Super-Agrobacterium ver. 4: Improving the Transformation Frequencies and Genetic Engineering Possibilities for Crop Plants

Nonaka, Satoko; Someya, T.; Kadota, Yasuhiro; Nakamura, Kouji; Ezura, Hiroshi

doi:10.3389/fpls.2019.01204

Cited by 27 publications

(23 citation statements)

References 63 publications

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“…Other desired Agrobacterium traits in the plant transformation and gene-editing field include auxotrophic strains for biocontainment and improved tissue culture (66), inactive transposons (67), reduced vector backbone integration (63,68,69), increased T-DNA transfer (70), and transient-only T-DNA delivery (71,72). Finally, agrobacteria that evade or subvert host plant factors that are detrimental to transformation efficiency, such as the plant defense system, can be critical to develop transformation systems for recalcitrant species or cultivars (73)(74)(75).…”

Section: Discussionmentioning

confidence: 99%

Efficient CRISPR-mediated base editing in Agrobacterium spp.

Rodrigues

Karimi

Impens

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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Agrobacteriumspp. are important plant pathogens that are the causative agents of crown gall or hairy root disease. Their unique infection strategy depends on the delivery of part of their DNA to plant cells. Thanks to this capacity, these phytopathogens became a powerful and indispensable tool for plant genetic engineering and agricultural biotechnology. AlthoughAgrobacteriumspp. are standard tools for plant molecular biologists, current laboratory strains have remained unchanged for decades and functional gene analysis ofAgrobacteriumhas been hampered by time-consuming mutation strategies. Here, we developed clustered regularly interspaced short palindromic repeats (CRISPR)-mediated base editing to enable the efficient introduction of targeted point mutations into the genomes of bothAgrobacterium tumefaciensandAgrobacterium rhizogenes. As an example, we generated EHA105 strains with loss-of-function mutations inrecA, which were fully functional for maize (Zea mays) transformation and confirmed the importance of RolB and RolC for hairy root development byA. rhizogenesK599. Our method is highly effective in 9 of 10 colonies after transformation, with edits in at least 80% of the cells. The genomes of EHA105 and K599 were resequenced, and genome-wide off-target analysis was applied to investigate the edited strains after curing of the base editor plasmid. The off-targets present were characteristic of Cas9-independent off-targeting and point to TC motifs as activity hotspots of the cytidine deaminase used. We anticipate that CRISPR-mediated base editing is the start of “engineering the engineer,” leading to improvedAgrobacteriumstrains for more efficient plant transformation and gene editing.

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Section: Discussionmentioning

confidence: 99%

Efficient CRISPR-mediated base editing in Agrobacterium spp.

Rodrigues

Karimi

Impens

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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“…Keeping in view the advantage of this phytopathogen, the complex Agrobacterium -plant interactions have been reviewed extensively [ 31 , 75 , 76 , 77 , 78 , 79 , 80 ]. Recently, Nonaka et al [ 81 ] have developed the Super- Agrobacterium ver. 4 that has major application in basic plant science research.…”

Section: Priority Towards Agrobacterium -Based Genetic Transformationmentioning

confidence: 99%

Agroinfiltration Mediated Scalable Transient Gene Expression in Genome Edited Crop Plants

Kaur

Manchanda

Kalia

et al. 2021

IJMS

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Agrobacterium-mediated transformation is one of the most commonly used genetic transformation method that involves transfer of foreign genes into target plants. Agroinfiltration, an Agrobacterium-based transient approach and the breakthrough discovery of CRISPR/Cas9 holds trending stature to perform targeted and efficient genome editing (GE). The predominant feature of agroinfiltration is the abolishment of Transfer-DNA (T-DNA) integration event to ensure fewer biosafety and regulatory issues besides showcasing the capability to perform transcription and translation efficiently, hence providing a large picture through pilot-scale experiment via transient approach. The direct delivery of recombinant agrobacteria through this approach carrying CRISPR/Cas cassette to knockout the expression of the target gene in the intercellular tissue spaces by physical or vacuum infiltration can simplify the targeted site modification. This review aims to provide information on Agrobacterium-mediated transformation and implementation of agroinfiltration with GE to widen the horizon of targeted genome editing before a stable genome editing approach. This will ease the screening of numerous functions of genes in different plant species with wider applicability in future.

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“…The comparative genomics of Agrobacterium strains, together with the functional analysis, will give insights into the synergetic effects, and the insights will contribute to developing better Agrobacterium strains than the existing ones. In addition, the disarmed Ti plasmids and/or the virulence helper plasmids can be further modified to develop the Super-Agrobacterium strains by introducing both the ACC deaminase (acdS) and GABA transaminase (gabT) genes (Nonaka et al, 2019). It was known that ethylene and gamma-aminobutyric acid (GABA) produced by plants are taken up into A. tumefaciens during AMGT and have negative effects on Agrobacterium-plant interactions (Nonaka et al, 2019).…”

Section: Amgt: the Progress In Agrobacterium Genomics And Genetic Engmentioning

confidence: 99%

“…In addition, the disarmed Ti plasmids and/or the virulence helper plasmids can be further modified to develop the Super-Agrobacterium strains by introducing both the ACC deaminase (acdS) and GABA transaminase (gabT) genes (Nonaka et al, 2019). It was known that ethylene and gamma-aminobutyric acid (GABA) produced by plants are taken up into A. tumefaciens during AMGT and have negative effects on Agrobacterium-plant interactions (Nonaka et al, 2019). Therefore, using these two genes to decrease the ethylene and GABA levels in Agrobacterium can improve AMGT frequency.…”

Section: Amgt: the Progress In Agrobacterium Genomics And Genetic Engmentioning

confidence: 99%

Agrobacterium-mediated delivery of CRISPR/Cas reagents for genome editing in plants enters an era of ternary vector systems

Zhang

Chen

2020

Sci. China Life Sci.

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Lack of appropriate methods for delivery of genome-editing reagents is a major barrier to CRISPR/Cas-mediated genome editing in plants. Agrobacterium-mediated genetic transformation (AMGT) is the preferred method of CRISPR/Cas reagent delivery, and researchers have recently made great improvements to this process. In this article, we review the development of AMGT and AMGT-based delivery of CRISPR/Cas reagents. We give an overview of the development of AMGT vectors including binary vector, superbinary vector, dual binary vector, and ternary vector systems. We also review the progress in Agrobacterium genomics and Agrobacterium genetic engineering for optimal strains. We focus in particular on the ternary vector system and the resources we developed. In summary, it is our opinion that Agrobacterium-mediated CRISPR/Cas genome editing in plants is entering an era of ternary vector systems, which are often integrated with morphogenic regulators. The new vectors described in this article are available from Addgene and/or MolecularCloud for sharing with academic investigators for noncommercial research.

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Super-Agrobacterium ver. 4: Improving the Transformation Frequencies and Genetic Engineering Possibilities for Crop Plants

Cited by 27 publications

References 63 publications

Efficient CRISPR-mediated base editing in Agrobacterium spp.

Efficient CRISPR-mediated base editing in Agrobacterium spp.

Agroinfiltration Mediated Scalable Transient Gene Expression in Genome Edited Crop Plants

Agrobacterium-mediated delivery of CRISPR/Cas reagents for genome editing in plants enters an era of ternary vector systems

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