カキ育種におけるマーカー選抜のための実用的 SCAR マーカーの設計 SCAR Markers for Practical Application of Marker-assisted Selection in Persimmon (Diospyros kaki Thunb.) Breeding

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Abstract

これまでの研究で,我々は果実の甘渋性を支配する <i>AST</i> 遺伝子座に連鎖した分子マーカーを単離しているが,これらのマーカーは'黒熊'由来の交雑集団に対して有効でなく,育種計画において実用的なマーカーではない.そこで本研究では,育種計画の中で簡便かつ確実に完全甘ガキ個体を選抜するための新しい sequence characterized amplified region(SCAR)マーカーの開発を試みた.完全甘ガキ個体と非完全甘ガキ個体の間で多型を示す新たな restriction fragment length polymorphism(RFLP)マーカーである 5R 領域を'西村早生','次郎'および'黒熊'由来交雑後代のゲノムライブラリーから単離して解析した結果,3 つの大きな欠損・挿入変異が <i>ast</i> 連鎖領域と <i>AST</i> 連鎖領域の間で確認された.そのうちの 1 つである Indel-3 の近隣にいくつかのプライマーを設計し,SCAR マーカーとしての有効性を検討したところ,2 つの forward primer(AST-F と PCNA-F)と 1 つの reverse primer(5R3R)の組み合わせで行う multiplex PCR が最も有効で確実な方法であることが分かった.<i>AST</i> 対立遺伝子に連鎖する 220 bp の断片は'黒熊','西村早生'および'会津身不知'のいずれの品種の後代でも共通に検出されることが示された.この multiplex PCR は,カキ育種における最も実用的で応用可能なマーカー選抜法であり,この方法によりカキ育種の進展は飛躍的に向上するものと考えられる.<br>

In previous studies, we have developed molecular markers linked to the <i>AST</i> locus that controls fruit astringency type in persimmon; however, these markers are not feasible for practical application to breeding programs since they are not fully effective for discriminating the pollination constant and non-astringent (PCNA) genotype from the non-PCNA genotype in a progeny derived from ‘Kurokuma’. Here we developed new sequence characterized amplified region (SCAR) markers that enable easy and reliable selection of the PCNA genotype from breeding populations. Genomic regions adjacent to probe 5R, which showed polymorphic fragments between PCNA and non-PCNA genotypes in restriction fragment length polymorphism (RFLP) analysis, were isolated from the genomic libraries of ‘Nishimura-wase’, ‘Jiro’, and ‘Kurokuma’-derived offspring. The isolated genomic regions were characterized and 3 insertion/deletion mutations were observed between <i>ast</i>- and <i>AST</i>-linked regions. Several primers were designed in the flanking region of Indel-3 and, in multiplex PCR, it was shown that using 2 forward primers, AST-F and PCNA-F and a reverse primer, 5R3R, is the most useful and reliable primer set. The <i>AST</i>-linked 220-bp fragment proved to be a common marker of ‘Kurokuma’-, ‘Nisimura-wase’- and ‘Aizumishirazu’-derived progenies. This multiplex PCR is considered the most practical tool for marker-assisted selection (MAS) and can enhance and accelerate progress in persimmon breeding.<br>

Journal

  • Journal of the Japanese Society for Horticultural Science

    Journal of the Japanese Society for Horticultural Science 79(2), 150-155, 2010

    THE JAPANESE SOCIETY FOR HORTICULTURAL SCIENCE

Codes

  • NII Article ID (NAID)
    130004951602
  • Text Lang
    ENG
  • ISSN
    1882-3351
  • Data Source
    J-STAGE 
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