Development and application of gene editing in citrus: how and what to do DOI Creative Commons
Shuai Wang,

Sailan Zhao,

Meixia Du

et al.

Horticulture Advances, Journal Year: 2024, Volume and Issue: 2(1)

Published: Dec. 10, 2024

Abstract Conventional breeding techniques have been effectively utilized for the enhancement of citrus varieties. Nonetheless, traits such as an extended juvenile phase, cross- or self-incompatibility, high genetic heterozygosity, and polyembryony posed significant challenges limitations to these methods. The clustered regularly interspaced short palindromic repeats (CRISPR) genome editing has progressively emerged a vital tool research. This article reviews array CRISPR/Cas systems, emphasizes recent advancements in using CRISPR/Cas, explores application this technology bolster resistance canker. review also covers development CRISPR/Cas-mediated transformation regeneration systems citrus, alongside approaches generating transgene-free germplasm. Moreover, regulatory landscape societal acceptance are examined. Lastly, potential applications proposed, with attention prospective challenges.

Language: Английский

A detailed landscape of CRISPR-Cas-mediated plant disease and pest management DOI Open Access
Subhasis Karmakar, Priya Das,

Debasmita Panda

et al.

Plant Science, Journal Year: 2022, Volume and Issue: 323, P. 111376 - 111376

Published: July 11, 2022

Language: Английский

Citations

63
Generation of the transgene-free canker-resistant Citrus sinensis using Cas12a/crRNA ribonucleoprotein in the T0 generation DOI Creative Commons
Hang Su, Yuanchun Wang, Jin Xu

et al.

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: July 4, 2023

Citrus canker caused by Xanthomonas citri subsp. (Xcc) is a destructive citrus disease worldwide. Generating disease-resistant cultivars the most effective, environmentally friendly and economic approach for control. However, traditional breeding lengthy laborious. Here, we develop transgene-free canker-resistant sinensis lines in T0 generation within 10 months through transformation of embryogenic protoplasts with Cas12a/crRNA ribonucleoprotein to edit susceptibility gene CsLOB1. Among 39 regenerated lines, 38 are biallelic/homozygous mutants, demonstrating 97.4% mutation rate. No off-target mutations detected edited lines. Canker resistance cslob1-edited results from both abolishing symptoms inhibiting Xcc growth. The C. have received regulatory approval USDA APHIS exempted EPA regulation. This study provides sustainable efficient control solution presents an genome-editing strategy other crops.

Language: Английский

Citations

43
A Review on Advanced CRISPR-Based Genome-Editing Tools: Base Editing and Prime Editing DOI
Ali Saber Sichani, Maryam Ranjbar,

Maryam Baneshi

et al.

Molecular Biotechnology, Journal Year: 2022, Volume and Issue: 65(6), P. 849 - 860

Published: Dec. 22, 2022

Language: Английский

Citations

40
Transgene-free genome editing of vegetatively propagated and perennial plant species in the T0 generation via a co-editing strategy DOI
Xiaoen Huang,

Hongge Jia,

Jin Xu

et al.

Nature Plants, Journal Year: 2023, Volume and Issue: 9(10), P. 1591 - 1597

Published: Sept. 18, 2023

Language: Английский

Citations

31
CRISPR technology towards genome editing of the perennial and semi-perennial crops citrus, coffee and sugarcane DOI Creative Commons
Guilherme Souza Prado,

Dhiôvanna Corrêia Rocha,

Lucas Nascimento dos Santos

et al.

Frontiers in Plant Science, Journal Year: 2024, Volume and Issue: 14

Published: Jan. 8, 2024

Gene editing technologies have opened up the possibility of manipulating genome any organism in a predicted way. CRISPR technology is most used tool and, agriculture, it has allowed expansion possibilities plant biotechnology, such as gene knockout or knock-in, transcriptional regulation, epigenetic modification, base editing, RNA prime and nucleic acid probing detection. This mostly depends on

Language: Английский

Citations

14
An Agrobacterium‐mediated base editing approach generates transgene‐free edited banana DOI
Senne Van den Broeck, Yvan Ngapout, Bart Panis

et al.

New Phytologist, Journal Year: 2025, Volume and Issue: unknown

Published: April 1, 2025

Genome editing for the development of improved varieties is supported by possibility segregating out editor T-DNA cassette after genome in many crop species. Removal prevents potential continuous activity transformed plant and furthermore facilitates regulatory approval. While transgene outcrossing exogenous sequences possible crops, this not case vegetatively propagated sterile such as Cavendish bananas. Therefore, gene techniques leading to transgene-free edited plants are essential untap those crops. Here, we present a method banana (Musa spp.) through co-editing strategy. A novel Agrobacterium tumefaciens-mediated approach combining embryogenesis chlorsulfuron selection was established validated whole sequencing. Editing acetolactate synthase (MaALS) genes using base allows effective plants. Moreover, plantlets were regenerated with mutations at two target sites, indicating that strategy can be used multiple genomic sites. The presented efficient represents first report

Language: Английский

Citations

1
Strategies for delivery of CRISPR/Cas-mediated genome editing to obtain edited plants directly without transgene integration DOI Creative Commons
Zuzana Kocsisova, Viktoriya Coneva

Frontiers in Genome Editing, Journal Year: 2023, Volume and Issue: 5

Published: July 20, 2023

Increased understanding of plant genetics and the development powerful easier-to-use gene editing tools over past century have revolutionized humankind's ability to deliver precise genotypes in crops. Plant transformation techniques are well developed for making transgenic varieties certain crops model organisms, yet reagent delivery regeneration remain key bottlenecks applying technology most Typical protocols produce transgenic, genetically modified (GM) rely on transgenes, chemical selection, tissue culture. make edited (GE) also use even though these may be undesirable final crop product. In some crops, transgenes routinely segregated away during meiosis by performing crosses, thus only a minor concern. other particularly those propagated vegetatively, complex hybrids, or with long generation times, such crosses impractical impossible. This review highlights diverse strategies CRISPR/Cas reagents regenerable cells recover plants without unwanted integration transgenes. Some examples include delivering DNA-free as ribonucleoproteins mRNA, relying expression from non-integrated DNA, using novel mechanisms viruses nanoparticles, unconventional selection methods avoid and/or avoiding culture altogether. These advancing rapidly already enabling scientists precision CRISPR tools.

Language: Английский

Citations

21
Promoter editing for the genetic improvement of crops DOI
Shi Lu, Jing Su, Myeong‐Je Cho

et al.

Journal of Experimental Botany, Journal Year: 2023, Volume and Issue: 74(15), P. 4349 - 4366

Published: May 19, 2023

Abstract Gene expression plays a fundamental role in the regulation of agronomically important traits crop plants. The genetic manipulation plant promoters through genome editing has emerged as an effective strategy to create favorable crops by altering pattern pertinent genes. Promoter can be applied directed manner, where nucleotide sequences associated with are precisely generated. Alternatively, promoter also exploited random mutagenic approach generate novel variations within designated promoter, from which elite alleles selected based on their phenotypic effects. Pioneering studies have demonstrated potential engineering well mining valuable for breeding. In this review, we provide update application increased yield, enhanced tolerance biotic and abiotic stresses, improved quality. We discuss several remaining technical bottlenecks how may better employed improvement future.

Language: Английский

Citations

18
Generation of transgene-free canker-resistant Citrus sinensis cv. Hamlin in the T0 generation through Cas12a/CBE co-editing DOI Creative Commons

Hongge Jia,

Ahmad A. Omar, Jin Xu

et al.

Frontiers in Plant Science, Journal Year: 2024, Volume and Issue: 15

Published: March 26, 2024

Citrus canker disease affects citrus production. This is caused by Xanthomonas citri subsp. (Xcc). Previous studies confirmed that during Xcc infection, PthA4, a transcriptional activator like effector (TALE), translocated from the pathogen to host plant cells. PthA4 binds binding elements (EBEs) in promoter region of susceptibility gene LOB1 (EBE -LOBP) activate its expression and subsequently cause symptoms. Previously, Cas12a/CBE co-editing method was employed disrupt EBE -LOBP pummelo, which highly homozygous. However, most commercial cultivars are heterozygous hybrids more difficult generate homozygous/biallelic mutants. Here, we edit Hamlin ( sinensis ), hybrid cultivar grown worldwide. Binary vector GFP-p1380N-ttLbCas12a:LOBP1-mPBE:ALS2:ALS1 constructed shown be functional via Xcc-facilitated agroinfiltration leaves. construct allows selection transgene-free regenerants GFP, edits ALS chlorsulfuron-resistant as marker for genome editing resulting transient T-DNA nCas9-mPBE:ALS2:ALS1, gene(s) interest (i.e., this study) through ttLbCas12a, thus creating citrus. Totally, 77 plantlets were produced. Among them, 8 transgenic plants (#Ham GFP 1 - #Ham 8), 4 NoGFP 4), rest wild type. plantlets, three lines 1, 2 3) contained biallelic mutations pthA4 , one line 4) had homozygous . We achieved 5.2% mutation efficiency –LOBP C. cv. Hamlin, compared 1.9% pummelo previous study. Importantly, four 3 survived resistant against canker. Taken together, has been successfully used canker‐resistant T0 generation biallelic/homozygous LOB1.

Language: Английский

Citations

7
Applications of CRISPR/Cas genome editing in economically important fruit crops: recent advances and future directions DOI Creative Commons

Zhimin Ma,

Lijing Ma, Junhui Zhou

et al.

Molecular Horticulture, Journal Year: 2023, Volume and Issue: 3(1)

Published: Jan. 28, 2023

Fruit crops, consist of climacteric and non-climacteric fruits, are the major sources nutrients fiber for human diet. Since 2013, CRISPR/Cas (Clustered Regularly Interspersed Short Palindromic Repeats CRISPR-Associated Protein) genome editing system has been widely employed in different plants, leading to unprecedented progress genetic improvement many agronomically important fruit crops. Here, we summarize latest advancements including efforts decipher mechanisms behind plant development immunity, We also highlight potential challenges improvements application tools optimizing expression cassette, improving delivery efficiency reagents, increasing specificity editing, transformation regeneration system. In addition, propose perspectives on crop breeding especially crops challenges. It is worth noting that manipulate with systems urgently needed demonstration.

Language: Английский

Citations

14