Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Phenotypic and molecular characteristics of second clone (T0V2) plants of the LeLs-antisense gene-transgenic chrysanthemum line exhibiting non-branching

무측지성 국화 형질전환 계통 영양번식 제2세대의 형태적 및 분자생물학적 특성

  • Lee, Su Young (National Institute of Horticultural and Herbal Science) ;
  • Kim, Jeong-Ho (National Institute of Horticultural and Herbal Science) ;
  • Cheon, Kyeong-Seong (National Institute of Horticultural and Herbal Science) ;
  • Lee, Eun Kyung (National Institute of Horticultural and Herbal Science) ;
  • Kim, Won Hee (National Institute of Horticultural and Herbal Science) ;
  • Kwon, O Hyeon (National Institute of Horticultural and Herbal Science) ;
  • Lee, Hye Jin (National Institute of Horticultural and Herbal Science)
  • Received : 2013.10.17
  • Accepted : 2013.11.02
  • Published : 2013.12.31
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Abstract

This study examined the phenotypic and molecular characteristics of the $2^{nd}$ clone ($T_0V_2$) plants of LeLs-antisense gene-transgenic chrysanthemum line (LeLs80) that exhibited non-branching, proving the relevance of these characteristics as a factor for use in environmental risk assessment. Results of the Southern blot analysis showed that three copies of the LeLs-antisense gene were introduced into the transgenic line, and northern analysis showed that the transcripted gene was normally expressed in the transgenic line. A flanking T-DNA sequencing method was used to determine that sequences of 184 and 464 bps flanked the LeLs-antisense gene in the transgenic line. These sequences, respectively, matched the 35S promoter for expression of the npt II gene and the NOS terminator for expression of the LeLs-antisense gene within the pCAMBIA 2300 vector.

환경위해성평가 연구 대상인 형질전환 이벤트로서의 자격을 확인하고자 형질전환세대($T_0V_0$)에서와 같이 영양번식 제1세대($T_0V_1$)에서도 도입유전자 LeLs-antisense의 발현 특성인 무측지성을 유지한 국화 무측지성 형질전환계통 LeLs80의 영양번식 제2세대($T_0V_2$)의 형태적 및 분자 생물학적 특성을 조사하였다. LeLs80 계통의 $T_0V_2$에서도 LeLs-antisense 유전자의 발현 특성인 무측지성이 안정적으로 유지되는 것을 확인하였다. 또한, Southern 및 Northern 분석에 의해 LeLs-antisense 유전자가 3 copy 도입되었으며, LeLs-antisense 유전자의 전사체가 정상적으로 발현되는 것도 확인할 수 있었다. 또한 flanking T-DNA sequencing method를 이용하여 LeLs-antisense 유전자의 주변 염기서열 분석 통해 LeLs80 계통의 genome내 LeLs-antisense 유전자 주변에 186 ~ 464 bp의 pCAMBIA2300 T-DNA right border 부근으로 추정되는 염기서열이 확인되었고, pCAMBIA2300 전 염기서열과의 비교 분석한 결과, pCAMBIA2300 T-DNA left border와 right border내 선발마커 유전자 NPT II의 발현 promoter 부분과 LeLs-antisense 유전자 발현 terminator 일부 염기서열과 일치하였다.

Keywords

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