Detection and Identification of Potato Soft Rot Pectobacterium carotovorum Subspecies carotovorum by PCR Analysis of 16S rDNA in Jordan

Abstract

Various bacterial species are known to be agents causing soft rot of potatoes. The results of this study showed that potato soft rot is widely spread in dif-ferent potato planting areas in Jordan. A survey was conducted through the years 2013-2015 to detect potato soft rot disease in Jordan, two hundred and four rotted potato samples were collected from different potato growing areas through different potato growing seasons. One hundred and thirty one bac-terial isolates were isolated, purified on selective media and identified as Pec-tobacterium carotovorum subspecies carotovorum (Pcc) by different bio-chemical and physiological tests. Furthermore, 131 Pcc Jordanian (Jo) isolates were identified by PCR analysis of total DNA extracted from isolates that were biochemically identified as Pcc using universal primer Fd1/Rd1. Cloning and sequencing of representative PCR products, amplifying the 16S rDNA region were done. Phylogenetic analysis of the Pcc Jo-isolates revealed other than 90% similarity with different reference Pcc strains available at the GenBank. Different rot causal agents also were detected by PCR amplification and fur-ther sequences. The sequencing data revealed similarities to Pseudomonas fluorescence, Enterobacteriaceae genera such as Enterobacter spp., Serratia spp. and Klebsiella spp., in addition to Pectobacterium carotovorum subsp. carotovorum. This study indicated that using molecular techniques such as amplification of 16S rDNA region is a sensitive and specific method for de-tecting Pcc as potato soft rot causal agent. So far this is the first study where Pcc has been identified by using PCR and sequencing approaches in Jordan. Keywords Pectobacterium carotovorum subsp. carotovorum (Pcc), Soft Rot of Potato, How to cite this paper: Abu-Obeid, I., Khlaif, H. and Salem, N. (2018) Detection and Identification of Potato Soft Rot Pec-tobacterium carotovorum Subspecies caro-tovorum by PCR Analysis of 16S rDNA in Jordan. Agricultural Sciences, 9, 546-556.