Faculty
Science
Supervisor Name
Dr. Mark Bernards
Keywords
ginseng, GRD, ginseng replant disease, DNA extraction, sandy-loam, soil
Description
Ginseng is a high value perennial crop grown for its roots. A four-year cultivation cycle is required to obtain marketable ginseng roots, during which a condition called ginseng replant disease (GRD) develops. GRD is characterized by a severe root rot disease, and prevents the growth of newly planted ginseng in a former ginseng garden1. Microbiome analysis of ginseng garden soil will help our understanding of the formation of GRD and its severity. However, extracting DNA from the sandy-loam soils required for ginseng cultivation is difficult. Commercial kits are either too expensive or not applicable to the large volumes of soil required to extract a suitable quality and quantity of DNA for Illumina Miseq next generation sequencing (NGS). Therefore, an optimized DNA extraction protocol is essential to identifying microbes that contribute to GRD. In this study, a DNA extraction method based on Method 6b from Kamble and Singh (2020)2 was used as the basis for an optimized protocol. The ZymoBIOMICS DNA Miniprep kit3 was used for comparison.
Acknowledgements
Funding acknowledgement: Western Undergraduate Summer Research Internship (WUSRI) and Natural Sciences and Engineering Research Council (NSERC).
I would like to thank Andrew Rabas, Dr. Dimitre Ivanov, Paul Wan, Dr. Greg Thorn, and Dr. Vera Tai for support on this project
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.
Document Type
Poster
Included in
Biology Commons, Plant Biology Commons, Plant Pathology Commons
Optimization of DNA Extraction from Difficult Sandy-loam Soils
Ginseng is a high value perennial crop grown for its roots. A four-year cultivation cycle is required to obtain marketable ginseng roots, during which a condition called ginseng replant disease (GRD) develops. GRD is characterized by a severe root rot disease, and prevents the growth of newly planted ginseng in a former ginseng garden1. Microbiome analysis of ginseng garden soil will help our understanding of the formation of GRD and its severity. However, extracting DNA from the sandy-loam soils required for ginseng cultivation is difficult. Commercial kits are either too expensive or not applicable to the large volumes of soil required to extract a suitable quality and quantity of DNA for Illumina Miseq next generation sequencing (NGS). Therefore, an optimized DNA extraction protocol is essential to identifying microbes that contribute to GRD. In this study, a DNA extraction method based on Method 6b from Kamble and Singh (2020)2 was used as the basis for an optimized protocol. The ZymoBIOMICS DNA Miniprep kit3 was used for comparison.