Chapter 19 : Analysis of Mating-Type Locus Organization and Synteny in Mushroom Fungi: Beyond Model Species

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In this chapter genes that are in or near the locus but do not function in determining mating-type specificity are referred to as non-mating-type MAT-linked genes. When trying to study the genes of mushroom fungi other than and , there are three major obstacles. First, the loci are likely to be complex and may comprise several genes. Second, the loci display high sequence variation among alleles and cannot be cloned by heterologous hybridization using probes from or . Finally, the absence of a transformation system in which to test the cloned fragments prevents definitive proof that an isolated DNA region carries . The chapter focuses on how MAT genes may be cloned in nonmodel mushroom species. Although most of this chapter has focused on how conserved gene order can be used to clone from nonmodel species, a more direct option is through PCR amplification of the homologues themselves. The authors have targeted the few conserved amino acids of the homeodomain region (HD1) by degenerate PCR and isolated the entire HD1 gene from this species by genome walking using cassette-mediated PCR.

Citation: James T. 2007. Analysis of Mating-Type Locus Organization and Synteny in Mushroom Fungi: Beyond Model Species, p 317-331. In Heitman J, Kronstad J, Taylor J, Casselton L (ed), Sex in Fungi. ASM Press, Washington, DC. doi: 10.1128/9781555815837.ch19
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Figure 19.1

Phylogeny of the Basidiomycota. The cladogram depicts the current knowledge of the relationships among the fungi discussed in the text. Data are derived from references , and , the Tree of Life Web project (http://tolweb.org/tree/phylogeny.html), and the Web project (http://mor.clarku.edu/).

Citation: James T. 2007. Analysis of Mating-Type Locus Organization and Synteny in Mushroom Fungi: Beyond Model Species, p 317-331. In Heitman J, Kronstad J, Taylor J, Casselton L (ed), Sex in Fungi. ASM Press, Washington, DC. doi: 10.1128/9781555815837.ch19
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Image of Figure 19.2
Figure 19.2

Schematic showing conserved gene order of the chromosomal region surrounding in mushroom fungi. Arrows indicate genes and their direction of transcription. Vertical lines connect homologous genes between species. Genes with vertical stripes or horizontal stripes represent homeodomain-type-1 () genes and homeodomain-type-2 () genes, respectively ( ). Genes in gray are found in all four species, and genes in black are restricted to a single species. This map includes a number of additional genes for that were missed in the previous annotation of the region ( ). , gene encoding a conserved hypothetical protein.

Citation: James T. 2007. Analysis of Mating-Type Locus Organization and Synteny in Mushroom Fungi: Beyond Model Species, p 317-331. In Heitman J, Kronstad J, Taylor J, Casselton L (ed), Sex in Fungi. ASM Press, Washington, DC. doi: 10.1128/9781555815837.ch19
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Figure 19.3

Cosegregation of and - in . Shown is an agarose gel (2%) of amplicons digested with MseI. In lane P is the parental dikaryon (Guy.21.2) that is heterozygous at the locus. The other lanes show the progeny of Guy.21.2. Lanes are of the mating type and possess a allele that lacks an MseI cut site. Lanes are of the mating type and possess the MseI cut site that digests the 297-bp amplicon into two fragments of 148 and 149 bp. Lanes marked M contain DNA marker.

Citation: James T. 2007. Analysis of Mating-Type Locus Organization and Synteny in Mushroom Fungi: Beyond Model Species, p 317-331. In Heitman J, Kronstad J, Taylor J, Casselton L (ed), Sex in Fungi. ASM Press, Washington, DC. doi: 10.1128/9781555815837.ch19
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Generic image for table
Table 19.1

Linkage of and to and in basidiomycetes

Citation: James T. 2007. Analysis of Mating-Type Locus Organization and Synteny in Mushroom Fungi: Beyond Model Species, p 317-331. In Heitman J, Kronstad J, Taylor J, Casselton L (ed), Sex in Fungi. ASM Press, Washington, DC. doi: 10.1128/9781555815837.ch19
Generic image for table
Table 19.2

Non-mating-type linked genes in basidiomycetes that encode proteins targeted to mitochondria

Citation: James T. 2007. Analysis of Mating-Type Locus Organization and Synteny in Mushroom Fungi: Beyond Model Species, p 317-331. In Heitman J, Kronstad J, Taylor J, Casselton L (ed), Sex in Fungi. ASM Press, Washington, DC. doi: 10.1128/9781555815837.ch19

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