Hall Megan E, Cadle-Davidson Lance, Fang Zhiwei, Wilcox Wayne F
Section of Plant Pathology and Plant-Microbe Biology, School of Integrative Plant Science, Cornell University, NYS Agricultural Experiment Station, Geneva, NY, 14456, United States.
Division of Plant Sciences, University of Missouri, Columbia, MO, 65211, United States.
Heliyon. 2019 Oct 5;5(10):e02597. doi: 10.1016/j.heliyon.2019.e02597. eCollection 2019 Oct.
Isolating DNA from microbes on the surface of a grape berry is a challenge due to their adhesion to the thick berry skin and cuticle, making studies of the grape microbiome challenging. We developed a field-to-lab DNA extraction procedure that starts in the vineyard, disrupts the grape berry surface while en route to the lab through agitation, and efficiently extracts microbial DNA from the surface of the grape. It is cost effective and utilizes commonly available laboratory chemicals with low toxicity (Table 1). This protocol allows researchers to extract DNA from the grape berry surface in the field, therefore undergoing minimal manipulation of those microbiomes before DNA extraction.
从葡萄浆果表面的微生物中分离DNA是一项挑战,因为它们会附着在厚厚的浆果表皮和角质层上,这使得对葡萄微生物组的研究颇具挑战性。我们开发了一种从田间到实验室的DNA提取程序,该程序始于葡萄园,在运往实验室的途中通过搅拌破坏葡萄浆果表面,并能有效地从葡萄表面提取微生物DNA。该方法具有成本效益,且使用的是常见的低毒实验室化学品(表1)。该方案使研究人员能够在田间从葡萄浆果表面提取DNA,因此在DNA提取之前对这些微生物组的操作降至最低。