Versatile DNA extraction from diverse plant taxa using ionic liquids and magnetic ionic liquids: a methodological...

Area of Science:

• Plant Biology
• Molecular Biology
• Biochemistry

Background:

• High-throughput plant biomolecular analyses require efficient DNA extraction.
• Conventional methods are time-consuming, require large samples, and involve multiple steps, hindering high-throughput applications.

Purpose of the Study:

• To develop a simple, rapid, and efficient protocol for DNA extraction from diverse plant tissues.
• To establish a method compatible with downstream molecular analyses.

Main Methods:

• Utilized an ionic liquid (IL)-based modified vortex-assisted matrix solid phase dispersion approach.
• Tested on milligram tissue fragments from 13 dicots and 4 monocots, including a century-old herbarium sample.
• DNA quality and quantity assessed via qPCR, DNA barcoding marker amplification (rbcL, nrITS), and Sanger sequencing.

Main Results:

• Successfully extracted DNA from a wide range of plant taxa using only 1.5 mg of tissue.
• Isolated DNA was of sufficient quality and quantity for sensitive analyses like qPCR.
• Extracted DNA remained stable at room temperature for up to 3 weeks, enabling downstream applications without immediate processing.
• No additional purification was needed before DNA amplification due to solvent compatibility with qPCR.

Conclusions:

• The presented method offers a rapid and simple solution for plant cell lysis and DNA extraction.
• The protocol is effective across diverse plant taxa and compatible with sensitive molecular techniques.
• This approach has significant potential for plant barcoding, agriculture, conservation, ecology, evolution, and forensics.