Application of the acetolactate synthase gene as a cisgenic selectable marker for Agrobacterium-mediated transformation in Chinese cabbage (Brassica rapa ssp. pekinensis)
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- Konagaya Ken-ichi
- National Institute of Agrobiological Sciences
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- Tsuda Mai
- National Institute of Agrobiological Sciences
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- Okuzaki Ayako
- National Institute of Agrobiological Sciences
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- Ando Sugihiro
- National Institute of Agrobiological Sciences
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- Tabei Yutaka
- National Institute of Agrobiological Sciences
書誌事項
- タイトル別名
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- Application of the acetolactate synthase gene as a cisgenic selectable marker for <i>Agrobacterium</i>-mediated transformation in Chinese cabbage (<i>Brassica rapa</i> ssp. <i>pekinensis</i>)
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抄録
Heading Chinese cabbage (Brassica rapa ssp. pekinensis) has been recalcitrant to regeneration and transformation. In particular, there are few reports concerning Japanese cultivars. We evaluated the factors that inhibit Agrobacterium-mediated transformation in heading Chinese cabbage. Then, we investigated the effects of selectable markers using heading Chinese cabbage cv. Chihiri 70 and compared the effects with those in broccoli cv. Ryokurei (B. oleracea var. italica) as a control for an easily transformable Brassica species. To utilize a selectable marker derived from a host plant cisgene, we cloned a genomic DNA fragment containing regulatory and coding sequences of the acetolactate synthase (ALS) gene from Chinese cabbage and mutagenized it to a herbicide resistant form. After transformation of Chinese cabbage and broccoli with this construct, transgenic plants were efficiently selected with the herbicide bispyribac sodium salt and screened by DNA gel blot analysis. The average transformation frequency of Chinese cabbage was 1.2±0.2%, which was similar to those in reports using antibiotic selectable markers and was lower than for broccoli (13.9±2.0%). Furthermore, the escape rate was restricted at a low level (about 35–50% lower than hygromycin selection), which is an advantage on practical transformation. We confirmed transgene inheritance and herbicide resistance of potted plants in the T1 generation. This report is the first to describe a selection system for the transformation of a Brassica crop that uses a herbicide-tolerant selectable marker derived from a cisgene.
収録刊行物
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- Plant Biotechnology
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Plant Biotechnology 30 (2), 125-133, 2013
日本植物バイオテクノロジー学会
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詳細情報 詳細情報について
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- CRID
- 1390282679304261248
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- NII論文ID
- 10031184116
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- NII書誌ID
- AA11250821
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- COI
- 1:CAS:528:DC%2BC3sXhtlCqsL3P
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- ISSN
- 13476114
- 13424580
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- NDL書誌ID
- 024676558
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
- Crossref
- CiNii Articles
- KAKEN
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- 抄録ライセンスフラグ
- 使用不可