Specific and Sensitive Detection of Enterobacter mori Using Reliable RT-PCR.

Area of Science:

• Plant Pathology
• Molecular Biology
• Agricultural Science

Background:

• Bacterial wilt, caused by Enterobacter mori, poses a significant threat to mulberry production in China.
• Current control strategies for Enterobacter mori are limited, necessitating effective disease detection methods.
• Latent infections of Enterobacter mori in mulberry plants complicate disease management.

Purpose of the Study:

• To develop a rapid and reliable polymerase chain reaction (PCR) assay for the detection of Enterobacter mori.
• To enable the screening of mulberry plant material for both symptomatic and latent infections of Enterobacter mori.

Main Methods:

• Primers were designed targeting the RNA polymerase β-subunit (rpoB) gene of Enterobacter mori.
• The developed PCR assay was validated using various bacterial strains, including Enterobacter type strains and other mulberry pathogens.
• Real-time PCR was employed to determine the sensitivity of the assay for detecting bacterial DNA and cells in plant tissues.

Main Results:

• The PCR assay demonstrated 100% sensitivity and 100% specificity against a range of bacterial pathogens and isolates.
• The assay successfully detected Enterobacter mori DNA at concentrations as low as 10 fg/μl and bacterial cells at 10^2 CFU/ml.
• Testing of symptomatic plants yielded 100% positive results, while 36.4% of asymptomatic plants showed latent infections.

Conclusions:

• The developed PCR assay is a fast, simple, reliable, and sensitive tool for detecting Enterobacter mori.
• This method is suitable for screening mulberry plant material, including asymptomatic plants, for latent infections.
• The assay will aid in managing bacterial wilt disease in mulberry orchards by enabling early and accurate diagnosis.