An efficient protocol for tissue sampling and DNA isolation from the stem bark of Leguminosae trees

Area of Science:

• Botany
• Molecular Biology
• Genetics

Background:

• Molecular studies traditionally rely on leaf DNA, posing challenges for tall tree species due to difficult and costly sampling.
• The Leguminosae family is crucial in the biodiverse and endangered Atlantic Forest and Cerrado biomes.
• Efficient DNA extraction is vital for studying these important tree species.

Purpose of the Study:

• To develop and evaluate a DNA isolation protocol using stem bark from Leguminosae trees.
• To provide a practical and cost-effective alternative to leaf sampling for molecular studies.
• To assess the applicability of the protocol across multiple Leguminosae species.

Main Methods:

• A modified CTAB (Cetyl trimethylammonium bromide) protocol was adapted for DNA isolation from stem bark.
• Procedures for stem bark sampling and storage (frozen or silica gel drying) were detailed.
• The protocol was initially developed for three species and tested on an additional 15 Leguminosae species.

Main Results:

• Satisfactory quality DNA was successfully isolated from the stem bark of all tested Leguminosae species.
• DNA yield from bark was slightly lower than from leaves, but purity was comparable.
• Both frozen and silica gel-dried bark samples yielded similar DNA quality and quantity.
• Successful polymerase chain reaction amplification was achieved for both plastid and nuclear genomes.

Conclusions:

• Stem bark is a viable and effective source of DNA for molecular studies in Leguminosae trees.
• The developed protocol simplifies field sampling, reducing costs and difficulties associated with accessing leaves of tall trees.
• This method facilitates genetic and molecular research on Leguminosae species in the Atlantic Forest and Cerrado.