Molecular and Physical characterization of the phage specific for bacterial-pathogen causing pink-rot inflorescence disease in date-palm

M, Lewaa L; B.A, Othman; M, Metwaly A; A, Dougdoug K

doi:10.21608/esjp.2023.319364

Molecular and Physical characterization of the phage specific for bacterial-pathogen causing pink-rot inflorescence disease in date-palm

Document Type : Researches

Authors

¹ The Central Laboratory of Date Palm Researches and Development, Agricultural Research Center, Giza, 12619, Egypt.

² Agricultural microbiology Dept. Faculty of Agriculture- Ain Shams University, Egypt.

10.21608/esjp.2023.319364

Abstract

Pink rot inflorescence disease in date palm (Phoenix dactylifera L.) is considered important disease in date palm in Egypt. The causal agent was isolated from trunk using specific media. The causal agent was identified as Serratia marcescens using 16s rRNA gene sequencing and was recorded in GenBank. Three different specific lytic phages of S. marcescens were isolated from soil and trunk of the diseased date palm tree using single plaque isolation technique and named SP1, SP2 and SS1. The morphological characters revealed that two bacteriophage isolates belong to Myoviridae family and one belongs to Siphoviridae family. Thermal inactivation point (TIP) of the different phages was 60, 80 and 60°C for SP1, SP2 and SS1 respectively. Stability to freezing and thawing were 3, 3 and 2 times respectively. pH range stability was 7-9, 6-9 and 5-9. Longevity In Vitro (LIV) of all phages appeared active more than 60 days. Sensitivity of the phages for UV were 20, 25 and 20min. respectively. SDS-PAGE showed molecular weight of protein of SP1 was ∼66, 35 and 29 KDa. SP2 was∼ 38, 35, 30 and 29 KDa. SS1 was ∼97, 38, 35, 30 and 29 KDa. RAPD-PCR showed SP1 amplified to 4 fragments with 2000, 1500, 1000 and 600 bp, phage SP2 amplified to 4 fragments with 1500, 1000, 700 and 600 bp, SS1 phage amplified to 5 fragments with 1500, 1000, 800, 700 and 600 bp.

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