A comparative analysis of karyotypes of three species of Macleaya—producers of a complex of isoquinoline alkaloids

Using a set of methods (C-banding, DAPI-staining, fluorescence hybridization in situ (FISH) with probes of 26S and 5S rDNA, and analysis of meiosis), the first comparative cytogenetic study of three species of Macleaya, producers of complex isoquinoline alkaloids, cordate Macleaya cordata (Willd.) R. Br. (2n = 20), small-fruited Macleaya microcarpa (Maxim.) Fedde (2n = 20) and Macleaya kewensis Turrill (2n = 20), was first carried out. On the basis of morphometric analysis, formulas of karyotypes were made for each species. Species ideograms for M. cordata, M. microcarpa, and M. kewensis were constructed taking into account the polymorphic variants of the C-banding patterns and indicating the location of 26S and 5S rDNA sites. A comparative study revealed that the karyotypes of M. microcarpa and M. kewensis have more in common with each other than with M. cordata. Analysis of meiotic chromosomes suggests of genetic stability of Macleaya genomes. The results of chromosome analysis were used to confirm the close relationship of Macleaya and to clarify their phylogenetic relationships.     

Effect of γ-Irradiation on Development of Lachrymator of Onion

A thermosensitive uracil requiring mutant of Bacillus subtilis Marburg 168 thy trp₂ ts42 was examined as to the colony forming ability at the permissive and nonpermissive temperatures. The viability of the mutant cells decreased rapidly at the restrictive temperature in the modified Woese’s (MW) medium. However, the cells retained viability when sodium succinate or potassium chloride was added to the medium at that temperature although uracil deficiency was unchanged. A little but significant incorporation of adenine-8-¹⁴C into RNA still continued even after the incorporation of N-acetyl-³H-d-glucosamine into acid insoluble fraction of the cells terminated in the MW medium at 48°C. Both incorporations as well as increase of absorbance were slowed down in the presence of sodium succinate at 48°C. This mutant, ts42, was more sensitive to deoxycholate (DOC) than the parent strain. The restoration of colony forming ability after the temperature shift back from 48 to 37°C was suppressed by the addition of DOC to the medium. However, the cell became resistant to DOC when uracil was added to the medium prior to the temperature shift.     

Mechanisms of action of the α-amylase of Micromonospora melanosporea

The -amylase of Micromonospora melanosporea was produced extracellularly during batch fermentation in a 5.0-1 fermentor. The absence of an organic nitrogen source in its growth medium facilitated subsequent purification of the enzyme by ammonium sulphate fractionation and two consecutive Superose-12 gel-filtration steps. The enzyme exhibited maxima for activity at pH 7.0 and 55° C and was 72% stable at pH 6.0–12.0 for 30 min at 40° C. It had a relative molecular mass of 45 000 and an isoelectric point at pH 7.6. The enzyme catalyses the conversion of starch to maltose (53%, w/w) as the predominant final end-product. Initial hydrolysis of this substrate, however, gave rise to the formation of maltooligosaccharides in the range maltotriose to maltohexaose. Maximum yields of these intermediate sugars accumulated to between 31 and 42% (w/w) as the reaction proceeded. The action of the M. melanosporea amylase on high concentrations of saccharides larger than maltotriose resulted in the formation of mainly maltose and maltotriose without concomitant glucose production. A combination of hydrolytic and transfer events is postulated to be responsible for this phenomenon and for the high maltose levels achieved. Correspondence to: C. T. Kelly     

Selection of optimal variants of Gō-like models of proteins through studies of stretching

The Gō-like models of proteins are constructed based on the knowledge of the native conformation. However, there are many possible choices of a Hamiltonian for which the ground state coincides with the native state. Here, we propose to use experimental data on protein stretching to determine what choices are most adequate physically. This criterion is motivated by the fact that stretching processes usually start with the native structure, in the vicinity of which the Gō-like models should work the best. Our selection procedure is applied to 62 different versions of the Gō model and is based on 28 proteins. We consider different potentials, contact maps, local stiffness energies, and energy scales—uniform and nonuniform. In the latter case, the strength of the nonuniformity was governed either by specificity or by properties related to positioning of the side groups. Among them is the simplest variant: uniform couplings with no i, i + 2 contacts. This choice also leads to good folding properties in most cases. We elucidate relationship between the local stiffness described by a potential which involves local chirality and the one which involves dihedral and bond angles. The latter stiffness improves folding but there is little difference between them when it comes to stretching.     

Chemistry of the “molecular trap” of protease-catalyzed splicing reaction of complementary segments of α-subunit of hemoglobin A

The complementary fragments of human Hb α, α_1–30, and α_31–141 are spliced together by V8 protease in the presence of 30%n-propanol to generate the full-length molecule (Hb α-semisynthetic reaction). Unlike the other protease-catalyzed protein/peptide splicing reactions of fragment complementing systems, the enzymic condensation of nonassociating segments of Hb α is facilitated by the organic cosolvent induced α-helical conformation of product acting as the “molecular trap” of the splicing reaction. The segments α_24–30 and α_31–40 are the shortest complementary segments that can be spliced by V8 protease. In the present study, the chemistry of the contiguous segment (product) α_24–40 has been manipulated by engineering the amino acid replacements to the positions α²⁷ and α³¹ to delineate the structural basis of the molecular trap. The location of Glu²⁷ and Arg³¹ residues in the contiguous segment α_24–40 (as well as in other larger segments) is ideal to generate (i, i+4) side-chain carboxylate-guanidino interaction in its α-helical conformation. The amino acid residue replacement studies have confirmed that the side chains at α²⁷ and α³¹ facilitate the semisynthetic reaction. The relative influence of the substitute at these sites on the splicing reaction depends on the chemical nature of the side chain and the location. The γ-carboxylate guanidino side-chain interaction appears to contribute up to a maximum of 85% of the thermodynamic stability of the molecular trap. The studies also demonstrate that the thermodynamic stability of the molecular trap is determined by two interdependent conformational aspects of the peptide. One is an amino acid-sequence-specific event that facilitates the induction of an α-helical conformation to the contiguous segment in the presence of organic cosolvent that imparts some amount of protease resistance to Glu³⁰-Arg³¹ peptide bond. The second structural aspect is a site-specific event, ani, i+4 side-chain interaction in the α-helical conformation of the peptide which imparts an additional thermodynamic stability to the molecular trap. The results suggest that conformationally driven “molecular traps” of protease-mediated ligation reactions of peptides could be designed into products to facilitate the modular assembly of peptides/proteins.     

Association of the subunits of the calcium-independent receptor of α-latrotoxin

CIRL-1 also called latrophilin 1 or CL belongs to the family of adhesion G protein-coupled receptors (GPCRs). As all members of adhesion GPSR family CIRL-1 consists of two heterologous subunits, extracellular hydrophilic p120 and heptahelical membrane protein p85. Both CIRL-1 subunits are encoded by one gene but as a result of intracellular proteolysis of precursor, mature receptor has two-subunit structure. It was also shown that a minor portion of the CIRL-1 receptor complexes dissociates, producing the soluble receptor ectodomain, and this dissociation is due to the second cleavage at the site between the site of primary proteolysis and the first transmembrane domain. Recently model of independent localization p120 and p85 on the cell surface was proposed. In this article we evaluated the amount of p120-p85 complex still presented on the cellular membrane and confirmed that on cell surface major amount of mature CIRL-1 presented as a p120-p85 subunit complex.     

Kinetics of renaturation and self-assembly of intermediates on the pathway of folding of the β2-subunit of Escherichia coli tryptophan-synthetase

The kinetics of renaturation of the β₂-subunit of Escherichia coli tryptophan-synthetase (l-serine hydrolyase (adding indole) E.C. 4.2.1.20) and those of its two proteolytic fragments F₁ and F₂ are studied and compared. Steps corresponding to the refolding of F₁, to the association of the folded F₁ and F₂ fragments, and to an isomerization of the associated protein are identified. These steps are ordered on the pathway of renaturation and some of their kinetic parameters are determined. This leads to a tentative kinetic model for the renaturation of nicked-β₂ starting from the denatured F₁ and F₂ fragments.The step corresponding to the refolding of the F₁ domain, as well as that corresponding to the last rate-limiting isomerization leading to the native protein, is shown to be the same in the refolding of the entire, uncleaved β₂-protein. It is concluded that the refolded F₁ fragment corresponds to a folding intermediate on the pathway of renaturation of the β₂-subunit.     

Inhibition of colony-spreading activity of Staphylococcus aureus by secretion of δ-hemolysin

Staphylococcus aureus spreads on the surface of soft agar, a phenomenon we termed "colony spreading." Here, we found that S. aureus culture supernatant inhibited colony spreading. We purified δ-hemolysin (Hld, δ-toxin), a major protein secreted from S. aureus, as a compound that inhibits colony spreading. The culture supernatants of hld-disrupted mutants had 30-fold lower colony-spreading inhibitory activity than those of the parent strain. Furthermore, hld-disrupted mutants had higher colony-spreading ability than the parent strain. These results suggest that S. aureus negatively regulates colony spreading by secreting δ-hemolysin.     

Spatial variation in species diversity and composition of flea assemblages in small mammalian hosts: geographical distance or faunal similarity?

Aim Spatial variation in the diversity of fleas parasitic on small mammals was examined to answer three questions. (1) Is the diversity of flea assemblages repeatable among populations of the same host species? (2) Does similarity in the composition of flea assemblages among populations of the same host species decay with geographical distance, with decreasing similarity in the composition of local host faunas, or with both? (3) Does the diversity of flea assemblages correlate with climatic variables? Location The study used previously published data on 69 species of small mammals and their fleas from 24 different regions of the Holarctic. Methods The diversity of flea assemblages was measured as both species richness and the average taxonomic distinctness of their component species. Similarity between flea assemblages was measured using both the Jaccard and Morisita–Horn indices, whereas similarity in the composition of host faunas between regions (host ‘faunal’ distance) was quantified using the Jaccard index. Where appropriate, a correction was made for the potentially confounding influence of phylogeny using the independent contrasts method. Results Flea species richness varied less within than among host species, and is thus a repeatable host species character; the same was not true of the taxonomic distinctness of flea assemblages. In almost all host species found in at least five regions, similarity in flea assemblages decreased with increases in either or both geographical and faunal distance. In most host species, the diversity of flea assemblages correlated with one or more climatic variable, in particular mean winter temperature. Main conclusions Spatial variation in flea diversity among populations of the same mammal species is constrained by the fact that it appears to be a species character, but is also driven by local climatic conditions. The results highlight how ecological processes interact with co‐evolutionary history to determine local parasite biodiversity.     

Global patterns in peatmoss biodiversity

DNA sequence data from the nuclear ribosomal internal transcribed spacers (ITS) and the trnL-trnF chloroplast DNA regions were used to quantify geographical partitioning of global biodiversity in peatmosses (Sphagnum), and to compare patterns of molecular diversity with patterns of species richness. Molecular diversity was estimated for boreal, tropical, Neotropical, nonboreal (tropical plus Southern Hemisphere), Old World and New World partitions, based on a total of 436 accessions. Diversity was partitioned among geographical regions in terms of combined nuclear and chloroplast sequence data and separately for the ITS and trnL-trnF data sets. Levels of variation were estimated using phylogenetic diversity (PD), which incorporates branch lengths from a phylogenetic tree, and the number of polymorphic nucleotide sites. Estimates of species richness suggest that peatmoss diversity is higher in New World than Old World regions, and that the Neotropics constitute a "hotspot" of diversity. Molecular estimates, in contrast, indicate that peatmoss biodiversity is almost evenly divided between New and Old World regions, and that the Neotropics account for only 20-35% of global peatmoss diversity. In general, levels of tropical and boreal peatmoss molecular diversity were comparable. Two species, S. sericeum from the Old World tropics and S. lapazense from Bolivia, are remarkably divergent in nucleotide sequences from all other Sphagna and together account for almost 20% of all peatmoss diversity, although they are represented by only three of the 436 accessions (0.7%). These species clearly demonstrate the nonequivalence of species biodiversity value.     

Chemotypes significance of lichenized fungi by structural characterization of heteropolysaccharides from the genera Parmotrema and Rimelia

Galactoglucomannans were isolated from the lichenized fungi of the genus Parmotrema (Parmotrema austrosinense, Parmotrema delicatulum, Parmotrema mantiqueirense, Parmotrema schindlerii, and Parmotrema tinctorum and that of Rimelia (Rimelia cetrata and Rimelia reticulata) via successive hot alkaline extraction and precipitation with Fehling solution. The structure of each polysaccharide was investigated using 13C NMR and HSQC-DEPT spectroscopy, methylation analysis, and HPSEC-MALLS. The galactoglucomannans had a (1-->6)-linked main chain of alpha-Manp units, substituted preferentially at O-2 and O-4 by alpha-Galp and beta-Galp nonreducing end-units, respectively. The C-1 region of the 13C NMR spectra of these heteropolysaccharides is typical of the lichen species, and is an additional tool in lichenized fungi classification.     

Biogeography of tuna and billfish communities

Aim The aims of this study were: (1) to identify global communities of tuna and billfish species through quantitative statistical analyses of global fisheries data; (2) to describe the spatial distribution, main environmental drivers and species composition of each community detected; and (3) to determine whether the spatial distribution of each community could be linked to the environmental conditions that affect lower trophic levels by comparing the partitions identified in this study with Longhurst’s biogeochemical provinces. Location The global ocean from 60° S to 65° N. Methods We implemented a new numerical procedure based on a hierarchical clustering method and a nonparametric probabilistic test to divide the oceanic biosphere into biomes and ecoregions. This procedure was applied to a database that comprised standardized data on commercial longline catches for 15 different species of tuna and billfish over a period of more than 50 years (i.e. 1953–2007). For each ecoregion identified (i.e. characteristic tuna and billfish community), we analysed the relationships between species composition and environmental factors. Finally, we compared the biogeochemical provinces of Longhurst with the ecoregions that we identified. Results Tuna and billfish species form nine well‐defined communities across the global ocean. Each community occurs in regions with specific environmental conditions and shows a distinctive species composition. High similarity (68.8% homogeneity) between the spatial distribution of the communities of tuna and billfish and the biogeochemical provinces suggests a strong relationship between these species and the physical and chemical characteristics of the global ocean. Main conclusions Despite their high tolerance for a wide range of environmental conditions, these highly migratory species are partitioned into clear geographical communities in the ocean at a global scale. The similarity between biogeochemical and biotic divisions in the ocean suggests that the global ocean is a mosaic of large biogeographical ecosystems, each characterized by specific environmental conditions that have a strong effect on the composition of the trophic web.     

Pronounced genetic diversity in tropical epiphyllous lichen fungi

Lowland tropical habitats harbour an unexplored genetic diversity of epiphyllous fungi. In the shade of rainforest understoreys, lichenized fungi are specialized to an ephemeral habitat where they produce little vegetative biomass and develop reproductive structures early. In a first population genetic study of epiphyllous lichen fungi, we analysed the intraspecific genetic diversity of five leaf-colonizing lichen mycobiont species. Sampling focused on a lowland perhumid forest plot in Costa Rica, with additional collections from other localities throughout the country. In all species we detected sympatric occurrence of highly diverged haplotypes. Haplotypes belonging to distinct clades in networks were also found on the same leaf, clearly indicating multiple independent colonization events on single leaves. Despite the unusually high genetic diversity of these leaf-colonizing tropical fungi, we did not detect pronounced spatial structure of the haplotype distribution between geographical regions. The observed patterns suggest that the diversity of foliicolous lichens could be much higher than expected, with several cryptic genetic lineages within each morphologically characterized species.     

Can large unmanaged trees replace ancient pollarded trees as habitats for lichenized fungi,non-lichenized fungi and bryophytes?

Management of ancient trees constitutes a major dilemma in the conservation of associated biodiversity. While traditional methods are often advocated, such practices may incur considerable costs and their effects have rarely been scientifically evaluated. We compared the communities of lichenized fungi, non-lichenized fungi, and bryophytes among equal number of coarse previously pollarded and unmanaged trees (n = 340). On 400 Ulmus glabra and 280 Fraxinus excelsior trees at 62 sites in Norway, we found 209 lichenized fungi, 128 non-lichenized fungi, and 115 bryophytes. Pollarded trees were richer in microhabitats than unmanaged trees and had significantly higher richness of bryophytes (ash) and non-lichenized fungi (ash and elm), the latter increasing with the availability of dead wood, cavities and coarse bark structure in pollarded trees. Further, the average total number of red-listed species, and red-listed lichenized fungi separately, were significantly higher on pollarded versus unmanaged trees, with diversity related to trunk circumference, depth of bark fissures and number of cavities. Our results underline the importance of microhabitats associated with old trees, but we cannot establish with certainty the importance of pollarding per se. Since we did not find any negative effect of canopy cover for community diversity, we assume that old trees with rich epiphytic communities can develop without management intervention. The high share (37 out of 49) of red-listed species occurring on unmanaged trees, and the fact that 11 red-listed species were found exclusively on unmanaged trees, may further indicate that unmanaged trees can with time replace the ancient pollarded trees as habitats for rich cryptogamic communities.     

Global Biogeography of Reef Fishes: A Hierarchical Quantitative Delineation of Regions

Delineating regions is an important first step in understanding the evolution and biogeography of faunas. However, quantitative approaches are often limited at a global scale, particularly in the marine realm. Reef fishes are the most diversified group of marine fishes, and compared to most other phyla, their taxonomy and geographical distributions are relatively well known. Based on 169 checklists spread across all tropical oceans, the present work aims to quantitatively delineate biogeographical entities for reef fishes at a global scale. Four different classifications were used to account for uncertainty related to species identification and the quality of checklists. The four classifications delivered converging results, with biogeographical entities that can be hierarchically delineated into realms, regions and provinces. All classifications indicated that the Indo-Pacific has a weak internal structure, with a high similarity from east to west. In contrast, the Atlantic and the Eastern Tropical Pacific were more strongly structured, which may be related to the higher levels of endemism in these two realms. The “Coral Triangle”, an area of the Indo-Pacific which contains the highest species diversity for reef fishes, was not clearly delineated by its species composition. Our results show a global concordance with recent works based upon endemism, environmental factors, expert knowledge, or their combination. Our quantitative delineation of biogeographical entities, however, tests the robustness of the results and yields easily replicated patterns. The similarity between our results and those from other phyla, such as corals, suggests that our approach may be of broad utility in describing and understanding global marine biodiversity patterns.     

黄精根际及药用部位内生真菌群落组成和生态功能分析

为寻找促进药用植物活性代谢产物合成的微生物,该文以黄精为研究对象,利用高通量测序技术和生态功能预测平台,测定根际土真菌、根茎和根内生真菌的ITS序列,分析其真菌多样性和群落组成,并预测根茎内生真菌的生态功能。结果表明:(1)测序得到1 023个可操作分类单元(OTUs),根际、根茎和根真菌OTU数分别为703、128和141,三种部位真菌群落组成差异显著,根际土存在特有的真菌类群,即壶菌门。(2)根际土、根茎及根共有OTU 41个,子囊菌门占共有真菌的58.15%,丰度最大。(3)根茎内生真菌被划分6个生态功能群,包括未定义腐生菌、菌寄生真菌、动物病原菌、植物病原菌、丛枝菌根真菌和地衣共生真菌,37个undefined种类(34.91%)在FUNGuild数据库中没有参考信息。研究认为根茎中优势菌属Setophoma、新赤壳属等内生真菌可能与活性代谢产物密切相关,可为黄精药用功能菌群的发掘提供数据参考。     

云南被子植物菊类分支的系统发育多样性及其分布格局

全球气候变化与人为活动等因素导致的生物多样性丧失,引起了全球各界对生物多样性保护的高度关注。传统生物多样性保护主要对物种、特有种、受威胁物种的种类组成及其分布模式开展研究,忽视了进化历史在生物多样性保护中的作用。云南是全球生物多样性热点地区的交汇区,生物多样性的保护历来受到广泛关注,为了更好地探讨云南生物多样性的保护措施,该研究以云南被子植物菊类分支物种为研究对象,基于物种间的演化关系,结合其地理分布,从进化历史的角度探讨物种、特有种、受威胁物种的种类组成及系统发育组成的分布格局,并整合自然保护地的空间分布,识别生物多样性的重点保护区域。结果表明:云南被子植物菊类分支的物种、特有种及受威胁物种的物种密度与系统发育多样性均显著正相关; 通过零模型分析发现,由南向北标准化系统发育多样性逐渐降低; 云南南部、东南部、西北部是云南被子植物菊类分支的重点保护区域,加强这些区域的保护,将最大化地保护生物多样性的进化历史和进化潜能。由此可见,融合进化历史信息的植物多样性格局分析不仅有助于更加深入地理解植物多样性的形成与演变,也为生物多样性保护策略的制定提供更多的思路。     

Long‐term experimental warming alters community composition of ascomycetes in Alaskan moist and dry arctic tundra

Arctic tundra regions have been responding to global warming with visible changes in plant community composition, including expansion of shrubs and declines in lichens and bryophytes. Even though it is well known that the majority of arctic plants are associated with their symbiotic fungi, how fungal community composition will be different with climate warming remains largely unknown. In this study, we addressed the effects of long‐term (18 years) experimental warming on the community composition and taxonomic richness of soil ascomycetes in dry and moist tundra types. Using deep Ion Torrent sequencing, we quantified how OTU assemblage and richness of different orders of Ascomycota changed in response to summer warming. Experimental warming significantly altered ascomycete communities with stronger responses observed in the moist tundra compared with dry tundra. The proportion of several lichenized and moss‐associated fungi decreased with warming, while the proportion of several plant and insect pathogens and saprotrophic species was higher in the warming treatment. The observed alterations in both taxonomic and ecological groups of ascomycetes are discussed in relation to previously reported warming‐induced shifts in arctic plant communities, including decline in lichens and bryophytes and increase in coverage and biomass of shrubs.