Figure 1. Sulfate deficiency induces accumulation of OsamiR395 in rice.

(a) Small RNA northern blotting analysis of mature OsamiR395 under different sulfate concentrations. Total RNA samples were prepared from leaf and root tissues of two weeks old rice grown in N6 medium with 0, 20, 1500 or 2000 μM (NH⁴⁺)₂SO₄ and used for small RNA northern blotting analysis. Antisense oligonucleotides of OsamiR395 was labeled with γ-[³²P]ATP and used as probe to detect the transcript level of mature OsamiR395. rRNA was used as a loading control. (b) Stem-loop real-time PCR analysis of mature OsamiR395 under different sulfate concentrations. Total RNA samples were prepared as in (a) and used for stem-loop real-time PCR analysis. OsaSIZ1 was used as a reference gene. Data are presented as means of three technique replicates, error bars represent SD (n = 3). (c) Real-time PCR analysis of rice pri-OsamiR395h under different sulfate concentrations. Total RNA samples were prepared as in (a) and used for real-time PCR analysis. OsaSIZ1 was used as a reference gene. Data are presented as means of three technique replicates, error bars represent SD (n = 3). The statistically significant difference between groups was determined by one-way ANOVA (F(df_between, df_within) = F ration, p = p-value, where df = degrees of freedom). Means not sharing the same letter are statistically significantly different (P < 0.05).