Grapevine genome editing: development of tools and implementation of the technology

Project Number
GENUS 17-02

Project title
Grapevine genome editing: development of tools and implementation of the technology

Project leader
Maree H J Campa, M

University of Stellenbosch. Department of Genetics

Team members
Young, P R
Korkie, M

Project description
The negative economic impact of biotic and abiotic stresses in vines are recognised by the international viticulture industry, and has prompted a multitude of research projects, spanning decades, and costing millions. To this day, natural tolerance, resistance or cures for these afflictions are rare, and best practises still rely on prevention of these stresses and diseases. In South Africa, promising leads from earlier GM-based approaches to address some of these problems were never allowed to mature, largely as a result of public acceptance issues dictated by the South African agricultural export markets.

The rapid developments in genome editing technologies over the last few years, and especially the versatility demonstrated in many applications of CRISPR/Cas9-based technology, may impact radically in the ongoing battle with most of these conditions in vineyards all over the world. As a first step to unlock the immense potential of this technology in the local industry, this project will aim to establish CRISPR technology in grapevine. During this proof-of-concept phase, we will establish a conventional CRISPR/Cas9-based protocol for editing the grapevine genome by targeting a single copy carotenoid gene that are central to grapevine secondary plant metabolism and will lead to a readily monitored phenotype. Considering the negative impact of viruses on the local and international grapevine industries, and the fact that they have RNA genomes, we will also attempt to silence an RNA transcript of an easily-monitored grapevine gene – the latter acting as proof-of-concept proxy to a virus like Grapevine leafroll-associated virus 3.

The project will comprise four major objectives, each comprising of a number of tasks with related milestones. The first objective will be the construction of all molecular vectors to introduce the different genetic elements required for CRISPR-based genome editing. The second objective will comprise the preparation of plant materials for routine genetic transformation protocols for Vitis cultivar(s), while the third will attempt to introduce the different genetic elements into appropriate grapevine tissues, and to regenerate grapevine plantlets for subsequent analysis. Finally, primary transformants will be evaluated, both phenotypically, and at a molecular level for efficacy and accuracy of the editing.

– Record end –

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