Development of a high-throughput method for examining the intra-genic recombination frequency in <i>Wx</i> locus in rice using an image analysis

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  • Uenoyama Anan
    Graduate School of Agriculture, Kyoto University
  • Koide Yohei
    Graduate School of Agriculture, Kyoto University Hakubi Center for Advanced Research, Kyoto University Research Faculty of Agriculture, Hokkaido University
  • Tsukiyama Takuji
    Graduate School of Agriculture, Kyoto University Faculty of Agriculture, Kindai University
  • Teraishi Masayoshi
    Graduate School of Agriculture, Kyoto University
  • Okumoto Yutaka
    Graduate School of Agriculture, Kyoto University

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Other Title
  • 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
  • イネWx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
  • イネ Wxザ ニ オケル イデンテキ クミカエ ヒンド カンイ ケイソクホウ ノ ガゾウ カイセキ ニ ヨル カンベンカ

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Abstract

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 wx 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, T65wx and 73wx1. These lines have different mutations on Wx gene, which controls starch synthesis in rice. Although these two lines produce only wx type pollen, F1 plants developed from a cross between them produce Wx type pollen in a low frequency because of an intra-genic recombination between the two mutations on Wx locus. Therefore, it is possible to analyze the recombination frequency from a frequency of Wx 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 Wx 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

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