Molecular characterisation and analysis to improve the efficiency of RNA interference as an agricultural pest control tool
Aline Pereira Rank, Aline Koch
University of Hohenheim, Institute of Phytomedicine, Germany
The world is demanding sustainable system production and one strategy that over the past years has earned a lot of attention is the use and application of technologies involving RNA interference. RNAi as it is called has been proved a benefic, specific, and promising technology. RNAi relativeness recent application still has a lot to improve and much basic laboratory knowledge to develop. One of the main strategies to determine its silence effect is to make use of molecular analysis, such as qPCRs. This analysis requires the use of some standard procedures and reactions to make it more accurate. One of the steps is to make use of reliable reference genes specific to our purpose. Reference genes are genes stable expressed in as many as possible situations avoiding effects under stress conditions or specific situations regarding our case of study. In our study was interesting to approach and select viable reference genes for Myzus persicae, Tetranychus urticae, and Plutella xylostella. Under specific stress conditions, the reference genes were selected based on material exposed to feeding assay with dsRNA solution, along with its developmental stages and temperature stress. All the data were analysed in four available platforms (Bestkeeper, GeNorm, Delta Cq, and NormFinder) and finally, two of the tested targets for all the pests and cases were selected using the RefFinder software analysis. The genes Actin and 18s rRNA were selected as the most promising for stability for all the insects. The results presented here will be useful for guaranteeing the quality of future RNAi and other specific molecular analyses.
Keywords: Biotechnology, QPCR, RNAi, sustainability
Contact Address: Aline Pereira Rank, University of Hohenheim, Institute of Phytomedicine, Otto-Sander-Straße 5, 70599 Stuttgart, Germany, e-mail: aline.pereirarankuni-hohenheim.de