Supplementary Figures
Figure S1. Proportion of small RNAs generated from different
categories of genomic loci in B_V0 and B_V7 small RNA libraries inBrassica rapa .
Figure S2. Cis-element analysis of bra-MIR1885 promoter.
G-box, GA-motif, P-box, TC, TCT, AAAC, ATCT, Box 4, I-box and LTR
represent cis -elements. A06: chrA06. 24219516 and 24221516:
Location information. Red or green bar represent position.
Figure S3. Analysis of the single nucleotide polymorphism (SNP)
density distribution, principal component analysis (PCA), and linkage
disequilibrium (LD) of B. napus population. (A ) SNP
density distribution (number of SNPs in 0.1 Mb sliding windows across
each chromosome). (B ) PCA analysis. Blue, green, and red points
represent winter (W), semi-winter (SW) and spring (S) ecotypes,
respectively. (C ) Genome-wide average LD decay estimated fromB. napus . (D ) LD heatmap of SNPs in bna-MIR1885promoter. Numbers indicate physical position where bna-MIR1885promoter locates. Deeper red color indicates stronger linkage
relationship. (E ) Comparison of the miR1885 relative expression
abundance between the two groups with extremely different phenotypes.
ES: Low-temperature sensitive accessions; ER: Low-temperature resistant
accessions. U6 was used as an internal control. Data are
presented as boxplot. Significant differences were determined by
Student’s t-test: * indicates p < 0.05 .
Figure S4. 5’-RACE assays of Bn. TIR.A09 (BnaA09g14980D)
and Bn. TNL.A03 (BnaA03g56180D). PCR products ofBn.A09.TIR and Bn.A03.TNL were separated in 2% agarose
gel. DNA ladders are labeled on the side. PCR products were cloned into
T vectors for Sanger sequencing. Red line represents target fragment;
black line represents inner primer.
Figure S5. Phenotypes of transgenic plants overexpressing
miR1885 after cold treatment. (A ) Phenotypes of transgenic
lines grown at 22 °C or 4 °C. Plants were grown 22 °C for 6 weeks (upper
panel), and then transferred to 4 °C for an additional 30 days (middle
panel), and then allowed to recover at 22 °C for 7 days. (B )
Post-germination phenotypes of miR1885-OE and WT after cold treatment.
Seeds were grown on the germination bed at 22 °C for 2 days and then
transferred to 6 °C for 7 days. (C ) Seeds were grown on the
germination bed at 22 °C for 7 days. (D ) Quantitative analysis
of net hypocotyl lengths from WT and MIR1885 -OE lines. n =
10-15 plants. Similar results were obtained in three independent
experiments.