Plant Breeding and Biotechnology

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Fig. 3.

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Fig. 3. Effect of heat stress (HS) on PVX-mediated CRISPR/Cas9 targeted mutagenesis of the NbPDS gene. (A) Analysis of leaf samples infiltrated with pPVX-Cas9::NbPDS-sgRNA1 for the presence of targeted modification using BslI recognition site loss assay. Expected bands for WT/mock: 237 bp and 670 bp; Expected bands for mutated: 237 bp, 670 bp and 903 bp. (B) Analysis of leaf samples infiltrated with pPVX-Cas9::NbPDS-sgRNA2 for the presence of targeted modification using StyI recognition site loss assay. Expected bands for WT/mock: 110 bp, 272 bp, and 521 bp; Expected bands for mutated: 110 bp, 272 bp, 382 bp, and 521 bp. (C) GE efficiencies of NbPDS-sgRNA1 and NbPDS-sgRNA2 constructs infiltrated samples. Infiltrated leaves were collected from plants treated with HS at 2, 3, 4, and 5 days after infiltration (DAI). Different letters indicate significant differences between HS treat--ments according to Duncan's multiple range test (P ≤ 0.05).
Plant Breed. Biotech. 2022;10:186-96 https://doi.org/10.9787/PBB.2022.10.3.186
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