Bacillus Aryabhattai is a gram-positive motile bacteria that was found in high altitudes in the air (Shivaji, 2009) and was also found to promote growth of soybean in South Korea (Park, 2017).

Data collected: December 12, 2016

References: Shivaji S, Chaturvedi P, Begum Z, Pindi P, Manorama R, Padmanaban A, Shouche Y, Pawar S, Vaishampayan P, Dutt C, Datta G, Manchanda R, Rao U, Bhargava P, Narlikar J. 2009. Janibacter hoylei sp. nov., Bacillus isronensis sp. nov. and Bacillus aryabhattai sp. nov., isolated from cryotubes used for collecting air from the upper atmosphere. International Journal of Systematic and Evolutionary Microbiology [Internet]. [cited 30 March 2017]; 59: 2977-2986.

 

Yun W, Lee I, Lee D, Oh Y, Lee U, Kim A, Seo C, Shahzad R, Hussain A, Kang S, Mun B, Park Y. 2017. Bacillus aryabhattai SRB02 tolerates oxidative and nitrosative stress and promotes the growth of soybean by modulating the production of phytohormones. PLoS ONE [Internet]. [cited 30 March 2017]; 12: 1-28.

 

Methods for isolation and identification:

• On a surface from a retention pond in Lancer Park, 5mm of surface water was collected ( Figure 1). A swab that was cleaned with sterile water was used to collect a water sample. It was then spread across a sterile LB agar plate. The plate was incubated at 37-degree celsius for 16 hours.
• A circular colony that was a lightish yellow color (Figure 2) was picked for 16s rRNA gene sequencing by PCR amplification.
• The genus and species of the bacteria was identified by the PCR product digesting with MspI and the sequence.

 

                                   Figure 1. Collection Site      

                                  Figure 2. Identified colony

 

Results:

• MspI digestion (Figure 3): The PCR experiment led to a 1,500 bp product. After the digestion of MspI, four bands were identified at 650 bp, 400 bp, 225 bp, and 175 bp.

               Figure 3. PCR amplificaton and MspI digestion results

 

• Seqquence Analysis (Figure 4): The sequences PCR product was able to read 914 bases that could infrom what the identity of the jenus and species of the colony was. The following pdf of our chromatogram is available through this link (rDNA_seq_1_PREMIX_KG2496_1). The analysis of the NCBI BLAST expresses that 99% identity between 16s rRNA gene Bacillus Aryabhattai with bases 14-914.

                     Figure 4. NCBI BLAST analysis of colony 10-1

 

Contributed by: Natalie A. Wood and Angelica M. Romero, BIOL Spring 2017, Group 10