La Trobe
1156743_Martoni,F_2021.pdf (1.64 MB)

Propylene Glycol and Non-Destructive DNA Extractions Enable Preservation and Isolation of Insect and Hosted Bacterial DNA

Download (1.64 MB)
journal contribution
posted on 24.08.2021, 07:06 by F Martoni, E Nogarotto, Alexander PiperAlexander Piper, R Mann, I Valenzuela, L Eow, L Rako, Brendan RodoniBrendan Rodoni, MJ Blacket
Plant bio-protection and biosecurity programs worldwide use trap-based surveillance for the early detection of agricultural pests and pathogens to contain their incursions and spread. This task is reliant on effective preservation in insect traps, which is required to maintain specimen quality for extended periods under variable environmental conditions. Furthermore, with traditional morphological examinations now increasingly paired with modern molecular diagnostic techniques, insect traps are required to preserve both the specimens’ morphology and the DNA of insects and vectored bacterial pathogens. Here, we used psyllids (Hemiptera) and their hosted bacteria as a model to test the preservative ability of propylene glycol (PG): a non-flammable, easily transportable preservative agent that could be used in pitfall, suction or malaise traps. We tested preservation using various PG concentrations, at different temperatures and for multiple time periods, paired with non-destructive DNA extraction methods, which allow isolation of both insect and arbobacterial DNA while retaining a morphological voucher of the insect host specimens. PG concentrations between 40% and 100% performed best for both insect and bacterial DNA preservation up to 28 days. Ultimately, given the viscous nature of PG at high concentrations, we recommend using it at a concentration between 40% and 60% to enable insects to sink into the solution, thus enhancing DNA preservation.

Funding

This research was funded by the Australian Government Department of Agriculture, Horticulture Innovation Australia (ST16010) and Grains Research and Development Corporation.

History

Publication Date

01/01/2021

Journal

Agriculture (Switzerland)

Volume

11

Issue

1

Article Number

ARTN 77

Pagination

(p. 1-16)

Publisher

MDPI

Rights Statement

Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/4.0/).