Abstract
The fungus Stachybotrys (S.) chartarum was isolated from culinary herbs, damp building materials, and improperly stored animal forage. Two distinct chemotypes of the fungus were described that produced either high-cytotoxic macrocyclic trichothecenes (S type) or low-cytotoxic atranones (A type). Recently, two distinct gene clusters were described that were found to be necessary for the biosynthesis of either macrocyclic trichothecenes (21 SAT (Satratoxin) genes) or atranones (14 ATR (Atranone) genes). In the current study, PCR primers were designed to detect SAT and ATR genes in 19 S. chartarum chemotype S and eight S. chartarum chemotype A strains. Our analysis revealed the existence of three different genotypes: satratoxin-producing strains that harbored all SAT genes but lacked the ATR gene cluster (genotype S), non-satratoxin-producing strains that possessed the ATR genes but lacked SAT genes (genotype A), and a hitherto undescribed hybrid genotype among non-satratoxin-producing strains that harbored all ATR genes and an incomplete set of SAT genes (genotype H). In order to improve the discrimination of genotypes, a triplex PCR assay was developed and applied for the analysis of S. chartarum and S. chlorohalonata cultures. The results show that genes for macrocyclic trichothecenes and atranones are not mutually exclusive in S. chartarum. Correlation of the new genotype-based concept with mycotoxin production data shows also that macrocyclic trichothecenes are exclusively produced by S. chartarum genotype S strains.
Highlights
Stachybotrys (S.) spp. were detected on dead plant materials and other cellulosic substratesSebastian Ulrich and Ludwig Niessen contributed to this work.(Biermaier et al 2015; El-Kady and Moubasher 1982)
Since S. chartarum strains cannot be further differentiated into chemotypes by this method (Ulrich et al 2016), the production of macrocyclic trichothecenes was analyzed to assign chemotypes to the respective strains
Further analysis revealed that all strains with a thymidine (T) at nucleotide position 279 of the TRI5 gene (S. chartarum type A) did not produce macrocyclic trichothecenes (Andersen et al 2003)
Summary
Stachybotrys (S.) spp. were detected on dead plant materials (e.g., herbs, straw, and hay) and other cellulosic substratesSebastian Ulrich and Ludwig Niessen contributed to this work.(Biermaier et al 2015; El-Kady and Moubasher 1982). Stachybotrys (S.) spp. were detected on dead plant materials (e.g., herbs, straw, and hay) and other cellulosic substrates. Gypsum materials as well as wall papers (Andersen et al 2011). On these materials, Stachybotrys spp. were associated with other fungi, i.e., Acremonium spp., Ulocladium spp., and Penicillium chrysogenum. S. chartarum and S. chlorohalonata were found to be the most frequently isolated species of the genus Stachybotrys (Wang et al 2015; Lombard et al 2016). The former species was sub-divided into two distinct chemotypes, the high-cytotoxic chemotype S and the low-cytotoxic chemotype A. Cultures of the two chemotypes were found to produce either macrocyclic trichothecenes (S type) or atranones (A type) (Jarvis et al 1995; Andersen et al 2003; Hinkley et al 2000, 2003)
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