Janthinobacterium agaricidamnosum is a purple, puntiform bacterial colony that can be found in the Buffalo Creek water. It has also been found in cultivated mushrooms (Lincoln SP. 1999)

References: 

1.  Lincoln SP, Fermor T, Tindall BJ. Janthinobacterium agaricidamnosum sp. nov., a soft rot pathogen of Agaricus bosporus. Int J Syst Bacteriol. 1999. 49(4): 1577-89.

Date Collected: February 8, 2017

Methods for Isolation and Identification:

• A water sample was taken from the top of the water of the Buffalo Creek (Figure 1). 100μl of the sample was placed on an agar plate and was incubated at 25 degrees Celsius for 48 hours. The plate was parafilmed and placed in the refrigerator for seven days.
• A purple, punctiform colony (Figure 2) was selected for 16S rRNA gene sequencing by PCR amplification.
• The PCR product was sent for DNA sequencing to identify the species of the bacteria.

Figure 1. site of collection (Buffalo Creek)

Figure 2. Selected colony for identification

Results:

• Msp1 digestion (figure 3): A light band was produced at 1000 bp by PCR.

Figure 3. Gel electrophoresis results of Buffalo Creek water PCR product and + MSP1.

• DNA sequence: The sequenced PCR product produced 861 quality base pairs that were used to identify the bacteria as Janthinobacterium agaricidamnosum. The chromatogram of the sequence is available as a pdf (HMX4_PREMIX_JF7562_30). NCBI BLAST revealed 99% similarity with 1 gaps out of 861 base pairs (Figure 4).

Figure 4. NCBI BLAST results of chosen colony.

Contributed by: Megan Bland and Hannah Hatke, BIOL 250 Spring 2017