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Gene editing in plants:progress and challenges

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National Science Review 2019年第3期6卷 421-437页

作者： Yanfei Mao jose ramon Botella Yaoguang Liu Jian-Kang Zhu Shanghai Center for Plant Stress Biology CAS Center of Excellence in Molecular Plant Sciences Chinese Academy of Sciences School of Agriculture and Food Sciences University of Queensland State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources College of Life Sciences South China Agricultural University Department of Horticulture and Landscape Architecture Purdue University

The clustered regularly interspaced short palindromic repeat(CRISPR)-associated protein 9(Cas9)genome editing system is a powerful tool for targeted gene modifications in a wide range of species,including plants. Over the last few years, this system has revolutionized the way scientists perform genetic studies and crop breeding, due to its simplicity, flexibility, consistency and high efficiency. Considerable progress has been made in optimizing CRISPR/Cas9 systems in plants, particularly for targeted gene mutagenesis. However, there are still a number of important challenges ahead, including methods for the efficient delivery of CRISPR and other editing tools to most plants, and more effective strategies for sequence knock-ins and replacements. We provide our viewpoint on the goals, potential concerns and future challenges for the development and application of plant genome editing tools.

关键词： CRISPR Cas9 genome editing base editing gene targeting crop breeding

Perspectives on the Application of Genome-Editing Technologies in Crop Breeding

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Molecular Plant 2019年第8期12卷 1047-1059页

作者： Kai Hua Jinshan Zhang jose ramon Botella Changle Ma Fanjiang Kong Baohui Liu Jian-Kang Zhu Shanghai Center for Plant Stress Biology CAS Center of Excellence in Molecular Plant SciencesChinese Academy of SciencesShanghai 200032China School of Agriculture and Food Sciences University of QueenslandBrisbaneQLD 4072Australia Shandong Provincial Key Lab of Plant Stress School of Life SciencesShandong Normal UniversityJinanChina School of Life Sciences Guangzhou UniversityGuangzhouChina Department of Horticulture and Landscape Architecture Purdue UniversityWest LafayetteIN 47907USA

Most conventional and modern crop-improvement methods exploit natural or artificially induced genetic variations and require laborious characterization of the progenies of multiple generations derived from time-consuming genetic crosses.Genome-editing systems,in contrast,provide the means to rapidly modify genomes in a precise and predictable way,making it possible to introduce improvements directly into elite varieties.Here,we describe the range of applications available to agricultural researchers using existing genome-editing tools.In addition to providing examples of genome-editing applications in crop breeding,we discuss the technical and social challenges faced by breeders using genome-editing tools for crop improvement.

关键词： genome editing crop breeding mutations base editing plants

Gene Targeting by Homology-Directed Repair in Rice Using a Geminivirus-Based CRISPR/Cas9 System

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Molecular Plant 2017年第7期10卷 1007-1010页

作者： Mugui Wang Yuming Lu jose ramon Botella Yanfei Mao Kai Hua Jian-kang Zhu Shanghai Center for Plant Stress Biology and Center for Excellence in Molecular Plant Sciences Chinese Academy of Sciences Shanghai 201602 China Plant Genetic Engineering Laboratory School of Agriculture and Food Sciences University of Queensland Brisbane QLD 4072 Austraria Department of Horticulture and Landscape Architecture Purdue University West Lafayette IN 47907 USA

Dear Editor, Rice （Oryza sativa） is the staple food for more than half of the world＇s population. Technologies enabling precise and efficient DNA knock-in or replacement, hereinafter referred to as KI, have the pot... 详细信息

关键词： DNA REPLICONS GENOME

A detailed procedure for CRISPR/Cas9-mediated gene editing in Arabidopsis thaliana

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Science Bulletin 2015年第15期60卷 1332-1347页

作者： Wenshan Liu Xiaohong Zhu Mingguang Lei Qingyou Xia jose ramon Botella Jian-Kang Zhu Yanfei Mao School of Life Sciences Chongqing University Shanghai Center for Plant Stress Biology Shanghai Institutes of Biological Sciences Chinese Academy of Sciences State Key Laboratory of Silkworm Genome Biology Southwest University Department of Horticulture and Landscape Architecture Purdue University School of Agriculture and Food Sciences University of Queensland

The newly developed CRISPR(Clustered Regularly Interspaced Short Palindromic Repeats)/Cas(CRISPR-associated) system has emerged as an efficient tool for genome-editing, providing an alternative to classical mutagenesis and transgenic methods to study gene function and improve crop traits. CRISPR/Cas facilitates targeted gene editing through RNA-guided DNA cleavage followed by cellular DNA repair mechanisms that introduce sequence changes at the site of cleavage. Here we describe a detailed procedure for our previously developed and highly efficient CRISPR/Cas9 method that allows the generation of heritable-targeted gene mutations andcorrections in Arabidopsis. This protocol describes the strategies and steps for the selection of targets, design of single-guide RNA(sg RNA), vector construction and analysis of transgenic lines. We also offer a method to target two loci simultaneously using vectors containing two different sg RNAs. The principles described in this protocol can be applied to other plant species to generate stably inherited DNA modifications.

关键词： RNA编辑基因组拟南芥转基因载体 DNA裂解介导程序协议描述

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Generation of new glutinous rice by CRISPR/Cas9-targeted mutagenesis of the Waxy gene in elite rice varieties

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Journal of Integrative Plant Biology 2018年第5期60卷 369-375页

作者： Jinshan Zhang Hui Zhang jose Ramón botella Jian-Kang Zhu Shanghai Center for Plant Stress Biology and Center for Excellence in Molecular Plant SciencesChinese Academy of SciencesShanghai 201602China University of Chinese Academy of Sciences Shanghai 201602China Department of Horticulture and Landscape Architecture Purdue UniversityWest LafayetteIN 47907USA Plant Genetic Engineering Laboratory School of Agriculture and Food SciencesUniversity of QueenslandBrisbane QLD 4072Australia

In rice, amylose content （AC） is controlled by a single dominant Waxy gene. We used Clustered Regularly Interspaced Short Palindromic Repeats （CRISPR）/CRISPR-associated 9 （Casg） to introduce a loss-of-function mutation into the Waxy gene in two widely cultivated elite japonica varieties. Our results show that mutations in the Waxy gene reduce AC and convert the rice into glutinous ones without affecting other desirable agronomic traits, offering an effective and easy strategy to improve glutinosity in elite varieties. Importantly, we successfully removed the transgenes from the progeny. Our study provides an example of generating improved crops with potential for commercialization, by editing a gene of interest directly in elite crop varieties.

关键词： Generation of new glutinous rice by CRISPR/Cas9-targeted mutagenesis of the Waxy gene in elite rice varieties Figure AC

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VERNAUZATION1 Modulates Root System Architecture in Wheat and Barley

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Molecular Plant 2018年第1期11卷 226-229页

作者： Kai P. Voss-Fels Hannah Robinson Stephen R. Mudge Cecile Richard Saul Newman Benjamin Wittkop Andreas Stahl Wolfgang Friedt Matthias Frisch lulian Gabur Anika Miller-Cooper Bradley C. Campbell Alison Kelly Glen Fox Jack Christopher Mandy Christopher Karine Chenu Jerome Franckowiak Emma S. Mace Andrew K. Borrell Howard Eagles David R. Jordan jose R. botella Graeme Hammer lan D. Godwin Ben Trevaskis Rod J. Snowdon Lee T. Hickey Department of Plant Breeding IFZ Research Centre for Biosystems Land Use and Nutrition Justus Uebig University Heinrich-Buff-Ring 26-32 35392 Giessen Germany Queensland Alliance for Agriculture and Food Innovation The University of Queensland St Lucia QLD 4072 Australia School of Agriculture and Food Sciences The University of Queensland Brisbane QLD 4072 Australia CSIRO Agriculture Canberra ACT 2601 Australia Department of Biometry and Population Genetics IFZ Research Centre for Biosystems Land Use and Nutrition Justus Liebig University Heinrich-Buff-Ring 26-32 35392 Giessen Germany Department of Agriculture and Fisheries Leslie Research FacilRy Toowoomba QLD 4350 Australia Queensland Alliance for Agriculture and Food Innovation The University of Queensland roowoomba QLD 4350 Australia Department of Agronomy and Plant Genetics University of Minnesota St Paul MN USA Department of Agriculture and Fisheries Hermitage Research Facility Warwick QLD 4370 Australia Queensland Alliance for Agriculture and Food Innovation Hermitage Research Facility The University of Queensland Warwick QLD 4370 Australia These authors contributed equally to this article.

Dear Editor Roots play a key role in plant growth regulation. It is well described that the below-ground plant architecture has a significant impact on plant performance under abiotic constraints and maintains stabili... 详细信息

关键词： TRAITS RESPONDS