Precise transcript targeting using RNAi for weed control

Abstract:

Background:

RNA interference (RNAi) is a natural post-transcriptional gene silencing (PTGS) system of eukaryotic cells and can be used as spray-induced gene silencing (SIGS) to target genes. Due to the complexity of plant cell structure and genome, using SIGS in plants has faced many barriers.

Objective:

We aimed to develop a delivery strategy and appropriate siRNA design to induce PTGS by SIGS.

Methods:

The rice Phytoene desaturase (OsPDS) gene was used as a model to test our SIGS. Three siRNAs (siPDS1, siPDS2, and siPDS3) targeting different regions of the OsPDS transcript were applied individually or mixed siPDS2+siPDS3 in solution with Silwet^TM adjuvant. Treatments were applied using a micropipette in rice seedlings (V2-stage), with the addition of proper control treatments: untreated-check and adjuvant-check. Seedlings were accessed phenotypically, physiologically, and molecularly.

Results:

The most prominent phenotype was observed for siPDS1 and siPDS2+siPDS3. Reduction in plant growth, chlorophyll index and shoot dry mass were reduced after siPDS2+siPDS3 application, while increasing anthocyanin content. OsPDS was downregulated after siPDS1 application with no change in the non-treated upper leaf.

Conclusions:

Our siRNA design and delivery strategy efficiently delivered the siRNA inside the plant and promoted target PTGS and compatible phenotype. The system used in this experiment did not show systematicity in the plant, probably due to the fast oxidative stress that hindered the systematic effect. This is an advance on the SIGS and can be a valuable tool for PTGS to find new targets for RNAi.

Keywords:
Post-transcriptional Gene Silencing; siRNAs; Exogenous Application; Gene Expression