Abstract
Recent investigations show that exogenously applied small interfering RNAs (siRNA) and long double-stranded RNA (dsRNA) precursors can be taken up and translocated in plants to induce RNA interference (RNAi) in the plant or in its fungal pathogen. The question of whether genes in the plant genome can undergo suppression as a result of exogenous RNA application on plant surface is almost unexplored. This study analyzed whether it is possible to influence transcript levels of transgenes, as more prone sequences to silencing, in Arabidopsis genome by direct exogenous application of target long dsRNAs. The data revealed that in vitro synthesized dsRNAs designed to target the gene coding regions of enhanced green fluorescent protein (EGFP) or neomycin phosphotransferase II (NPTII) suppressed their transcript levels in Arabidopsis. The fact that, simple exogenous application of polynucleotides can affect mRNA levels of plant transgenes, opens new opportunities for the development of new scientific techniques and crop improvement strategies.
Highlights
The increasing human population and discussions along the safety of transgenic plants promote the development of new strategies to regulate plant properties without genomic manipulations
The synthesized enhanced green fluorescent protein (EGFP)-double-stranded RNA (dsRNA) and neomycin phosphotransferase II (NPTII)-dsRNA were diluted in water to final concentrations of 0.1, 0.35, and 1 μg/μL
The results show that the exogenous application of dsRNA spread with soft brushes suppressed NPTII and EGFP transcript levels in A. thaliana
Summary
The increasing human population and discussions along the safety of transgenic plants promote the development of new strategies to regulate plant properties without genomic manipulations. Numerous investigations show that it is possible to switch off or decrease expression of particular genes for the regulation of plant stress tolerance, growth, and other processes via induction of RNA interference (RNAi or gene silencing process) [1]. It is known that RNAi serves for the regulation of various processes important for plants, such as growth and development, stress adaptation, or synthesis of biologically active compounds [2,3]. In the course of RNAi, double-stranded RNAs (dsRNAs) are processed by a ribonuclease into small interfering RNA (siRNA) or microRNA (miRNA). These small RNAs are incorporated in the RNA-induced silencing complex that provides cleavage, destabilization, or hindering translation of any homologous mRNAs [4]. The application of RNAi-based approaches, for gene regulation in plants, requires plant genomic modifications or application of the weakened plant viruses
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