Engineering canker‐resistant plants through CRISPR/Cas9‐targeted editing of the susceptibility gene CsLOB1 promoter in citrus

Plant Biotechnology Journal, Journal Year: 2017, Volume and Issue: 15(12), P. 1509 - 1519

Published: March 29, 2017

Summary Citrus canker, caused by Xanthomonas citri subsp. ( Xcc ), is severely damaging to the global citrus industry. Targeted editing of host disease‐susceptibility genes represents an interesting and potentially durable alternative in plant breeding for resistance. Here, we report improvement canker resistance through CRISPR /Cas9‐targeted modification susceptibility gene Cs LOB 1 promoter citrus. Wanjincheng orange sinensis Osbeck) harbours at least three copies G allele one copy − allele. The both alleles contains effector binding element EBE P thA4 which recognized main PthA4 activate expression promote development. Five pC as9/Cs 1sg RNA constructs were designed modify orange. Among these constructs, mutation rates 11.5%–64.7%. Homozygous mutants generated directly from explants. Sixteen lines that harboured modifications identified 38 mutant plants. Four (S2‐5, S2‐6, S2‐12 S5‐13), disrupted induction response infection, showed enhanced compared with wild type. No symptoms observed S2‐6 S5‐13 lines. Promoter alone was sufficient enhance Deletion entire sequence conferred a high degree canker. results demonstrate /Cas9‐mediated efficient strategy generation canker‐resistant cultivars.

Language: Английский

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The Enhancement of Plant Disease Resistance Using CRISPR/Cas9 Technology

Frontiers in Plant Science, Journal Year: 2018, Volume and Issue: 9

Published: Aug. 24, 2018

Genome editing technologies have progressed rapidly and become one of the most important genetic tools in implementation pathogen resistance plants. Recent years witnessed emergence site directed modification methods using meganucleases, zinc finger nucleases (ZFNs), transcription activator-like effector (TALENs), clustered regularly interspaced short palindrome repeats (CRISPR)/CRISPR-associated protein 9 (Cas9). Recently, CRISPR/Cas9 has largely overtaken other genome due to fact that it is easier design implement, a higher success rate, more versatile less expensive. This review focuses on recent advances plant protection technology model plants crops response viral, fungal bacterial diseases. As regards achievement viral disease resistance, main strategies employed species such as Arabidopsis Nicotiana benthamiana, which include integration CRISPR-encoding sequences target interfere with induction CRISPR-mediated targeted mutation host genome, will be discussed. Furthermore, based susceptibility genes crop rice, tomato, wheat citrus reviewed. After spending deciphering reading genomes, researchers are now rewriting them develop resistant specific pests pathogens.

Language: Английский

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Genome editing of the disease susceptibility gene CsLOB1 in citrus confers resistance to citrus canker

Plant Biotechnology Journal, Journal Year: 2016, Volume and Issue: 15(7), P. 817 - 823

Published: Dec. 10, 2016

Abstract Citrus is a highly valued tree crop worldwide, while, at the same time, citrus production faces many biotic challenges, including bacterial canker and Huanglongbing ( HLB ). Breeding for disease‐resistant varieties most efficient sustainable approach to control plant diseases. Traditional breeding of challenging due multiple limitations, polyploidy, polyembryony, extended juvenility long crossing cycles. Targeted genome editing technology has potential shorten varietal development some traits, disease resistance. Here, we used CRISPR /Cas9/sg RNA modify susceptibility gene Cs LOB 1 in Duncan grapefruit. Six independent lines, D 2, 3, 9, 10, 11 12, were generated. next‐generation sequencing six lines showed mutation rate was 31.58%, 23.80%, 89.36%, 88.79%, 46.91% 51.12% respectively, cells each line. 2 3 symptoms similar wild‐type grapefruit, when inoculated with pathogen Xanthomonas citri subsp. (Xcc). No observed on 12 4 days postinoculation DPI ) Xcc. Pustules caused by Xcc later stages, which much reduced compared that The pustules 9 10 did not develop into typical symptoms. side effects off‐target mutations detected mutated plants. This study indicates using will provide promising pathway generate varieties.

Language: Английский

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CRISPR/Cas System: Recent Advances and Future Prospects for Genome Editing

Trends in Plant Science, Journal Year: 2019, Volume and Issue: 24(12), P. 1102 - 1125

Published: Nov. 11, 2019

Language: Английский

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Applications and potential of genome editing in crop improvement

Genome biology, Journal Year: 2018, Volume and Issue: 19(1)

Published: Nov. 30, 2018

Genome-editing tools provide advanced biotechnological techniques that enable the precise and efficient targeted modification of an organism's genome. systems have been utilized in a wide variety plant species to characterize gene functions improve agricultural traits. We describe current applications genome editing plants, focusing on its potential for crop improvement terms adaptation, resilience, end-use. In addition, we review novel breakthroughs are extending genome-edited crops possibilities their commercialization. Future prospects integrating this revolutionary technology with conventional new-age breeding strategies also discussed.

Language: Английский

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The Candidatus Liberibacter–Host Interface: Insights into Pathogenesis Mechanisms and Disease Control

Annual Review of Phytopathology, Journal Year: 2017, Volume and Issue: 55(1), P. 451 - 482

Published: June 22, 2017

"Candidatus Liberibacter" species are associated with economically devastating diseases of citrus, potato, and many other crops. The importance these as well the proliferation new on a wider host range is likely to increase insects vectoring "Ca. expand their territories worldwide. Here, we review progress understanding pathogenesis mechanisms control approaches for they cause. We discuss Liberibacter virulence traits, including secretion systems, putative effectors, lipopolysaccharides (LPSs), important traits contribute disease development, e.g., flagella, prophages, salicylic acid hydroxylase. Liberibacters discussed. secrete Sec-dependent effectors (SDEs) or factors into phloem elements companion cells interfere targets (e.g., proteins genes), which cause cell death, necrosis, phenotypes cells, leading localized responses systemic malfunction phloem. Receptors remaining organelles in phloem, such plastid, vacuole, mitochondrion, endoplasmic reticulum, interact secreted SDEs and/or located outer membrane trigger responses. Some genes targeted by serve susceptibility that facilitate compatibility promoting pathogen growth suppressing immune responses) development. In addition, plant immunity response via pathogen-associated molecular patterns (PAMPs, lipopolysaccharides), leads premature callose deposition, protein accumulation, causing effect transportation. Physical presence metabolic activities may disturb function disrupting osmotic gradients, integrity conductivity. also management strategies, promising technologies. Citrus production Huanglongbing possible if most integrated. HLB discussed context local, area-wide, regional Huanglongbing/Asian Psyllid epidemiological zones. For zebra chip control, aggressive psyllid enables potato production, although insecticide resistance becoming an issue. Meanwhile, technologies clustered regularly interspaced short palindromic repeat (CRISPR)-derived genome editing provide unprecedented opportunity long-term solutions.

Language: Английский

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Use of CRISPR systems in plant genome editing: toward new opportunities in agriculture

Emerging Topics in Life Sciences, Journal Year: 2017, Volume and Issue: 1(2), P. 169 - 182

Published: Nov. 10, 2017

Initially discovered in bacteria and archaea, CRISPR–Cas9 is an adaptive immune system found prokaryotes. In 2012, scientists a way to use it as genome editing tool. 2013, its application plants was successfully achieved. This breakthrough has opened up many new opportunities for researchers, including the opportunity gain better understanding of plant biological systems more quickly. The present study reviews agricultural applications related CRISPR from 52 peer-reviewed articles published since 2014. Based on this literature review, main achieve improved yield performance, biofortification, biotic abiotic stress tolerance, with rice (Oryza sativa) being most studied crop.

Language: Английский

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Mechanistic insights into host adaptation, virulence and epidemiology of the phytopathogenXanthomonas

FEMS Microbiology Reviews, Journal Year: 2019, Volume and Issue: 44(1), P. 1 - 32

Published: Oct. 1, 2019

ABSTRACT Xanthomonas is a well-studied genus of bacterial plant pathogens whose members cause variety diseases in economically important crops worldwide. Genomic and functional studies these phytopathogens have provided significant understanding microbial-host interactions, virulence host adaptation mechanisms including microbial ecology epidemiology. In addition, several strains are as producers the extracellular polysaccharide, xanthan, used food pharmaceutical industries. This polymer has also been implicated phases disease cycle. this review, we summarise current knowledge on infection strategies regulatory networks controlling from species discuss novel opportunities that body work for control health.

Language: Английский

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Efficient Targeted Mutagenesis in Apple and First Time Edition of Pear Using the CRISPR-Cas9 System

Frontiers in Plant Science, Journal Year: 2019, Volume and Issue: 10

Published: Feb. 6, 2019

Targeted genome engineering has emerged as an alternative to classical plant breeding and transgenic methods improve crop plants. Among other (zinc finger nucleases or TAL effector nucleases) the CRISPR/Cas system proved be most effective, convenient least expensive method. In this study, we optimized conditions of application on apple explored its feasibility pear. As a proof concept, chose knock-out Phytoene Desaturase (PDS) Terminal Flower 1 (TFL1) genes. To edition efficiency, two different single guide RNAs (gRNAs) were associated Cas9 nuclease for each target gene. These gRNAs placed under control U3 U6 promoters. Characteristic albino phenotype was obtained 85 % lines targeted in MdPDS Early flowering observed 93 MdTFL1.1 gene 9 pear PcTFL1.1. Sequencing zones CRIPSR-PDS CRISPR-TFL1.1 showed that induced mutations but at variable frequencies. cases, cut DNA twenty base pairs near protospacer adjacent motif insertions more frequent than deletions substitutions. The profile PDS well TFL1.1 genes chimeric biallelic. Analysis sample potential off-target sequences construct indicated absence cases three mismatches. addition, transient transformation with CRISPR-PDS produced T-DNA free edited lines. Our overall results indicate that, despite occurrence chimerism, is powerful precise method induce mutagenesis first generation

Language: Английский

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CRISPR technology is revolutionizing the improvement of tomato and other fruit crops

Horticulture Research, Journal Year: 2019, Volume and Issue: 6(1)

Published: June 14, 2019

Fruits are major sources of essential nutrients and serve as staple foods in some areas the world. The increasing human population changes climate experienced worldwide make it urgent to production fruit crops with high yield enhanced adaptation environment, for which conventional breeding is unlikely meet demand. Fortunately, clustered regularly interspaced short palindromic repeat (CRISPR) technology paves way toward a new horizon crop improvement consequently revolutionizes plant breeding. In this review, mechanism optimization CRISPR system its application crops, including resistance biotic abiotic stresses, quality improvement, domestication highlighted. Controversies future perspectives discussed well.

Language: Английский

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