The phylogeny and systematics of the endemic Ethiopian Lophuromys flavopunctatus species complex based upon random amplified polymorphic DNA (RAPD) analysis

Random amplified polymorphic DNA (RAPD) analysis was performed to clarify systematics and phylogenetic relationships within the Ethiopian endemic representatives of Lophuromys flavopunctatus species complex. Data were analysed by both phenetic (UPGMA) and phylogenetic (neighbor-joining (NJ) and maximum parsimony) procedures. NJ and maximum parsimony analyses yielded identical phylogenetic trees that demonstrate the basal position of L. melanonyx with L. brevicaudus splitting next and sister-group relationship for L. brunneus–L. chrysopus. This phylogenetic pattern is congruent with inferences from allozymes for the considered species suggesting early divergence of Afroalpine species and recent origin of forest taxa. Thus, the results demonstrate that RAPD-PCR might be a useful technique for phylogenetic analysis at the species levels in vertebrates. Controversial taxonomy of L. brevicaudus, L. brunneus and L. chrysopus is clarified, with their specific ranks confirmed on the basis of tree topology and genetic distances.     

Lactate dehydrogenase in two digenetic trematodes and their host

Polyacrylamide gel electrophoresis of the two digenetic trematodes, Gigantocotyle explanalum from the liver and Gastrothylax crumenifer from the rumen of the water buffalo, Bubalus bubalis revealed the presence of at least six and seven isoenzymes of lactate dehydrogenase (LDH), respectively in a partially purified enzyme preparation. The respective host tissues showed five isoenzymes of LDH, which are characteristic to the vertebrates. Both parachloromercuribenzoate and iodoacetate affected the LDH activity of the parasites and host tissues differently. Spectrophotometric analysis also showed different specific activity and susceptibility to the action of thiol inhibitors. The host LDH was quite stable at 57°C for 30 min, but that of the parasites was less stable.     

Isoenzymes of creatine kinase and prostate-specific antigen in the monitoring of prostatic carcinoma]

We have valued serum levels of the prostatic specific antigen and of the creatine-kinase isoenzymes (MM-BB) in 30 patients with prostatic carcinoma at C and D stage. Our results demonstrate that these markers are useful for monitoring and for checking of metastasis. But the isoenzymes seem to us predictive in the monitoring of metastasis. The isoenzymes' determination was executed by electrophoretic method (Helena) which is sensitive, specific and swift.     

A genetic approach to species criteria in the amoeba genus Naegleria using allozyme electrophoresis

The present study employs allozyme electrophoresis to characterize and inter-relate 61 isolates of Naegleria. Diploidy was confirmed, with heterozygotes observed at 29 of the 33 loci established and in all but two isolates. With a single exception, isolates clustered at two levels of similarity, either below 21% or above 52%. It is argued that such a major discontinuity provides a sound biological basis for a species concept in Naegleria. On this basis the present species-level taxonomy does not reflect the genetic diversity of the genus. The study recognized 18 genetic groups of species rank. The subspecies N. australiensis italica deserves specific rank; additional thermophilic species not closely related to N. fowleri and N. lovaniensis are recognized; and N. gruberi as currently conceived is a complex of 10 species, at least five of which are represented in the formal culture collections. Most species are genetically too different for relationships to be elucidated by allozyme electrophoresis, supporting the view that some of the times of divergence within the genus are extremely ancient.     

Characterization of the pectin methylesterase-like gene AtPME3: a new member of a gene family comprising at least 12 genes in Arabidopsis thaliana

Pectin demethylesterification appears to be catalysed by a number of pectin methylesterase (PME) isoenzymes in higher plant species. In order to better define the biological role of these isoenzymes in plant cell growth and differentiation, we undertook molecular studies on the PME-encoding genes in Arabidopsis thaliana. In this paper, we report the characterization of AtPME3, a new PME-related gene of 4 kb in length that we have mapped on Chromosome III. AtPME3 encodes a putative mature PME-related isoenzyme of 34 kDa with a basic isoelectric point. Since the extent of the gene family encoding PME in higher plant species is still unknown, we resorted to the use of degenerate primers designed from several well-known consensus regions to identify new PME-related genes in the genome of Arabidopsis. Our results, in combination with several known expressed sequences tags (ESTs), indicate that the Arabidopsis genome contains at least 12 PME-related genes. Consequently, a method of systematic gene expression analysis has been applied in order to discern the expression pattern of these 12 genes throughout the plant at the floral stage. Whereas most of these genes appeared to be more or less ubiquitously expressed throughout the plant, several genes are distinguishable by their strikingly specific expression in certain organs. The present data bring a new insight into the role of specific PME-related genes in flower and root development.     

A survey of leaf flavonoids in the portulacaceae

A survey of 26 species in 12 genera of the Portulacaceae showed the presence of flavonoids in 24, and their absence in two, Hectorella caespitosa Hooker fil. and Lyallia kerguelensis Hooker fil. Of the 24 positive species, 16 are reported for the first time. In all 24 however, flavonols and flavones did not occur together suggesting that this dichotomy might be useful for classification at and below the generic level in the Portulacaceae, e.g. the genera Silvaea Philippi (syn. Philippiamara Kuntze), Ceraria Pearson & Stephens and Portulacaria Jacquin which were previously grouped together on palynological characteristics have now been shown to differ in their leaf flavonoids, subsidiary cells of the stomata and geographical location: Silvaea possesses flavones and is endemic to South America, while Ceraria and Portulacaria possess flavonols and are endemic to Africa. The dichotomy also occurs below the generic level. In Calandrinia H.B.K. and Portulaca L. the dichotomy amongst their species is supported by other new taxonomic characters from cytology, palynology, pubescence of surfaces, type of subsidiary cells surrounding the leaf stomata and seed morphology recently elucidated by the author.     

幕阜山脉石松类和蕨类植物多样性及生物地理学特征

通过资料收集及野外调查, 得出幕阜山脉地区共有石松类和蕨类植物26科72属261种, 具有一定的东西过渡性, 为鳞毛蕨-铁角蕨植物区系, 最大属为鳞毛蕨属(Dryopteris) (29种)。其中, 庐山的物种丰富度较高(224种), 以铁角蕨属(Asplenium)为主; 幕阜山的物种密度较大(2.09种/km ²), 以卷柏属(Selaginella)为主; 九宫山以瓦韦属(Lepisorus)为主; 三者共通种有95种, 新特有现象较丰富。该区属种分化限制明显, 表现在单种科、属及寡种科、属占总科数的60%及总属数的80%。从区系成分看, 该区科、属以热带成分为主, 科和属的热带性成分与温带性成分比值(R/T值)分别为2.6和2.3。与同纬度带山地石松类和蕨类植物属的R/T值比较, 中亚热带与北亚热带交界带的蕨类植物属的R/T值在2.18-2.36之间; 种的R/T值为0.2, 为热带成分的5倍, 表现出明显的温带性质, 是罗霄山脉植物区系温带成分的重要组成部分。该区石松类和蕨类植物区系与华东、华南植物区系联系比较紧密, 表现出华东与华南两区系成分的交汇。     

Purification of Cu, Zn-Superoxide Dismutase Isoenzymes from Fish Liver: Appearance of New Isoforms as a Consequence of Pollution

Liver cell-free extracts of fish (Mugil sp.) from polluted environments show new Cu, Zn-SOD isoenzymes when analyzed by polyacrylamide gel electrophoresis or isoelectrofocusing followed by in situ staining for SOD activity. The most active isoenzymes, with pI 6.1 and 5.1, were present both in control and problem samples while the isoenzymes of intermediate pI value showed significant differences. Fish from control areas showed three intermediate isoenzymes with pI 5.7, 5.5 and 5.4 (the last one quite faint) while polluted animals showed three bands of pI 5.9, 5.45 and 5.35, this last very intense. To further characterize their utility as biomarkers, Cu, Zn-SOD isoenzymes from polluted fish livers were purified to homogeneity. Five superoxide dismutase peaks were purified, named thereafter I (pI 6.1) to V (pI 5.1) respectively. Isoenzymes I and V displayed the highest specific activity. Upon incubation with moderate H₂O₂ concentrations, pure isoenzyme I yielded more acidic bands with pI 5.5, 5.45 and 5.35, this last being predominant. The pure isoenzyme V generated only a new band of pI 5.0. Concomitant with oxidation, the activity of peaks I and V was lost in a H₂O₂ concentration-dependent manner. The pattern of the new acidic bands generated upon the oxidixing treatment of isoenzyme I closely resembles that observed in crude extracts from polluted animals.     

Folk taxonomy and scientific nomenclature: Working together for conservation of fishery resources in Brazil

Folk taxonomies of fishers are important for understanding the dynamics and diversity of fishes throughout the world. This study analyzed the folk taxonomy of artisanal fishers of the coast of the state of Paraíba, Brazil. Data were collected through free lists, semi-structured interviews and participant observations. A total of 308 local names were recorded, corresponding to 127 scientific taxa. The broad folk categories ‘pescada’ (croakers) and ‘cação’ (shark) were the most common. The Cognitive Salience Index (CSI) was used to assess which fishes the fishers knew best; the mutton snapper (Lutjanus analis) had the highest CSI score. Four hierarchical levels of folk taxonomy (kingdom, life form, generic and specific) were recorded. The local ichthyofauna contained 16 species at risk at the national level and 34 at the global level. Our results suggest that fisher folk taxonomy can complement scientific knowledge, improve and update threatened species lists and in doing so benefit fisheries management and conservation actions.     

Genetic diversity of glucose phosphate isomerase from Entamoeba histolytica

To investigate the molecular basis of zymodeme analysis in the enteric protozoan parasite Entamoeba histolytica, genes encoding glucose phosphate isomerase (GPI) were isolated from four representative E. histolytica strains belonging to zymodeme II, II-, XIV, or XIX. Two alleles were obtained from each strain; six alleles with eight polymorphic nucleotide positions were identified among the four strains. Two of these eight polymorphic nucleotides resulted in non-conserved amino acid substitutions. Three GPI isoenzymes with distinct predicted isoelectric points were identified, which agrees well with the observed electrophoretic patterns of GPI from these strains. Amino acid comparisons of GPI from E. histolytica and other organisms revealed that all amino acid residues implicated for substrate binding and catalysis were conserved. Biochemical characterization of recombinant E. histolytica GPI confirmed that it possessed kinetic parameters similar to GPI from other organisms. The electrophoretic mobility of three GPI isoenzymes was examined by starch gel electrophoresis. Thus, we have established the molecular basis of the classical isoenzymes patterns that have been used for grouping E. histolytica isolates and for differentiation of E. histolytica from non-pathogenic Entamoeba dispar.     

Oltipraz-induced decrease in the activity of cytosolic glutathione S-transferase in Schistosoma mansoni.

The activity of glutathione S-transferase (GST) decreased progressively in Schistosoma mansoni from mice treated with oltipraz (OPZ). However, the peroxidase activity of GST (selenium-independent) and selenium-dependent glutathione peroxidase was not affected by OPZ treatment. Purification and quantification of GST from worms after OPZ treatment indicated that the decrease in enzyme activity was greater than could be accounted for by the decrease in GST protein content. SDS-polyacrylamide gel electrophoresis followed by Western blot analysis with GST isoenzyme specific antisera revealed a slight decrease in the quantity of both 26 and 28 kDa GSTs. Fractionation of cytosolic GSTs from male S. mansoni by chromatofocusing resolved three major isoenzymes (SmI, II and III) and a minor form which eluted first from the column. SmI, II and III all had a molecular weight of about 28 kDa on SDS-polyacrylamide gel electrophoresis. However, on electrophoresis in the absence of SDS, the three GST forms exhibited different mobilities. The pattern of SmI, II and III was similar in untreated and OPZ-treated worms, but the activities of the isoenzymes from treated worms were lower. The results suggest that OPZ interacts with the GST isoenzymes SmI, II and III in a similar manner; thus, the effects are not isoenzyme specific. Taken together, these results suggest that OPZ and/or its metabolites interact directly with GST resulting in inhibition of activity and reduction in total enzyme protein. This mechanism may be important in the antischistosomal action of OPZ.     

松属花粉的显微结构观察——对白皮松分类位置的探讨

对松属九个种花粉显微结构的光学显微镜和扫描电镜观察均表明,其本体腹侧纹饰是花粉粒一个非常稳定的形态特征,可作为松属分类的重要依据,据此探讨了白皮松的分类位置。     

甲基紫精对丹参培养细胞抗氧化防护系统的影响

为了探讨甲基紫精(MV)对丹参(Salvia miltiorrhiza)体内抗氧化防护系统的影响及其生理机制。以MV为诱导剂, 以敌草隆(DCMU)为抑制剂, 考察了MV与DCMU处理后丹参悬浮培养细胞中H₂O₂、丙二醛、还原型谷胱甘肽的含量以及抗氧化防护酶(超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT))活性变化和同工酶的表达差异。结果表明, MV处理显著提高了丹参培养细胞内H₂O₂、丙二醛以及还原型谷胱甘肽含量; MV处理使CAT、POD活性增强, 谱带颜色更亮, 条带增加。DCMU处理显著抑制了MV诱导的H₂O₂、丙二醛、还原型谷胱甘肽含量的增加, 抗氧化酶活性的升高和同工酶的表达。以上结果说明, MV可诱导丹参培养细胞叶绿体产生H₂O₂, H₂O₂激活了丹参培养细胞抗氧化防护系统以维持细胞正常的生理活动。     

Foliar anatomy of neotropical Salicaceae: potentially useful characters for taxonomy

The taxonomy of neotropical Salicaceae, a family that now includes the majority of the former Flacourtiaceae, has been problematic, especially because they display very diverse morphology and have several characteristics in common with many other families. Recent phylogenetic studies have proposed substantial changes at both family and generic levels. Considering the importance of anatomy as an aid for taxonomy, the gathering of anatomical data for the family is fundamental to help clarify the taxonomic problems. Leaves belonging to Abatia americana (four samples), Banara brasiliensis (2), Casearia arborea (4), C. decandra (5), C. gossypiosperma (2), C. obliqua (1), C. sylvestris (3), C. ulmifolia (3), Prockia crucis (3), and Xylosma prockia (4) and the closely related Carpotroche brasiliensis (3) from Achariaceae, were studied by standard microscopy techniques. The leaves were anatomically described, emphasizing their differences and similarities. Similar characters for the neotropical Salicaceae (former Flacourtiaceae) and Salicaceae strictu sensu were recognized, such as the presence of salicoid leaf teeth, brachyparacytic stomata, secondary growth of the petiole, abundance of crystals, collateral and arch-shaped vascular system at the midrib, and sclerenchyma accompanying the bundles. These data demonstrate that leaf anatomy can provide evidence to assist with the taxonomy of Salicaceae, at family, generic, and specific levels.     

蕨类植物孢子囊形态 I. 鳞始蕨科

孢子囊是蕨类植物的繁殖器官, 其形态在蕨类植物的分类和系统发育研究上具有重要意义。用次氯酸钠溶液处理新鲜成熟的孢子囊, 在光学显微镜下获得清晰的孢子囊图像, 系统研究了中国鳞始蕨科4属13种孢子囊的形态特征。结果表明, 鳞始蕨科孢子囊呈椭球形, 孢子囊柄由3列细胞构成, 环带类型为垂直环带。通过分析孢子囊形态数据探讨了中国鳞始蕨科属内及属间差异。结果表明, 乌蕨属(Odontosoria)、香鳞始蕨属(Osmolindsaea)、达边蕨属(Tapeinidium)和鳞始蕨属(Lindsaea)的孢子囊环带细胞数依次减少, 囊蒴体积、唇细胞数和囊壁细胞数的变化由大(多)到小(少)依次为乌蕨属、达边蕨属、香鳞始蕨属和鳞始蕨属。孢子囊属内差异最大的是阔片乌蕨(Odontosoria biflora)与乌蕨(O. chinensis)以及香鳞始蕨(Osmolindsaea odorata)与日本鳞始蕨(O. japonica); 而达边蕨属和鳞始蕨属的属内差异则很小。研究结果为揭示鳞始蕨科系统发育关系提供了形态基础, 特别是提出阔片乌蕨和乌蕨以及香鳞始蕨和日本鳞始蕨在孢子囊形态上的差异值得进一步研究。     

Effects of methyl viologen on the antioxidant system in cultured Salvia miltiorrhiza cells

为了探讨甲基紫精(MV)对丹参(Salvia miltiorrhiza)体内抗氧化防护系统的影响及其生理机制。以MV为诱导剂, 以敌草隆(DCMU)为抑制剂, 考察了MV与DCMU处理后丹参悬浮培养细胞中H₂O₂、丙二醛、还原型谷胱甘肽的含量以及抗氧化防护酶(超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT))活性变化和同工酶的表达差异。结果表明, MV处理显著提高了丹参培养细胞内H₂O₂、丙二醛以及还原型谷胱甘肽含量; MV处理使CAT、POD活性增强, 谱带颜色更亮, 条带增加。DCMU处理显著抑制了MV诱导的H₂O₂、丙二醛、还原型谷胱甘肽含量的增加, 抗氧化酶活性的升高和同工酶的表达。以上结果说明, MV可诱导丹参培养细胞叶绿体产生H₂O₂, H₂O₂激活了丹参培养细胞抗氧化防护系统以维持细胞正常的生理活动。     

Taxonomic revision of Lepisorus (J. Sm.) Ching sect. Lepisorus (Polypodiaceae) from China

The taxonomy of Lepisorus (J. Sm.) Ching sect. Lepisorus in China was revised based on herbarium specimen examinations, field observations, and microscopic study of rhizome scales, soral paraphyses, leaf epidermis and spores. As a result nine species were recognized: Lepisorus macrosphaerus (Baker) Ching, L. asterolepis (Baker) Ching, L. marginatus Ching, L. kuchenensis (Y. C. Wu) Ching, L. megasorus(C. Chr.) Ching, L. kawakamii (Hayata) Tagawa, L. subsessilis Ching & Y. X. Lin, L. affinis Ching and L. nudus (Hook.) Ching. Lepisorus kawakamii (Hayata) Tagawa was reinstated; L. gyirongensis Ching & S. K. Wu and L. longus Ching were reduced to synonyms of L. nudus and L. affinis respectively. The subdivision of L. macrosphaerusis was not accepted. Rhizome scales and paraphyses are the most useful characters for species delimitation as well as for infrageneric classification. Characteristics of the leaf epidermis and spore ornamentation are usually stable and thus of great significance in understanding the relationships among groups within the genus.     

A fast and gentle method for the isolation of myrosinase complexes from Brassicaceous seeds

Myrosinase is a β-thioglucosidase glucohydrolase that catalyses the hydrolysis of the thioglucoside bond in glucosinolates, allelochemicals present in Brassicaceous plants. These isoenzymes have been found to form complexes with other proteins; however, traditional isolation procedures involving ammonium sulphate precipitation and/or ion exchange chromatography do not allow for the isolation of these complexes. The present paper reports a fast and gentle procedure for the isolation of myrosinases in the complex form. Partial purification by Con A affinity chromatography followed by Sephadex G-200 gel filtration allowed for the isolation of myrosinase complexes from seeds of Brassica carinata, B. oleracea var. capitata, B. napus and Sinapis alba. Myrosinases in the Brassicas formed complexes of different molecular weight (500–600 kDa, 270–350 kDa and 140–200 kDa) whereas in seeds of S. alba it was only possible to isolate and detect 140–200 kDa complexes. In all species the complexes were formed by isoenzymes with isoelectric points between 4.8 and 5.6 and in some cases up to 6.8. SDS-PAGE confirmed that the myrosinase isoenzymes were composed by several protein subunits of molecular weights ranging between 10 and 110 kDa. The relative amount and enzymatic activity of the myrosinase complexes varied amongst the species studied. The isolation of myrosinase complexes in their native form is of great importance for the study of the hydrolysis of glucosinolates under autolysis conditions.     

Host adaptation and host-parasite co-evolution in Cryptosporidium: implications for taxonomy and public health

To assess the genetic diversity and evolution of Cryptosporidium parasites, the partial ssrRNA, actin, and 70 kDa heat shock protein (HSP70) genes of 15 new Cryptosporidium parasites were sequenced. Sequence data were analysed together with those previously obtained from other Cryptosporidium parasites (10 Cryptosporidium spp. and eight Cryptosporidium genotypes). Results of this multi-locus genetic characterisation indicate that host adaptation is a general phenomenon in the genus Cryptosporidium, because specific genotypes were usually associated with specific groups of animals. On the other hand, host–parasite co-evolution is also common in Cryptosporidium, as closely related hosts usually had related Cryptosporidium parasites. Results of phylogenetic analyses suggest that the Cryptosporidium parvum bovine genotype and Cryptosporidium meleagridis were originally parasites of rodents and mammals, respectively, but have subsequently expanded their host ranges to include humans. Understanding the evolution of Cryptosporidium species is important not only for clarification of the taxonomy of the parasites but also for assessment of the public health significance of Cryptosporidium parasites from animals.     

Amorosia littoralis gen. sp. nov., a new genus and species name for the scorpinone and caffeine-producing hyphomycete from the littoral zone in The Bahamas

The new generic and species name Amorosia littoralis gen. sp. nov. is introduced for the conidial dematiaceous hyphomycete isolated from the littoral zone in The Bahamas and reported in 2001 to produce the novel aza-anthraquinone scorpinone, and also caffeine. No satisfactory generic placement was found at the time, but subsequent morphological and molecular investigations reveal that a new generic name is required. The new genus has some similarity to several fungi described in Trichocladium, but differs substantially from the type species of that genus in the form of the conidia and the lack of ornamentation. BLAST studies using the 18S and 28S rDNA gene sequences place the new genus in the Sporormiaceae. In addition to the morphological studies, an ultrastructural examination of the conspicuous porate septa of hyphae showed that they do not belong to a basidiomycete.