Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Characterization of Oszinc626, knock-out in zinc finger RING-H2 protein gene, in Ac/Ds mutant lines of rice(Oryza sativar L.)

Zinc finger RING-H2 protein관련 Ac/Ds전이인자 삽입 변이체 Oszinc626 유전자의 특성 분석

  • Park, Seul-Ah (Cell and Genetics Division, National Institute of Agricultural Biotechnology, RDA) ;
  • Jung, Yu-Jin (Cell and Genetics Division, National Institute of Agricultural Biotechnology, RDA) ;
  • Ahn, Byung-Ohg (Cell and Genetics Division, National Institute of Agricultural Biotechnology, RDA) ;
  • Yun, Doh-Won (Cell and Genetics Division, National Institute of Agricultural Biotechnology, RDA) ;
  • Ji, Hyeon-So (Cell and Genetics Division, National Institute of Agricultural Biotechnology, RDA) ;
  • Park, Yong-Hwan (Cell and Genetics Division, National Institute of Agricultural Biotechnology, RDA) ;
  • Eun, Moo-Young (Cell and Genetics Division, National Institute of Agricultural Biotechnology, RDA) ;
  • Suh, Seok-Cheol (Cell and Genetics Division, National Institute of Agricultural Biotechnology, RDA) ;
  • Lee, Soon-Youl (Bioengineering, Hankyong National University) ;
  • Lee, Myung-Chul (Cell and Genetics Division, National Institute of Agricultural Biotechnology, RDA)
  • 박슬아 (농업생명공학연구원 세포유전과) ;
  • 정유진 (농업생명공학연구원 세포유전과) ;
  • 안병옥 (농업생명공학연구원 세포유전과) ;
  • 윤도원 (농업생명공학연구원 세포유전과) ;
  • 지현소 (농업생명공학연구원 세포유전과) ;
  • 박용환 (농업생명공학연구원 세포유전과) ;
  • 은무영 (농업생명공학연구원 세포유전과) ;
  • 서석철 (농업생명공학연구원 세포유전과) ;
  • 이순열 (한경대학교 응용생물과학과) ;
  • 이명철 (농업생명공학연구원 세포유전과)
  • Published : 2008.09.30
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Abstract

Ac/Ds mutant lines of this study were transgenic rice plants, each of which harbored the maize transposable element Ds together with a GUS coding sequence under the control of a promoterless(Ds-GUS). We selected the mutants that were GUS expressed lines, because the GUS positive lines will be useful for identifying gene function in rice. One of these mutants was identified knock-out at Oszinc626(NP_001049991) gene, encoding a RING-H2 zinc-finger protein, by Ds insertion. In this mutant, while primary root development is normal, secondary root development from lateral root was very poor and seed development was incomplete compare with normal plant. RING zinc-finger proteins play important roles in the regulation of development in a variety of organisms. In the plant kingdom, a few genes encoding RING zinc-finger proteins have been documented with visible effects on plant growth and development. The consensus of the RING-H2(C3-H2-C3 type) domain for this group of protein is $Cys-X_2-Cys-X_{28}-Cys-X-His-X_2-His-X_2-Cys-X_{14}-Cys-X_2-Cys$. Oszinc626 encodes a predicted protein product of 445 amino acids residues with a molecular mass of 49 kDa, with a RING-zinc-finger motif located at the extreme end of the C-terminus. RT-PCR analysis indicated that the expression of Oszinc626 gene was induced by IAA, cold, dehydration, high-salinity and abscisic acid, but not by 2,4-D, and the transcription of Oszinc626 gene accumulated primarily in rice immature seeds, root meristem and shoots. The gene accumulation patterns were corresponded with GUS expression.

본 연구는 동진벼 유래의 Ac/Ds 삽입변이집단의 GUS 분석을 통하여 뿌리 및 미숙종자에서 강하게 GUS가 발현한 개체를 선발하여 FST(flanking sequence tag) 분석 한 결과 Ds 전이 인자가 3번 염색체 zinc finger RING-H2 관련 Oszinc626 유전자의 첫 번째 exon 부위에 single copy로 삽입되어 있었으며, 선발변이체는 뿌리 및 종자 발달이 정상인 동진벼에 비해 매우 낮은 것으로 나타났다. Oszinc626 유전자는 RING-H2 type(C3-H2-C3)으로 $Cys-X_2-Cys-X_{28}-Cys-X-His-X_2-His-X_2-Cys-X_{14}-Cys-X_2-Cys$ 배열이 C-terminus 가장 말단에 위치하며, 49 kDa의 분자량을 가지고 있다. 또한 Southern blot 분석에서 Oszinc626 유전자는 벼 게놈상에 single copy로 존재하였다. RT-PCR을 통한 돌연변이 유전자의 발현분석 결과 250 mM의 염과, $4^{\circ}C$ 저온등과 같은abiotic stress에 의해 발현이 증가함을 보였고, 호르몬처리에 있어서 ABA와 IAA의 식물호르몬을 처리했을 경우 24시간까지 계속해서 발현양이 증가하는 것을 보이는 반면, 2,4-D 처리의 경우 30분 후에 발현이 일시적으로 증가되었으나 이후 발현이 급속히 감소한 것을 보였다. 벼의 조직 별 발현 검정에서 미성숙한 종자, 뿌리 분열조직 및 신초 등 주로 생장점 부위에서 강하게 발현되는 것을 보임에 따라 Oszinc626 유전자의 경우 식물의 생장에 관여하는 주동 유전자의 하나로 판단된다.

Keywords

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