Efficient disruption and replacement of an effector gene in the oomycetePhytophthora sojaeusingCRISPR/Cas9

• DOI: 10.1111/mpp.12318
• Publication Date: 2015-09-10T08:09:09Z
• Authors: Yufeng Fang; Brett M. Tyler
• Keywords: Oomycete; Phytophthora sojae

Phytophthora sojae is an oomycete pathogen of soybean. As a result its economic importance, P. has become model for the study genetics, physiology and pathology. The lack efficient techniques targeted mutagenesis gene replacement have long hampered genetic studies pathogenicity in species. Here, we describe CRISPR/Cas9 system enabling rapid genome editing sojae. Using RXLR effector Avr4/6 as target, observed that, absence homologous template, repair Cas9-induced DNA double-strand breaks (DSBs) was mediated by non-homologous end-joining (NHEJ), primarily resulting short indels. Most mutants were homozygous, presumably conversion triggered Cas9-mediated cleavage non-mutant alleles. When donor present, homology-directed (HDR) observed, which resulted with NPT II gene. By testing specific virulence several NHEJ HDR-mediated replacements soybean, validated contribution to recognition soybean R loci, Rps4 Rps6, but also uncovered additional contributions resistance these two loci. Our results establish powerful tool functional genomics Phytophthora, provides new avenues better control this pathogen.