Sensitive molecular diagnostic assays to mitigate the risks of asymptomatic bacterial diseases of plants.

摘要: Our highly concentrated monoculture makes crops vulnerable to pests and diseases. An increase in emerging non-indigenous bacterial diseases poses a real threat US agriculture. The United States has 100,000 miles of shoreline 6,000 border, making possible easy introduction crop Most threatening are the cross-domain enteric bacteria. In contrast animals, have hundreds major development molecular-based detection protocols for each pathogen is impossible with current technology. Rathayibacter toxicus, neurotoxin-producing bacterium transmitted by seed gall nematode, an example high-risk Select Agent. infects seeds grasses without showing any symptoms, often resulting death grazing cattle. A prerequisite control disease sensitive proper identification causal organism. Detecting bacteria samples plants symptoms relatively simple, whereas asymptomatic tissues difficult due extremely low numbers target present. Rapid serological assays work well symptomatic but not from tissue when levels below sensitivity limits. Classical agar-plating 1,000 fold more then serology or PCR. However, agar plating take 3 5 days require pathogenicity tests confirm identity. PCR-based allow rapid, accurate insensitive use 1 microL sample comparison 100 used plating. To overcome this disadvantage, enrichment technique termed BIO-PCR can be combination tissues. key developing successful protocol determine time required pin point-size colonies appear. For most plant pathogens 15 24 hours sufficient time, only 2 needed. greater sensitivity, combined 96-well microliter plates membranes detect single viable cell per 10 mL aqueous sample.