New Phytologist 184: 885-897 (2009)
The NADPH-Oxidase AtrbohB plays a role in Arabidopsis seed after-ripening [W]
Centro de Biotecnología y Genómica de Plantas (UPM-INIA), Universidad Politécnica de Madrid, 28223 Pozuelo de Alarcón (Madrid), Spain (M.A.T.)
Received April 30, 2009; accepted July 13, 2009; published online September 15, 2009
DOI 10.1111/j.1469-8137.2009.03005.x
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Figure 4. Alternative AtrbohB mRNA splicing is hormonally and developmentally regulated in Arabidopsis WT seeds. (a) Semiquantitative RT-PCR of fresh or after-ripened (ar) seeds. RNA was extracted from dry seeds and from seeds imbibed for 24 h in medium without (CON) or with 1 µM ABA added (24 ºC, continuous light). Two AtrbohB RT-PCR fragments were obtained: AtrbohB-a (485 bp) and AtrbohB-b (635 bp). The relative abundance of the two transcript types changed in the presence of ABA depending on the developmental state of the seeds. 5.8S rRNA was used as a loading control. Note that the RT-PCR results of dry and imbibed seeds can only be compared qualitatively, i.e. which bands are amplified. As we cannot expect 5.8 S rRNA to be constant between these two states a direct semiquantitative comparison of dry and imbibed seeds is not possible. (b) Diagram illustrating the alternative splicing of AtrbohB mediated by seed after-ripening. The upper parts show the location of the AtrbohB gene (At1g09090) on chromosome 1 and the first four exons and three introns of the transcribed pre-mRNA. In ABA-treated after-ripened seeds splicing of this pre-mRNA leads to the AtrbohB-a mRNA for which the predicted protein is drawn below. In ABA-treated fresh seeds, intron 1 of the pre-mRNA is retained in the AtrbohB-ß mRNA. Introns 2 and 3 are spliced out. Retention of intron 1 leads to a premature termination codon. |
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