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Figure 1. 

A heterograft model of A. annua scion and N. benthamiana rootstock. (a) The upper half of A.annua and bottom half of N. benthamiana were cut using a sharp knife. Through the apical wedge grafting technique, the two part were combined and co-grown for a month. (b) The density of gladular trichome in leaves and (c) artemisinin content were calculated as shown in the histogram, no less than three biological replicates were performed for each data.

Figure 2. 

DEGs and potential mobile mRNAs identified from the Aa/Nb heterograft model. The upper scion part belonged to A. annua, and the bottom rootstock part was N. benthamiana as separated with horizontal line. The left half of the information was the number of up-regulated and down-regulated of different expression genes related to comparing with the same plant. The right half of the photo exhibited the movement process of mRNAs and identification of non-homologous genes.

Figure 3. 

Volcano photo of differentially expressed miRNAs in scion and rootstock. Overall distribution of differentially expressed miRNA in (a) Aa scion and (b) Nb rootstock. The abscissa represents the differential expression multiple changes of miRNA in different samples. The ordinate represents the statistical significance of miRNA expression changes. Red dots represent significantly up-regulated differentially expressed genes, blue dots represent significantly down-regulated differentially expressed genes, and gray dots represent non-significantly differentially expressed genes.

Figure 4. 

Regulatory network research of transcription factors in differentially expression miRNAs and heatmap analysis of target gene among Aa scion and Nb rootstock. (a) Network plot of different expression miRNAs in A. annua scion. (b) Heatmap of different expression miRNAs targets in A. annua scion. (c) Network plot of different expression miRNAs in N. benthamiana rootstock. (d) Heatmap of different expression miRNAs targets in N. benthamiana rootstock. The legend at the bottom of the left figure marks the miRNA name, target gene name and gene type in different colors and shapes, respectively. Heatmap of target genes was used FPKM as expression quantity. Different colors indicate different gene expression levels, from blue to white to red, indicating low to high expression levels, with red indicating high expression genes and blue indicating low expression genes. The group of each gene corresponds to the color bar on the left.

Figure 5. 

Changes of genes involved in phytohormone signal transduction. (a) Phytohormone transduction pathway of A. annua scion. (b) Phytohormone transduction pathway of N. benthamiana rootstock. Changes in DEGs were mapped in boxes, green and red boxes represent down-regulated and up-regulated, respectively. Orange boxes represents genes in the pathway indicting bidirectional regulation. Purple boxes illustrate no DEG.

Figure 6. 

The content of measured ABA, JA and CK in A. annua scion and N. benthamiana rootstock. The abscissa represents the sampling site, and the ordinate represents the phytohormone concentration. The error bar represents the standard error for three independent experiments.