Characterizing the Conserved Wheat Seed Mycobiome: A Synergistic Integration of Culture-Dependent and Culture-Independent Methods

This research project aims to elucidate the diversity of fungal endophytes associated with wheat (Triticum aestivum L.) seeds, with a focus on non-pathogenic taxa that have been less explored in previous studies. Leveraging a combination of culture-dependent and culture-independent approaches, our study provides a comprehensive analysis of the wheat seed mycobiome. The study examined four winter wheat cultivars developed in the southeastern United States, chosen for their distinct phenotypic characteristics and disease resistance traits: Catawba, Hilliard, Shirley, and USG 3640. These cultivars were studied over a two-year period to assess the diversity and stability of their associated fungal communities.Our culture-dependent analysis utilized two different nutrient media to isolate and identify fungal taxa, yielding 645 fungal isolates across twelve genera. This methodological approach allowed for the detailed characterization of specific fungal taxa associated with the wheat seeds. Complementarily, metabarcoding analysis provided a broader view of the mycobiome, uncovering a wider range of fungal taxa and a higher number of unique sequences than what was observed through culture-dependent techniques alone.Notably, our findings indicate a reduction in fungal diversity in the year 2021, observed through both culture-dependent and independent methods. Despite this, the overall structure of the wheat seed fungal community exhibited remarkable stability across different cultivars and throughout the study period. This stability underscores the conserved nature of the wheat seed mycobiome, highlighting its potential resilience to temporal changes.The integration of culture-dependent and culture-independent methodologies has proven essential in capturing the full spectrum of fungal diversity. This synergistic approach not only enriches our catalog of wheat seed-associated fungal endophytes but also identifies areas for future research, particularly in enhancing culture-dependent techniques to further explore the wheat seed mycobiome.Our project contributes significant new insights into the complex interactions within the wheat seed ecosystem, offering valuable information for agricultural science, mycology, and crop improvement strategies. The data generated from this study have been submitted to the Sequence Read Archive (SRA), providing a valuable resource for researchers interested in the fungal biodiversity associated with crop seeds.