Print PageSaprotrophic fungi are one of the most important groups of organisms involved in the cycling of carbon in terrestrial ecosystems. This cycling of carbon depends upon the production of a complex suite of extracellular enzymes for degrading polymers in plant tissues such as lignin, hemicellulose, cellulose and pectin. Due to genes coding for extracellular enzymes being linked to important functional processes, their presence can be used to analyze microbial community diversity in varying environments. In this study primers were designed targeting the genes coding for pectinases of the Glycoside hydrolase family 28 (GH28) found in fungi. GH28 sequences in fungal genomes were aligned and four potential primer sites were identified. To test the primers DNA was extracted from multiple environmental samples and cultures of two saprotrophic fungal species, Aspergillus niger and Phanerochaete chrysosporium. Environmental samples consisted of leaf litter collected in late April 2010 from Manistee National Forest; MI and included the tree species Quercus velutina, Quercus alba, Acer saccharum and Tilia americana.
Results/Conclusions
Successful PCRs were cloned and sequenced. Using the most successful pair of primers, 1786F and 2089R, A. niger produced amplicons that were identified as GH28 polygalacturonase genes. P. chrysosporium however produced amplicons identified as P. chrysosporium scaffolding genes. Environmental samples from all leaf types produced amplicons of approximately 300 bp, which were characterized as members of the fungal GH28 genes similar to several taxa. These taxa included possible mycorrhizal fungi of the order Sebacinacea and several known plant pathogens, including Botryotinia fuckeliana and Colletotrichum lindemuthianum. The data gathered in this study indicates that the GH28 primers developed are able to amplify genes found in a wide variety of fungal taxa. Based upon this evidence they will be useful in characterizing the functional traits found in a broad range of microbial communities.
