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2019 Vol.37, Issue 6 Preview Page

Research Article

Development of Male-Sterile Elite Lines using Marker-Assisted Backcrossing (MABC) in Tomato

Marker-Assisted Backcrossing (MABC)을 이용한 토마토 웅성불임 엘리트 계통 육성

Dong Hyun Kim1†
Yu Jin Jung14†
Jong Hee Kim1
Hee Kyoung Kim1
Ki Hong Nam1
Hyo Ju Lee1
Myong-Kwon Kim2
Ill-Sup Nou3
Kwon Kyoo Kang14*
김 동현1†
정 유진14†
김 종희1
김 희경1
남 기홍1
이 효주1
김 명권2
노 일섭3
강 권규14*
1Department of Horticultural Life Science, Hankyong National University
2Tomato Research Center
3Department of Horticulture, Sunchon National University
3Institute of Genetic Engineering, Hankyong National University
1한경대학교 원예생명과학과
2(주)토마토연구소
3순천대학교 원예학과
3한경대학교 유전공학연구소
* Corresponding Author
† These authors contributed equally to this work.
December 2019. pp. 744-755
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Abstract

Recurrent backcrossing은 donor친으로부터 엘리트 품종에 하나 이상의 좌에서 대립유전자들을 도입하려고 할 때 일반적으로 사용되는 전통적인 육종 방법이다. 웅성불임 엘리트 토마토 계통을 육성하기 위해 마커 지원 foreground 및 background 선발은 여교잡 육종 과정에서 수행하였다. MABC(marker-assisted backcrossing)은 기존의 여교배와 비교할 때 반복친의 유전적 배경을 회복하는데 매우 유용하게 사용하고 있다. Foreground 선발을 위해서 육묘 단계에 녹색 배축을 보이는 안토시아닌 결손(aa) 토마토 변이체는 T2-517 계통 유래 InDel 마커를 사용하여 BC1F1 및 BC2F1 집단에서 선발하였다. 이들 특성은 웅성불임 ms1035 유전자와 2번 염색체상에 매우 밀접하게 연관되어 있으므로 MAS(marker-assisted selection)에 이용하였다. Background 선발을 위해서 MR10-3211과 T2-517 계통 간의 resequencing 데이터로부터 얻은 48개의 single nucleotide polymorphism(SNP) 마커는 토마토 게놈에 고르게 분포되도록 최종 선발하였다. 이형접합(Aa) 유전자형을 갖는 BC1F1 및 BC2F1 개체들은 48개의 SNP 마커 세트를 background 선발에 이용하였다. 다중 유전형 분석은 high-throughput genotyping system(EP1TM, Fluidigm®, USA)을 사용하여 수행되었다. 그 결과 BC1F1 세대에서 반복친 게놈과 87.5%가 유사한 1개의 식물체와 BC2F1 세대에서 반복친 게놈의 회복률이 95.5% 이상으로 보이는 3개의 식물체를 선발하였다. 선발된 개체들은 토마토 웅성불임 계통을 육성하기 BC2F2 및 BC2F3 세대로 고정하였다. 따라서 MABC 방법은 토마토 육종 프로그램에서 반복친 게놈을 회복시키기 위한 획기적인 방법으로 생각된다.

키워드
background selection
foreground selection
marker-assisted selection (MAS)
반복여교잡 단일염기다형성(SNP) 마커

Recurrent backcrossing is a traditional breeding method that is commonly employed to transfer alleles at one or more loci from a donor to an elite variety. In order to develop male-sterile elite tomato lines, marker-assisted foreground and background selections were performed during backcross breeding. Compared to conventional backcrossing, marker-assisted backcrossing (MABC) is extremely useful for recovery of a recurrent parent’s genetic background. For foreground selection, the tomato mutant anthocyanin absent (aa) presents a green hypocotyl during the seedling stage. Lines carrying the Aa genotype were selected from BC1F1 and BC2F1 populations using indel markers derived from the unique aa mutation of T2-517. This trait has been utilized in marker-assisted selection of male sterile 10 (ms10) at the seedling stage because their corresponding loci are closely linked on chromosome 2. For background selection, a total of 48 single nucleotide polymorphism (SNP) markers obtained from resequencing data between the MR10-3211 and T2-517 lines evenly distributed in the tomato genome were finally selected. BC1F1 and BC2F1 plants carrying the heterozygous (Aa) genotype were subjected to background selection using a set of 48 SNP markers. Multiple genotype analysis was done using a high-throughput genotyping system. As a result, one plant, 87.5% similar to the recurrent parent genome in the BC1F1 generation, and three plants with a 95.5% recovery rate of the recurrent parent genome in the BC2F1 generation, were selected. These selected plants were fixed in the BC2F2 and BC2F3 generations for the male sterile tomato elite line. We therefore demonstrate the utility of the MABC method for the recovery of recurrent parent genomes in tomato breeding strategies.

Keywords
background selection
foreground selection
marker-assisted selection (MAS)
recurrent backcrossing
single nucleotide polymorphism (SNP) marker
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Information
  • Publisher :KOREAN SOCIETY FOR HORTICULTURAL SCIENCE
  • Publisher(Ko) :원예과학기술지
  • Journal Title :Horticultural Science and Technology
  • Journal Title(Ko) :원예과학기술지
  • Volume : 37
  • No :6
  • Pages :744-755
  • Received Date :2019. 06. 10
  • Revised Date :2019. 08. 22
  • Accepted Date : 2019. 09. 02
  • DOI :https://doi.org/10.7235/HORT.20190074
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