Abstract
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.
Highlights
IntroductionPublisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations
Comparing the ten DNA extraction methods, we found that some methods resulted in lower cycle threshold (Ct) values while others resulted in a total lack of amplification
The Proteinase K (PK)+ATL buffer mix was used directly in the quantitative polymerase chain reaction (qPCR) without dilution or cleanup steps, resulting in a crude extraction buffer generated without using any laboratory steps to remove inhibitors
Summary
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Plant protection and plant biosecurity programs worldwide rely heavily on trap-based surveillance to detect invasive insects and their vectored bacterial pests [1,2]. A number of phloem- and xylem-feeding insects, such as psyllids (Psylloidea) or leafhoppers (Cicadellidae), are considered agricultural pests due to their ability to vector plant pathogenic bacteria causing significant economic losses to crops [3,4,5]. The tomato potato psyllid Bactericera cockerelli (Šulc 1909) vectors the bacterium Candidatus
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