Towards more accurate detection of pathogenic Gram-positive bacteria in waters

Medically important bacteria can persist in surface waters longer than was previously thought, by activating specific survival strategies and, thus, may represent a further threat to human health, in that they are non-detectable by the traditional culture methods currently used for the evaluation of microbiological quality. Combining microbial physiology, microbial biochemistry, microbial genetics, microbial ecology and molecular biology techniques allow us to achieve more accurate detection of human pathogens located in natural environments external to the human body.     

Towards more chemically robust polymer-supported chiral catalysts for the reactions of aldehydes with dialkylzincs

N-Methyl-alpha,alpha-diphenyl-L-prolinol derivatives with para-bromo substituents in one or both of the phenyl rings are easily bound to crosslinked polystyrene beads containing phenylboronic acid residues using Suzuki reactions. When the products were used as catalysts for the reactions of aldehydes with diethylzinc in toluene at 20 degrees C, the alcohols were produced in chemical yields >90% and with ees of upto 94%. The better of the two supported catalysts gave ees only 0-9% lower than those obtained with the corresponding soluble catalyst. One of the supported catalysts was recycled successfully nine times.     

Towards more biological mutation operators in gene regulation studies

Genetic regulation is often viewed as a complex system whose properties emerge from the interaction of regulatory genes. One major paradigm for studying the complex dynamics of gene regulation uses directed graphs to explore structure, behaviour and evolvability. Mutation operators used in such studies typically involve the insertion and deletion of nodes, and the insertion, deletion and rewiring of links at the network level. These network-level mutational operators are sufficient to allow the statistical analysis of network structure, but impose limitations on the way networks are evolved. There are a wide variety of mutations in DNA sequences that have yet to be analysed for their network-level effects. By modelling an artificial genome at the level of nucleotide sequences and mapping it to a regulatory network, biologically grounded mutation operators can be mapped to network-level mutations. This paper analyses five such sequence level mutations (single-point mutation, transposition, inversion, deletion and gene duplication) for their effects at the network level. Using analytic and simulation techniques, we show that it is rarely the case that nodes and links are cleanly added or deleted, with even the simplest point mutation causing a wide variety of network-level modifications. As expected, the vast majority of simple (single-point) mutations are neutral, resulting in a neutral plateau from which a range of functional behaviours can be reached. By analysing the effects of sequence-level mutations at the network level of gene regulation, we aim to stimulate more careful consideration of mutation operators in gene regulation models than has previously been given.     

Towards robust and repeatable sampling methods in eDNA‐based studies

DNA‐based techniques are increasingly used for measuring the biodiversity (species presence, identity, abundance and community composition) of terrestrial and aquatic ecosystems. While there are numerous reviews of molecular methods and bioinformatic steps, there has been little consideration of the methods used to collect samples upon which these later steps are based. This represents a critical knowledge gap, as methodologically sound field sampling is the foundation for subsequent analyses. We reviewed field sampling methods used for metabarcoding studies of both terrestrial and freshwater ecosystem biodiversity over a nearly three‐year period (n = 75). We found that 95% (n = 71) of these studies used subjective sampling methods and inappropriate field methods and/or failed to provide critical methodological information. It would be possible for researchers to replicate only 5% of the metabarcoding studies in our sample, a poorer level of reproducibility than for ecological studies in general. Our findings suggest greater attention to field sampling methods, and reporting is necessary in eDNA‐based studies of biodiversity to ensure robust outcomes and future reproducibility. Methods must be fully and accurately reported, and protocols developed that minimize subjectivity. Standardization of sampling protocols would be one way to help to improve reproducibility and have additional benefits in allowing compilation and comparison of data from across studies.     

Towards precise classification of cancers based on robust gene functional expression profiles

Background

Despite the continuous production of genome sequence for a number of organisms, reliable, comprehensive, and cost effective gene prediction remains problematic. This is particularly true for genomes for which there is not a large collection of known gene sequences, such as the recently published chicken genome. We used the chicken sequence to test comparative and homology-based gene-finding methods followed by experimental validation as an effective genome annotation method.

Results

We performed experimental evaluation by RT-PCR of three different computational gene finders, Ensembl, SGP2 and TWINSCAN, applied to the chicken genome. A Venn diagram was computed and each component of it was evaluated. The results showed that de novo comparative methods can identify up to about 700 chicken genes with no previous evidence of expression, and can correctly extend about 40% of homology-based predictions at the 5' end.

Conclusions

De novo comparative gene prediction followed by experimental verification is effective at enhancing the annotation of the newly sequenced genomes provided by standard homology-based methods.     

Towards more efficient large-scale DNA-based detection of terrestrial mammal predators from scats

The DNA detection of wildlife from environmental samples has the potential to contribute significantly to wildlife management and ecological research. In terrestrial ecosystems, much work has focused on the identification of mammal predators from faecal (scat) samples. However, the relatively high time and financial costs of collecting and analysing scat DNA remain barriers to more widespread implementation of such DNA detection methods, especially for high-throughput surveys. Here, we evaluate methods used for DNA extraction from scats, as applied to detection of the Australian red fox, an introduced predator. We compare the relative costs of two approaches: the method previously used to screen thousands of scat samples in surveys over several years, and a modified version which involves swabbing scats at the time of collection and using a mechanised liquid handling platform to extract DNA from the swabs. We demonstrate that mechanised DNA extraction from swabs is more efficient than manual DNA extraction from whole scats, in terms of both time and resources. This provides a means for rapid, high-throughput screening of scats for the presence of mammal predators, enabling time-effective management responses to non-invasive surveys.     

Towards the extremes: A critical analysis of pointer year detection methods

To study growth responses of trees to extreme events, analyses of so-called ‘pointer years’ are often performed. Thereby, the term pointer year refers to years in which a majority of trees shows extraordinary growth responses, like very narrow (or wide) ring widths. A wide variety of methods has been used to analyze pointer years, hampering comparisons between studies. The latter illustrates that there is a strong need to harmonize pointer year detection methods. This review contributes to that by describing and comparing the main methods (and variants thereof) found in the literature for the last two decades, both in a qualitative and quantitative way. We discuss methodological considerations and provide recommendations how to proceed with pointer year detection in future. Given that the individual methods for pointer year detection were found to highlight different aspects of extraordinary growth, the one method cannot completely substitute the other. Hence, we suggest to use multiple methods in a harmonized way to get the deepest insight into the nature of pointer years.     

Towards a more dynamic plant morphology

From the point of view of a dynamic morphology, form is not only the result of process(es) — it is process. This process may be analyzed in terms of two pairs of fundamental processes: growth and decay, differentiation and dedifferentiation. Each of these processes can be analyzed in terms of various modalities (parameters) and submodalities. This paper deals with those of growth (see Table 1). For the purpose of systematits and phylogenetic reconstruction the modalities and submodalities can be considered dynamic characters that have “states”. Each “state” of such a dynamic character is a more detailed process, hence not static. For example, determinate growth represents a “state” of the dynamic character (or modality) of growth duration. The processes of Table 1 can be applied to the whole plant kingdom (although in certain cases only some processes of the whole set may be applicable). Thus, the diversity of plant form is seen as a diversity of process combinations. From this point of view, change in form implies change in the process combination(s). Questions that arise are, for example, the following: Which process combinations actually occur? Which of these are the most frequent? How and why have process combinations changed during ontogeny and phylogeny? In comparative morphogenesis, process combinations are compared within an ontogeny or between ontogenies. The combinations may be repeated (i.e., conserved) or changed. Since repetition is limited, regularity that is the basis for structural categories is also limited or relative. With regard to change in process combinations, sequential change within an ontogeny and phylogenetic change between ontogenies can be distinguished. A large number of additional processes, such as heterochrony, that have been investigated by many zoologists and botanists, refer to these sequential and phylogenetic changes. General implications and consequences of the proposed approach are pointed out. As well, its limits, which are related to the language and concepts used, are discussed. The importance of a dynamic language is emphasized.     

Progress on methods of gene detection in preimplantation embryos

The advent of the polymerase chain reaction (PCR) and the development of fluorescence in situ hybridization (FISH) have had a tremendous impact on preimplantation genetic diagnosis (PGD). While PCR is a powerful tool in detecting genetic diseases or molecular markers affecting quantitative trait loci, the main use of FISH is screening for chromosomal aberrations. This presentation reviews the recent progress in preimplantation genetic diagnosis with an emphasis on bovine embryos. In particular the importance of biopsy size and strategies to avoid PCR contamination are discussed. Alternative DNA amplification and detection methods as well as methods to meet the challenge of multiple locus detection for marker assisted selection are presented.     

Phylogenetically poor plant communities receive more alien species, which more easily coexist with natives

Alien species can be a major threat to ecological communities, but we do not know why some community types allow the entry of many more alien species than do others. Here, for the first time, we suggest that evolutionary diversity inherent to the constituent species of a community may determine its present receptiveness to alien species. Using recent large databases from observational studies, we find robust evidence that assemblage of plant community types from few phylogenetic lineages (in plots without aliens) corresponds to higher receptiveness to aliens. Establishment of aliens in phylogenetically poor communities corresponds to increased phylogenetic dispersion of recipient communities and to coexistence with rather than replacement of natives. This coexistence between natives and distantly related aliens in recipient communities of low phylogenetic dispersion may reflect patterns of trait assembly. In communities without aliens, low phylogenetic dispersion corresponds to increased dispersion of most traits, and establishment of aliens corresponds to increased trait concentration. We conclude that if quantified across the tree of life, high biodiversity correlates with decreasing receptiveness to aliens. Low phylogenetic biodiversity, in contrast, facilitates coexistence between natives and aliens even if they share similar trait states.     

Towards robust evolutionary inference with integral projection models

Integral projection models (IPMs) are extremely flexible tools for ecological and evolutionary inference. IPMs track the distribution of phenotype in populations through time, using functions describing phenotype‐dependent development, inheritance, survival and fecundity. For evolutionary inference, two important features of any model are the ability to (i) characterize relationships among traits (including values of the same traits across ages) within individuals, and (ii) characterize similarity between individuals and their descendants. In IPM analyses, the former depends on regressions of observed trait values at each age on values at the previous age (development functions), and the latter on regressions of offspring values at birth on parent values as adults (inheritance functions). We show analytically that development functions, characterized this way, will typically underestimate covariances of trait values across ages, due to compounding of regression to the mean across projection steps. Similarly, we show that inheritance, characterized this way, is inconsistent with a modern understanding of inheritance, and underestimates the degree to which relatives are phenotypically similar. Additionally, we show that the use of a constant biometric inheritance function, particularly with a constant intercept, is incompatible with evolution. Consequently, current implementations of IPMs will predict little or no phenotypic evolution, purely as artefacts of their construction. We present alternative approaches to constructing development and inheritance functions, based on a quantitative genetic approach, and show analytically and through an empirical example on a population of bighorn sheep how they can potentially recover patterns that are critical to evolutionary inference.     

外来有害生物风险评估方法研究进展

外来有害生物给生态环境、农业生产、人类健康带来的影响和造成的经济损失越来越受到人们的关注。与此同时,对外来有害生物风险评估方法的研究及其应用也在逐步深入。通过对多指标综合评估法、农业气候相似距法、地理信息系统、生态气候模型评价、模糊综合评判等方法的分析,概述了外来有害生物风险评估方法的含义和利弊,探讨了不同外来物种在不同环境条件下的适生性,综述了国内外评估方法应用的研究进展,对该研究领域的发展趋势进行了展望。     

Towards alien plant prioritization in Italy: methodological issues and first results

In recent decades, multiple actions have been taken to counteract the relentless expansion of invasive alien species as well as to gain a better understanding of their effects on ecosystems. Here, we describe the approach designed by the Italian Botanical Society that is aimed at selecting a list of candidate alien plants to be subjected to a prioritization procedure. We selected a total of 96 species on the basis of data related to their occurrence on both a national and regional scale, their invasiveness and their potential to invade plant communities and/or habitats of community concern. This list represents the first result obtained by applying this standardized workflow and is a first step towards the identification of those alien species that should be included in the national list according to Regulation (EU) n. 1143/2014.     

植物外源基因组成分鉴定方法的研究进展

具外源基因组成分(外源染色体/染色体片段/基因)植株是目前进行基因组学研究以及作物改良的重要材料.迄今为止,已建立了基于性状观测、染色体分析、特异蛋白、DNA序列4种鉴定外源基因组成分的策略.其中,基于DNA序列的分子标记技术是当前鉴定外源基因组成分的主要手段,文中归纳了用于小麦、甘蓝等重要作物外源基因组成分的分子标记...     

Towards a more versatile alpha-glucan biosynthesis in plants

Starch is an important storage polysaccharide in many plants. It is composed of densely packed alpha-glucans, consisting of 1,4- and 1,4,6-linked glucose residues. The starch polymers are used in many industrial applications. The biosynthetic machinery for assembling the granule has been manipulated in many different ways to gain insight into the process of starch biosynthesis and to engineer starches with improved functionalities. With respect to the latter, two generic technologies with great potential have been developed: (i) introduction of new linkage types in starch polymers (1,3- and 1,6-linkages), and (ii) engineering granule-boundness. The toolbox to engineer this new generation of starch polymers is discussed.     

Use of artificial genomes in assessing methods for atypical gene detection

Parametric methods for identifying laterally transferred genes exploit the directional mutational biases unique to each genome. Yet the development of new, more robust methods—as well as the evaluation and proper implementation of existing methods—relies on an arbitrary assessment of performance using real genomes, where the evolutionary histories of genes are not known. We have used the framework of a generalized hidden Markov model to create artificial genomes modeled after genuine genomes. To model a genome, “core” genes—those displaying patterns of mutational biases shared among large numbers of genes—are identified by a novel gene clustering approach based on the Akaike information criterion. Gene models derived from multiple “core” gene clusters are used to generate an artificial genome that models the properties of a genuine genome. Chimeric artificial genomes—representing those having experienced lateral gene transfer—were created by combining genes from multiple artificial genomes, and the performance of the parametric methods for identifying “atypical” genes was assessed directly. We found that a hidden Markov model that included multiple gene models, each trained on sets of genes representing the range of genotypic variability within a genome, could produce artificial genomes that mimicked the properties of genuine genomes. Moreover, different methods for detecting foreign genes performed differently—i.e., they had different sets of strengths and weaknesses—when identifying atypical genes within chimeric artificial genomes.