Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Studies on nickel uptake in transgenic Arabidopsis thaliana introduced with TgMTP1 gene encoding metal tolerance protein

TgMTP1 과발현 애기장대에서 Nickel 흡수 연구

  • Received : 2015.11.30
  • Accepted : 2015.12.09
  • Published : 2015.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

To enhance phytoremediation, which removes heavy metal from soil, transgenic plants were applied to contaminated soil. We constructed a transformation vector expressing both $TgMTP_1$ (T. goesingense metal tolerance protein):HA and TgMTP:GFP genes. Transgenic plants were generated using an Agrobacterium-mediated transformation system that expressed the two vectors. Screening and analysis confirmed the incorporation of foreign genes into the Arabidopsis thaliana genome. Callus was induced in the 116 T3 line. These transgenic plants and calli were used for further analyses on the accumulation of Ni. The 116 T3-line plants and calli from selected lines were resistant to heavy metals and accumulated Ni in their leaves. The expression level of TgMTP RNA was equal in all leaves, but protein stability increased in the leaves with Ni treatment. According to these results, we suggest that $TgMTP_1$-overexpressing plants may be useful for phytoremediation of soil.

본 연구는 토양에 오염된 중금속을 제거하기 위한 식물정화공정에 사용할 식물체를 개발하기 위해 $TgMTP_1$ 유전자를 CaMV35S 상시 발현 할 수 있도록 Ti-plasmid 벡터를 구축하여 형질전환식물을 육성하였다. 유전자가 도입된 형질전환 애기장대에서 TgMTP유전자를 과발현하는 호모 TG-116 (T3 generation) 계통을 육성하여 그 특성을 조사하였다. 호모계통으로 육성한 TG-116 계통은 callus 및 식물체에서 중금속에 대한 저항성을 보였다. 특히 RT-PCR 및 Western 분석에서 유전자의 발현은 잎에서 높게 나타났으며, Ni 흡수 및 축적이 많이 일어났다. 따라서 MTP1 유전자가 발현되어 액포에 중금속을 축적하는 실험결과를 활용한다면 식물정화공정에 사용할 수 있는 다양한 유전자원으로 기대할 수 있다.

Keywords

References

  1. Baker AJ (1981) Accumulators and excluders-strategies in the response of plants to heavy metals. J. Plant Nutrit. 3:643- 654 https://doi.org/10.1080/01904168109362867
  2. Etim EE (2012) Phytoremediation and Its Mechanisms: International Journal of Environment and Bioenergy 2(3):120-136
  3. Gustin JL, Loureiro ME, Kim D, Na G, Tikhonova M, Salt DE (2009) MTP1-dependent Zn sequestration into shoot vacuoles suggests dual roles in Zn tolerance and accumulation in Zn-hyperaccumulating plants. Plant J. 57:1116-1127 https://doi.org/10.1111/j.1365-313X.2008.03754.x
  4. Kil EJ, Kim D (2015) Research of phytoremediation possibility using TgMTP1 overexpressing plants. Sungkyunkwan University Journal of Biotechnology and Bioengineering 21:1-5.
  5. Kim D, Gustin JL, Lahner B, Persns MW, Baek D, Yun DJ, Salt DE(2004) The plant CDF family member TgMTP1 from the Ni/Zn hyperaccumulator Thlaspi goesingense acts to enhance efflux of Zn at the plasma membrane when expressed in Saccharomyces cerevisiae. Plant J. 39:237-251 https://doi.org/10.1111/j.1365-313X.2004.02126.x
  6. Kobae Y, Uemura T, Sato MH, Ohnishi M, Mimura T, Nakagawa T, Maeshima TM (2004) Zinc transporter of Arabidopsis thaliana AtMTP1 is localized to vacuolar membranes and implicated in zinc homeostasis. Plant Cell Physiol. 45:1749-1758 https://doi.org/10.1093/pcp/pci015
  7. Kramer, U (2005) Phytoremediation: novel approaches to cleaning up polluted soils. Curr. Opin. Biotechnol. 16:133-141 https://doi.org/10.1016/j.copbio.2005.02.006
  8. Kupper H, Zhao FJ, McGrath SP (1999) Cellular compartmentation of zinc in leaves of the hyperaccumulator Thlaspi caerulescens. Plant Physiol. 119:305-311 https://doi.org/10.1104/pp.119.1.305
  9. Kupper H, Lombi E, Zhao FJ, McGrath SP (2000) Cellular compartmentation of cadmium and zinc in relation to other elements in the hyperaccumulator Arabidopsis halleri. Planta, 212:75-84 https://doi.org/10.1007/s004250000366
  10. Kupper H, Mijovilovich, A, Meyer-Klaucke W, Kroneck PMH (2004) Tissue- and age-dependent differences in the complexation of cadmium and zinc in the cadmium/zinc hyperaccumulator Thlaspi caerulescens (Ganges ecotype) revealed by X-ray absorption spectroscopy. Plant Physiol. 134:748-757 https://doi.org/10.1104/pp.103.032953
  11. Persans MW, Nieman K, Salt DE( 2001) Functional activity and role of cation-efflux family members in Ni hyperaccumulation in Thlaspi goesingense. Proc. Natl Acad. Sci. USA, 98:9995-10000 https://doi.org/10.1073/pnas.171039798
  12. Salt DE, Smith RD, Raskin I (1998) Phytoremediation. Annu. Rev.Plant physiol. Plant Mol. Biol. 49:643-668 https://doi.org/10.1146/annurev.arplant.49.1.643
  13. Weigel D, Glazebrook J (2002) Arabidosis A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.