Abstract
The red-necked longhorn beetle Aromia bungii (Faldermann, 1835) (Coleoptera: Cerambycidae) is native to east Asia, where it is a major pest of cultivated and ornamental species of the genus Prunus. Morphological or molecular discrimination of adults or larval specimens is required to identify this invasive wood borer. However, recovering larval stages of the pest from trunks and branches causes extensive damage to plants and is timewasting. An alternative approach consists in applying non-invasive molecular diagnostic tools to biological traces (i.e., fecal pellets, frass). In this way, infestations in host plants can be detected without destructive methods. This paper presents a protocol based on both real-time and visual loop-mediated isothermal amplification (LAMP), using DNA of A. bungii extracted from fecal particles in larval frass. Laboratory validations demonstrated the robustness of the protocols adopted and their reliability was confirmed performing an inter-lab blind panel. The LAMP assay and the qPCR SYBR Green method using the F3/B3 LAMP external primers were equally sensitive, and both were more sensitive than the conventional PCR (sensitivity > 103 to the same starting matrix). The visual LAMP protocol, due to the relatively easy performance of the method, could be a useful tool to apply in rapid monitoring of A. bungii and in the management of its outbreaks.
Highlights
Aromia bungii (Faldermann, 1835) (Coleoptera: Cerambycidae), the red-necked longhorn beetle, is an important pest of fruit and ornamental plants of the genus Prunus, both in native areas of east Asia and in newly invaded areas of Europe and Japan (EFSA 2019; EPPO 2020; CABI 2020)
A. bungii is in the list of priority pests in the European Union (EU 2019) and quarantine measures have been applied in Germany and Italy to eradicate this invasive pest (Hörren 2016) or to contain the risk of further outbreaks (Carella 2019)
The amplifiability of the DNA extracted from target and non-target frass samples (Table 3) gave satisfactory results
Summary
Aromia bungii (Faldermann, 1835) (Coleoptera: Cerambycidae), the red-necked longhorn beetle, is an important pest of fruit and ornamental plants of the genus Prunus, both in native areas of east Asia and in newly invaded areas of Europe and Japan (EFSA 2019; EPPO 2020; CABI 2020). A. bungii can infest healthy or weakened host species and complete several overlapping generations in the same tree (Ma et al 2007). A. bungii is in the list of priority pests in the European Union (EU 2019) and quarantine measures have been applied in Germany and Italy to eradicate this invasive pest (Hörren 2016) or to contain the risk of further outbreaks (Carella 2019). These quarantine measures can have a strong impact on nurseries and farmers
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