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Improvement of DNA/Metal Particle Adsorption in Tungsten-Based Biolistic Bombardment; Alkaline pH is Necessary for DNA Adsorption and Suppression of DNA Degradation

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Abstract

Tungsten particles have long been used as microcarriers in biolistic bombardment because of their cost-effectiveness compared to alternative gold particles—even if the former have several drawbacks, including their DNA-degrading activity. We characterized tungsten-induced DNA degradation to assess the value of this metal particle and to improve tungsten-based biolistic bombardment. Alkaline pH, low temperature, and high salt concentration were found to diminish tungsten-induced DNA breakdown. The pH was the most influential factor in this phenomenon, both in aqueous solutions and on the particles. Furthermore, alkaline pH greater than 9.4 of an adsorption mixture was found to be essential for DNA binding to metal particles. Based on these findings, we propose a new formula of DNA/tungsten adsorption by using TE buffers that keep alkaline pH (>9.4) of the mixture, in which tungsten-bound plasmid DNA cleavage was suppressed to half the level of that in the conventional DNA-binding condition.

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Abbreviations

cc:

closed circular

GFP:

green fluorescent protein

oc:

open circular

Spd:

spermidine-free base

TE:

Tris–HCl/EDTA

References

  • Breitler JC, Labeyrie A, Meynard D, Legavre T, Guiderdoni E (2002) Efficient microprojectile bombardment-mediated transformation of rice using gene cassettes. Theor Appl Genet 104:709–719

    Article  CAS  PubMed  Google Scholar 

  • Buchowicz J, Krysiak C (2003) Genotoxic effects of tungsten microprojectiles under conditions of biolistic transformation. In: Jacson JF, Linskens HF (eds) Genetic transformation of plants, Vol 23. Springer, New York, pp 175–188

    Google Scholar 

  • Carmody WR (1961) An easily prepared wide range buffer series. J Chem Educ 38:559–561

    Article  CAS  Google Scholar 

  • Chiu W, Niwa Y, Zeng W, Hirano T, Kobayashi H, Sheen J (1996) Engineered GFP as a vital reporter in plants. Curr Biol 6:325–330

    Article  CAS  PubMed  Google Scholar 

  • Grenoble DC, Kim CJ, Murrell LL (1983) Hydrocarbon conversion processes using improved transition metal oxide catalysts. Exxon Research and Engineering Co. Florham Park

    Google Scholar 

  • Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405

    Article  CAS  Google Scholar 

  • Kosugi S, Ohashi Y, Nakajima K, Arai Y (1990) An improved assay for β-glucuronidase in transformed cells: methanol almost completely suppresses a putative endogenous β-glucuronidase activity. Plant Sci 70:133–140

    Article  CAS  Google Scholar 

  • Krysiak C, Mazus B, Buchowicz J (1999a) Generation of DNA double-strand breaks and inhibition of somatic embryogenesis by tungsten microparticles in wheat. Plant Cell Tiss Organ Cult 58:163–170

    Article  CAS  Google Scholar 

  • Krysiak C, Mazus B, Buchowicz J (1999b) Relaxation, linearization and fragmentation of supercoiled circular DNA by tungsten microprojectiles. Transgenic Res 8:303–306

    Article  CAS  PubMed  Google Scholar 

  • Mazuś B, Krysiak C, Buchowicz J (2000) Tungsten particle-induced nicking of supercoiled plasmid DNA. Plasmid 44:89–93

    Article  PubMed  CAS  Google Scholar 

  • Nandadeva YL, Lupi CG, Meyer CS, Devi PS, Potrykus I, Bilang R (1999) Microprojectile-mediated transient and integrative transformation of rice embryogenic suspension cells: effects of osmotic cell conditioning and of the physical configuration of plasmid DNA. Plant Cell Rep 18:500–504

    Article  CAS  Google Scholar 

  • Rasco-Gaunt S, Barcelo P (1999) Immature inflorescence culture of cereals. A highly responsive system for regeneration and transformation. Methods Mol Biol 111:71–81

    CAS  PubMed  Google Scholar 

  • Russell JA, Roy MK, Sanford JC (1992) Physical trauma and tungsten toxicity reduce the efficiency of biolistic transformation. Plant Physiol 98:1050–1056

    Article  CAS  PubMed  Google Scholar 

  • Sanford JC, Smith FD, Russell JA (1993) Optimizing the biolistic process for different biological applications. Methods Enzymol 217:483–509

    Article  CAS  PubMed  Google Scholar 

  • Shapiro HS (1967) The preparation of pyrimidine oligonucleotides by acid hydrolysis of DNA. Methods Enzymol 12A:205–212

    Article  Google Scholar 

  • Southgate EM, Davey MR, Power JB, Marchant R (1995) Factors affecting the genetic engineering of plants by microprojectile bombardment. Biotechnol Adv 13:631–651

    Article  CAS  PubMed  Google Scholar 

  • Tanaka K, Asami T, Yoshida S, Nakamura Y, Matsuo T, Okamoto S (2005) Brassinosteroid homeostasis in Arabidopsis is ensured by feedback expressions of multiple genes involved in its metabolism. Plant Physiol 138:1117–1125

    Article  CAS  PubMed  Google Scholar 

  • Thomas R, Moulijn JA, De Beer VHJ, Medema J (1980) Structure/metathesis-activity relations of silica supported molybdenum and tungsten oxide. J Mol Cat 8:161–174

    Article  CAS  Google Scholar 

  • Veluthambi K, Gupta AK, Sharma A (2003) The current status of plant transformation technologies. Curr Sci 84:368–380

    CAS  Google Scholar 

Download references

Acknowledgements

We wish to express special thanks to Dr. Yasuo Niwa at Shizuoka Prefectural University, Japan for providing plasmid p35S-sGFP. We also thank to Dr. Yukiko Yokogawa at Kagoshima University, Japan for helpful discussion. This study is partially supported by a grant in aid of research from the president of Kagoshima University, Japan.

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Authors and Affiliations

  1. Department of Agricultural Sciences and Natural Resources, Kagoshima University, Kohrimoto 1-21-24, Kagoshima, 890-0065, Japan

    Yuya Yoshimitsu, Kiwamu Tanaka & Shigehisa Okamoto

  2. Department of Food Sciences and Nutritional Health, Kyoto Prefectural University, Shimogamo, Sakyo-ku, Kyoto, 606-8522, Japan

    Yasushi Nakamura

  3. Department of Biochemical Science and Technology, Kagoshima University, Kohrimoto 1-21-24, Kagoshima, 890-0065, Japan

    Takashi Tagawa & Tomoaki Matsuo

Authors
  1. Yuya Yoshimitsu
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  2. Kiwamu Tanaka
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  3. Takashi Tagawa
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  4. Yasushi Nakamura
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  5. Tomoaki Matsuo
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  6. Shigehisa Okamoto
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Correspondence to Shigehisa Okamoto.

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Yoshimitsu, Y., Tanaka, K., Tagawa, T. et al. Improvement of DNA/Metal Particle Adsorption in Tungsten-Based Biolistic Bombardment; Alkaline pH is Necessary for DNA Adsorption and Suppression of DNA Degradation. J. Plant Biol. 52, 524–532 (2009). https://doi.org/10.1007/s12374-009-9068-0

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  • DOI: https://doi.org/10.1007/s12374-009-9068-0

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