Rapid, field-deployable method for collecting and preserving plant metabolome for biochemical and functional characterization

Abstract

Study of plant metabolome is a growing field of science that catalogs vast biochemical and functional diversity of phytochemicals. However, collecting and storing samples of plant metabolome, sharing these samples across the scientific community and making them compatible with bioactivity assays presents significant challenges to the advancement of metabolome research. We have developed a RApid Metabolome Extraction and Storage (RAMES) technology that allows efficient, highly compact, field-deployable collection and storage of libraries of plant metabolome. RAMES technology combines rapid extraction with immobilization of extracts on glass microfiber filter discs. Two grams of plant tissue extracted in ethanol, using a specially adapted Dremel® rotary tool, produces 25–35 replicas of 10 mm glass fiber discs impregnated with phytochemicals. These discs can be either eluted with solvents (such as 70% ethanol) to study the metabolomic profiles or used directly in a variety of functional assays. We have developed simple, non-sterile, anti-fungal, anti-bacterial, and anti-oxidant assays formatted for 24-multiwell plates directly compatible with RAMES discs placed inside the wells. Using these methods we confirmed activity in 30 out of 32 randomly selected anti-microbial medicinal plants and spices. Seven species scored the highest activity (total kill) in the anti-bacterial (bacteria from human saliva) and two anti-fungal screens (Fusarium spp. and Saccharomyces cerevisiae), providing functional validation of RAMES technology. RAMES libraries showed limited degradation of compounds after 12 months of storage at -20°C, while others remained stable. Fifty-eight percent of structures characterized in the extracts loaded onto RAMES discs could be eluted from the discs without significant losses. Miniaturized RAMES technology, as described and validated in this manuscript offers a labor, cost, and time-effective alternative to conventional collection of phytochemicals. RAMES technology enables creation of comprehensive metabolomic libraries from various ecosystems and geographical regions in a format compatible with further biochemical and functional studies.

Metabolomics Study of Different Germplasm Resources for Three Polygonatum Species Using UPLC-Q-TOF-MS/MS

“…The metabolites of identical plants can vary when grown in different environments, for different organs, different producing areas, or through various processing methods ( Souza et al, 2019 ). Consequently, the quality of the medicinal components of the Polygonatum species (particularly in terms of their chemical compositions) is uneven due to the mixed germplasm, transregional introduction, and diverse planting and processing procedures ( Skubel et al, 2018 ). Moreover, traditional classification methods such as character ( Xu et al, 2018 ), microscopic ( Boiteau et al, 2018 ), and optical identification ( Ullah et al, 2017 ), cannot directly reflect the nuances in their chemical compositions.…”

Comparative Characteristics of Antibacterial Activity of Plants Growing in the Central Part of the Republic of Tajikistan

“…Preparation of plant extract, filter discs and nutrient agar to determine antibacterial activity. Working concentration and paper discs were prepared according to the method developed at Rutgers University [18]. Briefly, two grams of the plant material was weighed and cut into small pieces with a knife.…”

The Potential Use of Isothermal Amplification Assays for In-Field Diagnostics of Plant Pathogens

“…Rotorgrinding devices with blades such as food processors can homogenize even firm samples. The available grinding devices include portable models with disposable sample-grinding containers (see Dremel, Robert Bosch Gmbh, as an example) [29]. Homogenization of lignified tissue from wood samples is the most difficult to pre-treat.…”

“…However, these tools are fit only for sawn-off fragments. Bark grinding with homemade [36] or commercial [29] tools make it possible to test living trees. An increment borer enables wood samples to be obtained from living trees which can be homogenized by grinders or mortars [37,38].…”