Sequence database of 1172 T‐DNA insertion sites in <i>Arabidopsis</i> activation‐tagging lines that showed phenotypes in T1 generation

• Takanari Ichikawa

• Miki Nakazawa

• Mika Kawashima

• Shu Muto

• Kazushi Gohda

• Kumiko Suzuki

• Akie Ishikawa

• Hiroko Kobayashi

• Takeshi Yoshizumi

• Yuko Tsumoto

• Yumi Tsuhara

• Haruko Iizumi

• Yukiko Goto

• Minami Matsui

抄録

Plant genomic resources harbouring gain‐of‐function mutations remain rare, even though this type of mutation is believed to be one of the most useful for elucidating the function of unknown genes that have redundant partners in the genome. An activation‐tagging T‐DNA was introduced into the genome of Arabidopsis creating 55 431 independent transformed lines. Of these T1 lines, 1262 showed phenotypes different from those of wild‐type plants. We called these lines ‘AT1Ps’ ( a ctivation T1 p utants). The phenotypes observed include abnormalities in morphology, growth rate, plant colour, flowering time and fertility. Similar phenotypes re‐appeared either in dominant or semi‐dominant fashion in 17% of 177 AT2P plants tested. Plasmid rescue or an adaptor‐PCR method was used to identify 1172 independent genomic loci of T‐DNA integration sites in the AT1P plants. Mapping of the integration sites revealed that the chromosomal distribution of these sites is similar to that observed in conventional T‐DNA knock‐out lines, except that the intragenic type of integration is slightly lower (27%) in the AT1P plants compared to that observed in other random knock‐out populations (30–35%). Ten AT2P lines that showed dominant phenotypes were chosen to monitor expression levels of genes adjacent to the T‐DNA integration sites by RT‐PCR. Activation was observed in 7 out of 17 of the adjacent genes detected. Genes located up to 8.2 kb away from the enhancer sequence were activated. One of the seven activated genes was located close to the left‐border sequence of the T‐DNA, having an estimated distance of 5.7 kb from the enhancer. Surprisingly, one gene, the first ATG of which is located 12 kb away from the enhancer, showed reduced mRNA accumulation in the tagged line. Application of the database generated to Arabidopsis functional genomics research is discussed. The sequence database of the 1172 loci from the AT1P plants is available (<jats:ext-link xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="http://pfgweb.gsc.riken.go.jp/index.html">http://pfgweb.gsc.riken.go.jp/index.html ).

収録刊行物

• The Plant Journal

  The Plant Journal 36 (3), 421-429, 2003-10-13

  Wiley

キーワード

• Cell Biology
• Plant Science
• Genetics

詳細情報

• CRID: 1363107369307046912
• NII論文ID: 30014609365
• DOI: 10.1046/j.1365-313x.2003.01876.x
• ISSN: 1365313X
• ISSN: 09607412
• ISSN: http://id.crossref.org/issn/09607412
• Web Site: https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1046%2Fj.1365-313X.2003.01876.x
• Web Site: https://onlinelibrary.wiley.com/doi/pdf/10.1046/j.1365-313X.2003.01876.x
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