Articles
Grapevine genome editing for improving disease resistance and quality traits
Article number
1454_3
Pages
21 – 26
Language
English
Abstract
Conventional breeding of grapevine has limited applications for improving disease resistance and quality traits due to the heterozygous nature of the genome, incompatibility barriers among Vitis species and a long juvenile period.
Genome editing of grapevine has the potential to improve specific traits in elite cultivars without disrupting existing desirable characteristics.
In the current study, the grapevine Mildew Locus O (MLO) and Polyphenol Oxidase (PPO) genes were targeted using CRISPR/Cas 9-mediated genome editing to study plant response to Erysiphe necator infection, cluster/berry browning and fruit shelf life.
A binary vector containing dual guide RNA sequences targeting the MLO-6, MLO-7 and the PPO genes individually, along with Cas 9 and a hygromycin selectable marker, under the control of a CaMV 35S promoter was transferred to Agrobacterium. Embryogenic cultures were transformed with Agrobacterium harboring the binary vector to generate transgenic plant lines.
Molecular analyses including PCR and sequencing confirmed the presence of the Cas9 and hygromycin genes in regenerated plant lines.
Transgenic plant lines were asexually propagated to produce replicate clones and screened along with non-transgenic controls in a greenhouse for their response to infection by Erysiphe necator. Among the various plant lines studied, 3 exhibited a significant decrease in powdery mildew symptoms and infection severity compared to the non-trangenic controls.
We are currently growing transgenic plant lines with a CRISPR-Cas construct targeting PPO, to induce fruiting and evaluate cluster/berry browning and fruit shelf life.
The use of CRISPR/Cas-9 mediated genome editing can provide durable disease resistance and improved fruit quality traits with the potential to decrease production costs and ensure sustainability of the grape and wine industry.
Genome editing of grapevine has the potential to improve specific traits in elite cultivars without disrupting existing desirable characteristics.
In the current study, the grapevine Mildew Locus O (MLO) and Polyphenol Oxidase (PPO) genes were targeted using CRISPR/Cas 9-mediated genome editing to study plant response to Erysiphe necator infection, cluster/berry browning and fruit shelf life.
A binary vector containing dual guide RNA sequences targeting the MLO-6, MLO-7 and the PPO genes individually, along with Cas 9 and a hygromycin selectable marker, under the control of a CaMV 35S promoter was transferred to Agrobacterium. Embryogenic cultures were transformed with Agrobacterium harboring the binary vector to generate transgenic plant lines.
Molecular analyses including PCR and sequencing confirmed the presence of the Cas9 and hygromycin genes in regenerated plant lines.
Transgenic plant lines were asexually propagated to produce replicate clones and screened along with non-transgenic controls in a greenhouse for their response to infection by Erysiphe necator. Among the various plant lines studied, 3 exhibited a significant decrease in powdery mildew symptoms and infection severity compared to the non-trangenic controls.
We are currently growing transgenic plant lines with a CRISPR-Cas construct targeting PPO, to induce fruiting and evaluate cluster/berry browning and fruit shelf life.
The use of CRISPR/Cas-9 mediated genome editing can provide durable disease resistance and improved fruit quality traits with the potential to decrease production costs and ensure sustainability of the grape and wine industry.
Authors
S.A. Dhekney, P. Sardaru, P. Natarajan
Keywords
Vitis, gene editing, disease resistance, browning
Groups involved
- Division Plant Genetic Resources, Breeding and Biotechnology
- Working Group Genetic Transformation and Gene Editing
- Working Group Horticultural Biotechnology and Breeding
- Division Ornamental Plants
- Division Vegetables, Roots and Tubers
- Division Temperate Tree Fruits
- Division Vine and Berry Fruits
- Division Horticulture for Development
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