Fig. 6: A cis-NAT is induced by Pi starvation to regulate the alternative polyadenylation of SlSPX5.
a Quantitative PCR analysis of SlSPX5 full-length transcript and anti-sense transcript NATSPX5 in wild type (WT) grown under Pi-replete (P+) and –depleted (P-) conditions. After reverse transcription with gene specific primers, RT-qPCR was used to analyze the gene expression of SlSPX5 (left) and NATSPX5 (right). b Information on natspx5 mutants. CRISPR vectors containing two sgRNAs were constructed, with the upper panel showing the position of three sgRNAs designed for the first intron of SlSPX5, and the lower panel showing the two mutant forms of NATSPX5. c RT-qPCR analysis of SlSPX5 and NATSPX5 gene expression in WT and the two natspx5 mutants. d 3′RACE assay was used to analyze the abundance of SlSPX5 transcripts in WT, natspx5-1 and natspx5-2 under P+ and P- conditions. The gel image shows the RACE PCR products. e RT-qPCR analysis of PSI gene expression in WT and the two natspx5 mutants. In (a, c, e), data are presented as mean ± SD, with n = 3 biologically independent replicates. Statisticallly significant differences between WT and mutants were calculated using a two-sided unpaired Student’s t-test and are indicated by **(P < 0.01) and ***(P < 0.001). f Pi content quantifications in WT and the two natspx5 mutants under P+ and P- conditions. Data are represented as mean ± SD, with n = 6 biologically independent replicates. Statistical significance was assessed using a two-sided unpaired Student’s t-test. ns not significant. Source data are provided as a Source Data file.
