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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 Paenibacillus polymyxa strain NCT70 from agriculture
soil
Background
As a part of academic lab in bacteriology, agriculture soil sample was collected, serially diluted
and plated on to nutrient agar plates. The pure culture was preserved using glycerol stocking in
triplicates. This glycerol stocked cultures were 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
Results
Strain description: NCT70 grows luxuriantly in nutrient agar plate at 37°C in 24 hours. Colonies
are small, white, round with smooth edges. Colonies were on average 0.3mm in size (last quadrant
metrics). Cells are Gram positive, sporulative motile rods. Biochemical reaction implies Indole
negative, methyl red negative, voges proskauer negative and citrate negative. Triple Sugar Iron
implies heavy hydrogen sulfide gas production. Catalase positive and oxidase positive. The strain
NCT70 was sensitive to fluconazole (24mm), ampicillin (13mm) and resistant to neomycin,
streptomycin, clindamycin.
16S rRNA gene sequencing:
Based on the colony and biochemical characteristics, the strain NCT70 was identified as
Paenibacillus polymyxa. NCT70 was 99.94% similar to Paenibacillus polymyxa (GenBank
accession number: NR_037006). 16S rRNA gene sequence of NCT27 can be globally accessed
through genbank accession number OQ271437.
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 1359 positions in the
final dataset.
The strain NCT70 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