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Isolation, Characterization, and Total DNA Extraction to Identify Endophytic Fungi in Mycoheterotrophic Plants

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Mycobiome detection from a single subterranean gametophyte using metabarcoding techniques.

Ko-Hsuan Chen¹, Qiao-Yi Xie^1,2, Chiung-Chih Chang¹

¹Biodiversity Research Center, Academia Sinica Taipei Taiwan.

Applications in Plant Sciences

|May 2, 2022

Summary

This study reveals the diverse fungal communities (mycobiomes) within the achlorophyllous gametophytes of the epiphytic fern Ophioderma pendulum. The research establishes a new workflow for investigating these unique plant-microbe interactions.

Keywords:

Ophioderma Ophioglossaceae fungi gametophytes metabarcoding mycobiome

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Metagenomic Analysis of Silage

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Published on: January 13, 2017

Area of Science:

Plant Science
Microbiology
Mycology

Background:

Ferns and lycophytes, such as Ophioglossaceae and Lycopodiaceae, possess subterranean, achlorophyllous gametophytes that rely on fungal symbionts for nutrition.
Despite advances in high-throughput sequencing, the microbiomes of these inconspicuous gametophytes remain understudied.
Factors influencing the fungal communities (mycobiomes) of fern gametophytes require deeper investigation.

Purpose of the Study:

To develop and present a high-throughput metabarcoding workflow for analyzing the mycobiome of achlorophyllous fern gametophytes.
To characterize the fungal communities associated with the gametophytes of the epiphytic fern Ophioderma pendulum.
To provide methodological insights and precautions for studying fern gametophyte mycobiomes.

Main Methods:

A high-throughput metabarcoding approach was employed to study the mycobiome of Ophioderma pendulum gametophytes.
Nuclear ITS and small subunit ribosomal rDNA primers were used to target general fungi and arbuscular mycorrhizal fungi.
Microscopic examination was conducted concurrently to identify fungal structures within the gametophytes.

Main Results:

Both microscopic and metabarcoding analyses revealed a high diversity of fungi colonizing individual Ophioderma pendulum gametophytes.
The study successfully uncovered the mycobiome composition of an achlorophyllous gametophyte from an epiphytic fern species.
The applied workflow demonstrated efficacy in detecting diverse fungal inhabitants.

Conclusions:

The study establishes a robust methodology for exploring the mycobiomes of achlorophyllous gametophytes.
This research offers the first comprehensive insight into the mycobiome assembly of an epiphytic fern's achlorophyllous gametophyte.
The findings highlight the complex fungal associations crucial for the survival of these unique plant structures.