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Exogenous toluene gas removal improvement in recombinant rabbit cytochrome P450 2E1 (CYP2E1)-transgenic Ardisia pusilla DC

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A Correction to this article was published on 14 April 2021

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Abstract

The cytochrome P450 2E1 (CYP2E1) enzyme encoded by CYP2E1 plays an important role in the metabolism of organic compounds in mammalian liver cells. In this study, the plasmid pCAMBIA2300 harboring the rabbit CYP2E1 was transferred into Agrobacterium tumefaciens; this was then transferred into internodes of Ardisia pusilla. Shoot primordia, whose appearance resembles that of the in vitro protocorm-like body of orchids, regenerated from 121 of 332 internodes that were cultured in selection medium supplemented with 5 mg·L−1 kanamycin. PCR analysis confirmed that 25 of 89 putative transgenic plants (28.0%) stably harbored the CYP2E1 and NPTII genes, and Southern blot analysis revealed the presence of one to three copies of CYP2E1 within the genome of the 25 transgenic plants. The mRNA expression of the transgene was confirmed in 12 of 25 CYP2E1-transgenic plants by Northern blot analysis. Western blot analysis indicated the presence of a 53-kDa recombinant rabbit CYP2E1 protein in the 12 CYP2E1-transgenic A. pusilla plants. The aniline 4-hydroxylase activity of rabbit CYP2E1 in nine CYP2E1-transgenic A. pusilla plants was higher than that in non-transgenic A. pusilla plants. There was a statistically significant increase (5.1–6.1-fold) in the toluene removal ability of the transgenic line CYP2E1-2–2 compared to that observed in wild-type Ardisia plants at different times after exogenous toluene exposure.

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Acknowledgements

This study was supported by a grant from the Next-Generation BioGreen 21 Program (No. PJ01319803), Rural Development Administration, South Korea.

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Authors and Affiliations

  1. Floriculture Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, Wanju, 55365, South Korea

    Nan-Sun Kim, Ju Young Shin, Young Ah Lee, Hye Ryun An, Pue Hee Park, Pil Man Park & Su Young Lee

  2. Urban Agriculture Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, Wanju, 55365, South Korea

    Kwang Jin Kim

  3. Planning and Coordination Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, Wanju, 55365, South Korea

    Jeong Ho Kim & Pue Hee Park

  4. Metabolic Engineering Division, National Institute of Agricultural Sciences, Rural Development Administration, Wanju, 55365, South Korea

    Nan-Sun Kim

Authors
  1. Nan-Sun Kim
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  2. Ju Young Shin
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  3. Young Ah Lee
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  4. Kwang Jin Kim
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  8. Pil Man Park
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  9. Su Young Lee
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Contributions

Su Young Lee and Ju Young Shin obtained CYP2E1-transgenic Ardisia plants. YA Lee performed PCR analysis. Jeong Ho Kim performed Southern and Northern blot analyses. Nan-Sun Kim performed Western blot analysis and CYP2E1 activity assays. Kwang Jin Kim performed the experiment to measure the removal ability of exogenous toluene gas, and others (Hye Ryun An, Pil Man park, and Pue Hee Park) statistically analyzed the data.

Corresponding author

Correspondence to Su Young Lee.

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The authors declare that they have no conflicts of interest.

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Communicated by Tae-Ho Han, Ph.D.

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The original online version of this article was revised: The spelling of Su Young Lee’s name was incorrect.

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Kim, NS., Shin, J.Y., Lee, Y.A. et al. Exogenous toluene gas removal improvement in recombinant rabbit cytochrome P450 2E1 (CYP2E1)-transgenic Ardisia pusilla DC. Hortic. Environ. Biotechnol. 62, 619–627 (2021). https://doi.org/10.1007/s13580-021-00333-2

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