[논문]다양한 콩 자원들의 기내 조직배양 효율 및 형질전환

서미숙; 조철오; 정남희; 성순기; 최만수; 진민아; 김둘이

doi:10.7732/kjpr.2021.34.4.278

다양한 콩 자원들의 기내 조직배양 효율 및 형질전환
In Vitro Tissue Culture Frequency and Transformation of Various Cultivars of Soybean (Glycine max (L.) Merr.) 원문보기

韓國資源植物學會誌 = Korean journal of plant resources, v.34 no.4, 2021년, pp.278 - 286

서미숙 (국립식량과학원 작물기초기반과) , 조철오 (국립식량과학원 작물기초기반과) , 정남희 (국립원예특작과학원 과수과) , 성순기 ((주)팜한농, 종자.식량종자개발팀) , 최만수 (국립식량과학원 작물기초기반과) , 진민아 (국립식량과학원 작물기초기반과) , 김둘이 (국립식량과학원 작물기초기반과)

초록
AI-Helper

유전자 가위 기술 등 생명공학 기술을 콩에 적용하여 새로운 품종을 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, 외래 품종 2종과 함께 조직배양 효율을 조사하였다. 그 결과, 근연 관계가 높은 Kwangan, Anpyeong, Seonam은 발아율과 재분화 효율이 높았으며, Daepung, Daewon 품종은 발아율과 재분화율 모두 낮게 관찰되었다. 또한 3종의 외래 품종에서는 표준 유전체 해독에 사용된 Williams82와 Jack, Maverick 모두 높은 조직배양 효율을 보였다. 조직배양 효율이 높은 자원들을 대상으로 Agrobacterium법에 의한 형질전환을 수행하여 PCR 및 bar-strip 분석한 결과 Kwangan, Pungwon, Seonam, 그리고 Maverick 품종에서 제초제 저항성 유전자의 삽입을 확인할 수 있었다. 이들 결과를 바탕으로 농업적 가치가 높은 다양한 콩 품종들의 형질전환을 통한 새로운 품종 개발이 가능할 것이다.

Abstract ▼ AI-Helper

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 core-collection 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.

주제어

표/그림 (5)

그림 Fig. 1. Germination frequency from half-seed of total 23 soybean cultivars. Embryonic axis was counted at 10 days after culture. Bars represent the mean with standard deviation for three independent experiments. Some cultivars could not confirm the results from replicated data by seed contamination.
그림 Fig. 2. Phylogenetic analysis of the 21 cultivars belong to Korean soybean core collection. This tree was constructed using MEGA, version 6, software. Bootstrap values with 500 replicates are denoted as percentages.
그림 Fig. 3. Regeneration frequency from cotyledonary node in soybean cultivars. Bars represent the mean with standard deviation for three independent experiments. Some cultivars could not confirm the results from replicated data by seed contamination.
그림 Fig. 4. Agrobacterium-mediated transformation of pBAtc:tRNA vector with bar gene for gene editing in soybean cultivar 'Kwangan'. (A) Co-cultivated half-seed explants for three days, (B) Half seeds cotyledonary explants in SIM medium, (C) Regenerated shoot in SEM medium containing 5 mg/L glufosinate, (D) Rooted plantlet in RM medium containing 5 mg/L glufosinate.
그림 Fig. 5. Detection of the bar gene in transgenic soybean plants. (A) LibertyLink^® strip detection. The first line is control line, and the second line is test line. C; non-transgenic plant, T1~T7; transgenic plants (T1, T2; Kwangan, T3~T4; Maverick, T5~T6; Seonam, T7; Pungwon), (B) PCR amplification of the 506 bp bar gene fragment. M; 1 kb molecular marker, P; pBAtc:tRNA02 vector plasmid, C; non-transgenic plant, T1~T12; transgenic plants (T1~T2; Seonam, T3~T7; Kwangan, T8~T11; Maverick,T12; Pungwon).

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저자의 다른 논문 :

표제어: PCR

동의어: Packet Collision Rate

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