Scholarlink

<p>The majority of transformation protocols for <i>Arabidopsis thaliana</i> are based on <i>Agrobacterium</i>-mediated transformation of female reproductive cells <i>in vivo</i>, or somatic cells of leaf, stem, hypocotyl, or root explants <i>in vitro</i>. Here, <i>Agrobacterium</i>-mediated transformation systems using <i>in vitro</i> meristematic floral buds or apical meristems from precultured seedlings are reported. <i>Arabidopsis</i> floral cultures were established from inflorescences of seed-derived plants grown <i>in vitro</i>. On cytokinin-containing medium, chopped inflorescence segments quickly produced fast-growing cultures consisting mainly of real <i>de novo</i> inflorescences. Cytokinin-containing medium was also used for 1-wk preculture of seedlings for induction of multiple meristems from shoot apical buds. NPTII and GUS were used as selectable and visual markers for transformation of <i>in vitro</i> floral cultures, and aadA and RUBY were used for transformation of precultured seedlings. Transformation frequency for floral culture was about 15% (calculated as the number of independent transformants per initial number of explants). Transformation frequency for precultured seedlings using RUBY was roughly 12% (calculated as the number of RUBY positive plants per number of seedlings inoculated). Molecular and phenotypic analysis of T<sub>1</sub> progeny confirmed that the transgenes were stably integrated and inherited in progeny.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Floral and apical meristems as a target for Agrobacterium-mediated transformation of Arabidopsis thaliana

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年7月31日

查看全文
  • Vladimir Sidorov,
  • Peizhen Yang

摘要

The majority of transformation protocols for Arabidopsis thaliana are based on Agrobacterium-mediated transformation of female reproductive cells in vivo, or somatic cells of leaf, stem, hypocotyl, or root explants in vitro. Here, Agrobacterium-mediated transformation systems using in vitro meristematic floral buds or apical meristems from precultured seedlings are reported. Arabidopsis floral cultures were established from inflorescences of seed-derived plants grown in vitro. On cytokinin-containing medium, chopped inflorescence segments quickly produced fast-growing cultures consisting mainly of real de novo inflorescences. Cytokinin-containing medium was also used for 1-wk preculture of seedlings for induction of multiple meristems from shoot apical buds. NPTII and GUS were used as selectable and visual markers for transformation of in vitro floral cultures, and aadA and RUBY were used for transformation of precultured seedlings. Transformation frequency for floral culture was about 15% (calculated as the number of independent transformants per initial number of explants). Transformation frequency for precultured seedlings using RUBY was roughly 12% (calculated as the number of RUBY positive plants per number of seedlings inoculated). Molecular and phenotypic analysis of T1 progeny confirmed that the transgenes were stably integrated and inherited in progeny.