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Use of loop-mediated isothermal ampliflcation for detection of Ophiostoma clavatum, the primary blue stain fungus associated with Ips acuminatus

Lookup NU author(s): Professor Neil Boonham

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Abstract

Loop-mediated isothermal ampliflcation (LAMP) is an alternative ampliflcation technology which is highly sensitive and less time-consuming than conventional PCR-based methods. Three LAMP assays were developed, two for detection of species of symbiotic blue stain fungi associated with Ips acuminatus, a bark beetle infesting Scots pine (Pinus sylvestris), and an additional assay speciflc to I. acuminatus itself for use as a control. In common with most bark beetles, I. acuminatus is associated with phytopathogenic blue stain fungi involved in the process of exhausting tree defenses, which is a necessary step for the colonization of the plant by the insect. However, the identity of the main blue stain fungus vectored by I. acuminatus was still uncertain, as well as its frequency of association with I. acuminatus under outbreak and non-outbreak conditions. In this study, we employed LAMP technology to survey six populations of I. acuminatus sampled from the Southern Alps. Ophiostoma clavatum was detected at all sampling sites, while Ophiostoma brunneo-ciliatum, reported in part of the literature as the main blue stain fungus associated with I. acuminatus, was not detected on any of the samples. These results are consistent with the hypothesis that O. clavatum is the main blue stain fungus associated with I. acuminatus in the Southern Alps. The method developed in the course of this work provides a molecular tool by which it will be easy to screen populations and derive important data regarding the ecology of the species involved. © 2013, American Society for Microbiology.

Publication metadata

Author(s): Villari C, Tomlinson JA, Battisti A, Boonham N, Capretti P, Faccoli M

Publication type: Article

Publication status: Published

Journal: Applied and Environmental Microbiology

Year: 2013

Volume: 79

Issue: 8

Pages: 2527-2533

Print publication date: 01/04/2013

Online publication date: 08/02/2013

ISSN (print): 0099-2240

ISSN (electronic): 1098-5336

Publisher: American Society for Microbiology

URL: https://doi.org/10.1128/AEM.03612-12

DOI: 10.1128/AEM.03612-12

PubMed id: 23396326

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