• Non ci sono risultati.

Efficient and 'clean' CRISPR/CAS9 editing for fire blight resistance in apple

N/A
N/A
Protected

Academic year: 2021

Condividi "Efficient and 'clean' CRISPR/CAS9 editing for fire blight resistance in apple"

Copied!
1
0
0

Testo completo

(1)

Proceedings of the LXII SIGA Annual Congress Verona, Italy – 25/28 September, 2018

ISBN 978-88-904570-8-1

Poster Communication Abstract – 1.13

EFFICIENT AND ‘CLEAN’ CRISPR/CAS9 EDITING FOR FIRE BLIGHT

RESISTANCE IN APPLE

POMPILI V.*,**, PIAZZA S.*, DALLA COSTA L.*, LARGER S.*, STEFANI E.*, MALNOY M.*

*) Research and Innovation Centre, Fondazione Edmund Mach, San Michele a/A 38010 (Italy) **) Department of Agricultural Science and Biotechnology, University of Udine, Udine 33100 (Italy)

Malus x domestica, fire blight, DIPM, CRISPR/Cas9,T-DNA-free plants

Fire blight, caused by the bacterium Erwinia amylovora (E.a.), is one of the most economically important and invasive diseases affecting apple (Malus x domestica). Nowadays, the E.a.-apple interaction is widely described and the main molecular mechanisms underlying both plant host resistance and E.a. pathogenesis are elucidated. Nevertheless, the management of the disease remains still ardous in apple orchard. Indeed, the use of genetically modified plants is forbidden by the current GMO legislation and the application of antibiotic-based pesticides is prohibited due to their effects on human health and environment. Therefore, one of the hardest challenges for plant biotechnology is to provide apple varieties that are simultaneously disease-resistant and public-acceptable. In this study, conducted on two Malus x domestica susceptible varieties, ‘Royal Gala’ and ‘Golden Delicious’, a genome editing approach based on CRISPR/Cas9 via Agrobacterium tumefaciens (A.t.) was applied to produce the knock-out of MdDIPM4. In apple, evidences show that MdDIPM4 protein interacts specifically with the DspE effector, mandatory for the pathogenesis of Erwinia amylovora. About sixty transgenic lines were analyzed using an high throughput screening approach on the Illumina MiSeq platform in order to verify the CRISPR/Cas9-induced mutations. The eighty percent of the plants showed mutations, especially translation-terminated deletions resulting in MdDIPM4 knock-out. Moreover, with the aim of producing public-acceptale engineered apple plants, our strategy allowed to remove the entire exogenous T-DNA in those lines selected for the desired mutation. This mechanism relies on the heat schock-inducible expression of the Flp gene which induces a site-specific recombination at the two FRT sites flanking the A.t. left and right borders. Some genome edited lines were heat-treated and the removal of a 10.5 kb T-DNA cassette was proved. Currently, the selected genome edited and T-DNA-free apple plants are under in vivo investigation to test their resistance to Erwinia amylovora.

Riferimenti

Documenti correlati

This estimate is confirmed by an independent measurement on a separate sample of similarly-selected quiescent galaxies in the COSMOS field, using stacked far-infrared

To this end, an initial model, including about 100 sources, was obtained by analysing a Westerbork Synthesis Radio Telescope (WSRT) image at 139 MHz of the field of B1834+620

While the present article describes the techniques of signal reconstruction, event selection and detector calibration, details on the detector response model, the WIMP signal

The scientific applications of the survey range from DE stud- ies with clusters and AGNs to astrophysical investigations of low-mass groups at redshifts ∼ 0.5. The survey potential

The acronym stands for “Energy Return (ER) On Energy Invested (EI)” and is the ra- tio between the useful energy that could be extracted from a source and the investment

queste supposizioni. È comunque certo che Antistio Vetere curò il restauro della villa, in occasione del quale furono probabilmente scoperte le sorgenti di cui parla Plinio il

The results of the present study suggested that in a context where it may not be feasible to minimize job demands as the context-dependent job demand detected (i.e., having your

While establishing provisions that are not binding, in fact, the Global Compact, when expressly agreed upon, would still operate as a vehicle for international obligations, based