Genetic transformation and siRNA-mediated gene silencing for aphid resistance in tomato

Researchers explored the ability of RNA interference (RNAi) to silence the Acetylcholinesterase 1 (Ace 1) gene in aphid Myzus persicae and developed transgenic tomato plants resistant to aphid infestation.

Three plasmid constructs, T-449: a single Ace 1 fragment (forward orientation), T-452: two Ace 1 fragments (reverse and forward orientations), and T455: a single inverted Ace 1 fragment, were developed and transformed into two tomato cultivars, Jamila and Tomaland.

PCR, northern blotting, and small interfering RNAs (siRNA) analysis were performed to validate the success of Agrobacterium-mediated transformation. The efficiency of transformation was highest for the T-452 construct. In vivo effects of the transformed constructs were confirmed in feeding experiments, and there was significant downregulation of the Ace 1 gene. In addition, an aphid challenge assay was conducted to investigate the siRNA-mediated silencing of the target gene (Ace 1) in the inhibition of fecundity in M. persicae.

The researchers found that the plants that were transformed with the T-452 vector had 37.5% and 26.4% lower fecundity at 27 °C in the Jamila and Tomaland, respectively. The results strongly indicated that the plant-mediated silencing of aphid-RNA might be a robust and effective approach for developing pest and disease resistant in plants.

Access the full study at Agronomy.

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