イネ<i>Wx</i>座における遺伝的組換え頻度簡易計測法の画像解析による簡便化  [in Japanese] Development of a high-throughput method for examining the intra-genic recombination frequency in <i>Wx</i> locus in rice using an image analysis  [in Japanese]

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Author(s)

    • 小出 陽平 Koide Yohei
    • 京都大学大学院農学研究科|京都大学白眉センター|北海道大学農学研究院 Graduate School of Agriculture, Kyoto University|Hakubi Center for Advanced Research, Kyoto University|Research Faculty of Agriculture, Hokkaido University
    • 築山 拓司 Tsukiyama Takuji
    • 京都大学大学院農学研究科|近畿大学農学部 Graduate School of Agriculture, Kyoto University|Faculty of Agriculture, Kindai University

Abstract

作物の育種とは遺伝的組換えにより創出された多様な遺伝子組合せを持つ個体から有益な個体を選抜することであるため,組換え頻度の理解は育種にとって重要な課題である.本研究では,イネ<i>Wx</i>座をモデルとし,大規模な分離集団の代わりに花粉を用いた組換え頻度計測法を画像解析によって更に簡便にし,この方法を用いてレトロトランスポゾン(RT)挿入がイネの組換え頻度に与える影響を評価した.測定法の開発には<i>Wx</i>遺伝子座内の異なる部位に変異を持つ2種類のイネ系統(T65<i>wx</i>および73<i>wx</i>1)の交雑F<sub>1</sub>を用いた.このF<sub>1</sub>個体は変異部位間の組換えにより低頻度でウルチ型花粉を産出するため,ウルチ型花粉の出現頻度から組換え頻度を測定することができる.本研究では,F<sub>1</sub>個体の花粉を画像解析ソフトを用いて計数することで,組換え頻度を簡便かつ高い精度で測定することが可能となった.さらに,<i>Wx</i>遺伝子座第9エキソンに約7kbのRT挿入を持つヒメノモチを用いて組換え頻度を調査したところ,RT挿入は周辺の組換え頻度に影響を与えることが示唆された.

Plant breeding is to select agronomically valuable genotypes from a diverse set of gene combinations established through hybridizations and subsequent genetic recombinations. Therefore, it is important for the breeding science to understand how to control the recombination frequency. In the present study, we developed a new high-throughput method for analyzing the recombination frequency within <i>wx</i> locus in rice using an image processing software(ImageJ) and analyzed the effect of a retrotransposon(RT) on the recombination frequency around it. To develop the method, we used two near-isogenic rice lines, T65<i>wx</i> and 73<i>wx</i>1. These lines have different mutations on <i>Wx</i> gene, which controls starch synthesis in rice. Although these two lines produce only <i>wx</i> type pollen, F<sub>1</sub> plants developed from a cross between them produce <i>Wx</i> type pollen in a low frequency because of an intra-genic recombination between the two mutations on <i>Wx</i> locus. Therefore, it is possible to analyze the recombination frequency from a frequency of <i>Wx</i> type pollen. In this study, we used an image processing techniques for counting the total number of pollen grains. This enabled us to count the number of pollen grains more efficiently and precisely. By using the method described in this report, we also examined the effect of RT-insertion and found that an RT insertion on <i>Wx</i> gene reduces the intra-genic recombination frequency around the insertion.

Journal

  • Journal of Crop Research

    Journal of Crop Research 61(0), 45-49, 2016

    The Society of Crop Science and Breeding in Kinki, Japan

Codes

  • NII Article ID (NAID)
    130006900259
  • NII NACSIS-CAT ID (NCID)
    AA12344964
  • Text Lang
    JPN
  • ISSN
    1882-885X
  • NDL Article ID
    028918809
  • NDL Call No.
    Z18-705
  • Data Source
    NDL  J-STAGE 
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