Effectiveness of genotype-based selection in the production of marker-free and genetically fixed transgenic lineages : ectopic expression of a pistil chitinase gene increases leaf-chitinase activity in transgenic rice plants without hygromycin-resistance gene

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An extra copy of a rice pistil chitinase gene, termed<i> chip</i>, which is expressed predominantly in floral organs just before anthesis, was introduced into an economically important cultivar of rice (<i>Oryza sativa</i> L. cv. Koshihikari) together with the hygromycin-resistance gene (<i>hph</i>) as a marker gene by co-transformation. Over 100 regenerated rice plants (R<sub>0</sub>) and their self-pollinated first and second generations (R<sub>1</sub> and R<sub>2</sub>) were genotypically characterized by PCR-based genomic DNA analysis of the two transgenes to obtain stable transgenic lines, in which <i>chip</i> was inserted into a single locus separately from <i>hph</i>. We obtained homozygous <i>chip</i>-transgenic R<sub>1</sub> lines free of <i>hph</i> that stably expressed the <i>chip</i> mRNA and protein in rice leaf tissue. Chitin-hydrolytic activity in the leaves of these transgenic lines was higher than in those of non-transgenic controls. The resistance of the transgenic lines to rice blast disease fungus (<i>Magnaporthe grisea</i>) was examined by smearing water agar mixed with overcrowded conidia (race 007.0) on mechanically injured leaves, showing tendency to blast-disease resistance. Thus, the genotype-based selection was demonstrated to be effective at obtaining genetically stable transgenic plant lines without marker genes.

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  • Plant biotechnology  

    Plant biotechnology 23(4), 349-356, 2006-09-01 

    Japanese Society for Plant Cell and Molecular Biology

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各種コード

  • NII論文ID(NAID)
    10021908983
  • NII書誌ID(NCID)
    AA11250821
  • 本文言語コード
    ENG
  • 資料種別
    ART
  • ISSN
    13424580
  • NDL 記事登録ID
    8041054
  • 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  CJP引用  NDL  J-STAGE 
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