89 research outputs found

    Transposon-mediated gene search: finding a needle in a haystack

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    Sharing mutants and experimental information prepublication using FgMutantDb (https://scabusa.org/FgMutantDb)

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    There is no comprehensive storage for generated mutants of Fusarium graminearum or data associated with these mutants. Instead researchers relied on several independent and non-integrated databases. FgMutantDb was designed as a simple spreadsheet that is accessible globally on the web that will function as a centralized source of information on F. graminearum mutants. FgMutantDb aids in the maintenance and sharing of mutants within a research community. It will serve also as a platform for disseminating prepublication results as well as negative results that often go unreported. Additionally, the highly curated information on mutants in FgMutantDb will be shared with other databases such as FungiDB, Ensembl, PhytoPath, and PHI-base, through updating reports. Here we describe the creation and potential usefulness of FgMutantDb to the F. graminearum research community, and provide a tutorial on its use. This type of database could be easily emulated for collaborating and tracking research generated mutants in other fungal species

    Sequencing of 15 622 Gene-bearing BACs Clarifies the Gene-dense Regions of the Barley Genome

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    Barley (Hordeum vulgare L.) possesses a large and highly repetitive genome of 5.1 Gb that has hindered the development of a complete sequence. In 2012, the International Barley Sequencing Consortium released a resource integrating whole-genome shotgun sequences with a physical and genetic framework. However, because only 6278 bacterial artificial chromosome (BACs) in the physical map were sequenced, fine structure was limited. To gain access to the gene-containing portion of the barley genome at high resolution, we identified and sequenced 15 622 BACs representing the minimal tiling path of 72 052 physical-mapped gene-bearing BACs. This generated ~1.7 Gb of genomic sequence containing an estimated 2/3 of all Morex barley genes. Exploration of these sequenced BACs revealed that although distal ends of chromosomes contain most of the gene-enriched BACs and are characterized by high recombination rates, there are also gene-dense regions with suppressed recombination. We made use of published map-anchored sequence data from Aegilops tauschii to develop a synteny viewer between barley and the ancestor of the wheat D-genome. Except for some notable inversions, there is a high level of collinearity between the two species. The software HarvEST:Barley provides facile access to BAC sequences and their annotations, along with the barley–Ae. tauschii synteny viewer. These BAC sequences constitute a resource to improve the efficiency of marker development, map-based cloning, and comparative genomics in barley and related crops. Additional knowledge about regions of the barley genome that are gene-dense but low recombination is particularly relevant

    A single backcross effectively eliminates agronomic and quality alterations caused by somaclonal variation in transgenic barley

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    Transgenic crops have proven commercial utility but are created using processes known to produce undesirable variability known as somaclonal variation. This reduces the utility of transgenic germplasm to the plant breeder and complicates assessments of transgene-encoded phenotypes. Backcrossing transgenes into a wild-type genome is one solution, but producing near-isogenic lines requires a lengthy and resource-intensive process of multiple crosses. However, an abbreviated breeding scheme involving a single backcross to the wild-type parent used to produce a transgenic line, which would replace 75% of the variant alleles, should produce transgenic lines with improved performance. Comparisons were made of 'Conlon' barley (Hordeum vulgare L.), primary transgenic lines derived from Conlon, and lines derived from single backcrosses of primary transgenic lines to Conlon. The primary transgenic lines were different from Conlon for many agronomic and malting characteristics. Most of the backcross-derived lines did not differ significantly from Conlon for most agronomic characteristics. The backcross-derived lines were also similar to Conlon for malting quality traits but showed more differences than for agronomic characteristics. Differences between lines encoding TRI101 versus lines encoding PDR5 suggested that PDR5 insertion or expression may have affected malting quality. It is concluded that a single backcross is an effective, rapid, and inexpensive method for creating superior transgenic lines
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