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

MIRKIN, FEDERICO GABRIEL; BOCCARDO, NOELIA AYELEN; SEGRETIN, MARÍA EUGENIA; BRAVO-ALMONACID, FERNANDO FELIX

Congreso; XXXII Reunión Argentina de Fisiología Vegetal y XV Congreso Latinoamericano de Fisiología Vegetal; 2018

Insect pests represent a serious threat to food safety.Several strategies have shown effective in mitigatingcrop losses but unfortunately, due to the appearanceof resistant insects, new approaches are needed. Theexpression of double stranded RNAs (dsRNA) againstinsect targetsin plantsis an effectivestrategy for thismatter. In particular, theexpression of dsRNAsin plastids allows high level protection against coleopteransand lepidopterans due to the accumulation of longdsRNAs, which are more effective to trigger silencingin the insect than short interfering RNAs. With the objective of evaluating if this is a valid strategy againsthemipteran pests, we developed genetic constructs toobtain transplastomic and nuclear transgenic plantsto express dsRNAs with homology to Myzus persicaeMpc002 and v-atpase subunit E sequences, and Bemisia tabaci v-atpase subunit E sequence. For theplastometransformation we modified the plastid transformation vector pUTRGus previously developed in ourlaboratory, to adapt it for Golgen Gate cloning strategyfor fast and easy assemblyofvectors design toexpressintron containing hairpins in plastids. We obtainedtransplastomic and nuclear transgenic plantsthat arebeing characterized at the molecular level.