Particle bombardment – mediated gene transfer and GFP transient expression in <i>Setaria viridis</i>

Mookkan, Muruganantham

doi:10.1080/15592324.2018.1441657

Cited by 12 publications

(7 citation statements)

References 29 publications

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“…Agrobacterium tumefaciens infection have been demonstrated (Brutnell et al 2010;Van Eck et al 2017;Mookkan 2018;Van Eck 2018). Historically, much of the Setaria literature referred to A10 for phenotypic evaluation, which made selecting it for whole genome sequencing a logical choice.…”

Section: Introductionmentioning

confidence: 99%

“…Efficient genetic modification is a primary requirement for development of any model organism. Approaches for Setaria protoplasting, particle bombardment, and Agrobacterium tumefaciens infection have been demonstrated ( Brutnell et al 2010 ; Van Eck et al 2017 ; Mookkan 2018 ; Van Eck 2018 ). Historically, much of the Setaria literature referred to A10 for phenotypic evaluation, which made selecting it for whole genome sequencing a logical choice.…”

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confidence: 99%

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Reference Genome for the Highly TransformableSetaria viridisME034V

Thielen

Pendleton

Player

et al. 2020

G3 Genes|Genomes|Genetics

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Setaria viridis (green foxtail) is an important model system for improving cereal crops due to its diploid genome, ease of cultivation, and use of C4 photosynthesis. The S. viridis accession ME034V is exceptionally transformable, but the lack of a sequenced genome for this accession has limited its utility. We present a 397 Mb highly contiguous de novo assembly of ME034V using ultra-long nanopore sequencing technology (read N50=41kb). We estimate that this genome is largely complete based on our updated k-mer based genome size estimate of 401 Mb for S. viridis. Genome annotation identified 37,908 protein-coding genes and >300k repetitive elements comprising 46% of the genome. We compared the ME034V assembly with two other previously sequenced Setaria genomes as well as to a diversity panel of 235 S. viridis accessions. We found the genome assemblies to be largely syntenic, but numerous unique polymorphic structural variants were discovered. Several ME034V deletions may be associated with recent retrotransposition of copia and gypsy LTR repeat families, as evidenced by their low genotype frequencies in the sampled population. Lastly, we performed a phylogenomic analysis to identify gene families that have expanded in Setaria, including those involved in specialized metabolism and plant defense response. The high continuity of the ME034V genome assembly validates the utility of ultra-long DNA sequencing to improve genetic resources for emerging model organisms. Structural variation present in Setaria illustrates the importance of obtaining the proper genome reference for genetic experiments. Thus, we anticipate that the ME034V genome will be of significant utility for the Setaria research community.

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Section: Introductionmentioning

confidence: 99%

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confidence: 99%

Reference Genome for the Highly TransformableSetaria viridisME034V

Thielen

Pendleton

Player

et al. 2020

G3 Genes|Genomes|Genetics

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“…S3). In previous studies, particles coated with large amounts of DNA had low membrane permeability due to particle aggregation [21], and a high concentration of cationic peptide induced cytotoxicity in plant cells [14]. These results suggested that the expression levels induced by 16 and 24 µg of DNA-peptide complex per bombardment were remarkably decreased.…”

Section: Effect Of Dna-peptide Complex Load Per Bombardment In B Camp...mentioning

confidence: 82%

“…The cotyledons of B. campestris were bombarded once or twice to analyze the efficiency of Nluc expression. Several reports indicated that multiple bombardments affected gene transfer efficiency in various plant species [21][22][23]. Nluc expression induced by two bombardments was significantly lower than that induced by one bombardment (P = 0.0100, Fig.…”

Section: Effect Of the Number Of Bombardments In B Campestrismentioning

confidence: 95%

Particle bombardment-assisted peptide-mediated gene transfer for highly efficient transient assay

et al. 2023

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Objective A centrifugation-assisted peptide-mediated gene transfer (CAPT) method was recently developed as an efficient system for gene delivery into plant cells. However, the gene transfer efficiency of CAPT into plant cells was not entirely satisfactory for detecting transient expression of a transgene driven into mitochondria. Here, we report a new gene delivery system using a method called particle bombardment-assisted peptide-mediated gene transfer (PBPT). Results We investigated various parameters of the PBPT method to increase transient gene expression efficiency in Brassica campestris. The optimal conditions for PBPT were a single bombardment with gold particles coated with a DNA‒peptide complex (6 µg of DNA and 2 µg of peptide) at an acceleration pressure of 5 kg/cm2 and a target distance of 12.5 cm. Moreover, bombardment under the optimal conditions successfully transferred the transgene into the cells of other plant species, namely B. juncea and tomato. Thus, we developed a PBPT method for highly efficient delivery of a DNA‒peptide complex into plant mitochondria.

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“…Then, the particles are accelerated to very high velocity for bombardment. Such a high momentum of metal particles causes substantial damage to the chloroplasts after particle bombardment on explants (Russel et al, 1992;Mookkan, 2018). In this study, we contemplated establishing reliable and effective potato plastome engineering protocol through the incorporation of the marker gene into the potato plastome.…”

Section: Discussionmentioning

confidence: 99%

Rapid and efficient in vitro regeneration of transplastomic potato (Solanum tuberosum L.) plants after particle bombardment

Hossain¹,

Aksoy²,

Gökçe³

et al. 2021

Turk J Agric For

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In contrast with traditional nuclear gene transformation, transplastomic technology has opened a new horizon for the transgenic plant research that offers several beneficial aspects including the convenient use of transgene stacking and the generation of high expression levels of foreign proteins. However, this technology has been well adopted and established in tobacco, the introduction and adoption of cost-effective, swift, and reproducible protocol for in vitro regeneration of transplastomic potato is challenging and laborious. The present research aimed to develop such prompt and efficient protocol to instigate and revive the regeneration potential with the combinations of different plant growth regulators (PGRs). Leaves and internodal explants from four potato cultivars were transformed with chloroplast transformation vector via particle bombardment and cultured on MS media supplemented with suitable PGRs and selection agents. Leaf explants of cultivar Kuroda induced highest (92%) number of calli where cultivar Sante produced the highest (85.7%) transplastomic shoots. Thidiazuron was found more proficient (41%) for shoot regeneration. Finally, within only seven weeks, we got 21 spectinomycin resistant shoot, and 16 of those showed integration of target genes into the plastome in PCR screening.

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Particle bombardment – mediated gene transfer and GFP transient expression in Setaria viridis

Cited by 12 publications

References 29 publications

Reference Genome for the Highly TransformableSetaria viridisME034V

Reference Genome for the Highly TransformableSetaria viridisME034V

Particle bombardment-assisted peptide-mediated gene transfer for highly efficient transient assay

Rapid and efficient in vitro regeneration of transplastomic potato (Solanum tuberosum L.) plants after particle bombardment

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