OMICs, Epigenetics, and Genome Editing Techniques for Food and Nutritional Security DOI Creative Commons
Yuri Gogolev, Sunny Ahmar, Bala Anı Akpınar

et al.

Plants, Journal Year: 2021, Volume and Issue: 10(7), P. 1423 - 1423

Published: July 12, 2021

The incredible success of crop breeding and agricultural innovation in the last century greatly contributed to Green Revolution, which significantly increased yields ensures food security, despite population explosion. However, new challenges such as rapid climate change, deteriorating soil, accumulation pollutants require much faster responses more effective solutions that cannot be achieved through traditional breeding. Further prospects for increasing efficiency agriculture are undoubtedly associated with inclusion strategy knowledge obtained using high-throughput technologies tools future ensure design plant genomes predict desired phenotype. This article provides an overview current state research these areas, well study soil microbiomes, prospective use their potential a field microbiome engineering. In terms genomic phenomic predictions, we also propose integrated approach combines high-density genotyping phenotyping techniques, can improve prediction accuracy quantitative traits species.

Language: Английский

A simple and efficient CRISPR/Cas9 system permits ultra-multiplex genome editing in plants DOI Creative Commons
Suting Wu, Htin Kyaw, Zhijun Tong

et al.

The Crop Journal, Journal Year: 2024, Volume and Issue: 12(2), P. 569 - 582

Published: Feb. 15, 2024

The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics genetic improvement. Currently available genome-editing tools have limited number targets, restricting their application in research. In this study, we developed novel CRISPR/Cas9 ultra-multiplex consisting two template vectors, eight donor four destination one primer-design software package. By combining advantages Golden Gate cloning to assemble multiple repetitive fragments Gateway recombination large by changing structure amplicons used sgRNA expression cassettes, can single binary vector targeting more than 40 genomic loci. A rice knockout containing 49 cassettes was assembled high co-editing efficiency observed. This advances synthetic biology engineering.

Language: Английский

Citations

10
Crop Improvement: Comparison of Transgenesis and Gene Editing DOI Creative Commons
Natalya V. Permyakova, Е. В. Дейнеко

Horticulturae, Journal Year: 2024, Volume and Issue: 10(1), P. 57 - 57

Published: Jan. 6, 2024

The development and improvement of molecular biology methods have led to the creation new technologies that make it possible modify plant genomes by transferring integrating into genomes’ heterologous genes from various expression systems (genetic engineering), as well inducing knockouts one or more target interest (genomic editing). genome-editing is a milestone in modern breeding certainly relies on knowledge developed for transgenesis. This review will discuss issues related advantages disadvantages both improving economically valuable traits important crops.

Language: Английский

Citations

9
Impact of GAUT1 Gene Knockout on Cell Aggregation in Arabidopsis thaliana Suspension Culture DOI Creative Commons

Tatyana A. Frankevich,

Natalya V. Permyakova, Yuriy V. Sidorchuk

et al.

BioTech, Journal Year: 2025, Volume and Issue: 14(1), P. 2 - 2

Published: Jan. 2, 2025

The development of efficient producers recombinant pharmaceuticals based on plant cell suspension cultures is a pressing challenge in modern applied science. A primary limitation their relatively low yield the target protein. One strategy to enhance culture productivity involves reducing aggregation. In order minimize cell-to-cell adhesion culture, we used Cas9 endonuclease knock out GAUT1 gene, which key gene pectin biosynthesis genome Arabidopsis thaliana. resulting knockouts exhibited altered phenotypes and were unable form viable plants. induced from seedlings bearing homozygous deletion displayed darker coloration an increased number large aggregates compared control. biomass accumulation rate showed no difference control, while level GFP protein was significantly reduced. Thus, our findings indicate that disruptions synthesis formation larger adversely affect Alternative targets should be sought reduce aggregation levels through editing.

Language: Английский

Citations

1
A single-nuclei transcriptome census of the Arabidopsis maturing root identifies that MYB67 controls phellem cell maturation DOI Creative Commons
Charlotte Miller,

Sean Jarrell-Hurtado,

Michael Haag

et al.

Developmental Cell, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

The periderm provides a protective barrier in many seed plant species. development of the suberized phellem, which forms outermost layer this important tissue, has become trait interest for enhancing both resilience to stresses and plant-mediated CO

Language: Английский

Citations

1
Convenient, high-efficiency multiplex genome editing in autotetraploid alfalfa using endogenous U6 promoters and visual reporters DOI Creative Commons

Xiuzhi Xia,

Shihao Li, Na Wang

et al.

aBIOTECH, Journal Year: 2025, Volume and Issue: 6(1), P. 81 - 90

Published: Feb. 10, 2025

Abstract CRISPR/Cas is a simple, robust, versatile tool for plant biology studies and precision breeding. However, establishing high-efficiency gene editing system multiplex of the autotetraploid crop alfalfa ( Medicago sativa L.), most important forage legume worldwide, remains formidable challenge. Here, we systematically identified endogenous U6 promoters in through transient expression via Agrobacterium -mediated infiltration leaves. We further demonstrated efficacy three active genome using an optimized hairy root system. Subsequently, established improved CRISPR/Cas9 containing or four tandemly arrayed MsU6 -promoter-driven polycistronic tRNA-sgRNA (PTG) cassettes, each consisting units, to simultaneously edit genes, coupled with visual reporter RH1 RUBY . This toolkit showed efficient selection. successfully obtained regenerated, red-colored shoots resulting from stable transformation alfalfa. These results highlight potential application Our enables convenient, alfalfa, providing toolset facilitate functional multiple genes families basic research genetic improvement

Language: Английский

Citations

1
Transgene-free Genome Editing in Plants DOI Creative Commons

Xiaoyong Gu,

Lijing Liu, Huawei Zhang

et al.

Frontiers in Genome Editing, Journal Year: 2021, Volume and Issue: 3

Published: Dec. 2, 2021

Genome editing is widely used across plant species to generate and study the impact of functional mutations in crop improvement. However, transgene integration genomes raises important legislative concerns regarding genetically modified organisms. Several strategies have been developed remove or prevent gene editor constructs, which can be divided into three major categories: 1) elimination transgenic sequences via genetic segregation; 2) transient expression from DNA vectors; 3) DNA-independent delivery, including RNA preassembled Cas9 protein-gRNA ribonucleoproteins (RNPs). Here, we summarize main employed date discuss advantages disadvantages using these different tools. We hope that our work provide information concerning value alternative genome advance breeding.

Language: Английский

Citations

43
A peptide-mediated, multilateral molecular dialogue for the coordination of pollen wall formation DOI Creative Commons
Jekaterina Truskina,

Stefanie Brück,

Annick Stintzi

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2022, Volume and Issue: 119(22)

Published: May 24, 2022

Significance Pollen viability depends on a tough external barrier called the pollen wall. wall components are produced by tapetum cells, which surround developing grains within anther. Precise coordination of activity with grain development is required to ensure effective formation. Here, we reveal that this achieved through multidirectional dialogue involving three distinct cell types. We show peptide precursors from activated proteases stage specifically in grains. Unexpectedly, found peptides perceived not tapetum, but middle layer, encloses and grains, revealing an unsuspected role for enigmatic layer control development.

Language: Английский

Citations

30
High‐Complexity One‐Pot Golden Gate Assembly DOI Creative Commons
Andrew P. Sikkema, S. Kasra Tabatabaei, Yan‐Jiun Lee

et al.

Current Protocols, Journal Year: 2023, Volume and Issue: 3(9)

Published: Sept. 1, 2023

Abstract Golden Gate Assembly is a flexible method of DNA assembly and cloning that permits the joining multiple fragments in single reaction through predefined connections. The depends on cutting using Type IIS restriction enzyme, which cuts outside its recognition site therefore can generate overhangs any sequence while separating from generated fragment. By choosing compatible fusion sites, defined order reaction. Conventionally, this has been used to join five eight round, with yield accuracy dropping off rapidly for more complex assemblies. Recently, we demonstrated application comprehensive measurements ligation fidelity bias data data‐optimized design (DAD) enable high degree very assemblies simultaneous as many 52 one Here, describe methods applying DAD principles online tools evaluate existing sets standards, selecting new optimal sets, adding sites We further divide known sequences at points, including designing one‐pot small genomes. Using T7 bacteriophage genome an example, present protocol includes removal native (domestication) simultaneously parts generation by PCR. Finally, recommended cycling protocols medium complexity (12‐36 fragments), producing high‐quality parts, examples highlighting importance purity fragment stoichiometric balance outcomes, assessing success. © 2023 New England Biolabs, Inc. Current Protocols published Wiley Periodicals LLC. Basic Protocol 1 : Assessing overhang set NEBridge Ligase Fidelity Viewer 2 Generating high‐fidelity GetSet Tool Alternate Expanding 3 Dividing genomic SplitSet 4 One‐pot 12 into destination plasmid 24+ 5 12+ Support Generation high‐purity amplicons Cloning holding vector Quantifying concentration Qubit fluorometer Visualizing large via TapeStation Validating phage ONT long‐read sequencing

Language: Английский

Citations

21
Cleave and Rescue gamete killers create conditions for gene drive in plants DOI
Georg Oberhofer, Michelle L. Johnson, Tobin Ivy

et al.

Nature Plants, Journal Year: 2024, Volume and Issue: 10(6), P. 936 - 953

Published: June 17, 2024

Language: Английский

Citations

8
Receptor-like cytoplasmic kinases of different subfamilies differentially regulate SOBIR1/BAK1-mediated immune responses in Nicotiana benthamiana DOI Creative Commons
Wen Huang,

Ciska Braam,

Carola Kretschmer

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: May 21, 2024

Abstract Cell-surface receptors form the front line of plant immunity. The leucine-rich repeat (LRR)-receptor-like kinases SOBIR1 and BAK1 are required for functionality tomato LRR-receptor-like protein Cf-4, which detects secreted effector Avr4 pathogenic fungus Fulvia fulva . Here, we show that kinase domains directly phosphorylate each other residues Thr522 Tyr469 domain Nicotiana benthamiana its activity interacting with signalling partners, respectively. By knocking out multiple genes belonging to different receptor-like cytoplasmic (RLCK)-VII subfamilies in N. benthamiana:Cf-4 , members RLCK-VII-6, −7, −8 differentially regulate Avr4/Cf-4-triggered biphasic burst reactive oxygen species. In addition, RLCK-VII-7 play an essential role resistance against oomycete pathogen Phytophthora palmivora Our study provides molecular evidence specific roles RLCKs downstream SOBIR1/BAK1-containing immune complexes.

Language: Английский

Citations

7