Recent Advances in the Citrus Genetic Engineering for Stress Tolerance/Resistance DOI
Pankaj Kumar,

Komaljeet Gill,

Shagun Sharma

et al.

Published: Jan. 1, 2023

Citrus is an economically important and commercial fruit crop grown in the tropical subtropical regions of world. There a growing need for adequate performance production, only way to meet this via implementation effective reliable breeding programs while also combating negative effects rapidly spreading diseases other abiotic stresses. biology complicated, leading polyembryony, self-incompatibility, sterility, parthenocarpy, etc., all which make conventional practices complex tedious. Besides, citrus genotypes, whether cultivated or wild, lack several desirable traits. Therefore, genetic engineering technologies provide possible solution addressing challenges traditional operations. This chapter provides systematic, updated, comprehensive overview progress over last two decades biotic stress tolerance. The alternative emerging transformation strategies, especially tissue-localized transient transformation, have been reviewed. For developing stress-resilient through plant engineering, modern such as transgenics, intragenesis, cisgenesis, genome-editing, particularly clustered regularly interspaced short palindromic repeats (CRISPR)/Cas, must be encouraged.

Language: Английский

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
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
Base Editors for Citrus Gene Editing DOI Creative Commons
Xiaoen Huang, Yuanchun Wang, Nian Wang

et al.

Frontiers in Genome Editing, Journal Year: 2022, Volume and Issue: 4

Published: Feb. 28, 2022

Base editors, such as adenine base editors (ABE) and cytosine (CBE), provide alternatives for precise genome editing without generating double-strand breaks (DSBs), thus avoiding the risk of instability unpredictable outcomes caused by DNA repair. Precise gene mediated in citrus has not been reported. Here, we have successfully adapted ABE to edit TATA box promoter region canker susceptibility

Language: Английский

Citations

31
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
The potential of CRISPR-Cas genome editing technologies to mitigate biotic stress in plants DOI Creative Commons
Israt Jahan, Prodipto Bishnu Angon,

Sharah Jabeen Mou

et al.

CABI Agriculture and Bioscience, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 9, 2025

Language: Английский

Citations

0
CRISPR plants against oxidative stress: Methods and applications DOI
Kavita Ghosal

Elsevier eBooks, Journal Year: 2025, Volume and Issue: unknown, P. 259 - 293

Published: Jan. 1, 2025

Language: Английский

Citations

0
Recent advances in innovative strategies for plant disease resistance breeding DOI Creative Commons
Dekun Wang, Rong Yang, Mengting Liu

et al.

Frontiers in Plant Science, Journal Year: 2025, Volume and Issue: 16

Published: May 21, 2025

Plant disease poses a great threat to crop production. The mechanisms underlying plant-pathogen interactions are critical research topics worldwide. In recent years, significant breakthrough studies have been reported, broadening our understanding of plant immunity. Based on these findings, many strategies developed improve defense against various diseases. Here, we summarize and their applications in aimed at promoting resistance. Besides domain swapping, gene shuffling, random mutation, three additional the last decade. first strategy is editing host susceptibility ( S ) genes prevent pathogen infection. Editing Mlo DMR6 species good examples this approach. second promoters or resistance R genes. This widely used counteract Xanthomonas , such as modifying LOB1 SWEET several crops enhance third designing products, especially nucleotide-binding leucine-rich repeat (NLR) receptors. approach based growing knowledge structural features NLRs, which seen advances recently. To date, all NLR-engineering attempts focused rice paired Pikp-1/Pikp-2 (allelic Pikm-1/Pikm-2) RGA4/RGA5. bioengineering NLRs provides promising method combat diverse pathogens. Detailed also discussed review, organized around strategies. summary, with progresses immune mechanism, innovative molecular available mitigate pathogens future.

Language: Английский

Citations

0
CRISPR-Cas Genome Editing for Horticultural Crops Improvement: Advantages and Prospects DOI Creative Commons
Е. Б. Рукавцова, Н. С. Захарченко, Vadim G. Lebedev

et al.

Horticulturae, Journal Year: 2022, Volume and Issue: 9(1), P. 38 - 38

Published: Dec. 30, 2022

Horticultural plants, in particular fruit trees, berry crops, and ornamentals, are valuable objects for studying their genetic biochemical properties. Along with traditional methods of these modern molecular technologies emerging, genome editing using CRISPR/Cas9 nucleases. In this review, we have analyzed advances horticultural plants. To date, it has become possible to improve many plant characteristics technology, e.g., making plants resistant biotic abiotic stress factors, changing the time flowering ripening, growth as well taste properties fruits. been successfully carried out Dozens genes from modified by means technology. We considered main ways delivering constructs limitations that complicate target genes. The article reviews prospects important humans.

Language: Английский

Citations

15
Efficient transgene-free genome editing in plants in the T0 generation based on a co-editing strategy DOI Open Access
Xiaoen Huang,

Hongge Jia,

Jin Xu

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: March 2, 2023

Abstract Transgene-free genome editing of plants in the T0 generation is highly desirable but challenging, especially perennials and vegetatively propagated plants. Here, we investigated co-editing strategy for generating transgene-free, gene-edited via Agrobacterium -mediated transient expression cytosine base editor (CBE)/gRNA-Cas12a/crRNA-GFP planta . Specifically, CBE/gRNA was used to edit ALS gene confer resistance herbicide chlorsulfuron as a selection marker, which has no negative effects on plant phenotypes; Cas12a/crRNA genes(s) interest; GFP selecting transgene-free transformants. Using this approach, genome-edited were efficiently generated various genes (either individual or multiplex) tomato, tobacco, potato, citrus generation. The biallelic/homozygous mutation rates target among herbicide-resistant transformants ranged from 8% 50%. Whole sequencing further confirmed absence off-target mutations edited efficient generation, thus being potent tool genetic improvement.

Language: Английский

Citations

8
Improvement of Crops Using the CRISPR/Cas System: New Target Genes DOI

Yulia Ukhatova,

M. V. Erastenkova,

E. S. Korshikova

et al.

Молекулярная биология, Journal Year: 2023, Volume and Issue: 57(3), P. 387 - 410

Published: May 1, 2023

The success of genome editing crops using the CRISPR/Cas system largely depends on correct choice target genes, for which directed changes will increase yield and improve quality plant raw materials resistance to biotic abiotic stress factors. This work systematizes catalogs data genes used cultivated plants. latest systematic review examined articles indexed in Scopus database published before August 17, 2019. Our covers period from 18, 2019 March 15, 2022. A search according given algorithm allowed us identify 2090 articles, among only 685 contain results gene 28 species plants (the was carried out 56 crops). significant part these papers considered either previously similar works, or studies related field reverse genetics, 136 new whose modification is aimed at improving traits important breeding. In total, 287 were subjected order properties breeding over entire application. presents a detailed analysis genes. most often increasing productivity disease resistance, as well materials. It noted whether it possible obtain stable transformants time publication applied non-model cultivars. range modified cultivars number has been significantly expanded, particular, wheat, rice, soybean, tomato, potato, rapeseed, grape, maize. vast majority cases, constructs delivered agrobacterium-mediated transformation, less commonly, biolistics, protoplast transfection, haploinducers. desired change achieved by knockout. some knockdown nucleotide substitutions out. To plants, base-editing prime-editing technologies are increasingly used. emergence convenient contributed development specific molecular genetics many crop species.

Language: Английский

Citations

7