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Becker, Lindsey [1], Cubeta, Marc A. [2].

Elucidating the seed mycobiome of four wheat cultivars: a culture-based approach.

The occurrence of pathogenic fungal taxa associated with wheat (Triticum aestivum L.) seeds is well studied, but less is known about beneficial taxa of the wheat seed mycobiome. The goal of our research is to characterize wheat seed fungal endophyte and ectophyte diversity via a culture-based approach. Four publicly available wheat cultivars developed in the southeastern United States with varying phenotypic and disease resistance traits were examined: Hilliard, Shirley, Catawba, and USG3640. We hypothesize that there will be greater alpha diversity of fungi on the exterior of the wheat seed (ectophytes) compared to fungi residing inside wheat seeds (endophytes). To isolate ectotrophic fungi, wheat seeds were immersed in sterile water and vortexed gently for 60 seconds. A 1:100 diluted suspension was transferred to Malt Yeast Extract Agar (MYEA) amended with streptomycin and tetracycline (50 mg/L each), and cyclosporin (4 mg/L). Preliminary experiments suggest that cyclosporin reduces individual fungal colony growth, thus preventing rapid overgrowth that can lead to lower diversity estimates for culture-based approaches. To isolate endophytic fungi, wheat seeds were sequentially submerged in 95% EtOH for 60 seconds, half-strength household bleach (3% active NaOCl) for 2 minutes, 95% EtOH for 30 seconds, sterile water for 30 seconds, followed by a final rinse in sterile water. Post surface sterilization, seeds were air dried, macerated, and sieved to obtain < 177 ┬Ám fragments, followed by plating on amended MYEA media as described above. By utilizing these methods, we are developing a catalog of fungal taxa associated with the wheat seed exterior and interior. Comparison of fungal taxa associated with each wheat cultivar will allow for greater understanding of the wheat seed mycobiome and diversity. The culture library generated by this study will help validate amplicon sequencing of wheat seeds and contribute to the design of synthetic microbial communities.

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1 - North Carolina State University, Department of Entomology and Plant Pathology, 851 Main Campus Drive, 263 Partners III, Raleigh, NC, 27606, USA
2 - North Carolina State University, Entomology and Plant Pathology, Center for Integrated Fungal Research, Raleigh, NC, 27606, USA

Seed Mycobiome

Presentation Type: Oral Paper
Session: MY6, Mycology: Systematics and Biodiversity
Location: /
Date: Thursday, July 22nd, 2021
Time: 11:00 AM(EDT)
Number: MY6005
Abstract ID:494
Candidate for Awards:MSA Best Oral Presentation Award by a Graduate Student

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