Biotic Stress/Applied Plant Bio

Abstract

CS-8-1 - Plastid Transformation in Arabidopsis thaliana

Sunday, July 15
3:18 PM - 3:38 PM

Deletion of the acc2 nuclear gene makes Arabidopsis hypersensitive to spectinomcyin, the selective agent for the identification of transplastomic events. We have found that plastid transformation efficiency is 100x higher in the acc2 knockout background than in wild type Arabidopsis. However, it was difficult to obtain fertile transplastomic plants in the available Col-0 acc2-1 and Sav0 hypersensitive accessions (Yu et al. Plant Physiol. 175: 186, 2017). We deleted the ACC2 gene using CRISPR/Cas9 in the regenerable RLD and Ws ecotypes and have shown that deletion of ACC2  makes the seedlings hypersensitive to spectinomycin. Selection for spectinomycin resistance in bombarded leaf cultures of the hypersensitive RLD and Ws backgrounds yielded putative transplastomic events at a high frequency. Classification of the clones as transplastomic lines or spontaneous spectinomycin resistant mutants and demonstration of seed transmission of spectinomycin resistance is in progress.


 

Co-Authors

Lisa Lamanna – Rutgers University; Qiguo Yu – Rutgers University; Megan Kelly – Rutgers University; Kerry Lutz – Farmingdale State College

Pal Maliga, PhD

Prof.
Rutgers University

Pal Maliga (Rutgers University) developed a method for stable transformation of land plant chloroplast genomes. Chloroplast genome engineering in higher plants has led to an explosion of research concerning the chloroplast genome’s role in photosynthesis, functional analysis of plastid genes by reverse genetics, and mechanisms of plastid gene regulation. His current research interest lies in tissue specific regulation of plastid transgene expression using PPR RNA binding proteins. His research group is also working on a protocol for plastid transformation in Arabidopsis, where they aim to provide a readily applicable research tool for the plant community.

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CS-8-1 - Plastid Transformation in Arabidopsis thaliana



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