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Fig. 2 | BMC Plant Biology

Fig. 2

From: Reduced expression of AtNUP62 nucleoporin gene affects auxin response in Arabidopsis

Fig. 2

Analysis of the atnup62 mutant. a Upper panel: Structure of the AtNUP62 gene and position of the T-DNA insertion in the SALK_037337 (atnup62-1) mutant. Exons are represented by rectangles (9 exons) and introns by black lines. 5‘ and 3‘-UTR are thick red lines. The protein is composed of 739 amino acids. The ORF regions encoding the FG repeat, serine-rich domain (approximately 480 amino acids) and nsp1-C-like domain (amino acids 565 to 647) are figured in pale green and yellow, respectively. The T-DNA insertion site is indicated (LB and RB: left and right borders) with the sequence of the 15 bases just upstream of the insertion site, coming from our own sequencing of the junction region. The two pairs of primers (a and b) used for PCR are indicated. Lower panel: RT-PCR amplification of RNAs prepared from inflorescences of wild-type Col-0 (Col) and atnup62-1 mutant (nup). Left lanes (amplification of cDNAs): after reverse transcription, PCR amplification was carried out using two couples of primers, a or b, for AtNUP62, or a couple of primers targeting the EF1-α gene. Right lanes (labelled Genomic DNA): amplifications of wild-type and mutant genomic DNA with primers B. Ladder: 1 kb plus from Promega, arrow at 1 kb. All amplified DNA fragments are present in the gel at a position consistent with in silico prediction. b Q-PCR analysis of AtNUP62 expression in different genotypes. Results are expressed as fold changes compared to wild type. Values are means and standard errors of two biological samples (three technical replicates per sample). c Phenotype of atnup62-1 mutant and complemented plants, at the adult stage (upper panels, zooms on flowers on top right) and on in vitro germinations on MS/2 medium (lower panels, 6 day-old seedlings). Lower panels, from left to right: wild type, plantlets with 3 cotyledons and 1 cotyledon from the atnup62 mutant, plantlet with abnormal cotyledons from the 35S::AtNUP62-GFP line, close to a plant (on the right) displaying a normal appearance. Arrows indicate abnormal plantlets. Cotyledons of complemented plants are similar to those of wild-type plants

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