Construction of probe of the plant growth-promoting bacteria Bacillus subtilis useful for fluorescence in situ hybridization

Strains of Bacillus subtilis are plant growth-promoting bacteria (PGPB) of many crops and are used as inoculants. PGPB colonization is an important trait for success of a PGPB on plants. A specific probe, based on the 16s rRNA of Bacillus subtilis, was designed and evaluated to distinguishing, by fl...

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Bibliographic Details
Published in:Journal of microbiological methods 2016-09, Vol.128, p.125-129
Main Authors: Posada, Luisa F., Alvarez, Javier C., Hu, Chia-Hui, de-Bashan, Luz E., Bashan, Yoav
Format: Article
Language:English
Subjects:
B. amyloliquefaciens
Bacillus subtilis
Fluorescence in situ hybridization
Specific probe
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Summary:Strains of Bacillus subtilis are plant growth-promoting bacteria (PGPB) of many crops and are used as inoculants. PGPB colonization is an important trait for success of a PGPB on plants. A specific probe, based on the 16s rRNA of Bacillus subtilis, was designed and evaluated to distinguishing, by fluorescence in situ hybridization (FISH), between this species and the closely related Bacillus amyloliquefaciens. The selected target for the probe was between nucleotides 465 and 483 of the gene, where three different nucleotides can be identified. The designed probe successfully hybridized with several strains of Bacillus subtilis, but failed to hybridize not only with B. amyloliquefaciens, but also with other strains such as Bacillus altitudinis, Bacillus cereus, Bacillus gibsonii, Bacillus megaterium, Bacillus pumilus; and with the external phylogenetic strains Azospirillum brasilense Cd, Micrococcus sp. and Paenibacillus sp. The results showed the specificity of this molecular probe for B. subtilis. [Display omitted] •A probe that can differentiate between Bacillus subtilis and B. amyloliquefaciens was constructed.•The probe is based on nucleotides 465 to 483 of 16s rRNA gene.•The probe did not detect several other Bacillus species and other bacteria.•The probe provides clear visualization by Fluorescent in situ hybridization.
ISSN:0167-7012
1872-8359
DOI:10.1016/j.mimet.2016.05.029