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Fig. 1 | Phytopathology Research

Fig. 1

From: Gene editing with an oxathiapiprolin resistance selection marker reveals that PuLLP, a loricrin-like protein, is required for oospore development in Pythium ultimum

Fig. 1

Transcript levels and CRISPR/Cas9-mediated gene knockout of PuLLP. a Transcript levels of PuLLP in the Py. ultimum wild type and PuM90 knockout mutant strains detected using RNA-seq (upper) and qRT-PCR (bottom) when mycelia were inoculated in V8 liquid medium for 12, 24, 36, 48, or 96 h. b Schematic diagram of homology-directed repair (HDR)-mediated modification of the PuLLP gene. Top: an ‘all-in-one’ plasmid (pYF515) harboring both Cas9 and sgRNA cassettes was co-transformed with a plasmid (pBS-SK II +) containing homologous donor DNA, hph with PuLLP flanking sequences. Locations of the primers used to screen the HDR mutants and Sanger sequencing traces of junction regions confirming that the PuLLP ORF was precisely replaced are shown. c Analysis of genomic DNA from the wild-type (WT), an empty-vector control line (EV), and PuLLP knockout mutants (∆PuLLP) using the primers shown above and actin primers as a positive control

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Phytopathology Research

ISSN: 2524-4167