Additional file 2 of Screening of differentially expressed microRNAs and target genes in two potato varieties under nitrogen stress

Additional file 2: Figure 1. Analysis of variability among groups of differential miRNAsHN: excessive N application; LN: no N application; Y: Yanshu4; D: Atlantic;S: seedling stage; A: budding stage (A) LN_YLSvsLN_DLS (B) LN_YLAvsLN_DLA (C) HN_YLSvsHN_DLS. (D)HN_YLAvsHN_DLA (E) HN_DLSvsLN_DLS (F) HN_DLAvsLN_DLA. (G) HN_YLSvsLN_YLS (H)HN_YLAvsLN_YLA (I) LN_DLAvsLN_DLS. (J)HN_DLAvsHN_DLS (K)LN_YLAvsLN_YLS (L)HN_YLAvsHN_YLS.(M)LN_YRSvsLN_DRS (N)LN_YRAvsLN_DRA (O)HN_YRSvsHN_DRS. (P)HN_YRAvsHN_DRA (Q)HN_DRSvsLN_DRS(R)HN_DRAvsLN_DRA. (S)HN_YRSvsLN_YRS (T)HN_YRAvsLN_YRA (U)LN_DRAvsLN_DRS. (V)HN_DRAvsHN_DRS(W)LN_YRAvsLN_YRS (X)HN_YRAvsHN_YRS. Using the values of log2(fold change) as the horizontalcoordinate and -log10(p-value) as the vertical coordinate, volcanoplots were constructed for all miRNAs during differential expression analysis.The horizontal coordinate represents the fold change in the differentialexpression of miRNAs in different samples, and the vertical coordinaterepresents the statistical significance of the difference in the change inexpression levels of miRNAs. The red color represents up-regulatedsignificantly differentially expressed genes, the blue color representsdown-regulated significantly differentially expressed genes, and the grey dots representnon-significant differentially expressed genes. Figure 2. Comparison between groups of differential miRNAs. HN: excess N; LN:no N; Y: Yanshu4; D: Atlantic; S: seedling stage; A: budding stage. Thehorizontal coordinates indicate the data obtained after the comparison ofgroups, and the vertical coordinates indicate the number of up- anddown-regulated miRNAs. The red color represents up-regulated miRNAs, the bluecolor represents down-regulated miRNAs, and the numbers represent the number ofup- and down-regulated miRNAs. Figure 3. Results of clustering analysis ofdifferential miRNAs. HN: excess N; LN: no N; Y: Yanshu4; D: Atlantic; S: seedling;A: bud onset. (A)LN_YLSvsLN_DLS (B)LN_YLAvsLN_DLA (C)HN_YLSvsHN_DLS. (D)HN_YLAvsHN_DL.(E)HN_DLSvsLN_DLS (F)HN_DLAvsLN_DLA. (G)HN_YLSvsLN_YLS(H)HN_YLAvsLN_YLA (I)LN_DLAvsLN_DLS.(J)HN_DLAvsHN_DLS (K)LN_YLAvsLN_YLS (L)HN_YLAvsHN_YLS. (M)LN_YRSvsLN_DRS (N)LN_YRAvsLN_DRA(O)HN_YRSvsHN_DRS. (P)HN_YRAvsHN_DRA (Q)HN_DRSvsLN_DRS (R)HN_DRAvsLN_DRA. (S)HN_YRSvsLN_YR.(T)HN_YRAvsLN_YRA (U)LN_DRAvsLN_DRS. (V)HN_DRAvsHN_DRS (W)LN_YRAvsLN_YRS (X)HN_YRAvsHN_YRS

附加文件2：图1 差异微小RNA（miRNA）组间变异分析。HN：过量施氮；LN：不施氮；Y：烟薯4号；D：大西洋；S：苗期；A：现蕾期。(A) LN_YLSvsLN_DLS (B) LN_YLAvsLN_DLA (C) HN_YLSvsHN_DLS。(D) HN_YLAvsHN_DLA (E) HN_DLSvsLN_DLS (F) HN_DLAvsLN_DLA。(G) HN_YLSvsLN_YLS (H) HN_YLAvsLN_YLA (I) LN_DLAvsLN_DLS。(J) HN_DLAvsHN_DLS (K) LN_YLAvsLN_YLS (L) HN_YLAvsHN_YLS。(M) LN_YRSvsLN_DRS (N) LN_YRAvsLN_DRA (O) HN_YRSvsHN_DRS。(P) HN_YRAvsHN_DRA (Q) HN_DRSvsLN_DRS (R) HN_DRAvsLN_DRA。(S) HN_YRSvsLN_YRS (T) HN_YRAvsLN_YRA (U) LN_DRAvsLN_DRS。(V) HN_DRAvsHN_DRS (W) LN_YRAvsLN_YRS (X) HN_YRAvsHN_YRS。本研究以log₂(倍数变化)为横轴、-log₁₀(p值)为纵轴，对所有差异miRNA进行差异表达分析并构建火山图。其中，横轴代表不同样本中miRNA差异表达的倍数变化，纵轴代表miRNA表达水平变化差异的统计学显著性。红色圆点代表显著上调的差异表达miRNA，蓝色圆点代表显著下调的差异表达miRNA，灰色圆点代表无显著差异的miRNA。

图2 差异miRNA组间比较。HN：过量施氮；LN：不施氮；Y：烟薯4号；D：大西洋；S：苗期；A：现蕾期。横轴为组间比较所得数据，纵轴为上调与下调miRNA的数量。红色代表上调miRNA，蓝色代表下调miRNA，图中数字标注上调和下调miRNA的具体数目。

图3 差异miRNA聚类分析结果。HN：过量施氮；LN：不施氮；Y：烟薯4号；D：大西洋；S：苗期；A：现蕾期。(A) LN_YLSvsLN_DLS (B) LN_YLAvsLN_DLA (C) HN_YLSvsHN_DLS。(D) HN_YLAvsHN_DLA (E) HN_DLSvsLN_DLS (F) HN_DLAvsLN_DLA。(G) HN_YLSvsLN_YLS (H) HN_YLAvsLN_YLA (I) LN_DLAvsLN_DLS。(J) HN_DLAvsHN_DLS (K) LN_YLAvsLN_YLS (L) HN_YLAvsHN_YLS。(M) LN_YRSvsLN_DRS (N) LN_YRAvsLN_DRA (O) HN_YRSvsHN_DRS。(P) HN_YRAvsHN_DRA (Q) HN_DRSvsLN_DRS (R) HN_DRAvsLN_DRA。(S) HN_YRSvsLN_YRS (T) HN_YRAvsLN_YRA (U) LN_DRAvsLN_DRS。(V) HN_DRAvsHN_DRS (W) LN_YRAvsLN_YRS (X) HN_YRAvsHN_YRS