Gene Assembly in Agrobacterium via Nucleic Acid Transfer Using Recombinase Technology (GAANTRY)

Hathwaik, Leyla T.; Thomson, James G.; Thilmony, Roger

doi:10.1007/978-1-0716-1068-8_1

Cited by 4 publications

(6 citation statements)

References 16 publications

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“…This method enables high efficiency production of T0 plants, and the MiniMax variety resulted in rapid generation of T1 seeds. Using the GAANTRY system [6] containing the mCherry3 reporter gene [9] in the transformation improves identification initial transformed lines, as well as rapid selection of transformed seeds. Transformed T1 seeds were obtained at an efficiency of 4.0% in 145 days.…”

Section: Discussionmentioning

confidence: 99%

“…Agrobacterium strain JGT44 was used for soybean transformation. This is a GAANTRY [6] strain-modified version of EHA105 (unpublished). The JGT44 strain was engineered to provide positive selection for spectinomycin.…”

Section: Construct Genotype and Expected Phenotypementioning

confidence: 99%

“…Here we report the development of a transformation protocol involving organogenic regeneration in the MiniMax variety that requires ~145 days for T1 seed generation (Figure 1). For plant transformation we utilized the GAANTRY system [6] to facilitate introduction of a selectable marker, a visible reporter gene, and an assayable reporter gene on a single T-DNA (Figure 2). The combination of simple transformation and regeneration in an early maturing variety enables accelerated genetic research for soybean improvement.…”

Section: Introductionmentioning

confidence: 99%

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Agrobacterium-Mediated Transformation of the Dwarf Soybean MiniMax

Shao,

McCue,

Thomson

2024

Plants

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This study aims to establish an Agrobacterium-mediated transformation system for use with the ‘MiniMax’soybean cultivar. MiniMax is a mutant soybean whose growth cycle is around 90 days, half that of most other soybean varieties, making it an optimal model cultivar to test genes of interest before investing in modification of elite lines. We describe an efficient protocol for Agrobacterium-mediated transformation using MiniMax seeds. It uses a modified ‘half seed’ regeneration protocol for transgenic soybean production, utilizing the rapid generation MiniMax variety to obtain T1 seeds in approximately 145 days. Addition of phloroglucinol (PG) to the regeneration protocol was key to obtaining high-efficiency rooting of the regenerated shoots. Transfer to soil was accomplished using an organic soil amendment containing nutrients and mycorrhiza for plants to thrive in the greenhouse. This combination of genotype and stimulants provides a transformation protocol to genetically engineer MiniMax seeds with a transgenic lab-to-greenhouse production efficiency of 4.0%. This is the first report of MiniMax soybean whole plant transformation and heritable T1 transmission. This protocol provides an ideal resource for enhancing the genetic transformation of any soybean cultivar.

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Section: Discussionmentioning

confidence: 99%

Section: Construct Genotype and Expected Phenotypementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Agrobacterium-Mediated Transformation of the Dwarf Soybean MiniMax

Shao,

McCue,

Thomson

2024

Plants

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“…Aimed to further enhance the PAHs conversion, integrated multiple omics with macrogenome-scale metabolic network model will provide theoretical guidance for screening key metabolic pathways and rate-limiting targets in artificial MMS. In addition, new gene/cluster discovery, upgrading of data analysis tools (such as KEGG and WGCNA), and gene engineering tools (such as CRISPR, DNA assembly, and enzyme engineering) would provide a large number of supports for modifying artificial MMS (Lapinaite et al, 2020 ; Hathwaik et al, 2021 ).…”

Section: Future Perspectivesmentioning

confidence: 99%

Artificial mixed microbial system for polycyclic aromatic hydrocarbons degradation

Cui¹,

He²,

Ntakirutimana³

et al. 2023

Front. Microbiol.

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Polycyclic aromatic hydrocarbons (PAHs) are environmental pollutants with major risks to human health. Biological degradation is environmentally friendly and the most appealing remediation method for a wide range of persistent pollutants. Meanwhile, due to the large microbial strain collection and multiple metabolic pathways, PAH degradation via an artificial mixed microbial system (MMS) has emerged and is regarded as a promising bioremediation approach. The artificial MMS construction by simplifying the community structure, clarifying the labor division, and streamlining the metabolic flux has shown tremendous efficiency. This review describes the construction principles, influencing factors, and enhancement strategies of artificial MMS for PAH degradation. In addition, we identify the challenges and future opportunities for the development of MMS toward new or upgraded high-performance applications.

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“…Gene Assembly in Agrobacterium by Nucleic Acid Transfer using Recombinase Technology (GA A NTRY) uses multiple unidirectional site-specific recombinases to stack multiple genes in vivo. The GA A NTRY ArPORT1 strain of A. rhizogenes was demonstrated to be effective at stacking up to ten genes in rice (Hathwaik et al 2021 ). As A. rhizogenes has proven effective at transforming woody plants (Gomes et al 2019 ), the GA A NTRY system could prove useful for reconstituting metabolic pathways into woody plant chasses.…”

Section: Concluding Remarks and Future Perspectivesmentioning

confidence: 99%

Metabolic engineering in woody plants: challenges, advances, and opportunities

2021

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Woody plant species represent an invaluable reserve of biochemical diversity to which metabolic engineering can be applied to satisfy the need for commodity and specialty chemicals, pharmaceuticals, and renewable energy. Woody plants are particularly promising for this application due to their low input needs, high biomass, and immeasurable ecosystem services. However, existing challenges have hindered their widespread adoption in metabolic engineering efforts, such as long generation times, large and highly heterozygous genomes, and difficulties in transformation and regeneration. Recent advances in omics approaches, systems biology modeling, and plant transformation and regeneration methods provide effective approaches in overcoming these outstanding challenges. Promises brought by developments in this space are steadily opening the door to widespread metabolic engineering of woody plants to meet the global need for a wide range of sustainably sourced chemicals and materials.

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Gene Assembly in Agrobacterium via Nucleic Acid Transfer Using Recombinase Technology (GAANTRY)

Cited by 4 publications

References 16 publications

Agrobacterium-Mediated Transformation of the Dwarf Soybean MiniMax

Agrobacterium-Mediated Transformation of the Dwarf Soybean MiniMax

Artificial mixed microbial system for polycyclic aromatic hydrocarbons degradation

Metabolic engineering in woody plants: challenges, advances, and opportunities

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