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Title:
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Development of a SCAR marker and a strain-specific genomic marker for the detection of the biocontrol agent strain CPA-8 Bacillus amyloliquefaciens (formerly B. subtilis)
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Author:
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Gotor Vila, Amparo María; Teixidó i Espasa, Neus; Usall i Rodié, Josep; Dashevskaya, Svetlana; Torres Sanchis, Rosario
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Notes:
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In this work, reliable tools were developed to detect and identify the biocontrol
strain CPA-8 using DNA amplification techniques. As a first approach, the
RAPD (random amplified polymorphic DNA) technique was applied to a
collection of 77 related Bacillus species. Among the primers tested, the primer
pair OPG1/OPG6 amplified a 668 bp specific product to the strain CPA-8 that
was sequenced and used to design SCAR (sequence-characterised amplified
regions) primer pairs. The SCAR-4 marker amplified a semi-specific fragment
of 665 bp not only for the strain CPA-8 but also for other 12 strains whose
morphology was completely different from CPA-8. Another approach was
developed to obtain a strain-specific genomic marker related to ecological
adaptations of Bacillus amyloliquefaciens species. The primer pair F2/R2 obtained
from RBAM 007760, a gene involved in surface adhesion, amplified a 265 bp
fragment unique for strain CPA-8. Our results revealed that these two molecular
markers, SCAR-4 and RBAM 007760 F2/R2 provide suitable monitoring
tools to specifically identify the biocontrol CPA-8 when applied against brown
rot caused by Monilinia spp. in stone fruit. Moreover, our findings demonstrate
that the strain CPA-8 is affiliated with B. amyloliquefaciens species that was
formerly designated as Bacillus subtilis.
This research was supported by the European project BIOCOMES FP7-612713 and by the Catalan government (Generalitat de Catalunya) for the PhD grant 2015-FI-B100166 (Amparo M. Gotor). No conflict of interest declared. |
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Subject(s):
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-Bacillus spp
-Biological control
-Molecular marker
-Monitoring
-SCAR |
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Rights:
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© Association of Applied Biologists, 2016
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Document type:
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article
acceptedVersion |
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Published by:
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Wiley;
Association of Applied Biologists
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