Agrobacterium tumefaciens-mediated transformation of the lichen fungus, Umbilicaria muehlenbergii

Sook Young Park, Min Hye Jeong, Hai Ying Wang, Jung A. Kim, Nan Hee Yu, Sungbeom Kim, Yong Hwa Cheong, Seogchan Kang, Yong Hwan Lee, Jae Seoun Hur

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Transformation-mediated mutagenesis in both targeted and random manners has been widely applied to decipher gene function in diverse fungi. However, a transformation system has not yet been established for lichen fungi, severely limiting our ability to study their biology and mechanism underpinning symbiosis via gene manipulation. Here, we report the first successful transformation of the lichen fungus, Umbilicaria muehlenbergii, via the use of Agrobacterium tumefaciens. We generated a total of 918 transformants employing a binary vector that carries the hygromycin B phosphotransferase gene as a selection marker and the enhanced green fluorescent protein gene for labeling transformants. Randomly selected transformants appeared mitotically stable, based on their maintenance of hygromycin B resistance after five generations of growth without selection. Genomic Southern blot showed that 88% of 784 transformants contained a single T-DNA insert in their genome. A number of putative mutants affected in colony color, size, and/or morphology were found among these transformants, supporting the utility of Agrobacterium tumefaciens-mediated transformation (ATMT) for random insertional mutagenesis of U. muehlenbergii. This ATMT approach potentially offers a systematic gene functional study with genome sequences of U. muehlenbergii that is currently underway.

Original languageEnglish (US)
Article numbere83896
JournalPloS one
Volume8
Issue number12
DOIs
StatePublished - Dec 30 2013

Fingerprint

Umbilicaria
Lichens
Agrobacterium tumefaciens
Fungi
Agrobacterium radiobacter
lichens
Genes
fungi
Hygromycin B
hygromycin B
hygromycin-B kinase
genes
Mutagenesis
Genome
insertional mutagenesis
genome
Symbiosis
Insertional Mutagenesis
site-directed mutagenesis
Southern Blotting

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

Park, S. Y., Jeong, M. H., Wang, H. Y., Kim, J. A., Yu, N. H., Kim, S., ... Hur, J. S. (2013). Agrobacterium tumefaciens-mediated transformation of the lichen fungus, Umbilicaria muehlenbergii. PloS one, 8(12), [e83896]. https://doi.org/10.1371/journal.pone.0083896
Park, Sook Young ; Jeong, Min Hye ; Wang, Hai Ying ; Kim, Jung A. ; Yu, Nan Hee ; Kim, Sungbeom ; Cheong, Yong Hwa ; Kang, Seogchan ; Lee, Yong Hwan ; Hur, Jae Seoun. / Agrobacterium tumefaciens-mediated transformation of the lichen fungus, Umbilicaria muehlenbergii. In: PloS one. 2013 ; Vol. 8, No. 12.
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abstract = "Transformation-mediated mutagenesis in both targeted and random manners has been widely applied to decipher gene function in diverse fungi. However, a transformation system has not yet been established for lichen fungi, severely limiting our ability to study their biology and mechanism underpinning symbiosis via gene manipulation. Here, we report the first successful transformation of the lichen fungus, Umbilicaria muehlenbergii, via the use of Agrobacterium tumefaciens. We generated a total of 918 transformants employing a binary vector that carries the hygromycin B phosphotransferase gene as a selection marker and the enhanced green fluorescent protein gene for labeling transformants. Randomly selected transformants appeared mitotically stable, based on their maintenance of hygromycin B resistance after five generations of growth without selection. Genomic Southern blot showed that 88{\%} of 784 transformants contained a single T-DNA insert in their genome. A number of putative mutants affected in colony color, size, and/or morphology were found among these transformants, supporting the utility of Agrobacterium tumefaciens-mediated transformation (ATMT) for random insertional mutagenesis of U. muehlenbergii. This ATMT approach potentially offers a systematic gene functional study with genome sequences of U. muehlenbergii that is currently underway.",
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Park, SY, Jeong, MH, Wang, HY, Kim, JA, Yu, NH, Kim, S, Cheong, YH, Kang, S, Lee, YH & Hur, JS 2013, 'Agrobacterium tumefaciens-mediated transformation of the lichen fungus, Umbilicaria muehlenbergii', PloS one, vol. 8, no. 12, e83896. https://doi.org/10.1371/journal.pone.0083896

Agrobacterium tumefaciens-mediated transformation of the lichen fungus, Umbilicaria muehlenbergii. / Park, Sook Young; Jeong, Min Hye; Wang, Hai Ying; Kim, Jung A.; Yu, Nan Hee; Kim, Sungbeom; Cheong, Yong Hwa; Kang, Seogchan; Lee, Yong Hwan; Hur, Jae Seoun.

In: PloS one, Vol. 8, No. 12, e83896, 30.12.2013.

Research output: Contribution to journalArticle

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AU - Kim, Jung A.

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AU - Kim, Sungbeom

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AU - Lee, Yong Hwan

AU - Hur, Jae Seoun

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