Ruiz-Pérez Carlos A, Zambrano María Mercedes
School of Biological Sciences, Georgia Institute of Technology, Ford Environmental Sciences & Technology Building, Atlanta, USA.
Molecular Genetics, Corporación Corpogen, Bogotá, Colombia.
Bio Protoc. 2017 Feb 20;7(4):e2142. 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.
植物叶际是指植物地上部分的所有组织，它被认为是微生物定殖的最广泛的生态系统之一，这些微生物既可以在植物表面作为附生植物定殖，也可以作为内生植物定殖于植物内部。这些与植物相关的微生物群落是微生物多样性的储存库，对植物健康至关重要。对不同植物（如安第斯山脉帕拉莫生态系统特有的植物）中的微生物群落进行表征，有助于了解微生物之间可能的相互作用以及微生物的功能特性。从植物内生微生物群落中获取DNA需要多个步骤，以确保样本不受植物表面（附生植物）存在的微生物污染。首先对植物叶片进行表面消毒，切成小块，用玻璃珠匀浆，然后使用市售试剂盒进行DNA提取。然后使用Qubit 2.0对DNA样本进行定量和分析，以用于16S rRNA基因的PCR扩增。
