Endophytic Microbial Community DNA Extraction from the Plant Phyllosphere

• Published in: Bio-protocol Vol. 7; no. 4; p. e2142
• Main Authors: Ruiz-Pérez, Carlos A, Zambrano, María Mercedes
• Format: Journal Article
• Language: English
• Published: United States Bio-Protocol 20-02-2017; Bio-protocol LLC
• Subjects: Biology; Methods; 16S rRNA; Metagenomics; Endophyte; Epiphyte; Phyllosphere
• ISSN: 2331-8325; 2331-8325
• DOI: 10.21769/BioProtoc.2142

The plant phyllosphere, which represents all plant parts that are above the ground, is considered one of the most extensive ecosystems to be colonized by microorganisms, both at the surface as epiphytes or as endophytes within the plant. These plant-associated microbial communities are reservoirs of microbial diversity and they can be important for plant health. The characterization of microbial communities in diverse plants, such as
plants that are endemic to the Paramo ecosystem in the Andes Mountains, can shed light regarding possible interactions among microorganisms and microbial functional properties. Obtaining DNA from plant endophytic microbial communities involves various steps to ensure that samples are free of contamination from microorganisms present on the plant surface (epiphytes). Plant leaves are first surface sterilized, cut into pieces, homogenized using glass beads, and then used for DNA extraction using a commercially available kit. DNA samples are then quantified and analyzed using Qubit
2.0 for use in PCR amplification of 16S rRNA genes.