Agrobacterium tumefaciens-Mediated Transformation of Setaria viridis

Eck, Joyce Van; Swartwood, Kerry; Pidgeon, Kaitlin; Maxson-Stein, Kimberly

doi:10.1007/978-3-319-45105-3_20

Cited by 22 publications

(24 citation statements)

References 35 publications

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“…The time from infection of callus to recovery of mature T1 seed was approximately 4 months. Depending on the composition of the selectable marker gene cassette in the vectors used for transformation, the transformation efficiency, based on infection of the total number of mature seed-derived calli per experiment, ranged from 5 to 15% (Van Eck et al, 2017). Transformation efficiency is defined as the percentage of infected material that gives rise to at least one independent transgenic line.…”

Section: Agrobacterium Tumefaciens- Mediated Transformation Of Maturementioning

confidence: 99%

“…The promoter driving hpt was also found to be key for efficient transformation. Vectors that contained hpt driven by a monocot promoter resulted in recovery of a greater number of transgenic lines compared to vectors where hpt expression was under the control of promoters best suited for dicots (Van Eck et al, 2017). In addition, those vectors that had an intron in the hpt gene resulted in a greater number of transgenic lines than when an intron was not present in the gene.…”

Section: Agrobacterium Tumefaciens- Mediated Transformation Of Maturementioning

confidence: 99%

Section: Agrobacterium Tumefaciens- Mediated Transformation Of Maturementioning

confidence: 99%

“…Martins et al (2015b) followed similar methodology as reported by Van Eck et al (2017). A comparison of the primary components that make up the various reported transformation methods (Martins et al, 2015b; Van Eck and Swartwood, 2015; Van Eck et al, 2017) is outlined in Table 1 . The vectors used for transformation, more specifically the type of selectable marker gene and its promoter contained in the vector, were the key parameters that influenced transformation efficiency in the methods described by Martins et al (2015b).…”

Section: Agrobacterium Tumefaciens- Mediated Transformation Of Maturementioning

confidence: 99%

“…Efforts by researchers worldwide led to advancement of Agrobacterium and biolistics approaches for monocot transformation that allowed genetic engineering of the major cereal crops (maize, rice, wheat, barley, sorghum, oats, and millets), which represents roughly two-thirds of the world food supply (Ji et al, 2013). For S. viridis transformation methods, there are reports of Agrobacterium -mediated transformation of mature seed-derived callus (Brutnell et al, 2010; Van Eck and Swartwood, 2015; Van Eck et al, 2017) and non-tissue culture methods ( in planta ) through infection of immature inflorescences followed by recovery of transgenic seed (Martins et al, 2015a; Saha and Blumwald, 2016). …”

Section: Introductionmentioning

confidence: 99%

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The Status of Setaria viridis Transformation: Agrobacterium-Mediated to Floral Dip

Eck

2018

Front. Plant Sci.

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Setaria viridis has many attributes, including small stature and simple growth requirements, that make it attractive as a model species for monocots. Genetic engineering (transformation) methodology is a key prerequisite for adoption of plant species as models. Various transformation approaches have been reported for S. viridis including tissue culture-based and in planta by Agrobacterium tumefaciens infection of floral organs referred to as the floral dip method. The tissue culture-based method utilizes A. tumefaciens infection of mature seed-derived callus with subsequent recovery of stable transgenic lines. Vectors found to be most effective contain the hygromycin phosphotransferase selectable marker gene driven by either Panicum virgatum or Zea mays ubiquitin promoters. As for the floral dip method, there are two reports based on Agrobacterium infection of young S. viridis inflorescences. Plants were allowed to mature, seeds were collected, and analysis of the progeny verified the presence of transgenes. Each transformation approach, tissue culture-based and floral dip, has advantages and disadvantages depending on the expertise of personnel and resources available. While the tissue culture-based method results in a higher transformation efficiency than floral dip, implementation requires a specific technical skillset that limits availability of experienced personnel to successfully perform transformations. Less technical experience is required for floral dip; however, a lack of high-quality growth chambers or greenhouses that provide the necessary optimum growing conditions would reduce an already low transformation efficiency or would not result in recovery of transgenic lines. An overview of transformation methods reported for S. viridis is presented in this review.

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Section: Agrobacterium Tumefaciens- Mediated Transformation Of Maturementioning

confidence: 99%

Section: Agrobacterium Tumefaciens- Mediated Transformation Of Maturementioning

confidence: 99%

Section: Agrobacterium Tumefaciens- Mediated Transformation Of Maturementioning

confidence: 99%

Section: Agrobacterium Tumefaciens- Mediated Transformation Of Maturementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The Status of Setaria viridis Transformation: Agrobacterium-Mediated to Floral Dip

Eck

2018

Front. Plant Sci.

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Agrobacterium‐Mediated Transformation of Setaria viridis, a Model System for Cereals and Bioenergy Crops

2021

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Setaria viridis is an emerging model system for the genetic and molecular characterization of cereals and bioenergy crops. Here, we describe a detailed procedure for genetic transformation of the S. viridis accession line ME034V-1. This method utilizes callus generated from mature seeds for infection with Agrobacterium tumefaciens strain AGL1 to regenerate hygromycin-resistant stable transgenic plants. It takes approximately 7 weeks to generate callus from mature seeds, 11-17 weeks from infection to the regeneration of transgenic lines, and an additional 3-4 weeks for plant growth in the greenhouse for seed collection. The protocol as presented consistently results in transformation frequency of approximately 25% for the generation of transgenic plants, with fewer escapes and higher survivability in soil for optimal seed collection.

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Protein expression and gene editing in monocots using foxtail mosaic virus vectors

Beernink

Ellison

et al. 2019

Plant Direct

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Plant viruses can be engineered to carry sequences that direct silencing of target host genes, expression of heterologous proteins, or editing of host genes. A set of foxtail mosaic virus (FoMV) vectors was developed that can be used for transient gene expression and single guide RNA delivery for Cas9‐mediated gene editing in maize, Setaria viridis, and Nicotiana benthamiana. This was accomplished by duplicating the FoMV capsid protein subgenomic promoter, abolishing the unnecessary open reading frame 5A, and inserting a cloning site containing unique restriction endonuclease cleavage sites immediately after the duplicated promoter. The modified FoMV vectors transiently expressed green fluorescent protein (GFP) and bialaphos resistance (BAR) protein in leaves of systemically infected maize seedlings. GFP was detected in epidermal and mesophyll cells by epifluorescence microscopy, and expression was confirmed by Western blot analyses. Plants infected with FoMV carrying the bar gene were temporarily protected from a glufosinate herbicide, and expression was confirmed using a rapid antibody‐based BAR strip test. Expression of these proteins was stabilized by nucleotide substitutions in the sequence of the duplicated promoter region. Single guide RNAs expressed from the duplicated promoter mediated edits in the N. benthamiana Phytoene desaturase gene, the S. viridis Carbonic anhydrase 2 gene, and the maize HKT1 gene encoding a potassium transporter. The efficiency of editing was enhanced in the presence of synergistic viruses and a viral silencing suppressor. This work expands the utility of FoMV for virus‐induced gene silencing (VIGS), virus‐mediated overexpression (VOX), and virus‐enabled gene editing (VEdGE) in monocots.

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Agrobacterium tumefaciens-Mediated Transformation of Setaria viridis

Cited by 22 publications

References 35 publications

The Status of Setaria viridis Transformation: Agrobacterium-Mediated to Floral Dip

The Status of Setaria viridis Transformation: Agrobacterium-Mediated to Floral Dip

Agrobacterium‐Mediated Transformation of Setaria viridis, a Model System for Cereals and Bioenergy Crops

Protein expression and gene editing in monocots using foxtail mosaic virus vectors

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