Abstract
Truffle fungi are esteemed for their aromatic qualities and are among the most widely cultivated edible ectomycorrhizal fungi. Here we document a successful method for establishing Tuber lyonii, the pecan truffle, on pecan (Carya illinoinensis) seedlings in a field setting. We assessed the impacts of soil fumigation and varying concentrations of truffle spore inoculum on the ectomycorrhizal fungal and the complete fungal communities as well as the colonization of T. lyonii on pecan roots at three nurseries in Georgia, United States. To identify fungal communities on pecan seedlings, we performed high-throughput amplicon sequencing of the fungal ITS1 rDNA region. Our 5-year long field experiment demonstrates that fumigation and inoculation together resulted in the highest persistence of T. lyonii on pecan roots. While fungal OTU numbers fluctuated over the years of our experiments, there was no statistical support to demonstrate diversification of communities when Shannon diversity metrics were used. However, we did find that older seedlings were less likely to be dominated by T. lyonii compared to younger ones, suggesting successional changes in the fungal community over time. This suggests that transplanting inoculated seedlings after 2 or 3 years post-inoculation is optimal for future truffle propagation efforts. Our results demonstrate that T. lyonii can be established in situ with methods that are compatible with current pecan nursery industry practices and that fungal communities on pecan seedlings vary depending on the experimental treatments used during planting. While the pecan truffle is not yet widely cultivated, our results provide insights for future large-scale cultivation of this and perhaps other Tuber species.
Highlights
Truffle fungi are characterized by closed sporocarps in which the fertile, spore-bearing layer is totally encased by sterile tissue at maturity
For all fungi we recovered a total of 10,070,714 reads from our Illumina MiSeq runs after data filtering
We identified a total of 47 ECM fungi operational taxonomic units (OTUs) of the 1,322 total fungi OTUs (Supplementary Table 2)
Summary
Truffle fungi are characterized by closed sporocarps in which the fertile, spore-bearing layer is totally encased by sterile tissue at maturity. The use of animals for dispersal is facilitated by volatile organic compounds (VOCs) released by truffle fruiting bodies through the activity of endogenous bacterial communities at maturity (Splivallo et al, 2015). Truffles in the genus Tuber (Tuberaceae, Ascomycota), release VOCs that are gastronomically enticing and may mimic animal pheromones (Claus et al, 1981; Splivallo et al, 2011). Members of the genus Tuber grow in a mutually beneficial symbiotic relationship, known as ectomycorrhizae (ECM), with the roots of certain host trees (Bonito et al, 2010; Tedersoo and Smith, 2013). The fungus receives carbohydrates from the host and in exchange, the mycobiont provides greater access to water, essential nutrients, added protection from fungal root pathogens and stress from drought (Smith and Read, 2008)
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