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Characterization and microsatellite marker development for a common bark and ambrosia beetle associate, Geosmithia obscura
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  • Grace Pietsch,
  • William Klingeman,
  • Matthew Huff,
  • Miroslav Kolarik,
  • Denita Hadziabdic,
  • Romina Gazis,
  • Margaret (Meg) Staton
Grace Pietsch
The University of Tennessee

Corresponding Author:[email protected]

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William Klingeman
The University of Tennessee
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Matthew Huff
The University of Tennessee
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Miroslav Kolarik
Czech Academy of Sciences
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Denita Hadziabdic
The University of Tennessee
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Romina Gazis
University of Florida
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Margaret (Meg) Staton
University of Tennessee
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Background. Symbioses between Geosmithia fungi and wood-boring and bark beetles seldom result in disease induction within the plant host. Yet exceptions exist such as Geosmithia morbida, the causal agent of Thousand Cankers Disease (TCD) of walnuts and wingnuts, and Geosmithia sp. 41, the causal agent of Foamy Bark Canker disease of oaks. Isolates of G. obscura were recovered from black walnut trees in eastern Tennessee and at least one isolate induced cankers following artificial inoculation. Due to the putative pathogenicity and lack of recovery of G. obscura from natural lesions, a molecular diagnostic screening tool was developed using microsatellite markers mined from the G. obscura genome. Results. A total of 3,256 candidate microsatellite markers were identified (2236, 789, 137 di-, tri-, and tetra- motifs were identified, respectively), with 2011, 703, 101 di-, tri-, and tetra- motifs containing markers with primers. From these, 75 microsatellite markers were randomly selected, screened, and optimized, resulting in 28 polymorphic markers that yielded single, consistently recovered bands which were used in downstream analyses. Five of these microsatellite markers were found to be specific to G. obscura and did not cross-amplify into other, closely related species. Although the remaining tested markers could be useful, they cross-amplified within different Geosmithia species, making them not reliable for G. obscura detection. Conclusion. Five novel microsatellite markers (GOBS9, GOBS10, GOBS41, GOBS43, GOBS50) were developed based on the G. obscura genome. These species-specific microsatellite markers are available as a tool for use in molecular diagnostics and can assist future surveillance studies.
10 Jan 2022Submitted to MicrobiologyOpen
11 Jan 2022Submission Checks Completed
11 Jan 2022Assigned to Editor
11 Jan 2022Reviewer(s) Assigned
16 Feb 2022Review(s) Completed, Editorial Evaluation Pending
16 Feb 2022Editorial Decision: Revise Major
08 Mar 20221st Revision Received
09 Mar 2022Assigned to Editor
09 Mar 2022Submission Checks Completed
09 Mar 2022Review(s) Completed, Editorial Evaluation Pending
10 Mar 2022Reviewer(s) Assigned
18 Mar 2022Editorial Decision: Revise Minor
08 Apr 20222nd Revision Received
25 Apr 2022Assigned to Editor
25 Apr 2022Submission Checks Completed
25 Apr 2022Review(s) Completed, Editorial Evaluation Pending
27 Apr 2022Editorial Decision: Accept
Jun 2022Published in MicrobiologyOpen volume 11 issue 3. 10.1002/mbo3.1286