Amplificação isotérmica mediada por Loop: Uma técnica rápida e eficiente para a detecção de Trialeourodes vaporariorum e Sclerotinia sclerotiorum em sementes de soja

Abstract

The demand for food is increasing due to the great changes that the world has been going through. Brazil stands out in the production and export of food and with each passing year the production of grains has been breaking a record. However, losses caused by diseases and pests are a major limitation of productivity and, to avoid these losses, we need to carry out the control quickly and efficiently. The first step towards successful control is the detection of the pest / pathogen in the cultivation area and this must be early and correct. Loop-mediated isothermal amplification is a molecular technique that has been used in several biological fields. It does not need sophisticated equipment for its application and therefore it can be used in the field or in the laboratory. In addition, the results can be visualized directly with the naked eye by changing the color of the reaction. In this study, we used the technique to quickly identify the whitefly Trialeurodes vaporariorum, with or without DNA extraction. Our result showed that for the identification, only 60 minutes of incubation are necessary, and the DNA extraction step is unnecessary. Also, a LAMP test was developed to detect Sclerotinia sclerotiorum in soybean seeds. In this study, our results demonstrate that with a 48-hour inoculum potential, it is possible to detect the pathogen when only 1% of soybean seeds are infected with the fungus. Both works provide valuable results for the areas of entomology and phytopathology. And they demonstrate the versatility of the technique that can be applied with different biological materials.