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)

The results of the meetings of the International Committee on Taxonomy of Viruses, held in Madrid, September 1975, are briefly reported: rules of viral nomenclature, composition of the new Executive Committee, and a list of the names so far officially agreed. Introduction The International Committee on the Taxonomy of Viruses (ICTV), which is a committee of the Section on Virology of the International Association of Microbiological Societies (IAMS), completed a round of meetings during the Third International Congress for Virology that was held in Madrid from September 10--17, 1975. Since ICTV only meets during these conferences, which are held every four years, the meetings are important occasions for reviewing the classification and nomenclature of viruses. Decisions on new names, which encapsulate the recognition of natural "groups" of viruses, evolve slowly. Official approval for new names depends upon a series of sequential steps ; recommendations by one or more of the subcommittees of the Executive Committee of ICTV (subcommittees on Bacterial, Invertebrate, Plant and Vertebrate Viruses respectively, and for some of the larger viral groups which span several kinds of hos t s the Coordination Subcommittee), which are considered by the Executive Committee of ICTV and may finally be submitted for approval by ICTV itself. Only after this final approval does a name become "official". The results of the last five years of work by ICTV and its committees will be published early this year as a separate volume of "Intervirology", the official journal of the Section on Virology of IAMS, and additional copies will be produced for sale as separates. This "Second P~eport" will include, besides "approved" and common names of virus groups, a brief description of the properties of each

Classification and Nomenclature of Viruses

Panama disease of banana, caused by the fungus Fusarium oxysporum f. sp. cubense, is a serious constraint both to the commercial production of banana and cultivation for subsistence agriculture. Previous work has indicated that F. oxysporum f. sp. cubense consists of several clonal lineages that may be genetically distant. In this study we tested whether lineages of the Panama disease pathogen have a monophyletic origin by comparing DNA sequences of nuclear and mitochondrial genes. DNA sequences were obtained for translation elongation factor 1α and the mitochondrial small subunit ribosomal RNA genes for F. oxysporum strains from banana, pathogenic strains from other hosts and putatively nonpathogenic isolates of F. oxysporum. Cladograms for the two genes were highly concordant and a partition-homogeneity test indicated the two datasets could be combined. The tree inferred from the combined dataset resolved five lineages corresponding to “F. oxysporum f. sp. cubense” with a large dichotomy between two taxa represented by strains most commonly isolated from bananas with Panama disease. The results also demonstrate that the latter two taxa have significantly different chromosome numbers. F. oxysporum isolates collected as nonpathogenic or pathogenic to other hosts that have very similar or identical elongation factor 1α and mitochondrial small subunit genotypes as banana pathogens were shown to cause little or no disease on banana. Taken together, these results indicate Panama disease of banana is caused by fungi with independent evolutionary origins.

https://www.pnas.org/content/pnas/95/5/2044.full.pdf

Multiple evolutionary origins of the fungus causing Panama disease of banana: Concordant evidence from nuclear and mitochondrial gene genealogies

Pretreatment technologies are aimed to increase enzyme accessibility to biomass and yields of fermentable sugars. In general, pretreatment methods fall into four different categories including physical, chemical, physico-chemical, and biological. This paper comprehensively reviews the lignocellulosic wastes to bioethanol process with a focus on pretreatment methods, their mechanisms, advantages and disadvantages as well as the combinations of different pretreatment technologies. Moreover, the new advances in plant “omics” and genetic engineering approaches to increase cellulose composition, reduce cellulose crystallinity, produce hydrolases and protein modules disrupting plant cell wall substrates, and modify lignin structure in plants have also been expansively presented.

Lignocellulosic biomass to bioethanol, a comprehensive review with a focus on pretreatment

The present invention provides a method of identifying a base at a target position in a sample nucleic acid sequence, said method comprising: subjecting a primer hybridised to said sample nucleic acid immediately adjacent to the target position, to a polymerase primer extension reaction in the presence of a nucleotide, whereby the nucleotide will only become incorporated if it is complementary to the base in the target position, and determining whether or not said nucleotide is incorporated by detecting whether Ppi is released, the identity of the target base being determined from the identity of any nucleotide incorporated, wherein, where said nucleotide comprises an adenine base, an α-thio triphosphate analogue of said nucleotide is used, ant the Rp isomer of said analogue and/or the degradation products of said analogue are eliminated from the polymerase reaction step.

Method of sequencing dna

A method for detecting a specific polynucleotide sequence in a material is disclosed. The method includes exposing said material to an oligonucleotide primer having a sequence complementary to part of said specific polynucleotide sequence wherein said primer binds to part of said polynucleotide sequence when present in said material. Primer bound to the polynucleotide sequence is extended wherein any extended primer includes a detectable element and/or a separation element. Any extended primer is then separated into a fraction wherein said fraction does not have detectable element not included in said extended primer. One then determines whether any extended primer is present in said fraction by assaying said fraction for said extended primer wherein the presence of said extended primer is indicative of the presence of the specific polynucleotide sequence in said material and the absence of said extended primer in said fraction is indicative of the absence of the specific polynucleotide sequence in said material.

Detection of a nucleic acid sequence or a change therein

We have cloned, sequenced and analysed an additional five circular ssDNA components of banana bunchy top virus (BBTV) which we have called components 2, 3, 4, 5 and 6. These components were present in all BBTV infections tested. Four of these components (components 3, 4, 5 and 6) had one large open reading frame (ORF) in the virion sense located 3′ of a stem-loop structure. Each ORF had a potential TATA box and one or two potential polyadenylation signals associated with it and each polyadenylation signal had an associated GC-rich region containing the trinucleotide sequence TTG. A number of ORFs were identified in component 2 but none of these had appropriately located potential TATA boxes and polyadenylation signals associated with them. None of the ORF amino acid sequences nor the full DNA sequences of any of the components had significant sequence identity with any known protein or nucleic acid sequences. However, the ORF of component 4 encoded a 30 residue hydrophobic domain which may indicate that this ORF encoded a trans-membrane protein. Further, the ORFs of components 3 and 5 potentially encoded proteins of about 20 kDa, the size of the BBTV coat protein. There were two regions of sequence identity between the five components described here and the previously described component 1. Each component contained a conserved stem-loop structure and a nonanucleotide potential TATA box which was 5′ of the large virion-sense ORF in five of the components. The stem-loop structures were incorporated in a common region (CR-SL) of 69 nucleotides which was 62% identical between components. All six BBTV components also contained a major common region (CR-M) which was located 5′ of the CR-SL in each component, in the non-coding region and was 76% identical over 92 nucleotides. Each CR-M contained a near-complete 16 nucleotide direct repeat and a GC-box which was similar to the rightward promoter element found in wheat dwarf geminivirus. From these results, BBTV appears to belong to an undescribed plant virus group which could also include subterranean clover stunt virus, coconut foliar decay virus, faba bean necrotic yellows virus and milk vetch dwarf virus.

The genome organization of banana bunchy top virus: analysis of six ssDNA components

Publisher Summary This chapter explores the possible reasons for the negligence of banana bunchy top virus (BBTV), the way in which it has been controlled in Australia, and the possible directions for future research that could reduce the often devastating effects of the disease The economic effects of the disease were as dramatic as its spread In New South Wales, practically no plantation was free of BBTV in the Tweed and Brunswick areas Most attempts to control or eradicate banana bunchy top virus have been made in commercial banana-producing countries In Australia, a control program has successfully contained the disease at a very low level in southeast Queensland and the north coast of New South Wales for about 50 years However, the disease still requires continual monitoring The basis for control came from the discovery that (1) banana bunchy top was caused by a virus, (2) this virus was transmitted by the aphid, P nigronervosa, (3) the virus could be disseminated in infected vegetative planting material, and (4) there were no known non-musaceous alternate hosts For the campaign to be successful, little more information about the virus was required and in the ensuing 60 years little more has been discovered Most research has been confined to verification of the identity of the disease in particular areas, varietal reactions to infection, some epidemiology and more recently isolation of the dsRNAs associated with infection

Banana bunchy top: an economically important tropical plant virus disease.

An effective method has been developed for the stable transformation and regeneration of Cavendish banana (Musa spp. AAA group) cv 'Grand Nain' by microprojectile bombardment. Embryogenic cell suspensions were initiated using immature male flowers as the explant. Cells were co-bombarded with the neomycin phosphotransferase (nptII) selectable marker gene under the control of a banana bunchy top virus (BBTV) promoter or the CaMV 35S promoter, and either the β-glucuronidase (uidA) reporter gene or BBTV genes under the control of the maize polyubiquitin promoter. Plants were regenerated, under selection with kanamycin, that were co-transformed with nptII and either the uidA or BBTV genes. Molecular characterisation of transformants demonstrated that the transgenes had been stably integrated into the banana genome.

/pdf/genetic-transformation-of-cavendish-banana-musa-spp-aaa-jgfxpusxsn.pdf

Genetic transformation of Cavendish banana (Musa spp. AAA group) cv 'Grand Nain' via microprojectile bombardment.

Two pairs of degenerate primers were designed from sequences within the potyviral CI (CIFor/CIRev) and HC-Pro-coding regions (HPFo/HPRev), and these were shown to be highly specific to members of the genus Potyvirus. Using the CIFor and CIRev primers, three novel potyviruses infecting crop and weed species from Vietnam were detected, namely telosma mosaic virus (TelMV) infecting telosma (Telosma cordata, Asclepiadaceae), peace lily mosaic virus (PeLMV) infecting peace lily (Spathiphyllum patinii, Araceae) and wild tomato mosaic virus (WTMV) infecting wild tomato (Solanum torvum, Solanaceae). The fragments amplified by the two sets of primers enabled additional PCR and complete genomic sequencing of these viruses and a banana bract mosaic virus (BBrMV) isolate from the Philippines. All four viruses shared genomic features typical of potyviruses. Sequence comparisons and phylogenetic analyses indicated that WTMV was most closely related to chilli veinal mottle virus (ChiVMV) and pepper veinal mottle virus (PVMV), while PeLMV, TelMV and BBrMV were related to different extents to members of the bean common mosaic virus (BCMV) subgroup.

James L. Dale

Papers

Detection of a nucleic acid sequence or a change therein

The genome organization of banana bunchy top virus: analysis of six ssDNA components

Banana bunchy top: an economically important tropical plant virus disease.

Genetic transformation of Cavendish banana (Musa spp. AAA group) cv 'Grand Nain' via microprojectile bombardment.

Design and application of two novel degenerate primer pairs for the detection and complete genomic characterization of potyviruses