CRISPR/Cas-Mediated Genome Editing for the Improvement of Oilseed Crop Productivity

Subedi, Udaya; Ozga, Jocelyn A.; Chen, Guanqun; Foroud, Nora A.; Singer, Stacy D.

doi:10.1080/07352689.2020.1782568

Cited by 12 publications

(8 citation statements)

References 277 publications

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“…This platform requires a Cas nuclease, which produces a double-stranded DNA break, and a single guide RNA (sgRNA), which includes a short user-defined sequence that guides the Cas nuclease to a highly specific chromosomal locus of choice immediately upstream of a protospacer-adjacent motif (PAM). Due to the unlinked nature of the resulting edit and the introduced transgene, this technology allows for the rapid production of non-transgenic germplasm bearing mutations that are identical in nature to those achieved spontaneously or through conventional breeding approaches such as chemical mutagenesis ( Subedi et al, 2020a , b ). While the regulatory status of crop varieties derived from genome editing is still uncertain in some countries, many others, including the United States, have concluded that in the absence of foreign DNA they are not “GM,” and will therefore not be subjected to costly and burdensome regulatory processes ( Schmidt et al, 2020 ; Singer et al, 2021a ).…”

Section: Introductionmentioning

confidence: 99%

The CRISPR/Cas9-Mediated Modulation of SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE 8 in Alfalfa Leads to Distinct Phenotypic Outcomes

et al. 2022

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Alfalfa (Medicago sativa L.) is the most widely grown perennial leguminous forage and is an essential component of the livestock industry. Previously, the RNAi-mediated down-regulation of alfalfa SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE 8 (MsSPL8) was found to lead to increased branching, regrowth and biomass, as well as enhanced drought tolerance. In this study, we aimed to further characterize the function of MsSPL8 in alfalfa using CRISPR/Cas9-induced mutations in this gene. We successfully generated alfalfa genotypes with small insertions/deletions (indels) at the target site in up to three of four MsSPL8 alleles in the first generation. The efficiency of editing appeared to be tightly linked to the particular gRNA used. The resulting genotypes displayed consistent morphological alterations, even with the presence of up to two wild-type MsSPL8 alleles, including reduced leaf size and early flowering. Other phenotypic effects appeared to be dependent upon mutational dosage, with those plants with the highest number of mutated MsSPL8 alleles also exhibiting significant decreases in internode length, plant height, shoot and root biomass, and root length. Furthermore, MsSPL8 mutants displayed improvements in their ability to withstand water-deficit compared to empty vector control genotypes. Taken together, our findings suggest that allelic mutational dosage can elicit phenotypic gradients in alfalfa, and discrepancies may exist in terms of MsSPL8 function between alfalfa genotypes, growth conditions, or specific alleles. In addition, our results provide the foundation for further research exploring drought tolerance mechanisms in a forage crop.

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Section: Introductionmentioning

confidence: 99%

The CRISPR/Cas9-Mediated Modulation of SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE 8 in Alfalfa Leads to Distinct Phenotypic Outcomes

et al. 2022

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“…In addition, the large‐scale interspecific hybridization has accumulated unique resources with rich genetic diversity and genomic structural variants (Song et al ., 1995 ; Udall et al ., 2005 ). However, this germplasm was created based on traditional breeding techniques that are often imprecise and long‐term processes (Subedi et al ., 2020 ), and their value in trait improvement and heterosis utilization will be improved and fully exploited with the assistance of omics analysis, genome‐wide selection, speed breeding, and gene editing technology. First, it is very important to comprehensively dissect the complex genomic structures and pangenome diversity across the Brassica species, which could help build a genome‐wide atlas for genome‐based improvement.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, gene editing has become the most popular and promising technique for functional genomics, and it promises to revolutionize crop improvement by creating new non‐transgenic varieties in a fast, efficient, and technically simple way without the use of transgenes (Schenke and Cai, 2020 ; Shelake et al ., 2019 ; Songstad et al ., 2017 ; Subedi et al ., 2020 ). Gene editing makes it possible to generate targeted deletions, insertions, gene knockouts, and point mutations, to modulate gene expression by targeting transcription factors or epigenetic machinery to DNA, or to target and modify RNA (Broeders et al ., 2020 ).…”

Section: Challenges and Approaches For Further Exploring And Expanding The Rapeseed Gene Poolmentioning

confidence: 99%

Exploring the gene pool ofBrassica napusby genomics‐based approaches

Jing

Snowdon

et al. 2021

Plant Biotechnology Journal

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Summary De novo allopolyploidization in Brassica provides a very successful model for reconstructing polyploid genomes using progenitor species and relatives to broaden crop gene pools and understand genome evolution after polyploidy, interspecific hybridization and exotic introgression. B. napus (AACC), the major cultivated rapeseed species and the third largest oilseed crop in the world, is a young Brassica species with a limited genetic base resulting from its short history of domestication, cultivation, and intensive selection during breeding for target economic traits. However, the gene pool of B. napus has been significantly enriched in recent decades that has been benefit from worldwide effects by the successful introduction of abundant subgenomic variation and novel genomic variation via intraspecific, interspecific and intergeneric crosses. An important question in this respect is how to utilize such variation to breed crops adapted to the changing global climate. Here, we review the genetic diversity, genome structure, and population‐level differentiation of the B. napus gene pool in relation to known exotic introgressions from various species of the Brassicaceae, especially those elucidated by recent genome‐sequencing projects. We also summarize progress in gene cloning, trait‐marker associations, gene editing, molecular marker‐assisted selection and genome‐wide prediction, and describe the challenges and opportunities of these techniques as molecular platforms to exploit novel genomic variation and their value in the rapeseed gene pool. Future progress will accelerate the creation and manipulation of genetic diversity with genomic‐based improvement, as well as provide novel insights into the neo‐domestication of polyploid crops with novel genetic diversity from reconstructed genomes.

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“…Depending on the type of Cas9, Cas9 recognizes a PAM sequence that is different from NGG ( Leenay and Beisel, 2017 ). Since the introduction of CRISPR/Cas9 technology, genome editing research has been conducted in various organisms, such as plants, humans, and microalgae ( Jeon et al, 2017 ; Jaganathan et al, 2018 ; Subedi et al, 2020 ; Li et al, 2020b ).…”

Section: Crispr/cas9 and Lipid Metabolic Engineeringmentioning

confidence: 99%

Applications and prospects of genome editing in plant fatty acid and triacylglycerol biosynthesis

Park

Kim²

2022

Front. Plant Sci.

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Triacylglycerol (TAG), which is a neutral lipid, has a structure in which three molecules of fatty acid (FA) are ester-bonded to one molecule of glycerol. TAG is important energy source for seed germination and seedling development in plants. Depending on the FA composition of the TAG, it is used as an edible oil or industrial material for cosmetics, soap, and lubricant. As the demand for plant oil is rising worldwide, either the type of FA must be changed or the total oil content of various plants must be increased. In this review, we discuss the regulation of FA metabolism by Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9, a recent genome-editing technology applicable to various plants. The development of plants with higher levels of oleic acid or lower levels of very long-chain fatty acids (VLCFAs) in seeds are discussed. In addition, the current status of research on acyltransferases, phospholipases, TAG lipases, and TAG synthesis in vegetative tissues is described. Finally, strategies for the application of CRISPR/Cas9 in lipid metabolism studies are mentioned.

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CRISPR/Cas-Mediated Genome Editing for the Improvement of Oilseed Crop Productivity

Cited by 12 publications

References 277 publications

The CRISPR/Cas9-Mediated Modulation of SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE 8 in Alfalfa Leads to Distinct Phenotypic Outcomes

The CRISPR/Cas9-Mediated Modulation of SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE 8 in Alfalfa Leads to Distinct Phenotypic Outcomes

Exploring the gene pool ofBrassica napusby genomics‐based approaches

Applications and prospects of genome editing in plant fatty acid and triacylglycerol biosynthesis

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