The discovery of the CRISPR/Cas genome-editing system has revolutionized our understanding of the plant genome. CRISPR/Cas has been used for over a decade to modify plant genomes for the study of specific genes and biosynthetic pathways as well as to speed up breeding in many plant species, including both model and non-model crops. Although the CRISPR/Cas system is very efficient for genome editing, many bottlenecks and challenges slow down further improvement and applications. In this review we discuss the challenges that can occur during tissue culture, transformation, regeneration, and mutant detection. We also review the opportunities provided by new CRISPR platforms and specific applications related to gene regulation, abiotic and biotic stress response improvement, and de novo domestication of plants.

Centre for Agricultural Research Loránd Eötvös Research Network Martonvásár Hungary

Consiglio Nazionale delle Ricerche Portici Italy; CREA Research Centre for Vegetable and Ornamental Crops Pontecagnano Italy

CREA Research Centre for Vegetable and Ornamental Crops Pontecagnano Italy

Department of Agricultural Genetic Engineering Faculty of Agricultural Sciences and Technologies Nigde Omer Halisdemir University Nigde Turkey; Centre of Excellence in Molecular Biology University of the Punjab Lahore Pakistan

Department of Chemistry and Biotechnology Tallinn University of Technology Tallinn Estonia

Department of Genetics Faculty of Biological and Veterinary Sciences Nicolaus Copernicus University Toruń Poland; Centre for Modern Interdisciplinary Technologies Nicolaus Copernicus University Toruń Poland

Department of Molecular Biology and Genetics Faculty of Science Ondokuz Mayis University Samsun Turkey

Flanders Research Institute for Agricultural Fisheries and Food Melle Belgium

Heinrich Heine University Institute of Plant Biochemistry Centre for Plant Genome Engineering Düsseldorf Germany; Division of Molecular Biology Centre of the Region Hana for Biotechnological and Agriculture Research Faculty of Science Palacký University Olomouc Czech Republic

Molecular Biology and Genetics Department Institute of Biological Sciences Faculty of Biology and Environmental Sciences Cardinal Stefan Wyszyński University Warsaw Poland

Molecular Plant Breeding Institute of Agricultural Sciences Eidgenössische Technische Hochschule Zurich Switzerland; Plant Biochemistry Institute of Molecular Plant Biology ETH Zurich Switzerland

National Institute of Biology Department of Biotechnology and Systems Biology Ljubljana Slovenia

Plant Breeding and Acclimatization Institute National Research Institute Błonie Poland

Rede de Química e Tecnologia Faculdade de Farmácia Universidade do Porto Porto Portugal

Thünen Institute of Forest Genetics Grosshansdorf Germany

University of Ljubljana Biotechnical Faculty Ljubljana Slovenia

University of Novi Sad Institute of Lowland Forestry and Environment Novi Sad Serbia

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