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Development of an Efficient In Vitro Screening Method for Selection of Resistant Lily Cultivars Against Fusarium oxysporum f. sp. lilii

백합 구근부패병 저항성 검정을 위한 기내 간편 검정법 개발

  • Jang, Ji-Young (Sustainable Resources Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Moon, Ki-Beom (Sustainable Resources Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Ha, Jang-Ho (Sustainable Resources Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Park, Ji-Sun (Sustainable Resources Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Mi-Jin (Sustainable Resources Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Jeon, Jae-Heung (Sustainable Resources Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Lee, Geung-Joo (Department of Horticulture, Chungnam National University) ;
  • Kim, Hyun-Soon (Sustainable Resources Research Center, Korea Research Institute of Bioscience and Biotechnology)
  • 장지영 (한국생명공학연구원 지속가능자원연구센터) ;
  • 문기범 (한국생명공학연구원 지속가능자원연구센터) ;
  • 하장호 (한국생명공학연구원 지속가능자원연구센터) ;
  • 박지선 (한국생명공학연구원 지속가능자원연구센터) ;
  • 김미진 (한국생명공학연구원 지속가능자원연구센터) ;
  • 전재흥 (한국생명공학연구원 지속가능자원연구센터) ;
  • 이긍주 (충남대학교 원예학과) ;
  • 김현순 (한국생명공학연구원 지속가능자원연구센터)
  • Received : 2015.05.15
  • Accepted : 2015.07.16
  • Published : 2015.12.31

Abstract

The soilborne fungus Fusarium oxysporum f. sp. lilii (Fol) is a serious threat to all lily cultivars, especially infecting bulbs and flowers. It has become increasingly important to develop varieties resistant against the bulb rot disease. Genetic diversity of cultivars and reliable screening methods are required for this purpose. Here, an efficient in vitro screening system for evaluating resistance to Fol in 38 in vitro-grown lily plants was investigated. Various factors including culture conditions of Fol, inoculum density, appropriate plant materials, inoculation method and duration, and incubation period of plant materials after inoculation were combined to optimize the screening method. As a result, we suggest optimal conditions for an in vitro screening system for the selection of Fol-resistant lily cultivars as follows. Fol was grown on potato dextrose agar (PDA) medium for 6 days at $25^{\circ}C$ in darkness and used as working inoculation. Spore suspensions were prepared (inoculum density: $1.0{\times}10^4$ $spores{\cdot}mL^{-1}$), and then leaf segments $1.5{\times}2.0cm^2$ were inoculated by dipping for 22 hours at $25^{\circ}C$ in dark. Later, leaves were cultured on 0.6% agar plates at $25^{\circ}C$ and 50% humidity with a photoperiod of 16 hours light/8 hours dark (fluence rate of $40{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) to examine the progress of bulb rot. After 7 days, disease levels were classified into indices 1 (no symptom) to 6 (serious bulb rot). Soil inoculation of Fol carried out with resistant or susceptible lily cultivars that had been selected through in vitro screening confirmed the reproducibility of results. Therefore, the in vitro screening method established in this study is efficient and reliable for selection of lily cultivars resistant against bulb rot disease.

토양 전염균인 Fusarium oxysporum f. sp. lilii(Fol)은 식물에 심각한 위협을 주는 균으로 특히 백합의 구근과 꽃에 심각한 영향을 준다. 따라서 저항성 품종의 육성이 중요시되고 있는데, 저항성 품종이 육성되기 위해서는 신속, 간편한 검정법 체계와 유전적 다양성이 요구되는 상황이다. 본 실험에서는 기내배양시스템을 이용하여 백합의 Fol에 대한 저항성 정도를 효율적으로 검정하는 방법을 확립하고자 하였다. 병원균의 배양조건, 병원균의 식물체 접종 농도, 대상 절편체 종류, 접종 후 적정 발병 기간 등과 같은 조건을 단독, 혹은 조합하여 실험 한 결과 다음의 기내검정조건을 확립하였다. Fol균을 6일 동안 PDA 배지에서 배양한 후 포자 현탁액을 $1.0{\times}10^4$ $spores{\cdot}mL^{-1}$농도로 조절하여 기내배양 중인 식물체의 잎을 $1.5{\times}2.0cm^2$의 크기로 절단하여 $25^{\circ}C$, 암조건에서 22시간 침지시켜 접종하였다. 접종이 끝난 잎절편체는 0.6% 한천배지에 치상하여 온도 $25^{\circ}C$, 습도 50%, 16시간 광조건(광도 $40{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$)을 유지하면서 병의 발달을 관찰하였다. 발병 정도를 지수화하여 통계처리를 하였다. 기내검정을 통하여 저항성 및 민감성으로 선발한 백합종들을 토양검정법으로 실험한 결과 동일한 저항성 정도를 얻을 수 있었으며, 이는 본 연구에서 확립한 Fol에 대한 구근부패병 기내검정법이 효율적으로 품종 선발에 이용될 수 있음을 보여준다고 하겠다.

Keywords

References

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