다양한 콩 자원들의 기내 조직배양 효율 및 형질전환
In Vitro Tissue Culture Frequency and Transformation of Various Cultivars of Soybean (Glycine max (L.) Merr.)
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유전자 가위 기술 등 생명공학 기술을 콩에 적용하여 새로운품종을 개발하기 위해서는 효율적인 조직배양 기술이 필수적이다. 식물의 유전형은 조직배양 효율에 의존하는 형질전환 기술의 성공 여부를 결정짓는 중요한 요소로 알려져 있다. 본 연구에서는 우리나라 콩 핵심 집단 내 21개 자원들을 선발하여, 외래품종 2종과 함께 조직배양 효율을 조사하였다. 그 결과, 근연 관계가 높은 Kwangan, Anpyeong, Seonam은 발아율과 재분화효율이 높았으며, Daepung, Daewon 품종은 발아율과 재분화율 모두 낮게 관찰되었다. 또한 3종의 외래 품종에서는 표준 유전체 해독에 사용된 Williams82와 Jack, Maverick 모두 높은조직배양 효율을 보였다. 조직배양 효율이 높은 자원들을 대상으로 Agrobacterium법에 의한 형질전환을 수행하여 PCR 및bar-strip 분석한 결과 Kwangan, Pungwon, Seonam, 그리고Maverick 품종에서 제초제 저항성 유전자의 삽입을 확인할 수있었다. 이들 결과를 바탕으로 농업적 가치가 높은 다양한 콩 품종들의 형질전환을 통한 새로운 품종 개발이 가능할 것이다.
Abstract
Efficient in vitro regeneration system is essential for the successful crop breeding of soybean (Glycine max (L.) Merr.) using the new biotechnology. The genotype of donor plants strongly influences the establishment of tissue culture system. Therefore, the screening of genotypes with excellent tissue culture ability is very important for soybean genetic improvement. In this study, we report the tissue culture efficiency of 21 soybean cultivars belong to Korean soybean corecollection and two foreign cultivars (Jack and Maverick). The Kwangan, Anpyeong and Seonam are share close genetic relationship in 21 cultivars and these three cultivars were observed the high frequency of germination and regeneration. Furthermore, the high tissue culture abilities were also observed in the Williams 82 used in reference genome sequencing and the two foreign cultivars. The transformation of pBAtc:tRNA with bar gene was performed by Agrobacterium tumefaciens in the cultivars with high tissue culture ability. Transformation of the bar gene was identified by PCR analysis in Kwangan, Pungwon, Seonam, and Maverick. Our results provide useful information for the breeding of various soybean cultivars by plant biotechnology such as, genome editing.
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