The lion's share of bacteria in various environments cannot be cloned in the laboratory and thus cannot be sequenced using existing technologies. A major goal of single-cell genomics is to complement gene-centric metagenomic data with whole-genome assemblies of uncultivated organisms. Assembly of single-cell data is challenging because of highly non-uniform read coverage as well as elevated levels of sequencing errors and chimeric reads. We describe SPAdes, a new assembler for both single-cell and standard (multicell) assembly, and demonstrate that it improves on the recently released E+V-SC assembler (specialized for single-cell data) and on popular assemblers Velvet and SoapDeNovo (for multicell data). SPAdes generates single-cell assemblies, providing information about genomes of uncultivatable bacteria that vastly exceeds what may be obtained via traditional metagenomics studies. SPAdes is available online ( http://bioinf.spbau.ru/spades ). It is distributed as open source software.

SPAdes, a new genome assembly algorithm and its applications to single-cell sequencing ( 7th Annual SFAF Meeting, 2012)

So far in this course we have dealt entirely with the evolution of characters that are controlled by simple Mendelian inheritance at a single locus. There are notes on the course website about gametic disequilibrium and how allele frequencies change at two loci simultaneously, but we didn’t discuss them. In every example we’ve considered we’ve imagined that we could understand something about evolution by examining the evolution of a single gene. That’s the domain of classical population genetics. For the next few weeks we’re going to be exploring a field that’s actually older than classical population genetics, although the approach we’ll be taking to it involves the use of population genetic machinery. If you know a little about the history of evolutionary biology, you may know that after the rediscovery of Mendel’s work in 1900 there was a heated debate between the “biometricians” (e.g., Galton and Pearson) and the “Mendelians” (e.g., de Vries, Correns, Bateson, and Morgan). Biometricians asserted that the really important variation in evolution didn’t follow Mendelian rules. Height, weight, skin color, and similar traits seemed to

Human biochemical genetics

Long-term dietary intake influences the structure and activity of the trillions of microorganisms residing in the human gut, but it remains unclear how rapidly and reproducibly the human gut microbiome responds to short-term macronutrient change. Here we show that the short-term consumption of diets composed entirely of animal or plant products alters microbial community structure and overwhelms inter-individual differences in microbial gene expression. The animal-based diet increased the abundance of bile-tolerant microorganisms (Alistipes, Bilophila and Bacteroides) and decreased the levels of Firmicutes that metabolize dietary plant polysaccharides (Roseburia, Eubacterium rectale and Ruminococcus bromii). Microbial activity mirrored differences between herbivorous and carnivorous mammals, reflecting trade-offs between carbohydrate and protein fermentation. Foodborne microbes from both diets transiently colonized the gut, including bacteria, fungi and even viruses. Finally, increases in the abundance and activity of Bilophila wadsworthia on the animal-based diet support a link between dietary fat, bile acids and the outgrowth of microorganisms capable of triggering inflammatory bowel disease. In concert, these results demonstrate that the gut microbiome can rapidly respond to altered diet, potentially facilitating the diversity of human dietary lifestyles.

/pdf/diet-rapidly-and-reproducibly-alters-the-human-gut-drvn0pi8vp.pdf

Diet rapidly and reproducibly alters the human gut microbiome

Six DNA regions were evaluated as potential DNA barcodes for Fungi, the second largest kingdom of eukaryotic life, by a multinational, multilaboratory consortium. The region of the mitochondrial cytochrome c oxidase subunit 1 used as the animal barcode was excluded as a potential marker, because it is difficult to amplify in fungi, often includes large introns, and can be insufficiently variable. Three subunits from the nuclear ribosomal RNA cistron were compared together with regions of three representative protein-coding genes (largest subunit of RNA polymerase II, second largest subunit of RNA polymerase II, and minichromosome maintenance protein). Although the protein-coding gene regions often had a higher percent of correct identification compared with ribosomal markers, low PCR amplification and sequencing success eliminated them as candidates for a universal fungal barcode. Among the regions of the ribosomal cistron, the internal transcribed spacer (ITS) region has the highest probability of successful identification for the broadest range of fungi, with the most clearly defined barcode gap between inter- and intraspecific variation. The nuclear ribosomal large subunit, a popular phylogenetic marker in certain groups, had superior species resolution in some taxonomic groups, such as the early diverging lineages and the ascomycete yeasts, but was otherwise slightly inferior to the ITS. The nuclear ribosomal small subunit has poor species-level resolution in fungi. ITS will be formally proposed for adoption as the primary fungal barcode marker to the Consortium for the Barcode of Life, with the possibility that supplementary barcodes may be developed for particular narrowly circumscribed taxonomic groups.

/pdf/nuclear-ribosomal-internal-transcribed-spacer-its-region-as-9rhgb6gvih.pdf

Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi

The RefSeq project at the National Center for Biotechnology Information (NCBI) maintains and curates a publicly available database of annotated genomic, transcript, and protein sequence records (http://www.ncbi.nlm.nih.gov/refseq/). The RefSeq project leverages the data submitted to the International Nucleotide Sequence Database Collaboration (INSDC) against a combination of computation, manual curation, and collaboration to produce a standard set of stable, non-redundant reference sequences. The RefSeq project augments these reference sequences with current knowledge including publications, functional features and informative nomenclature. The database currently represents sequences from more than 55,000 organisms (>4800 viruses, >40,000 prokaryotes and >10,000 eukaryotes; RefSeq release 71), ranging from a single record to complete genomes. This paper summarizes the current status of the viral, prokaryotic, and eukaryotic branches of the RefSeq project, reports on improvements to data access and details efforts to further expand the taxonomic representation of the collection. We also highlight diverse functional curation initiatives that support multiple uses of RefSeq data including taxonomic validation, genome annotation, comparative genomics, and clinical testing. We summarize our approach to utilizing available RNA-Seq and other data types in our manual curation process for vertebrate, plant, and other species, and describe a new direction for prokaryotic genomes and protein name management.

Reference sequence (RefSeq) database at NCBI: current status, taxonomic expansion, and functional annotation

The value of dried fungal cultures for taxonomic comparisons using PCR and RFLP analysis

The ascomycete genus Huntiella (Microascales) has a cosmopolitan distribution and occurs on a wide range of woody plants. Little is known regarding the identity, diversity, origin, or impact of these fungi in China. Recently, isolates of Huntiella spp. were collected from stumps of freshly felled trees or wounds on plantation-grown Eucalyptus in Guangdong, Guangxi, Fujian, and Hainan provinces of southern China. Additional isolates were obtained from stumps of Acacia confusa near Eucalyptus plantations in Hainan Province. The aim of this study was to identify these Huntiella species and to test their pathogenicity on Eucalyptus seedlings. Morphology and multigene phylogenies of the nuclear rDNA internal transcribed spacer (ITS1-5.8S-ITS2 = ITS) region and partial β-tubulin (BT1) and translation elongation factor 1α (TEF1α) genes revealed nine previously unknown Huntiella species, eight from Eucalyptus and one from A. confusa. The mating types of these species were determined, showing that seven are heterothallic, one is homothallic, and one is unisexual (MAT1-2-1 gene). Pathogenicity tests showed that the nine Huntiella species can produce lesions on Eucalyptus seedlings, larger than wounds caused by controls on these plants. This study provides a basic understanding of the distribution, diversity, and pathogenicity of Huntiella species in southern China.

Nine novel species of Huntiella from southern China with three distinct mating strategies and variable levels of pathogenicity.

Linde, c., Wingfield, ~. J., and Kemp, G. H. J. 1994. Root and root collar disease of Eucalyptus gramlis caused by Pythium spJendens. Plant Dis. 78:1006-1009. A serious root and root collar disease of Eucal}ptus grandis occurred in the Kwambonambi area of northern ~'ata] during the past 4 yr. This disease typically occurs on young trees that rapidly \I.'ill and die. Pythium splendens was consistently isolated from foots and root collars of dying trees. P. splendens inoculated on two different clones of E. grandis and on Eucalyptus fastigata ",..ashighly pathogenic. Isolates of P. splendens showed the same or an even higher degree of virulence than PhYlOphlhora cinnamomi, a well-known Eucalyplus root pathogen, on the E grandis clones tested. Virulence of P. splendens on E. Jasligara was, however, less than that of PhYlOphlhura cinnamomi.

/pdf/root-and-root-collar-disease-of-eucalyptus-grandis-caused-by-24skvdogac.pdf

Root and root collar disease of Eucalyptus grandis caused by Pythium splendens

Ras2 is important for growth and pathogenicity in Fusarium circinatum.

SYNOPSIS DNA was isolated from 12 clones of Eucalyptus grandis, and subsequently amplified with arbitrary primers (RAPD-PCR). It was possible to distinguish between the different clones on the basis of the resultant fingerprints. We conclude that the RAPD technique is suited for the identification of E. grandis clones, and should also be useful in genetic studies.

Michael J. Wingfield

Papers

The value of dried fungal cultures for taxonomic comparisons using PCR and RFLP analysis

Nine novel species of Huntiella from southern China with three distinct mating strategies and variable levels of pathogenicity.

Root and root collar disease of Eucalyptus grandis caused by Pythium splendens

Ras2 is important for growth and pathogenicity in Fusarium circinatum.

RAPD-fingerprinting to Identify Eucalyptus grandis Clones