Methods for preservation and transcriptomic analysis of Lactobacillus Plantarum P1, P2 phage

Munguntsetseg Battur1, Tserenkhand Zorigtbaaatar2, Bolormaa Chuluun-Otgon3, Chen Xia4, Nant Nyein Zar Ni Naing5, Tian Yang6*

* Corresponding Author: Tian Yang

1, 4, 6 College of Food Science and Technology, Yunnan Agricultural University, Kunming, P.R.China. Mla1220@yahoo.com
2 School of Animal Science and Biotechnology, Mongolian University of Life Sciences, Ulaanbaatar, Mongolia. Tserenkhand.z@muls.edu.mn
3 College of Food Science, Northeast Agricultural University, Harbin, P.R.China 2976813175@qq.com
5 College of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot, P.R.China chenxia8280@163.com

Digital Object Identifier: 

https://doi.org/10.53468/mifyr.2024.04.03.1

Abstract – The survival of Lactobacillus plantarum virulent phages P1 and P2 in three cryoprotectants: glycerol(15%), chloroform(20%), and dimethyl sulfoxide(7%) under different temperatures(4℃, -20℃, -80℃) were evaluated every half month. The study was undertaken to establish suitable preservation methods that could be used to study phage biological properties and genomic characteristics further. Transcriptomic properties of phage activity in different stages of lysis were also analyzed. This information may be used further to explore the relationship between bacteriophage and host bacteria. The results of this study showed that Lactobacillus plantarum virulent phage P1, stored for 60 days in all cryoprotectives at all temperatures, exhibited a survival titer greater than 107 PFU/mL (initial titer 108 PFU/mL). The optimum cryoprotectant used was glycerol at -80℃. In this respect, the phage could be preserved for 10 months with a 105 PFU/mL titer. Maintenance of Lactobacillus plantarum virulent phage P2 under different storage conditions was evaluated. Preservation of the phage for up to three months was achieved at 4°C using all cryoprotectants. In this respect, the phage titer was greater than 108 PFU/mL. For long-term phage maintenance (10 months), the optimum preservation conditions at -80°C included 15% glycerol or 7% dimethyl sulfoxide as cryoprotectants. Under these conditions, the phage titer was greater than 107 PFU/mL. The infective phases of phage P2 and its host strain were investigated using transcriptomic analyses. It was observed that when phage P2 infects its host strain, it can control the replication and assembly process of phage by regulating key enzymes (such as prolong factor EF-Tu, EF-G, EF-Ts; translation starting factors IF1, IF2, IF3; RNA synthesis enzyme rpoA, rpoB, rpoC; signaling recognizing particles Fts Y, Ffh; as well as releasing factor RF1).

Keywords – L. plantarum IMAU10120, Bacteriophage, Phage preservation, Transcriptomic Analysis

Article History: Received 22 March 2024, Received in revised form 25 March 2024, Accepted 30 August 2024

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