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National College Culture Collection Centre
PG & Research Department of Biotechnology and Microbiology
National College, Tiruchirappalli- 620 001, Tamil Nadu, India
1
Isolation and characterization of Virgibacillus pantothenticus strain NCT120 from air
collected from Lab-II
Background
As a part of academic lab in bacteriology, settle plate method was used to sample the air
microflora in nutrient agar. Morphologically distinct colonies were purified using quadrant
streaking. The pure culture was preserved using glycerol stocking in triplicates. One of these
glycerol stocks was revived and used for further morphological and molecular characterization.
Work plan
Bergey’s manual was used to determine the lowest possible taxonomic hierarchy and the strain
was commercially outsourced for bidirectional 16S rRNA gene sequencing for species level
resolution.
National College Culture Collection Centre
PG & Research Department of Biotechnology and Microbiology
National College, Tiruchirappalli- 620 001, Tamil Nadu, India
2
Strain description: NCT120 readily grows in nutrient agar plate at 37°C in 24 hours. Colonies
appeared white, round and shiner with smooth edges. Colonies are 0.4mm in size (last quadrant
metrics). Cells are Gram positive, sporulative, motile bacilli. Biochemical reaction implies Indole
positive, methyl red positive, voges proskauer negative and citrate negative reactions. Triple
Sugar Iron implies acid butt and slant with no gas production. Catalase and oxidase were positive.
Primary enzyme screening implies positive for amylase and lipase synthesis. NCT21 was
sensitive to streptomycin (21mm), ampicillin (13mm), ciprofloxacin (12mm), rifampicin (12mm)
and resistant to amoxicillin.
16S rRNA gene sequencing:
Based on the colony, biochemical and molecular characteristics, the strain NCT120 was identified
as Virgibacillus pantothenticus. NCT120 was 99.51% similar to Virgibacillus pantothenticus
(GenBank accession number:
NR_114091). 16S rRNA gene sequence of
NCT27 can be globally accessed through
genbank accession number OQ271439.
The Neighbor-Joining method tree with
percentage of replicate trees in which the
associated taxa clustered together in the
bootstrap test (500 replicates) are shown.
The tree is drawn to scale, with branch
lengths in the same units as those of the
evolutionary distances used to infer the
phylogenetic tree. The evolutionary
distances were computed using the
Maximum Composite Likelihood method
and are in the units of the number of base
substitutions per site. This analysis
involved 11 nucleotide sequences. All
ambiguous positions were removed for
each sequence pair (pairwise deletion
option). There were a total of 1402
positions in the final dataset.
The strain NCT120 was deposited in
National College Culture Collection Centre (ncccc) and are freely available for academic and
research purposes. The strain can be obtained here www.ncccc.in