1. Volume 7, Issue 1, Pages 218-230 (January 2014)
Cloning and Characterization of miRNAs and Their Targets, Including a Novel miRNA- Targeted NBS–LRR Protein Class Gene in Apple (Golden Delicious) 
Chao Ma, You Lu, Songlin Bai, Wennan Zhang, Xuwei Duan, Dong Meng, Zhigang Wang, Aide Wang, Zongshan Zhou, Tianzhong Li 
Molecular Plant 
Volume 7, Issue 1, Pages (January 2014)
DOI: /mp/sst101
Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

2. Figure 1
Complexity of the Known miRNA Families Identified in Malus × Domestica.
The horizontal axis indicates the Md-miRNA family, and the vertical axis indicates the number of members of those families. There are nine family members in Md-miR156 which is the largest miRNA family, but the Md-miR319, Md-miR397, Md-miR408, Md-miR482, Md-miR530, and Md-miR846 had only one family member.
Molecular Plant 2014 7, DOI: ( /mp/sst101)
Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

3. Figure 2 The Genomic Location of Malus × Domestica miRNA.
The cloned miRNA was located in the apple genome; the target numbers are marked with parentheses. miRLn11 is underlined in red.
Molecular Plant 2014 7, DOI: ( /mp/sst101)
Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

4. Figure 3
The Real-Time PCR Results of 41 Mature miRNA in Five Different Apple Tissues.
The miRNAs showed different abundance in leaf, flower, phloem, peel, and sarcocarp. miRNAs had low expression level in the leaves and flower tissues; many kinds of miRNA were weakly detected, but many miRNAs were highly expressed in the fruit tissue; the difference in miRNA expression level was relatively small in phloem, with most miRNAs expressed moderately. Three technical replicates were preformed simultaneously for each transgenic plant line. miRNA expression levels are expressed relative to the (constitutive and constant) level of the 5s rRNA.
Molecular Plant 2014 7, DOI: ( /mp/sst101)
Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

5. Figure 4 Experimental Validation of Predicted Targets of miRNAs.
The target gene 5′ RACE-PCRs were performed with gene-specific primers and the resulting PCR products were sequenced. The contig numbers are listed at the upper left corner of each figure. The top numbers give the site of the miRNA in that contig. The arrows indicate the positions of inferred cleavage sites and numbers above indicate the detected cleavage site of independent clones.
Molecular Plant 2014 7, DOI: ( /mp/sst101)
Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

6. Figure 5
The Different Abundance of Md-miRLn11 and Md-NBS in Different Resistance Apple Varieties.
For the disease index standard, see Supplemental Table 1. HR-1 to HR-10, 10 highly resistant apple varieties; R-1 to R-10, 10 resistant apple varieties; S, 10 susceptible apple varieties; HS, 10 highly susceptible apple varieties.
Molecular Plant 2014 7, DOI: ( /mp/sst101)
Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

7. Figure 6 Northern Blot Analysis of the Infiltrated Plant.
(A) Expression analysis of the Md-miRLn11 and Md-NBS gene in non-infiltrated wild-type plant, SC (scratch control) plant, GFP gene-infiltrated, and Md-miRLn11-infiltrated plant (JiGuan apple). The Actin gene probe served as a control.
(B) Expression analysis of the Md-miRLn11 and Md-NBS gene in non-infiltrated wild-type plant, SC plant, GFP gene-infiltrated, and Md-NBS gene-infiltrated plant (FuJi apple). The 5.8s rRNA probe served as a control.
Molecular Plant 2014 7, DOI: ( /mp/sst101)
Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

8. Figure 7
The Gene Expression and the Leaf Phenotypes after Infiltration.
(A) The infiltration of Md-miRLn11 in resistance (JiGuan) apple. The WT1–WT3 indicate the non-infiltrated JiGuan apple plant; the SC1–SC3 indicate three scratch controls; the GFP1–GFP3 indicate the GFP-gene-infiltrated plant; miRLn11 Lv-0–1 to miRLn11 Lv-7–3 indicate the different incidence rates of Md-miRLn11-infiltrated plants.
(B) The infiltration of Md-NBS gene in susceptible (FuJi) apple. The WT1–WT3 indicate the non-infiltrated FuJi apple plant; the SC1–SC3 indicate three scratch controls; the GFP1–GFP3 indicate the GFP gene-infiltrated plant; NBS Lv-0–1 to NBS Lv-7–3 indicate the different incidence rates of Md-NBS gene-infiltrated plants.
Molecular Plant 2014 7, DOI: ( /mp/sst101)
Copyright © 2013 The Authors. All rights reserved. Terms and Conditions