A multivariate approach to the selection of biological reserves

Multivariate analysis provides an effective context for the examination of some significant aspects of biodiversity and conservation. The framework is a multidimensional space that integrates sample sites, taxa and environments. This approach enables terms such as representativeness, complementarity and irreplaceability to be integrated within an intuitive and practical framework for reserve design. Cluster analysis is proposed to determine what is there by defining a set of complementary clusters. These clusters are sampled in a representative manner; from the core outward. The degree of irreplaceability of a site is defined as the multivariate distance of each potential reserve site to its nearest neighbour.     

The importance of urban reserves for conserving beetle communities: a case study from New Zealand

Urban reserves provide a major opportunity for conservation of indigenous biodiversity in the heavily urbanised landscape of Waitakere City (Auckland), New Zealand. However, there is little documented information on what indigenous fauna survives in these reserves. Beetle (Coleoptera) communities associated with two small, isolated urban reserves and two sites in a larger forest area within the Waitakere Ranges were sampled using pitfall traps and analysed. A total of 887 beetles, from 23 families and 89 recognisable taxonomic units were caught. The urban reserves had a marked reduction in species richness and abundance of beetles compared with the sites within the larger forest areas. Various environmental factors influencing the distribution of beetles across the sites were investigated. The most important factors were size of fragment, local forest cover and soil moisture. Common species (>5 specimens in the total dataset) found in the Waitakere Ranges and small urban reserves, were either endemic or indigenous to New Zealand. Therefore, even though these reserves may be isolated from a larger, more continuous forest tract, they have considerable potential as reservoirs of beetle diversity in highly modified landscapes and the contribution of urban reserves to the local sustainability of beetle assemblages emphasises the importance of maintaining green areas in and around cities.     

Snake communities of moist rainforest and derived savanna sites of Nigeria: biodiversity patterns and conservation priorities

The taxonomic composition and the abundance of two communities of snakes were studied in two different areas of southern Nigeria. One community was studied in a derived savanna area (environs of Ejule, 06°54N, 07°23E), and one community was studied in a moist rainforest area (environs of Eket, 04°50N, 07°59E). Both the specific diversity and the mean frequency of observation of snakes were significantly higher in the forest area than in the savanna area, and the dominance index was higher in the savanna than in the forest site. However, most of the species were found in only one of the two areas, depending on their habitat requirements (e.g. Gastropyxis smaragdina, Dispholidus typus, Thelotornis kirtlandii, Dendroaspis jamesoni, Bitis nasicornis, Causus maculatus, etc). The forest community housed a significant number of arboreal and semiaquatic taxa, but in both sites most of the species were terrestrial. The commonest species in the forest area was an oviparous, semiaquatic, batracophagous natricine snake (Afronatrix anoscopus), whereas the commonest species in the savanna area was an oviparous terrestrial lacertophagous psammophine snake (Psammophis phillipsi). Some conservation implications of our biodiversity analyses are presented. It is suggested that the moist rainforest represents a critically endangered habitat, and should deserve special attention by the international scientific community. Oil industry activity is especially dangerous for snake communities, especially in the southernmost regions of Nigeria.     

Arabidopsis consensus intron sequences

We have analysed 998 Arabidopsis intron sequences in the EMBL database. All Arabidopsis introns to adhere to the :GU...AG: rule with the exception of 1% of introns with :GC at their 5 ends. Virtually all of the introns contained a putative branchpoint sequence (YUNAN) 18 to 60 nt upstream of the 3 splice site. Although a polypyrimidine tract was much less apparent than in vertebrate introns, the most common nucleotide in the region upstream of the 3 splice site was uridine. Consensus sequences for 5 and 3 splice sites and branchpoint sequences for Arabidopsis introns are presented.     

Aldimine to ketoamine isomerization (Amadori rearrangement) potential at the individual nonenzymic glycation sites of hemoglobin a: Preferential inhibition of glycation by nucleophiles at sites of low isomerization potential

The relative roles of the two structural aspects of nonenzymic glycation sites of hemoglobin A, namely the ease with which the amino groups could form the aldimine adducts and the propensity of the microenvironments of the respective aldimines to facilitate the Amadori rearrangement, in dictating the site selectivity of nonenzymic glycation with aldotriose has been investigated. The chemical reactivity of the amino groups of hemoglobin A forin vitro reductive glycation with aldotriose is distinct from that in the nonreductive mode. The reactivity of amino groups of hemoglobin A toward reductive glycation (i.e., propensity for aldimine formation) decreases in the order Val-1(), Val-1(), Lys-66(), Lys-61(), and Lys-16(). The overall reactivity of hemoglobin A toward nonreductive glycation decreased in the order Lys-16(), Val-1(), Lys-66(), Lys-82(), Lys-61(), and Val-1(). Since the aldimine is the common intermediate for both the reductive and nonreductive modification, the differential selectivity of protein for the two modes of glycation is clearly a reflection of the propensity of the microenvironments of nonenzymic glycation sites to facilitate the isomerization reaction (i.e., Amadori rearrangement). A semiquantitative estimate of this propensity of the microenvironment of the nonenzymic glycation sites has been obtained by comparing the nonreductive (nonenzymic) and reductive modification at individual glycation sites. The microenvironment of Lys-16() is very efficient in facilitating the rearrangement and the relative efficiency decreases in the order Lys-16(), Lys-82(), Lys-66(), Lys-61(), Val-1(), and Val-1(). The propensity of the microenvironment of Lys-16() to facilitate the Amadori rearrangement of the aldimine is about three orders of magnitude higher than that of Val-1() and is about 50 times higher than that of Val-1(). The extent of nonenzymic glycation at the individual sites is modulated by various factors, such as thepH, concentration of aldotriose, and the concentration of the protein. The nucleophiles—such as tris, glycine ethyl ester, and amino guanidine—inhibit the glycation by trapping the aldotriose. The nonenzymic glycation inhibitory power of nucleophile is directly related to its propensity to form aldimine. Thus, the extent of inhibition of nonenzymic glycation at a given site by a nucleophile directly reflects the relative role ofpK _a of the site in dictating the glycation at that site. The nonenzymic glycation of an amino group of a protein is an additive/synergestic consequence of the propensity of the site to form aldimine adducts on one hand, and the propensity of its microenvironment to facilitate the isomerization of the aldimines to ketoamines on the other. The isomerization potential of microenvironment plays the dominant role in dictating the site specificity of the nonenzymic glycation of proteins.     

Leaf litter inhabiting beetles as surrogates for establishing priorities for conservation of selected tropical montane cloud forests in Honduras, Central America (Coleoptera; Staphylinidae, Curculionidae)

Thirteen tropical montane cloud forests (TMCFs) in Honduras and adjacent El Salvador were evaluated for species diversity and endemism in leaf litter inhabiting Curculionidae (weevils) and Staphylinidae in June and August of 1994. Totals of 26,891 specimens representing 293 species of Curculionidae, and 7349 specimens representing 224 species of Staphylinidae, were collected. Regional endemism was high with 173 species (58.7% of total) of Curculionidae and 126 species (56.3% of total) of Staphylinidae, restricted to single sites. Measures of diversity (number of observed species [Sobs]; number of endemic species) and estimates of biodiversity (Chao 2, first and second order Jackknife and Bootstrap richness estimators) are given for each site for Curculionidae and Staphylinidae. Priority-areas analyses based on Sobs (Greedy), Sobs, Chao 2, number of endemic species, site complementarity and higher taxonomic diversity (Curculionidae only) are presented. Weak or lack of correlation was noted between site area and site diversity for either Curculionidae or Staphylinidae. The optimum sequence for site conservation was determined based on Sobs (Greedy). Four sites are identified as the highest conservation priorities; Parque Nacional Montecristo, P.N. La Muralla, P.N. Santa Barbara and P.N. Comayagua. Identification as high priority sites supports their designation as Parques Nacionales rather than the less diverse Reservas Biologias or Reservas Vidas Silvestres. While closely approximating performance of Sobs (Greedy), no one of Sobs, number of endemics, Chao 2 or site complementarity give results equivalent to the optimum sequence based on Sobs (Greedy) and the latter is concluded to be the best method for establishing conservation priorities in TMCF. Results of the analyses based on one of Curculionidae or Staphylinidae differ in the ordering of site priorities based on each of Sobs, Chao 2 or number of endemics at each site, with at best, weak positive correlations between results based on each taxon. Data from other taxa are necessary to determine if one of Curculionidae or Staphylinidae emerges as the better surrogate for general patterns of biodiversity in TMCF. Conservation strategies in Central America should emphasize the importance of TMCF particularly in view of high regional endemism. In formulating these strategies, consideration needs to be given to the preservation of many small regional preserves rather than fewer larger preserves. Data from inventories of other taxa should be included where available and all biological data should be integrated with social and cultural issues of regional importance.     

Environmental diversity: on the best-possible use of surrogate data for assessing the relative biodiversity of sets of areas

The conservation goal of representation of biodiversity (in the broad sense of all species) in protected areas requires best-possible use of available surrogate information. One standard approach is based on indicator groups of taxa. A minimum set of areas having at least one representation of each indicator species is taken to be representative of other organisms. This same minimum-set approach is adapted to other attributes of biodiversity, for example, derived environmental clusters. A weakness of these approaches is that useful information is lost; for example, for environmental clusters, there is no distinction made either among or within clusters. A more powerful surrogate approach can use some expression of environmental and/or biotic pattern so that variation among areas is seen as part of a continuum rather than partitioned into arbitrary clusters/attributes. The challenge in using pattern effectively is to adopt a robust model for the relationship between pattern and the underlying units of biodiversity, i.e. species. An environmental space (a continuum or ordination pattern), combined with the standard ecological continuum model relating species to environmental space, has advantages over other patterns based on hierarchy or distance matrices. Because an environmental space can be estimated either directly (observed environmental data) or indirectly (data on indicator groups), the corresponding surrogate-measure of biodiversity, environmental diversity (ED) makes best-possible use of either kind of data. We conclude that the arbitrariness of the attribute approach can be replaced by a robust surrogate pattern approach that is flexible and avoids unwarranted assumptions.     

Full 1H NMR assignment of a 24-nucleotide RNA hairpin: Application of the 1H 3D-NOE/2QC experiment

The subject RNA models the binding site for the coat protein of the R17 virus, as well as the ribosome recognition sequence for the R17 replicase gene. With an RNA of this size, overlaps among the sugar protons complicate assignments of the ¹H NMR spectrum. The cross peaks that overlap significantly in 2D-NOE spectra can frequently be resolved by introducing a third, in our approach the double-quantum, frequency axis. In particular the planes in a 3D-NOE/2QC spectrum perpendicular to the 2Q axis are extremely useful, showing a highly informative repeating NOE-2Q pattern. In this experiment substantial J-coupling confers special advantages. This always occurs for geminal pairs (H5/H5 for RNA plus H2/H2 for DNA), as well as for H5/H6, for H3/H4 in sugars with substantial populations of the N-pucker, for H1/H2 for S-puckered sugars, and usually for H2/H3. For the 24-mer RNA hairpin the additional information from the 3D-NOE/2QC spectrum allowed assignment of all of the non-exchangeable protons, eliminating the need for stable-isotope labeling.     

The mutational spectrum of single base-pair substitutions in mRNA splice junctions of human genes: Causes and consequences

Summary A total of 101 different examples of point mutations, which lie in the vicinity of mRNA splice junctions, and which have been held to be responsible for a human genetic disease by altering the accuracy of efficiency of mRNA splicing, have been collated. These data comprise 62 mutations at 5 splice sites, 26 at 3 splice sites and 13 that result in the creation of novel splice sites. It is estimated that up to 15% of all point mutations causing human genetic disease result in an mRNA splicing defect. Of the 5 splice site mutations, 60% involved the invariant GT dinucleotide; mutations were found to be non-randomly distributed with an excess over expectation at positions +1 and +2, and apparent deficiencies at positions –1 and –2. Of the 3 splice site mutations, 87% involved the invariant AG dinucleotide; an excess of mutations over expectation was noted at position -2. This non-randomness of mutation reflects the evolutionary conservation apparent in splice site consensus sequences drawn up previously from primate genes, and is most probably attributable to detection bias resulting from the differing phenotypic severity of specific lesions. The spectrum of point mutations was also drastically skewed: purines were significantly overrepresented as substituting nucleotides, perhaps because of steric hindrance (e.g. in U1 snRNA binding at 5 splice sites). Furthermore, splice sites affected by point mutations resulting in human genetic disease were markedly different from the splice site consensus sequences. When similarity was quantified by a consensus value, both extremely low and extremely high values were notably absent from the wild-type sequences of the mutated splice sites. Splice sites of intermediate similarity to the consensus sequence may thus be more prone to the deleterious effects of mutation. Regarding the phenotypic effects of mutations on mRNA splicing, exon skipping occurred more frequently than cryptic splice site usage. Evidence is presented that indicates that, at least for 5 splice site mutations, cryptic splice site usage is favoured under conditions where (1) a number of such sites are present in the immediate vicinity and (2) these sites exhibit sufficient homology to the splice site consensus sequence for them to be able to compete successfully with the mutated splice site. The novel concept of a potential for cryptic splice site usage value was introduced in order to quantify these characteristics, and to predict the relative proportion of exon skipping vs cryptic splice site utilization consequent to the introduction of a mutation at a normal splice site.     

Viability and activity in readily culturable bacteria: a review and discussion of the practical issues

In microbiology the terms viability and culturability are often equated. However, in recent years the apparently self-contradictory expression viable-but-nonculturable (VBNC) has been applied to cells with various and often poorly defined physiological attributes but which, nonetheless, could not be cultured by methods normally appropriate to the organism concerned. These attributes include apparent cell integrity, the possession of some form of measurable cellular activity and the apparent capacity to regain culturability. We review the evidence relating to putative VBNC cells and stress our view that most of the reports claiming a return to culturability have failed to exclude the regrowth of a limited number of cells which had never lost culturability. We argue that failure to differentiate clearly between use of the terms viability and culturability in an operational versus a conceptual sense is fuelling the current debate, and conclude with a number of proposals that are designed to help clarify the major issues involved. In particular, we suggest an alternative operational terminology that replaces VBNC with expressions that are internally consistent.     

A model for the mechanism of precise integration of a microinjected transgene

A unique transgenic mouse line has undergone transgene integration in a very precise fashion. The phenotype displayed by mice of the line followed the predicted inheritance patterns for X-linked transgene insertion which has been confirmed. In order to investigate the mechanism of integration the DNA sequence of the transgene and cellular junctions have been determined. A comparison between wild type and transgenic mutant sequences at the site of insertion revealed that there was no loss or rearrangement of cellular DNA upon integration of the transgene. The cellular sequences at the transgene 5 and 3 joins are contiguous in the wild type. The integrant exists as a head to tail tandem dimer with minimal loss of sequence compared with the injected monomer. Analysis of the site of insertion has revealed a 5 bp homology between the 5 end of the transgene and the cellular sequences. In addition, adjacent to the site of insertion within the cellular sequences, there are several sequence motifs implicated in recombination events including a clustering of strong consensus sites for DNA topoisomerase type I and a region of homology to the human minisatellite consensus core sequence, theEscherichia coli Chi site and the meiotic recombination hotspot within the E gene of the murine major histocompatibility complex. This clustering of features is likely to have been factorial in the integrity of the insertion event. A model depicting the mechanism of this precise integration is proposed.     

Spatial and temporal variation in soil CO2 efflux in an old-growth neotropical rain forest, La Selva, Costa Rica

Our objectives were to quantify and compare soil CO₂ efflux of two dominant soil types in an old-growth neotropical rain forest in the Atlantic zone of Costa Rica, and to evaluate the control of environmental factors on CO₂ release. We measured soil CO₂ efflux from eight permanent soil chambers on six Oxisol sites. Three sites were developed on old river terraces (old alluvium) and the other three were developed on old lava flows (residual). At the same time we measured soil CO₂ concentrations, soil water content and soil temperature at various depths in 6 soil shafts (3 m deep). Between old alluvium sites, the two-year average CO₂ flux rates ranged from 117.3 to 128.9 mg C m^–2 h^–1. Significantly higher soil CO₂ flux occurred on the residual sites (141.1 to 184.2 mg C m^–2 h^–1). Spatial differences in CO₂ efflux were related to fine root biomass, soil carbon and phosphorus concentration but also to soil water content. Spatial variability in CO₂ storage was high and the amount of CO₂ stored in the upper and lower soil profile was different between old alluvial and residual sites. The major factor identified for explaining temporal variations in soil CO₂ efflux was soil water content. During periods of high soil water content CO₂ emission decreased, probably due to lower diffusion and CO₂ production rates. During the 2-year study period inter-annual variation in soil CO₂ efflux was not detected.     

The use of ground beetles (Coleoptera: Carabidae) in conservation assessments of exposed riverine sediment habitats in Scotland and northern England

Exposed riverine sediments (ERS) are an important invertebrate habitat. Ground beetle data from pitfall trap surveys of 165 sites in the catchments of the Rivers Carron, Nith, Spey, Tweed and Eden and from 48 sites in the catchments of the Rivers Blyth, Breamish, Coquet, Glen, Pont, South Tyne, Till, Tweed and Tyne were used to produce ERS habitat classifications in Scotland and northern England respectively. Within each habitat type the quality of sites were assessed using indices based on rarity and fidelity values and using species richness as a measure of diversity. Considerable variation in these indices was seen within and between habitat types, showing potential for their use in assessing site quality and for ranking sites within habitats. The use of standardised invertebrate survey and distribution data, together with species assemblage information, for the conservation of invertebrate habitat types as an alternative to the use of selected indicator species, is discussed.     

How natural is a nature reserve?: an ideological study of British nature conservation landscapes

Areas set apart for nature conservation in Britain are broadly categorised according to their cultural purpose, and names are assigned to these in this paper. Nature reserves may be similar to zoos and botanic gardens in aiming to maintain the diversity of species and if so are termed biodiversity reserves. This tradition understands nature as a static collection of entities apart from humans. Maintaining traditional management at a site is arguably a good way to sustain species, it also retains old ways in which humans and nature were integrated in the life of the nation and so are called historic countryside parks. There is growing interest in wilderness areas, where nature is seen as primarily processes protected from human interference. Despite the strength of each of these, they suffer from attempting to restrict nature to a ghetto, a process that is economically and environmentally costly. Companion places are places which set sustainable examples of integrating human life and economic activity with maintaining biodiversity and offering an opportunity to encounter wild processes at the heart of life. The language of these four types, or vectors, of nature reserves is offered to help the discussion of our place in nature.     

A method for the deletion of restriction sites in bacterial plasmid deoxyribonucleic acid

Summary A general method has been developed for the deletion of restriction endonuclease sites in bacterial plasmid DNA. The procedure involves partial digestion of the covalently closed circular plasmid DNA with an appropriate restriction endonuclease under conditions which allow accumulation of unit-length linear DNA molecules, controlled digestion of the exposed 5 ends with the 5-exonuclease, and in vivo recircularization of the resulting linear DNA in a bacterial host cell. The method has been used for the deletion of one of the two EcoRI sites in the plasmid pML2 (colE1-Km). Two of the resulting plasmids, pCR1 and pCR11, have a single EcoRI cleavage site, but retain genetic determinants specifying resistance to colicin E1 and kanamycin, and thus may be useful as vectors for the cloning and amplification of DNA in bacteria.     

Carbon and nitrogen isotope ratios in different compartments of a healthy and a declining Picea abies forest in the Fichtelgebirge,NE Bavaria

Summary Natural carbon and nitrogen isotope ratios were measured in different compartments (needles and twigs of different ages and crown positions, litter, understorey vegetation, roots and soils of different horizons) on 5 plots of a healthy and on 8 plots of a declining Norway spruce (Picea abies (L.) Karst.) forest in the Fichtelgebirge (NE Bavaria, Germany), which has recently been described in detail (Oren et al. 1988a; Schulze et al. 1989). The ¹³C values of needles did not differ between sites or change consistently with needle age, but did decrease from the sun-to the shade-crown. This result confirms earlier conclusions from gas exchange measurements that gaseous air pollutants did no long-lasting damage in an area where such damage was expected. Twigs (¹³C between-25.3 and-27.8) were significantly less depleted in ¹³C than needles (¹³C between-27.3 and-29.1), and ¹³C in twigs increased consistently with age. The ¹⁵N values of needles ranged between-2.5 and-4.1 and varied according to stand and age. In young needles ¹⁵N decreased with needle age, but remained constant or increased in needles that were 2 or 3 years old. Needles from the healthy site were more depleted in ¹⁵N than those from the declining site. The difference between sites was greater in old needles than in young ones. This differentiation presumably reflects an earlier onset of nitrogen reallocation in needles of the declining stand. ¹⁵N values in twigs were more negative than in needles (-3.5 to-5.2) and showed age- and stand-dependent trends that were similar to the needles. ¹⁵N values of roots and soil samples increased at both stands with soil depth from-3.5 in the organic layer to +4 in the mineral soil. The ¹⁵N values of roots from the mineral soil were different from those of twigs and needles. Roots from the shallower organic layer had values similar to twigs and needles. Thus, the bulk of the assimilated nitrogen was presumably taken up by the roots from the organic layer. The problem of separation of ammonium or nitrate use by roots from different soil horizons is discussed.     

Microspore cultures as donor tissue for the initiation of embryogenic cell suspensions in barley

We have initiated embryogenic cell suspension cultures of barley (Hordeum vulgare L.) Igri from isolated microspore cultures. Data were obtained on the time required for establishment, frequency of establishment, i.e. number of calluses out of the total number of initiations giving rise to suspensions, and embryogenic capacity of the suspension cultures. For comparison, establishment of embryogenic cell suspensions from callus derived from immature zygotic embryos of Igri, Dissa and Golden Promise was also carried out. The results revealed that embryogenic suspension cultures were established in half the time and with a seven-fold higher frequency from microspore cultures than from zygotic embryo-derived calluses. The suspension cultures were still capable of embryo formation after two years. However, only albino plantlets were regenerated. For comparison, long term callus cultures derived from microspores, anthers and zygotic embryos were established. From the anther and zygotic embryo-derived callus cultures green plants were continuously regenerated, whereas the microspore-derived callus cultures lost this ability after the second subculture.     

Prebiotic Formation of ADP and ATP from AMP,Calcium Phosphates and Cyanate in Aqueous Solution

Adenosine-5-triphosphate was synthesized by the phosphorylation of adenosine-5-diphosphate in aqueous solution containing cyanate as a condensing reagent and insoluble calcium phosphate produced from phosphate and calcium chloride. In a similar manner, adenosine-5-diphosphate was synthesized from adenosine-5-monophosphate. When the experiment was carried out in the conditions of 4 °C and pH 5.75, the formation of adenosine-5-diphosphate and adenosine-5-triphosphate from adenosine-5-monophosphate was observed in the yields of 19 and 7%, respectively. The other nucleoside-5-triphosphates were also produced from their respective diphosphates.     

The catalysis of nucleotide polymerization by compounds of divalent lead

Summary Soluble lead salts and a number of lead-containing minerals catalyze the formation of oligonucleotides from nucleoside 5-phosphorimidazolides. The effectiveness of lead compounds correlates strongly with their solubility. Under optimal conditions we were able to obtain 18% of pentamer and higher oligomers from ImpA. Reactions involving ImpU gave smaller yields.Abbreviations A adenosine - U uridine - Im imidazole - MeIm 1-methyl-imidazole - EDTA ethylenediaminetetraacetic acid - pA adenosine 5-phosphate - pU uridine 5-phosphate - Ap adenosine cyclic 2:3-phosphate - ATP adenosine 5-triphosphate - AppA P₁,P₂-diadenosine 5-diphosphate - pNp (N = A,U) nucleotide 2(3), 5-diphosphate - ImpA adenosine 5-phosphoreimidazolide - ImpU uridine 5-phosphorimidazolide - A ²pA adenylyl-[25]-adenosine - A ³pA adenylyl-[35]-adenosine - pA ²pA 5-phospho-adenylyl-[25]-adenosine - pA ³pA 5-phospho-adenylyl-[35]-adenosine - pUpU 5-phospho-uridylyl-uridine - pApU 5-phospho-adenylyl-uridine - pUpA 5-phospho-uridylyladenine - (pA)n (n, 2,3,4,) oligoadenylates with 5 terminal phosphate - ImpApA 5-phosphorimidazolide of adenylyl adenosine - (pA) ₅₊ pentamer and higher oligoadenylates with 5 terminal phosphate - (Ap)nA (n = 2,3,4) oligoadenylates without terminal phosphates In the following we do not specify the nature of the internucleotide linkageIn the following we do not specify the nature of the internucleotide linkage