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This package contains Perl programs/scripts that perform frequently needed operations on FASTA format files.

GRAbB (Genomic Region Assembly by Baiting) is a new program that is dedicated to assemble specific genomic regions from NGS data. This approach is especially useful when dealing with multi copy regions, such as mitochondrial genome and the rDNA repeat region, parts of the genome that are often neglected or poorly assembled, although they contain interesting information from phylogenetic or epidemiologic perspectives, but also single copy regions can be assembled. The program is capable of targeting multiple regions within a single run. Furthermore, GRAbB can be used to extract specific loci from NGS data, based on homology, like sequences that are used for barcoding. To make the assembly specific, a known part of the region, such as the sequence of a PCR amplicon or a homologous sequence from a related species must be specified. By assembling only the region of interest, the assembly process is computationally much less demanding and may lead to assemblies of better quality. In this study the different applications and functionalities of the program are demonstrated such as: exhaustive assembly (rDNA region and mitochondrial genome), extracting homologous regions or genes (IGS, RPB1, RPB2 and TEF1a), as well as extracting multiple regions within a single run. The program is also compared with MITObim, which is meant for the exhaustive assembly of a single target based on a similar query sequence. GRAbB is shown to be more efficient than MITObim in terms of speed, memory and disk usage. The other functionalities (handling multiple targets simultaneously and extracting homologous regions) of the new program are not matched by other programs. The program is available with explanatory documentation at https://github.com/b-brankovics/grabb. GRAbB has been tested on Ubuntu (12.04 and 14.04), Fedora (23), CentOS (7.1.1503) and Mac OS X (10.7). Furthermore, GRAbB is available as a docker repository: brankovics/grabb (https://hub.docker.com/r/brankovics/grabb/).

Sporothrix globosa is a thermo-dimorphic fungus belonging to a pathogenic clade that also includes Sporothrix schenckii, which causes human and animal sporotrichosis. Here, we present the first genome assemblies of two S. globosa strains providing data for future comparative genomic studies in pathogenic Sporothrix species.

In recent years increasing demands and the relatively low-care cultivation of the crop have resulted in an enormous expansion of the acreage of maize in China. However, particularly in China, Fusarium ear rot forms an important constraint to maize production. In this study, we showed that members of both the Fusarium fujikuroi species complex (FFSC) and the Fusarium graminearum species complex are the causal agents of Fusarium ear rot in the main maize producing areas in China. Fumonisin producing Fusarium verticillioides was the most prevalent species, followed by fumonisin producing Fusarium proliferatum and 15-acetyldeoxynivalenol producing F. graminearum. Both Fusarium temperatum and Fusarium boothii were identified for the first time in the colder regions in China, extending their known habitats to colder environments. Mating type analysis of the different heterothallic FFSC species, showed that both types co-occur in each sampling site suggestive of the possibility of sexual recombination. Virulence tests with F. boothii (from maize) and F. graminearum from maize or wheat showed adaptation to the host. In addition, F. graminearum seems to outcompete F. boothii in wheat-maize rotations. Based on our findings and previous studies, we conclude that wheat/maize rotation selects for F. graminearum, while a wheat/rice rotation selects for F. asiaticum. In contrast, F. boothii is selected when maize is cultivated without rotation. A higher occurrence of F. temperatum is observed on maize in colder climatological regions in China, while Fusarium meridionale seems restricted to mountain areas. Each of these species has their characteristic mycotoxin profile and deoxynivalenol and fumonisin are the potential threats to maize production in Northern China.

Sporothrix pallidais considered to be a mostly avirulent environmental fungus, phylogenetically closely related to the well-known pathogenSporothrix schenckii Here, we present the first assembly of its genome, which provides a valuable resource for future comparative genomic studies between nonpathogenic and pathogenicSporothrixspp.

In recent years increasing demands and the relatively low-care cultivation of the crop have resulted in an enormous expansion of the acreage of maize in China. However, particularly in China, Fusarium ear rot forms an important constraint to maize production. In this study, we showed that members of both the Fusarium fujikuroi species complex (FFSC) and the Fusarium graminearum species complex are the causal agents of Fusarium ear rot in the main maize producing areas in China. Fumonisin producing Fusarium verticillioides was the most prevalent species, followed by fumonisin producing Fusarium proliferatum and 15-acetyldeoxynivalenol producing F. graminearum. Both Fusarium temperatum and Fusarium boothii were identified for the first time in the colder regions in China, extending their known habitats to colder environments. Mating type analysis of the different heterothallic FFSC species, showed that both types co-occur in each sampling site suggestive of the possibility of sexual recombination. Virulence tests with F. boothii (from maize) and F. graminearum from maize or wheat showed adaptation to the host. In addition, F. graminearum seems to outcompete F. boothii in wheat-maize rotations. Based on our findings and previous studies, we conclude that wheat/maize rotation selects for F. graminearum, while a wheat/rice rotation selects for F. asiaticum. In contrast, F. boothii is selected when maize is cultivated without rotation. A higher occurrence of F. temperatum is observed on maize in colder climatological regions in China, while Fusarium meridionale seems restricted to mountain areas. Each of these species has their characteristic mycotoxin profile and deoxynivalenol and fumonisin are the potential threats to maize production in Northern China.

The occurrence resistance to methyl benzimidazole carbamates (MBC)-fungicides in the Fusarium graminearum species complex (FGSC) is becoming a serious problem in the control of Fusarium head blight in China. The resistance is caused by point mutations in the ß2-tubulingene. So far, five resistant genotypes (F167Y, E198Q, E198L, E198K and F200Y) have been reported in the field. To establish a high-throughput method for rapid detection of all the five mutations simultaneously, an efficient single-nucleotide-polymorphism-based genotyping method was developed based on the Luminex xMAP system. One pair of amplification primers and five allele specific primer extension probes were designed and optimized to specially distinguish the different genotypes within one single reaction. This method has good extensibility and can be combined with previous reported probes to form a highly integrated tool for species, trichothecene chemotype and MBC resistance detection. Using this method, carbendazim resistant FGSC isolates from Jiangsu, Anhui and Sichuan Province in China were identified. High and moderate frequencies of resistance were observed in Jiangsu and Anhui Province, respectively. Carbendazim resistance in F. asiaticum is only observed in the 3ADON genotype. Overall, our method proved to be useful for early detection of MBC resistance in the field and the result aids in the choice of fungicide type.

Infections caused by the genus Fusarium have emerged over the past decades and range from onychomycosis and keratitis in healthy individuals to deep and disseminated infections with high mortality rates in immune-compromised patients. As antifungal susceptibility can differ between the different Fusarium species, identification at species level is recommended. Several clinical observations as hyaline hyphae in tissue, necrotic lesions in the skin and positive blood tests with fungal growth or presence of fungal cell wall components may be the first hints for fusariosis. Many laboratories rely on morphological identification, but especially multi-locus sequencing proves better to discriminate among members of the species complexes involved in human infection. DNA-based diagnostic tools have best discriminatory power when based on translation elongation factor 1-α or the RNA polymerase II second largest subunit. However, assays based on the detection of other fusarial cell compounds such as peptides and cell wall components may also be used for identification. The purpose of this review is to provide an overview and a comparison of the different tools currently available for the diagnosis of fusariosis.

GRAbB (Genome Region Assembly by Baiting) is program designed to assemble selected regions of the genome or transcriptome using reference sequences and NGS data.

The structure of the Fusarium culmorum mitogenome is similar to that of closely related Fusarium spp.: it has a total length of 103,844 bp, the base composition of the genome is the following: A (35.4%), T (32.9%), C (14.6%), and G (17.1%). The mitogenome contains 13 protein-coding genes, 2 ribosomal RNA (rRNA), and 28 transfer RNA (tRNA) genes, all coded on the same strand of DNA. The gene order is identical to that of the other Fusarium and Hypocreales mitogenomes. Maximum likelihood and Bayesian analysis based on the concatenated amino acid dataset of mitochondrial protein-coding genes confirmed close genetic relationship of F. culmorum to the other type B trichothecene producers F. graminearum and F. gerlachii.

Abstract The structure of the Fusarium gerlachii mitogenome is similar to that of closely related Fusarium graminearum; it has a total length of 93,428 bp, the base composition of the genome is: A (35.3%), T (32.8%), C (14.7%) and G (17.2%). The mitogenome contains 13 protein-coding genes, 2 ribosomal RNA (rRNA) and 28 transfer RNA (tRNA) genes. The tRNA genes range in size from 62 bp to 88 bp. The gene order is identical to that of the other Fusarium mitogenomes.