Horticultural Science & Technology (원예과학기술지)

Korean Society of Horticultural Science (한국원예학회)
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Construction of Tomato yellow leaf curl virus Clones for Resistance Assessment in Tomato Plants

토마토 작물의 TYLCV 저항성 평가에 이용할 수 있는 감염성 클론 개발

  • Choi, Seung Kook (National Institute of Horticultural & Herbal Science, Rural Development Administration) ;
  • Choi, Hak Soon (National Institute of Horticultural & Herbal Science, Rural Development Administration) ;
  • Yang, Eun Young (National Institute of Horticultural & Herbal Science, Rural Development Administration) ;
  • Cho, In Sook (National Institute of Horticultural & Herbal Science, Rural Development Administration) ;
  • Cho, Jeom Deog (National Institute of Horticultural & Herbal Science, Rural Development Administration) ;
  • Chung, Bong Nam (National Institute of Horticultural & Herbal Science, Rural Development Administration)
  • 최승국 (농촌진흥청 국립원예특작과학원) ;
  • 최학순 (농촌진흥청 국립원예특작과학원) ;
  • 양은영 (농촌진흥청 국립원예특작과학원) ;
  • 조인숙 (농촌진흥청 국립원예특작과학원) ;
  • 조점덕 (농촌진흥청 국립원예특작과학원) ;
  • 정봉남 (농촌진흥청 국립원예특작과학원)
  • Received : 2012.08.29
  • Accepted : 2012.12.04
  • Published : 2013.04.30
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Abstract

Five isolates of Tomato yellow leaf curl virus (TYLCV) collected from various regions of Korea were amplified using PCR and determined the sequences of full-length genome, respectively. The PCR-amplified DNA of each TYLCV isolate was introduced into a binary vector to construct infectious clone containing 1.9 copies of the corresponding viral genome. Various cultivars and breeding lines of tomato were inoculated with Agrobacterium tumefaciens harboring infectious clone of each TYLCV isolate to assess resistance against TYLCV. Susceptible cultivar 'Super-sunread' revealed typical yellowing and narrowing of the upper leaves. In contrast, breeding linesTY12, GC9, GC171, and GC173, which contained the TY-1 and/or TY-3 genes that confer resistance against TYLCV in nature, were completely symptomless, suggesting that the lines were resistant to challenging TYLCV isolates. Symptoms of TYLCV in susceptible tomato cultivars are significantly different from those of TYLCV in the resistant tomato cultivars at 30 days after agroinfiltration. Although genomic DNAs of TYLCV were detected from the breeding lines TY12, GC9, GC171, and GC173 using real-time PCR analysis with specific primers, levels of TYLCV DNA accumulation in the resistant breeding lines were much lower than those of TYLCV DNA accumulation in susceptible tomato cultivars. Similar symptom severity and levels of TYLCV DNA accumulation were observed from TYLCV infections mediated by Bemisia tabaci in the resistant and susceptible tomato cultivars. Concentration of agrobacterium did not affect the response of tomato cultivars against TYLCV inoculation. Taken together, these results suggest that TYLCV inoculation via agroinfiltration is as effective as inoculation through Bemisia tabaci and is useful for breeding programs of TYLCV-resistant tomato.

국내에서 채집한 5종의 토마토황화잎말림바이러스(Tomato yellow leaf curl virus, TYLCV) 분리주들에 대하여 PCR법을 이용하여 염기서열을 결정하였다. 전체 유전체의 1.9배 복제수를 가지는 감염성 클론을 제작하기 위하여, PCR로 증폭된 TYLCV DNA를 식물형질전환용 벡터에 각각 도입시켰다. TYLCV 저항성 평가를 위하여 저항성 토마토 시판 품종과 육성 계통을 각각 TYLCV 분리주를 가지고 있는 Agrobacterium tumefaciens로 접종하였다. 감수성 품종인 'Super-sunroad' 품종은 접종 30일 후에 상엽에 황화, 위축 증상을 나타낸 반면, TYLCV에 대한 저항성유전자들로 알려진 Ty-1 또는 Ty-3 유전자를 포함하고 있는 TY12, GC9, GC171, GC173 등의 토마토 육성 계통은 전신 잎에 TYLCV 병징 발현이 없었으므로 이들 계통은 TYLCV 저항성으로 판별되었다. Agroinfiltration을 통한 접종 30일에 감수성 품종과 저항성 품종 간에 TYLCV 병징 차이가 뚜렷하였다. 특이적 프라이머들을 이용한 실시간 PCR법에 의해 TY12, GC9, GC171, GC173 육성 계통에서 TYLCV DNA가 검출되었지만, 저항성 계통에서 TYLCV DNA 축적 수준은 감수성 토마토 품종에서의 TYLCV DNA 축적 수준보다 매우 낮았다. 유사한 병징 세기 및 TYLCV DNA 축적 수준이 담배가루이를 통하여 감수성과 저항성 토마토 품종 및 육성 계통에 TYLCV를 감염시켰을 때에도 관찰되었다. Agroinfiltration 시 아그로박테리움의 농도는 토마토 품종의 TYLCV 저항성 반응에 영향을 미치지 않았다. 따라서 본 연구결과들은 agroinfiltration을 통한 TYLCV 접종이 담배가루이를 통한 TYLCV 감염처럼 효과적이며 TYLCV 저항성 토마토 육성 프로그램에 활용 가능하다는 것으로 보여주었다.

Keywords

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