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. 2000 Jan 1;14(1):28-33.

Identification and disruption of an Arabidopsis zinc finger gene controlling seed germination

M Papi  1 , S Sabatini, D Bouchez, C Camilleri, P Costantino, P Vittorioso
Affiliations

Identification and disruption of an Arabidopsis zinc finger gene controlling seed germination

M Papi et al. Genes Dev. .

Abstract

We describe here the Arabidopsis gene DAG1, encoding a zinc finger transcription factor of the Dof family, and show that it is involved in the control of seed germination. By a reverse genetics approach, we isolated an Arabidopsis mutant line with one T-DNA insertion in DAG1. Seeds from homozygous knockout dag1-1 plants do not develop dormancy and germinate also in the absence of light. Segregation analysis indicates that the effect of the mutation is maternal. Accordingly, in situ mRNA hybridizations reveal expression of DAG1 in the vascular tissue of the flower and maturing fruit but not in the seed.

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Figures

Figure 1
Figure 1
Structure of the DAG1 gene and position of the T-DNA insertion, which is not drawn to scale. (Solid rectangles) exons; (arrows) primers utilized for the isolation of the mutant line. The DAG1 cDNA fragment was utilized as a probe for Northern analysis and to derive the sense and antisense riboprobes for in situ mRNA hybridizations.
Figure 2
Figure 2
Siliques from dag1-1 mutant and WS plants.
Figure 3
Figure 3
Germination rates of WS and dag1-1 mutant seeds. (A) Freshly harvested seeds; (B) seeds dry-stored for 4 weeks after harvest; (C) stored seeds, germination in the presence of ABA, scored at day 7; (D) stored seeds, germination in the presence of paclobutrazol, scored at day 7; (E) stored seeds, germination in total darkness [(□) WS seeds; (●) dag1-1 seeds]; (F) stored seeds, germination in total darkness after a red (R) and a far-red (FR) light pulse [(open bars) WS seeds; (solid bars) dag1-1 seeds].
Figure 4
Figure 4
Analysis of DAG1 expression in different organs and developmental stages. (A) RT–PCR with DAG1-specific primers on RNA from WS and dag1-1: (F12, F13) flowers at stages 12 and 13; (S16, S17) siliques at stages 16 and 17. (−) RT–PCR negative control; (+) PCR performed with WS DNA as a template. (B) In situ mRNA hybridizations with tritium-labeled antisense DAG1 riboprobe on sections of WS flowers at stage 12 (F12, longitudinal section), gynoecium of F12 flowers (Gy12, transverse section), siliques at stages 16 (Si16, transverse section) and 17 (Si17, transverse section), late globular-stage embryos (GE, longitudinal section), and torpedo-stage embryos (TE, longitudinal section). (Left) Dark-field images of the hybridizations; (right) corresponding fluorescence micrographs. Arrows point to the vasculature (v), gynoecium (gy), funiculus (fu) and ovule (ov). Bar 250 μm in F12 panels, 200 mm in Gy12, 200 μm in Si16, 400 μm in Si17, 60 μg in GE and TE.
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