Baldi, P.

Gualandri, V.

Komjanc, M.

Malnoy, M.

Nicolussi Golo, G.

Patocchi, A.

Piazza, S.

Yousaf, A.

English

Archivio istituzionale della ricerca - Fondazione Edmund Mach

Proceedings of the LXVI SIGA Annual CongressBari, 5/8 September, 2023ISBN: 978-88-944843-4-2Poster Communication Abstract – 7.30 IMPROVE APPLE SCAB RESISTANCE VIA CISGENIC APPROACHBALDI P.*, YOUSAF A.**, PIAZZA S.***, NICOLUSSI GOLO G.****, GUALANDRI V.*****, MATTEO K.******, PATOCCHI A.*******, MALNOY M.*********) Foundation Edmund Mach**) AgroscopeNew breeding technology, Malus, cisgenesis, biotic stress resistanceApple scab is one of the most common and most serious diseases that effectsapple trees worldwide, caused by Venturia inaequalis which can be reducedby unique resistance genes. At present, we are cloning several apple scabgenes. One of them, the apple scab Rvi 12 (Vb), originating from Siberiancrab apple 'Hansen's baccata #2 has been mapped previously and a candidategene, named Rvi12_Cd5, belonging to LRR receptor-like serine/threonine-protein kinase family was identified. In this study, the DNA sequence ofnative promoter was extended and characterized by in-silico analysis whichshowed homology with major defense related transcription factor bindingsites such as ERF, bZIP, bHLH, MYBs, NAC and WRKY. The coding sequence ofRvi12_Cd5 controlled by the native promoter and 35S promoter along theselectable marker gene nptII was introduced by agrobacterium-mediatedtransformation in the apple scab susceptible ‘Gala’ cultivar. Successful 45independent transgenic lines were characterized for copy number andtranscription of the candidate resistance gene. The transformed lines with1 or 2 integrations, expressing the candidate gene were acclimatized to thegreenhouse for scab inoculations. The phenotypic characterization showedsignificant reduction in scab susceptibility in several transformed linescompared to control ‘Gala’ plants. Additionally, the NGS method wasperformed for the identification of T-DNA integration points in transformed‘lines carrying the vector with the native promoter, resulting in 5chromosomal positions in the apple genome. Moreover, these plants weresubjected to T-DNA removal using the heat-shock inducible FLP/FRTrecombination system and confirmed by quantifying marker gene nptII. Thereal-time PCR showed complete exogenous DNA elimination in severalindependent lines. Currently, these results are being validated bysequencing.

Improve apple scab resistance via cisgenic approach

https://openpub.fmach.it/bitstream/10449/81737/3/2023%20SIGA%20Malnoy%207.30.pdf

Improve apple scab resistance via cisgenic approach

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Archivio istituzionale della ricerca - Fondazione Edmund Mach