Isolation and characterization of Paenibacillus polymyxa strain NCT82 from a agri soil

Abstract

As a part of academic lab in microbiology, agri soil samples were collected, serially diluted and spread plated onto a nutrient agar media. 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.

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National College Culture Collection Centre www.ncccc.in
PG & Research Department of Biotechnology and Microbiology
National College, Tiruchirappalli- 620 001, Tamil Nadu, India
1
N4C
Isolation and characterization of Paenibacillus polymyxa strain NCT82 from a agri soil
Background
As a part of academic lab in microbiology, agri soil samples were collected, serially diluted and
spread plated onto a nutrient agar media. 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 www.ncccc.in
PG & Research Department of Biotechnology and Microbiology
National College, Tiruchirappalli- 620 001, Tamil Nadu, India
2
N4C
Strain description: NCT82 grows luxurantly in nutrient agar plate at 37°C in 24 hours. Colonies
are small, round, milky white, shiny with smooth edges and are 4mm in diameter (last quadrant
metrics). Cells are Gram positive, non-motile rods. Biochemical reaction implies Indole negative,
methyl red negative, voges proskauer negative and citrate negative. The strain is positive for
amylase production and negative for lipases. Triple Sugar Iron implies weakly acid butt and
alkaline slant with no gas production. Catalase negative and oxidase negative. The strain NCT82
was sensitive to rifampicin and resistant to penicillin and streptomycin and erythromycin.
16S rRNA gene sequencing:
Based on the colony, biochemical
and molecular characteristics,
the strain NCT82 was identified
as Paenibacillus polymyxa. The
16S rRNA gene sequences of
NCT82 was 99.51% similar
Paenibacillus polymyxa
(GenBank accession number:
NR_037006) and can be globally
accessed through genbank
accession number OQ255783.
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 1387 positions in the final dataset.
The strain NCT82 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