Agrobacterium tumefaciens-mediated transformation of the soybean pathogen Phomopsis longicolla

Li, Shuxian; Ridenour, John B.; Kim, Hun; Hirsch, Robert; Rupe, J. C.; Bluhm, Burton H.

doi:10.1016/j.mimet.2012.12.015

Cited by 11 publications

(9 citation statements)

References 18 publications

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“…RTO4_16381 is depicted as localized to the lipid droplet in Figure 6, along with eleven other lipid droplet-associated proteins with high confidence lipid accumulation phenotypes or significant fitness defects on fatty acids. The products of these genes were observed in proteomic analysis of R. toruloides lipid droplets by Zhu et al (55), except for RTO4_11043 (similar to human BSCL2 ) and DGA1 which have been localized to the lipid droplet in many other species (100-106).…”

Section: Resultsmentioning

confidence: 99%

Functional genomics of lipid metabolism in the oleaginous yeastRhodosporidium toruloides

Coradetti

Pinel

Geiselman

et al. 2017

Preprint

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Section: Resultsmentioning

confidence: 99%

Functional genomics of lipid metabolism in the oleaginous yeastRhodosporidium toruloides

Coradetti

Pinel

Geiselman

et al. 2017

Preprint

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“…Mutants of P. longicolla , including strain PL343, were created via ATMT from wild‐type strain PL2010AR as previously described (Li et al . ). Agrobacterium tumefaciens strain AGL‐1 (Lazo et al .…”

Section: Methodsmentioning

confidence: 97%

“…Recently, new molecular resources have become available, including a protocol for genetic transformation (Li et al . ) and a draft genome sequence of P. longicolla (Li et al . ).…”

Section: Introductionmentioning

confidence: 99%

MoNSTR‐seq, a restriction site‐associated DNA sequencing technique to characterize Agrobacterium ‐mediated transfer‐DNA insertions in Phomopsis longicolla

Zaccaron

Sharma

Bluhm

2017

Lett Appl Microbiol

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Significance and Impact of the Study: This study describes MoNSTR-seq (Mutation analysis via Next-generation DNA Sequencing of T-DNA Regions), an adaptation of restriction site-associated DNA sequencing (RAD-seq) to identify the position of transfer-DNA (T-DNA) insertions in the genome of Phomopsis longicolla, an important pathogen of soybean. The technique enables high-throughput characterization of mutants generated via Agrobacterium tumefaciens-mediated transformation (ATMT), thus accelerating gene discovery via forward genetics. This technique represents a significant advancement over existing approaches to characterize T-DNA insertions in fungal genomes. With minor modifications, this technique could be easily adapted to taxonomically diverse fungal pathogens and additional mutagenesis cassettes. AbstractPhomopsis longicolla (Hobbs) causes Phomopsis seed decay and stem lesions in soybean (Glycine max). In this study, a novel, high-throughput adaptation of RAD-seq termed MoNSTR-seq (Mutation analysis via Next-generation DNA Sequencing of T-DNA Regions) was developed to determine the genomic location of T-DNA insertions in P. longicolla mutants. Insertional mutants were created via Agrobacterium tumefaciens-mediated transformation, and one mutant, strain PL343, was further investigated due to impaired stem lesion formation. Mutation analysis via Next-generation DNA Sequencing of T-DNA Regions, in which DNA libraries are created with two distinct restriction enzymes and customized adapters to simultaneously enrich both T-DNA insertion borders, was developed to characterize the genomic lesion in strain PL343. MoNSTR-seq successfully identified a T-DNA insertion in the predicted promoter region of a gene encoding a cellobiose dehydrogenase (CDH1), and the position of the T-DNA insertion in strain PL343 was confirmed by Sanger sequencing. Thus, MoNSTR-seq represents an effective tool for molecular genetics in P. longicolla, and is readily adaptable for use in diverse fungal species.

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“…Moreover, transgenetic integration may cause unintended alterations of the genome by deletions, insertions, or rearrangement, which is responsible for the pleio-tropic effects (Kuiper et al, 2001;Cellini et al, 2004;Garcí a-Cañas et al, 2010). Agrobacterium tumefaciens-mediated transformation (ATMT) is an efficient transgenetic method developed in recent 20 years (Michielse et al, 2005;Moon et al, 2008;Li et al, 2013). Because ATMT has many advantages, such as simple manipulations, high transformation efficiency and high single-copy rates, it has been employed as a routine procedure for a large number of plant species (Li et al, 2017) including Poplar (Maheshwari and Kovalchuk, 2016).…”

Section: Introductionmentioning

confidence: 99%

Light and Dark Apporaches in Agrobacterium-Mediated Transformation in 84K Poplar

Wang¹,

Ding²,

Mao³

2017

mpb

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High efficiency transformation and strong regeneration ability of plants are prerequisites to the establishment of good plant transgenic system. This paper describes two Agrobacterium-mediated transformation protocols for poplar, they are Light-induced method and dark-induced method by A. tumefaciens to introduce target genes into Populus alba X Populus glandulosa and comparatively analyzed two transgenetic systems. We summarized the advantages and disadvantages of the two methods. However, the dark-induced method had higher transformation efficiency and light-induced method has shorter period. So, a combination of the two methods can help maximize on efficiency and high levels of gene expression in transgenic plants. This study provides a useful foundation for further research toward the development of 84K Poplar.

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Agrobacterium tumefaciens-mediated transformation of the soybean pathogen Phomopsis longicolla

Cited by 11 publications

References 18 publications

Functional genomics of lipid metabolism in the oleaginous yeastRhodosporidium toruloides

Functional genomics of lipid metabolism in the oleaginous yeastRhodosporidium toruloides

MoNSTR‐seq, a restriction site‐associated DNA sequencing technique to characterize Agrobacterium ‐mediated transfer‐DNA insertions in Phomopsis longicolla

Light and Dark Apporaches in Agrobacterium-Mediated Transformation in 84K Poplar

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