A recent paper describes the discovery of 9 new introns in Saccharomyces cerevisiae by Ron Davis’s group at Stanford, using high density tiling arrays from Affymetrix. The arrays are designed for both strands allow the detection of transcripts transcribed from both strands. The arrays were also put to work by the Davis and Steinmetz labs to create a high density map of transcription in yeast and for polymorphism mapping from the Kruglyak lab.
Whole genome tiling arrays have also been employed in other fungi. For example, Anita Silâ€™s group at UCSF constructed a random tiling array for Histoplasma capsulatum and used it to identify genes responding to reactive nitrogen species. A similar approach was used in Cryptococcus neoformans to investigate temperature regulated genes using random sequencing clones.
As the technology has become cheaper, it may become sensible to use a tiling array to detect transcripts rather than ESTs when attempting to annotate a genome. In the Histoplasma work transcriptional units could be identified from hybridization alone. Some of the algorithms will need some work to correct incorporate this information, and the sensitivity and density of the array will influence this. These techniques can be part of a resequencing approaches or fast genotyping progeny from QTL experiments when the sequence from both parents is known (or at least enough of the polymorphims for the genetic map).
What is superior about the current Affymetrix yeast tiling array is the inclusion of both strands. This allows detection of transcripts from both strands. Several anti-sense transcripts in yeast have been discovered recently including in the IME4 locus through more classical approaches, but perhaps many more await discovery with high resolution transcriptional data from whole genome tiling arrays.
It seems intuitive enough that the size of an organism’s genome should be related to its evolutionary complexity. As a general rule, this tends to be true. But look within a class of organisms and you’ll find a great deal of genome size – also known as a C-value – variation. A newt’s genome, for example, is ten times the size of a frogâ€™s.
This discrepancy between genome size and evolutionary complexity is known as the C-value paradox and it has long captured the imagination of biologists. Genome sequencing and annotation have revealed that a great amount of an organism’s genome is non-coding, suggesting that a great deal of genetic content may be gained or lost without affecting the so-called “evolutionary complexity” of the organism (though whether this non-coding DNA is truly “junk” is still up for debate).
In a recent Nucleic Acids Research paper, Gregory et al introduce another toolset to aid in our understand of genome size: the genome size databases. Three separate databases catalog the genome size statistics for various Plants, Animals and Fungi respectively, collectively covering >10,000 species. While various methods of estimating genome size may produce conflicting estimates of genome size (caveat emptor!), these tools should serve to help guide analyses and experiments of genome size evolution. Specifically, by enabling comparisons of genome size across multiple phylogenetic levels, these datasets should facilitate a better understanding of where the genome size/complexity relationship falls off.
As an interesting side note, the authors mention a few particular findings in fungi. The histogram of genome size in Fungi (see the figure) tends to be tighter than in Plants and Animals, with almost all taxa within the range of 1C or 10-60 Mb of DNA. That said, a few species appear to exhibit considerable intraspecific variation. While this may be due to the aforementioned methodological errors, the authors consider that dikaryotic hybrids and heterokaryotes may contribute to this observation. It seems that we may only be scratching the surface of genome size variation in Fungi and if genome size is indeed rapidly evolving in Fungi, they may serve to as good models to study this evolutionary phenomenon.
A paper in Nature this week describes how a few mutations can alter the interactions between species in a biofilm from competitive to cooperative system. This is a great study that goes from start to finish on studying community interactions, looking at an evolved phenotype, and understanding the genetic and physiological basis for the adaptation.
Acinetobacter sp. and Pseudomonas putida were raised in a carbon-limited environment with only benzyl alcohol as the carbon source. Acinetobacter can processes the benzyl alcohol, while P. putida is unable to. Acinetobacter takes up the bezyl alcohol and secretes benzoate that P. putida can then use as a carbon source. The research group propagated these in chemostats and looked at different starting concentrations of the organisms. They found that evolved P. putida had a different morphology and did several experiments to determine the relative fitness of the derived and ancestral genotype.
They went on to also map the mutations in P. putida and found two independent mutations in wapH (I think this is the right gene)â€”a gene involved in lipopolysaccharide (LPS) biosynthesis. They then engineered the ancestral strain to have a mutation in P. putida and found the rough colony phenotype morphology indistinguishable from the strain derived from experimental evolution.
There are various evolutionary and niche adaptation implications arising from this study. One application to mycology is to how lichens evolved in that an algael cell and a fungal cell must communicate and cooperate.
The current contributors to this blog are
Jason Stajich maintains this blog site, a wiki for collaboration, and software and is an Assistant Professor University of California, Riverside in the Department of Plant Pathology and Microbiology and Institute for Integrative Genome Biology. He also provides some genome browsers for fungal genomes as part of his research and in collaboration with the community.
Thomas Sharpton is a postdoctoral fellow at University of California, San Francisco in the Gladstone Institute
Chris Villalta, grad student at UC Berkeley
Balaji Rajashekar was previously at Lund University
We welcome other participants. If you would like to contribute content to this site or to our wiki, please sign up for an account and contact Jason by email.
In a recent Microbiology Mini-Review, Meriel Jones catalogs both the potential benefits and problems that arise from fungal genome sequencing. Using the nine genomes (being) sequenced from the Aspergillus clade, Jones addresses several issues tied to a singular theme: if we are to unlock the potential that fungal genome sequencing holds, both academically and entrepreneurially, then a more robust infrastructure that enables comparative and functional annotation of genomes must be established.
Fortunately, like any good awareness advocate, Jones points us in the direction of e-Fungi, a UK based virtual project aimed at setting up such an infrastructure. Anyone can navigate this database to either compare the stored genomic information or evaluate any fungus of interest in the light of the e-Fungi genomic data. The data appears to be precomputed, similar to IMG from JGI, so there are inherent limitations on the data that one can obtain. However, tools such as these put important data in the hands of expert mycologists that can turn the information into something biologically meaningful.
As Jones points out, this is just the beginning. If fungal genomes are to live up to their promise, they must engage more than just experts at reading genomes.
The JGI has previously released A. niger strain ATCC 1015 sequence in November 2005. ATCC 1015 is used in industrial production of citric acid as it is one of the best producers of citric acid. In Nature Biotechnology a Dutch team has published the sequence of another strain, CBS 513.88 which is an early ancestor of ATCC 1015 used in industrial enzyme production.
The Baylor sequencing center has published the genome of two honey bee pathogens. Recently Baylor and collaborators published a slew of honey bee genome papers and it is great that they have also chosen to follow up on the parasites as well.
The group published the genomes of the bacteria pathogen Paenibacillus larvae and fungal pathogen Ascosphaera apis. A. apis is in the Onygenales clade which also includes the fungal human pathogens Coccidioides, Histoplasma, and Blastomyces.
Currently the genome annotation is limited to the bacterial genome where many ab initio gene prediction programs exist and no annotation is provided for A. api. We should be able to apply gene prediction parameters trained from other Onygenales fungi to get a resonable annotation. Study of this pathogenic genome may also provide insight into the evolution of this clade of fungi which contains most of the primary fungal pathogens of humans.
Ants, fungi, and bacteria
I have to admit that I am fascinated by co-evolution of symbiotic and mutalistic systems. A review by Richard Robinson gives an overview. A great example is the mutalism between ants and fungi where the ants cultivate the fungi for food. There are more layers to the relationship as a fungal parasite (Escovopsis) attacks the cultivated fungi, and a bacteria. Several researchers have studied the coevolution of these studies including Ulrich Mueller and Cameron Currie. Currie and Mueller have published several great studies describing the patterns of coevolution and the nature of the cooperation.
Continue reading Tripartate symbioses with fungi
The FGI and the Broad Institute have released the 7X genome assembly of Puccinia graminis f. sp tritici in roughly 4500 contigs. This represents the first rust fungus to be sequenced and the second Urediniomycete that has been sequenced, Sporobolomyces roseus being the first. This rust fungus is “the causal agent of stem rust, has caused serious disease of small cereal grains (wheat, barley, oat, and rye) worldwide.”
Sally Otto and colleagues have identified that populations of laboratory yeast strains convereged on diploidy in this study. This is nicely consistent with the observation that most wild strains isolated from the environment are diploid.
For example Robert Mortimer describes some properties of wild wine strains and most are found to be diploid.