Fig. 1 | Communications Biology

Fig. 1

From: CRISPR/Cas9 editing of endogenous banana streak virus in the B genome of Musa spp. overcomes a major challenge in banana breeding

Schematic representation of banana streak virus (BSV) strain Obino l’Ewai (BSOLV) and molecular analysis of Gonja Manjaya plants. a Episomal BSOLV showing three open-reading frames (ORF) and gene annotation in ORF3 (GenBank accession KJ013506). The colors purple, light blue, brown, and black indicate ORF1, ORF2, ORF3, and the intragenic region of BSOLV, respectively; P1–P4 show the position of primer pairs; S1–S3 indicate target sites for gRNA1, gRNA2, and gRNA3, respectively. b Endogenous BSV strain Obino l’Ewai (eBSOLV) integrated in banana genome (adapted from ref. 5). The duplication of BSV sequences, either complete or partial in the same or opposite direction, within the integrated eBSOLV are shown in the diagram. Green indicates banana genome; SBP indicates position of probe used for Southern hybridization; P1–P4 represent PCR primer pair locations. c PCR amplification to confirm the presence of BSOLV and/or eBSOLV in Gonja Manjaya plantlets. 1‒4, in vitro mother plantlets of Gonja Manjaya; PCR products using P1–P3 primers are labeled on the top of the gel picture; M molecular marker. d PCR analysis to confirm the absence of episomal BSOLV in in vitro mother plantlets of Gonja Manjaya used for genome editing. PC1 and PC2, symptomatic field plants of plantain Agbagba; PC3, symptomatic Gonja Manjaya plant from glasshouse under stress condition; CW, asymptomatic in vitro plantlet of Cavendish Williams as negative control; NTC, no template control. e Southern blot hybridization to confirm the presence of eBSOLV in Gonja Manjaya plantlets. Genomic DNA for Southern blot was digested with HindIII. The same four individual Gonja Manjaya plants (1‒4) were used for PCR analyses and Southern hybridization

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