Generation of transplastomic tobacco plants expressing dsRNAs against hemipteran targets for plant protection

MIRKIN FG; NOELIA BOCCARDO; SEGRETIN ME; BRAVO ALMONACID F

Congreso; XXXII Reunión Argentina XVI Congreso Latinoamericano de Fisiología Vegetal; 2018

Insect pests represent a serious threat to food safety. Several strategies have shown effective in mitigating crop losses but unfortunately, due to the appearance of resistant insects, new approaches are needed. The expression of double stranded RNAs (dsRNA) against insect targets in plants is an effective strategy for this matter. In particular, the expression of dsRNAs in plastids allows high level protection against coleopterans and lepidopterans due to the accumulation of long dsRNAs, which are more effective to trigger silencing in the insect than short interfering RNAs. With the objective of evaluating if this is a valid strategy against hemipteran pests, we developed genetic constructs to obtain transplastomic and nuclear transgenic plants to express dsRNAs with homology to Myzus persicae Mpc002 and V-Atpase subunit E sequences, and Bemisia tabaci v-atpase subunit E sequence. For plastome transformation we modified the plastid transformation vector pUTRGus previously developed in our laboratory, to adapt it for Golgen Gate cloning strategy for fast and easy assembly of vectors design to express intron containing hairpins in plastids. We obtained transplastomic and nuclear transgenic plants that are being characterized at the molecular level.