Gene trap lines identify Arabidopsis genes expressed in stomatal guard cells

被引:53
作者
Galbiati, Massimo [1 ]
Simoni, Laura [1 ]
Pavesi, Giulio [1 ]
Cominelli, Eleonora [1 ]
Francia, Priscilla [1 ]
Vavasseur, Alain [2 ]
Nelson, Timothy [3 ]
Bevan, Michael [4 ]
Tonelli, Chiara [1 ]
机构
[1] Univ Milan, Dipartimento Sci Biomol & Biotecnol, I-20133 Milan, Italy
[2] CEN Cadarache, Direct Sci Vivant, Lab Echanges Membranaires & Signalisat, F-13108 St Paul Les Durance, France
[3] Yale Univ, Dept Mol Cellular & Dev Biol, New Haven, CT 06520 USA
[4] John Innes Ctr Plant Sci Res, Dept Cell & Dev Biol, Norwich NR4 7UH, Norfolk, England
关键词
guard cell; gene trap; laser capture microdissection; cis-acting elements; Arabidopsis thaliana;
D O I
10.1111/j.1365-313X.2007.03371.x
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
We employed a gene trap approach to identify genes expressed in stomatal guard cells of Arabidopsis thaliana. We examined patterns of reporter gene expression in approximately 20 000 gene trap lines, and recovered five lines with exclusive or preferential expression in stomata. The screen yielded two insertions in annotated genes, encoding the CYTOCHROME P450 86A2 (CYP86A2) mono-oxygenase, and the PLEIOTROPIC DRUG RESISTANCE 3 (AtPDR3) transporter. Expression of the trapped genes in guard cells was confirmed by RT-PCR experiments in purified stomata. Examination of homozygous mutant lines revealed that abscisic acid (ABA)-induced stomatal closure was impaired in the atpdr3 mutant. In three lines, insertions occurred outside transcribed units. Expression analysis of the genes surrounding the trapping inserts identified two genes selectively expressed in guard cells, corresponding to a PP2C PROTEIN PHOSPHATASE and an unknown expressed protein gene. Statistical analyses of the chromosomal regions tagged by the gene trap insertions revealed an over-represented [A/T]AAAG motif, previously described as an essential cis-active element for gene expression in stomata. The lines described in this work identify novel genes involved in the modulation of stomatal activity, provide useful markers for the study of developmental pathways in guard cells, and are a valuable source of guard cell-specific promoters.
引用
收藏
页码:750 / 762
页数:13
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