Abstract
Plant pathogenic bacteria, phytoplasmas, viruses and viroids are difficult to control, and preventive measures are essential to minimize the losses they cause each year in different crops. In this context, rapid and accurate methods for detection and diagnosis of these plant pathogens are required to apply treatments, undertake agronomic measures or proceed with eradication practices, particularly for quarantine pathogens. In recent years, there has been an exponential increase in the number of protocols based on nucleic-acid tools being those based on PCR or RT-PCR now routinely applied worldwide. Nucleic acid extraction is still necessary in many cases and in practice inhibition problems are decreasing the theoretical sensitivity of molecular detection. For these reasons, integrated protocols that include the use of molecular techniques as screening methods, followed by confirmation by other techniques supported by different biological principles are advisable. Overall, molecular techniques based on different types of PCR amplification and very especially on real-time PCR are leading to high throughput, faster and more accurate detection methods for the most severe plant pathogens, with important benefits for agriculture. Other technologies, such as isothermal amplification, microarrays, etc. have great potential, but their practical development in plant pathology is still underway. Despite these advances, there are some unsolved problems concerning the detection of many plant pathogens due to their low titre in the plants, their uneven distribution, the existence of latent infections and the lack of validated sampling protocols. Research based on genomic advances and innovative detection methods as well as better knowledge of the pathogens' lifecycle, will facilitate their early and accurate detection, thus improving the sanitary status of cultivated plants in the near future.
Highlights
Plant pathogenic bacteria, phytoplasmas, viruses and viroids cause harmful, widespread and economically important diseases in a very broad range of plant species worldwide (Agrios, 2001; Janse, 2007)
For these reasons, integrated protocols that include the use of molecular techniques as screening methods, followed by confirmation by other techniques supported by different biological principles are advisable
Molecular techniques based on different types of polymerase chain reaction (PCR) amplification and very especially on real-time PCR are leading to high throughput, faster and more accurate detection methods for the most severe plant pathogens, with important benefits for agriculture
Summary
Phytoplasmas, viruses and viroids cause harmful, widespread and economically important diseases in a very broad range of plant species worldwide (Agrios, 2001; Janse, 2007). The prevention measures demand pathogen detection methods of high sensitivity, specificity and reliability, because many phytopathogenic bacteria and viruses can remain latent in “subclinical infections”, and/or in low numbers, and/or in some special physiological states in propagative plant material and in other reservoirs (Helias et al, 2000; Grey and Steck, 2001; Janse et al, 2004; Biosca et al, 2006; Ordax et al, 2006). Accurate detection of phytopathogenic organisms is crucial for virtually all aspects of plant pathology, from basic research on the biology of pathogens to the control of the diseases they cause. The need for rapid techniques of high accuracy is especially necessary for quarantine pathogens, because the risk of the disease and the spread of the inoculum must be reduced to nearly zero (López et al, 2003)
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