1. DCL-suppressedNicotiana benthamianaplants: valuable tools in research and biotechnology
- Author
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Elena Dadami, Anastasis Oulas, Kriton Kalantidis, Eleni Mitta, Konstantina Katsarou, and Eirini Bardani
- Subjects
0106 biological sciences ,0301 basic medicine ,Small RNA ,Soil Science ,Nicotiana benthamiana ,Plant Science ,Biology ,01 natural sciences ,03 medical and health sciences ,chemistry.chemical_compound ,RNA interference ,RNA polymerase ,Molecular Biology ,fungi ,food and beverages ,RNA ,Argonaute ,biology.organism_classification ,Cell biology ,RNA silencing ,030104 developmental biology ,chemistry ,biology.protein ,Agronomy and Crop Science ,010606 plant biology & botany ,Dicer - Abstract
RNA silencing is a universal mechanism involved in development, epigenetic modifications and responses to biotic and abiotic stresses. The major components of this mechanism are Dicer-like (DCL), Argonaute (AGO) and RNA-dependent RNA polymerase (RDR) proteins. Understanding the role of each component is of great scientific and agronomic importance. Plants, including Nicotiana benthamiana, an important plant model, usually possess four DCL proteins, each of which has a specific role, namely being responsible for the production of an exclusive small RNA population. Here, we used RNA interference (RNAi) technology to target DCL proteins and produced single and combinatorial mutants for DCL. We analysed the phenotype for each DCL knockdown plant, together with the small RNA profile, by next-generation sequencing (NGS). We also investigated transgene expression, as well as viral infections, and were able to show that DCL suppression results in distinct developmental defects, changes in small RNA populations, increases in transgene expression and, finally, higher susceptibility in certain RNA viruses. Therefore, these plants are excellent tools for the following: (i) to study the role of DCL enzymes; (ii) to overexpress proteins of interest; and (iii) to understand the complex relationship between the plant silencing mechanism and biotic or abiotic stresses.
- Published
- 2018