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. 2020 Nov 30;11:585561. doi: 10.3389/fpls.2020.585561

FIGURE 4.

FIGURE 4

Tissue preferential RNA yield, RNase activity, and RNA integrity assessment of 21 days after germination rice grown under phosphate-sufficient (0.32 mM) or phosphate-starvation (0 mM) conditions. RNA yield of the samples from shoots and roots under phosphate-sufficient or -deficient conditions for 21 days using NanoDrop 2000 (A). Four biological replicates were prepared and analyzed independently. Statistical significance was analyzed using Student’s t-test (∗∗∗P < 0.001; ∗∗P < 0.01). RNase activity was analyzed by agarose gel electrophoresis: 1 μg of total RNA from 21-day-old rice shoot tissues was incubated with 0.5 μg (left) and 0.1 μg (right) of protein from crude extracts from shoot and root samples grown in phosphate-sufficient or -deficient conditions for 21 days. The incubated total RNAs were equally measured (1 μg) and loaded (B). Although we conducted experiments using three biological replicates, we presented only one dataset. Electropherograms of RNA samples from shoots (C,D) and roots (E,F) grown under phosphate-sufficient (C,E) or -deficient (D,F) conditions for 21 days. A standard size ladder in nucleotides (nt) is shown in the X-axis at the bottom of each graph. FU, fluorescent units at the Y-axis. The electrophoresis column generated by the program is located on the right side of each sample graph. The purple arrow indicates the peak of small RNAs. The positions of the 18S and 25S ribosomal RNAs are indicated by a green and red arrow, respectively. Compared with the root RNAs, the leaf RNAs exhibited additional fragments corresponding to the 23S and 16S chloroplastic rRNA fragments, respectively. The positions of the 16S and 23S ribosomal RNAs are indicated by a yellow and blue arrow, respectively.