G-to-A mutation at a 5′ splice site of fad3c caused impaired splicing in a low linolenic mutant of canola (Brassica napus L.)

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Two genes (<i>fad3a and fad3c</i>) encode for endoplasmic delta-15 linoleate desaturase, which is responsible for the desaturation of linoleic acid (C18:2) into linolenic acid (C18:3) in canola (<i>Brassica napus</i>). The canola mutant line DMS100 carries a G-to-A base substitution at the 5′ splice site, located at +1 G, of <i>fad3c</i> intron 6 and contains reduced C18:3 content in oil seeds. Reverse transcription (RT) PCR analysis was used to amplify part of the <i>fad3c</i> transcript spanning intron 5, exon 6 and intron 6 using the primers specific to the exon 5 and 7 to investigate whether the mutation caused any abnormal splicing. The RT-PCR amplified a fragment of larger size in the <i>fad3c</i> mutant than in wild-type. Sequencing of the RT-PCR fragments revealed that the mutation activated an impaired splicing which retained the entire intron 6 in the mature <i>fad3c</i> transcript of the mutant. The impaired splicing could result in early termination of translation and synthesis of a shorter <i>fad3c</i> polypeptide because the intron 6 contained stop codons in all three possible reading frames. The incomplete translation could produce an inactive enzyme which blocked the desaturation of linoleic acid (C18:2) to linolenic acid (C18:3), resulting in the decrease of C18:3 accumulation in canola seeds. This is consistent with the observation that the <i>fad3c</i> mutant has significantly lower C18:3 content (<3%) than the wild type (~7%).

収録刊行物

  • Plant biotechnology  

    Plant biotechnology 24(4), 397-400, 2007-09-01 

    Japanese Society for Plant Cell and Molecular Biology

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各種コード

  • NII論文ID(NAID)
    10021911864
  • NII書誌ID(NCID)
    AA11250821
  • 本文言語コード
    ENG
  • 資料種別
    SHO
  • ISSN
    13424580
  • NDL 記事登録ID
    8904930
  • NDL 雑誌分類
    ZR3(科学技術--生物学--植物)
  • NDL 請求記号
    Z54-J126
  • データ提供元
    CJP書誌  NDL  J-STAGE 
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