassⅠ亚家族包括其他11个基因编码的蛋白质,这11个基因在各组织中的表达总体上较低.研究结果显示:葡萄Actin基因家族的表达具有组织特异性.     

分子生物学技术在双翅目实蝇科昆虫系统发育研究中的应用

分子生物学技术如同工酶电泳、RFLP、RAPD、核酸序列分析、微卫星DNA和探针杂交等,在实蝇科昆虫系统发育研究中具有重要作用。利用这些技术对实蝇种群进行系统发育研究,揭示其亲缘及进化关系,从生命本质上寻找实蝇种群间的内在联系。文章综述上述几种分子生物学技术在实蝇科昆虫核酸结构、种内和种间的亲缘及进化关系等方面的研究进展,分析在应用中存在的问题,展望这些分子生物学技术在实蝇科昆虫系统发育中的应用前景。     

Confirmation of soil-feeding termites (Isoptera; Termitidae; Termitinae) in Australia using stable isotope ratios

1. Carbon (δ¹³C) and nitrogen (δ¹⁵N) stable isotope ratios of termites (Isoptera) were examined in Darwin, northern Australia. It is suggested that the stable isotope technique, together with phylogenetics, is a useful tool to understand the evolution of functional groups in detritivores.
2. A high δ¹⁵N value was observed in the Termes-Capritermes branch of the subfamily Termitinae and the genus Amitermes , two distinct taxonomic groups that evolved from wood-feeding to soil-feeding in Australia. Among eight Termes-Capritermes branch species, only two species ( Xylochomitermes melvillensis and Ephelotermes melachoma ) were discernible as wood/soil interface feeders, the remaining six species analysed were soil-feeders, where the diet preference was identified by using δ¹⁵N of workers.
3. The Termes-Capritermes group in Australia contains both wood/soil interface feeders and soil-feeders, whereas wood/soil interface feeders in Cameroon are from the Termes-Capritermes branch while soil-feeders are from Cubitermes group. The result confirmed that soil-feeding forms evolved both in Australia and Africa, but from different phylogenetic groups.     

Morphological evidence for the evolutionary origin of Astigmata (Acari: Acariformes)

A century ago, Antonio Berlese first discussed the close phylogenetic relationship between the large mite groups Oribatida and Astigmata. Since then, information having phylogenetic value has greatly increased and the paradigms within which we interpret it have changed. Herein I refine the general hypothesis that Astigmata originated within oribatid mites and suggest Malaconothridae as a possible sister group. Among the 14 apomorphies used to support the origin of Astigmata within oribatid mites are possession of lateral opisthosomal glands, regression of hysterosomal setal pair f1, paired prelarval denticles, partially internalized chelicerae with incomplete adaxial walls, an atelobasic rutellum, pretarsal condylophores that articulate posteriorly with the tarsus, a dorsally fused palp tibia and tarsus and transdehiscent ecdysis. A further 13 apomorphies support the origin of Astigmata at some level within Malaconothroidea. These include absence of an oblique labiogenal articulation, presence of a distal rutellar lamella, shortening of the palp tarsus, larval regression of hysterosomal seta f2, loss of the bothridial seta in all instars, and several losses and modifications of leg setae. The hypothesis brings to light evolutionary questions that were previously obscured by incorrect or inappropriate classifications. The nomenclatural problems that arise from it are best solved by considering Astigmata as a subgroup within Oribatida.     

Trichoderma harzianum及其近缘种的分子系统学研究

Thichoderma harzianum是木霉属内最常见的一个“集合种”。本研究对来源不同的T.harzianum及其相似种的46个菌株进行了ITS序列测定,将其ITSl—5．8S—ITS2序列与来自EMBL的参考菌株的序列进行比较,并进行系统发育分析,此外对其中的18个菌株进行了RAPD多态性分析,试图明确T.harzianum的多样性以及与其相似种之间的关系。ITS结果表明,T.harzianum及其相似种可分成2个群(A、B)：A群由T.hamatum、T.asperellum、T.at-roviride、T.koningii和T.viride组成,并形成2个分支,表明T.viride和T.koningii、T.atroviride的亲缘关系较近,而与T.hamatum、T.asperellum较远;B群由T.spirale、T.hamatum、T.inhamatum、T.harzianum和T.anam。Hypocrea vinosa组成,并形成6个分支。T.inhamatum可分成2个群(Ti1、Ti2)、T.harzianum至少可分成5个群(Thl、Th2、Th4、Th5、Th6)。结果还表明T.hamatum的遗传差异较大,T.hamatum的模式菌株归属于A群,而其他的T.hamatum的菌株归属于B群。RAPD结果与ITS的结果基本一致。