Horticultural Science & Technology (원예과학기술지)

Korean Society of Horticultural Science (한국원예학회)
권호 표지

Development of Gene-based DNA Marker for Verticillium Wilt Resistance in Tomato

토마토 반신위조병 저항성 선발용 DNA 마커 개발

Park, Young-Hoon;Lee, Yong-Jae;Kang, Jum-Soon;Choi, Young-Whan;Son, Beung-Gu
박영훈;이용재;강점순;최영환;손병구

  • Published : 20080900
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Abstract

Ve-SCAR (sequence characterized amplified region) tightly linked to verticillium wilt resistance genes (Ve1 and Ve2) in tomato has potential to be used for marker-assisted selection (MAS) in breeding programs. In our MAS program using this SCAR marker, it was revealed that the cultivar ‘Mountain Crest’ and several individual plants in a segregating population were susceptible to verticillium wilt, but carried marker genotypes of homozygous resistance. To develop a gene sequence-based marker that would universally work for diverse tomato breeding materials, the Ve1 and Ve2 genes were amplified from the resistance check ‘Hayslip’, susceptible check ‘Ace’, and ‘Mountain Crest’ and the PCR amplicons were tested for RFLP (restriction fragment length polymorphism) by using 14 restriction enzymes. Only the restriction enzyme Hinc II digestion revealed polymorphisms among those cultivars. In addition, sequence alignment of the Ve1 gene of those cultivars clearly showed that ‘Mountain Crest’ carried a second susceptible allele distinguished from other susceptible lines. The polymorphic Hinc II sites were confirmed by the gene sequence analysis and a CAPS (cleaved amplified polymorphic site) marker was developed. Compared to the previous SCAR marker, our new CAPS marker can be more reliable and efficient for the selection of verticillium wilt resistance in tomato.

기존 토마토 반신위조병(verticillium wilt) 저항성 분자표지인 Ve-SCAR를 이용한 분자표지이용선발(MAS) 과정에서 표현형은 감수성인 반면 저항성 마커 유전자형을 보이는 한 품종(‘Mountain Crest’)과 분리집단이 발견된 바, 보고된 저항성 유전자(Ve1, Ve2)의 염기서열을 이용하여 더욱 효과적인 마커를 개발하고자 하였다. Ve1과 Ve2 유전자 특이적 PCR 프라이머를 합성하여 저항성 표본 품종인 ‘Hayslip’과 감수성 표본 품종인 ‘Ace’, 그리고 ‘Mountain Crest’로부터 유전자 단편을 증폭시켜, 총 14개 제한효소의 RFLP 양상을 조사하였다. 이 중 Hinc Ⅱ로부터 각 품종 간 다형성이 확인되었으며, 각 PCR 증폭산물의 염기서열 비교분석을 통해 ‘Mountain Crest’가 PCR증폭산물의 염기서열 비교분석을 통해 ‘Mountain Crest’가 Hinc Ⅱ 제한효소자리에 대해 다른 감수성 식물재료와는 상이한 Ve1 유전자 염기서열을 지니고 있음을 밝혔다. 이를 기반으로 저항성과 두 감수성 대립유전자들의 감별이 동시에 가능한 공우성 CAPS 마커를 개발하여 본 육종재료에 대한 유효성을 검정하였다. 본 연구에서 개발된 CAPS 마커는 기존의 Ve-SCAR 마커에 비해 더욱 정확한 선발효과를 지닐 뿐 아니라, 저항성 유전자 자체의 염기서열분석에 의해 개발되어 연관성 소실(linkage break)의 문제가 없다는 점에서 분자표지이용 선발을 통한 내병성 품종육성에 효과적으로 활용될 수 있다.

Keywords

References

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