DATA AVAILABILITY: All data is available in the manuscript and Electronic Supplementary Material. The MaYMV RSA BR1A and MaYMV RSA SCM genome sequences have been deposited in Genbank (Accessions MG570476; MN943641, respectively). RNA-seq data has been deposited at the NCBI GEO (Gene Expression Omnibus) repository (Accessions GSE94442, GSE99005).
Online Resource 1. Bioinformatics pipeline based on de novo assembly of
unmapped reads used for discovery of maize yellow mosaic virus in maize RNA‐seq data. (a) Raw reads were assessed with FastQC, trimmed with
Trimmomatic based on the FastQC results, and aligned to the reference genomes of maize and C. zeina to collect unmapped reads. (b)Unmapped reads were assembled using Trinity, their protein sequences predicted
with TransDecoder and compared against the NCBI nr database using
BLASTP.
Online Resource 2. Oligonucleotide primers used to amplify and Sanger sequence the complete genomes of
Maize yellow mosaic virus isolates RSA BR1A and RSA SCM.
Online Resource 3. Percent nucleotide identity for complete genomes between maize yellow mosaic virus from South Africa and other worldwide isolates.
Online Resource 4. Comparison of predicted maize yellow mosaic virus proteins of RSA BR1A and RSA SCM from South Africa and MaYMV Yunnan 11 from China.
Online Resource 5. RNA-dependent RNA polymerase-based phylogenetic analysis of maize yellow mosaic virus and related Poleroviruses. The
evolutionary history was inferred from the RNA-dependent RNA polymerasenucleotide sequences by using the Maximum Likelihood method with the
GTRGAMMA model. Bootstrap consensus values are shown at the nodes. The sequences were extracted from complete genome sequences with the
following NCBI accession numbers: RSA BR1A (MG570476), RSA SCM (MN943641), Kenya KALRO (MH205607), Kenya MYDV-like (MF974579),
Tanzania 76 (MG664790.1), Ethiopia (MF684369), Nigeria (KY684356.1), China Y11 (KU248489.1), China Y1 (KU179221.1), China MYDV-RMV2
(KT992824.1), China SC (MK652149), Brazil (KY940544.1), Ecuador (KY052793), BVG Gimje (KT962089.1) and MYDV-RMV (KC921392.1). The latter
two were used as outgroups. The scale bar indicates the number of nucleotide substitutions per site.
Online Resource 6. Confirmation by Sanger sequencing that the expected maize yellow mosaic virus
RT-PCR product was amplified from maize inbred B73 sample BR1B. Sequences derived from sequencing
the 753bp RT-PCR product with the MaYMV-F or MaYMV-R primers were named BR1B.MaYMV.F or
BR1B.MaYMV.R, respectively. These sequences were aligned to the corresponding sequence from the
assembled MaYMV RSA BR1A genome sequence determined by 5’RACE, 3’RACE, RT-PCR and Sanger
sequencing [BR1A (MG570476)], and the reference sequence MaYMV Yunnan 11 from China [KU248489].
The MaYMV-F and MaYMV-R primer sequences are reported in Chen et al. (2016). Non-consensus sites
are shown by an asterisk.
Online Resource 7. Representative maize leaf symptoms of samples that were positive for maize yellow
mosaic virus with the RT‐PCR assay using MaYMV‐F and MaYMV‐R primers. A, B: Mosaic symptoms;
C‐D: Yellow streaks; E‐H: Narrow yellow streaks. A‐H samples: 17‐4263, 17‐4267, 17‐4245, 17‐4135,
17‐4275, 17‐4261, 17‐4284, 17‐4172, respectively. Maizegenotypes are not known.
Online Resource 8. Actin RT-PCR to confirm RNA and cDNA integrity for samples that were RT-PCR negative for
maize yellow mosaic virus. RT-PCR products amplified using the primer pair actinF and actinR were visualized by
agarose gel electrophoresis, with an expected 169 bp cDNA actin product. Lane M, O’GeneRuler 100 bp DNA Ladder
(Thermo-Fischer, Waltham, USA); lane 1, RT no template control; lane 2, PCR no template control; lane 3, maize
healthy control/RNA positive control; lane 4, 16-3308; lane 5, 16-3328; lane 6, 16-3224; lane 7, 16-3252; lane 8,
16-3256. The additional 277 bp product in lanes 3-8 is the actin gDNA product, since the primers flank an intron,
indicating presence of some gDNA in the samples.
Online Resource 9. Maize B73 reads corresponding to RNA viruses.