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. 2009 Sep 29;122(20):3749–3758. doi: 10.1242/jcs.050625

Fig. 5.

Fig. 5.

RAB-D1 [N121I] inhibits Arabidopsis growth and cannot be rescued by RAB-D2a. (A) Seedlings exhibiting inducible expression of RAB-D1 [N121I] (D1 NI) but not wild type (wt) grown in the presence (+Dex) or absence (–Dex) of the inducer dexamethasone. Seedlings are shown from one wild-type and two N121I lines in which inducible expression of the transgene was confirmed by RNA gel blot (not shown). (B) As in A, but with seedlings expressing RAB-D1 [N121I] alone (D1 NI) compared with the F1 progeny of a cross to lines expressing wild-type RAB-D1 or RAB-D2a from the same inducible promoter system. (C) Quantitative analysis of rosette diameter in 18-day-old plants expressing RAB-D1 [N121I] only (D1 NI) in the presence (+) or absence (–) of dexamethasone (dark grey and white bars, respectively) or in the F1 progeny of crosses to lines expressing wild-type RAB-D1 (black bars) or RAB-D2a (pale grey bars). Each black and pale grey bar represents F1 progeny from an independent cross. Sets of data whose means are not significantly different (P<0.001) from D1 NI in either the absence or presence of dexamethasone are marked by single and double asterisks respectively. (D) RT-PCR analysis of transcript accumulation from the RAB-D1 wild-type and N121I mutant transgenes in the parental N121I line (NI) and the F1 progeny from crosses between lines carrying each transgene (NI×wt) in the presence (+) or absence (–) of dexamethasone (Dex). The upper panel shows total RAB-D1 transcript accumulation. The second panel shows RT-PCR products digested with HindIII to distinguish transcripts from the transgenes expressing the N121I and wild-type sequences; the upper band is characteristic for the mutant sequence and shows that the mutant is transcribed in the rescued plants that exhibit normal growth rates. The third panel shows that transcripts from the endogenous (Endo.) RAB-D1 locus are unaffected by expression of the transgenes. The lower panel shows transcripts for GAPDH, which served as a loading control. RNA was extracted after 18 days of growth.