Detection of local Erwinia Isolates Causing Diseases in Potato by Using DNA Amplification by Polymerase Chain Reaction Technique (PCR)

Abstract

Seventy samples of infected potato's tubers were collected from Baghdad city during the period from September 2011 to February 2012. These samples were collected from Alradwaniyah potato's store/ Albyaa market and from Awrad Alnahar potato's store in Abu ghraib and two bacterial strains of Erwinia were obtained from Plant Protection department/ College of Agriculture/ Baghdad University. The Crystal Violet Pectate (CVP) used as a selective-diagnostic medium to isolate and purify Erwinia. The strains and isolates of Erwinia were characterized according to the morphology and microscopic characteristics, along with the biochemical tests and API 20E tests. The Erwinia isolates have a positive reaction for Catalase, Raffinose and Cellobiose and a negative reaction for Oxidase and Sorbitol. By using Reagent Genomic DNA Kit method for Total DNA extraction, during electrophoresis process, the results showed that some of selected isolates and the two standard strains of Erwinia contained at least two bands of small plasmid DNA. The identification was confirmed by using DNA amplification by polymerase chain reaction technique (PCR) and Erwinia carotovora subsp. carotovora specific primer such as (Ec001), the selected local isolates of Erwinia and Erwinia carotovora subsp. carotovora ATCC 15713 potato's pathogen had a positive reaction (one band 312 bp) for the former primer.

Bacteriophage As a Promising Biocontrol Agent for Pectobacterium Carotovorum, Inciting Potato Soft Rot Disease

“…It was found that Pcc belonged to the Enterobacteriaceae family that possessed circular, creamy, convex, smooth and entire margin, gram-negative, rod-shaped, none forming spores, motile with peritrichous flagella and cannot produce pigments. They grow and form pits on CVP media, creamy colored colonies with no fluorescent pigment on King B medium and produce small and dark red colonies on raffinose medium, the results of the present study are in accordance with the finding obtained by (Kettani-Halabi et al, 2013 andMohamed &Selman, 2013). Biochemical tests on Pcc isolates gave positive reactions for KOH, pectinase, catalase, methyl red, voges proskaur, casein and gelatin hydrolysis, H2S production, Citrate and acid production from glucose, sucrose, fructose, lactose, cellobiose and raffinose tests.…”

“…Biochemical tests on Pcc isolates gave positive reactions for KOH, pectinase, catalase, methyl red, voges proskaur, casein and gelatin hydrolysis, H2S production, Citrate and acid production from glucose, sucrose, fructose, lactose, cellobiose and raffinose tests. Conversely, all Pcc isolates can grow at 37°C, 5% and 7% NaCl, but all isolates gave negative reactions for oxidase, indole, urease, starch hydrolysis, and acid production from maltose tests these results are compatible with (Mohamed andSelman, 2013 andNazerian et al, 2013). The biochemical results were confirmed by the API system that revealed that the tested isolates belong to Pectobacterium carotovorum, these results are compatible with (Mohamed andseleman, 2013 andHassan,2017).…”

The Efficacy of Local Streptomyces Diastatochromogenes Isolate as a Biological Control Agent of Potato Tuber Soft Rot Disease Caused by Pectobacterium carotovorum in Iraq

“…Whereas control tubers were inoculated with distilled water rather than the bacterial suspension, and then all experiment tubers were kept at 28 °C ± 2 for 5 days. The size of the rotted areas created by each isolate on potato tubers was a significant indication of the rotting ability of the isolates [24].…”

Isolation and Diagnosis of the Bacterium Pectobacterium Caratovorium, the Causative Agent of Soft Rot Disease on Potatoes, Macroscopically and Biochemically

“…The damage to the potato slice was measured by measuring the diameter of the lesion. Isolates with high pathogenicity in the potato slice assay were selected and stored in sterile test tubes in the refrigerator for use in subsequent experiments [13][14][15][16][17].…”