Development of rapid mapping system, identifing mutant gene loci with the soybean mutant library.

2020 Fiscal Year Final Research Report

• Project/Area Number: 17K07604
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Science in genetics and breeding
• Research Institution: Saga University
• Principal Investigator: Watanabe Satoshi 佐賀大学, 農学部, 講師 (40425541)
• Co-Investigator(Kenkyū-buntansha): 穴井 豊昭 佐賀大学, 農学部, 教授 (70261774)
• Project Period (FY): 2017-04-01 – 2021-03-31
• Keywords: ダイズ / DNAマーカー / 突然変異集団 / 次世代シークエンス解析 / マッピング

Outline of Final Research Achievements

High-resolution melting (HRM) analysis and optimization of the nearest neighboring nucleotide (NNN) of an SNP by nucleotide substitution in the primer can detect a single nucleotide polymorphism (SNP) in two polymerase chain reaction (PCR) fragments as a melting temperature (Tm) difference without additional experimental steps, such as gel electrophoresis. We also identified some of the responsible genes for mutant lines. To identify the gene responsible for the mutant phenotype, we applied NNNs-HRM genotyping analysis to a small number of F2 plants with the mutant phenotype. This analysis was followed by alignment of short reads obtained by NGS analysis to the identified QTL (the locus of the mutated gene) region and identification of functional SNP for the mutant phenotype. This procedure is time and cost effective and DNA marker developed by this procedure is very effective to select a line having favorable allele during the breeding programs using these mutant alleles.

Free Research Field

植物遺伝育種学

Academic Significance and Societal Importance of the Research Achievements

本課題で開発した技術は突然変異集団から表現型選抜によって見出された突然変異系統の変異遺伝子について、表現型の変化の原因となるDNA多型を選択的遺伝子型決定法と次世代シークエンス解析技術を組み合わせることで、比較的容易に同定することを可能にした。またその変異遺伝子を新品種の育成に利用するにあたり、遺伝子型判別の容易なDNAマーカーと併せて育種現場に提供することが可能となった。