RESUMEN
Common in Fungal Extracellular Membrane (CFEM) domain proteins are considered to be unique to fungi and closely related to pathogenicity. However, the Puccinia striiformis f. sp. tritici (Pst) effector containing the CFEM domain has not been reported. Here, we obtained an effector, PstCFEM1, containing a functional N-terminal signal peptide sequence and the CFEM domain from Pst race CYR31. qRT-PCR assay indicated that the transcript levels of PstCFEM1 were highly induced during the early stages of infection. Overexpression of PstCFEM1 suppressed Pst322 (an elicitor-like protein of Pst)-trigged cell death, reactive oxygen species (ROS) accumulation and callose deposition. Host-induced gene silencing (HIGS) experiments showed that knockdown of PstCFEM1 decreased the virulence of Pst, while ROS accumulation in silenced plants increased near the infection site. In addition, wheat containing the PstCFEM1-silenced construct increased resistance to multiple races of Pst. Our data suggest that PstCFEM1 suppresses wheat defense by inhibiting ROS accumulation and contributes to increased virulence of Pst.
RESUMEN
Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most destructive diseases of wheat and has been largely managed using demethylation inhibitor (DMI) fungicide triadimefon in China. To determine the sensitivity of Pst, a Chinese Pst isolate and its sexually produced progeny isolates were tested with triadimefon using the detached leaf method. The half maximal effective concentration (EC50) values varied greatly among the progeny isolates, ranging from 0.06 mg L-1 to 7.89 mg L-1. Twenty-six of the 56 tested progeny isolates were less sensitive to triadimefon than the parental isolate. A single-nucleotide mutation at the 401 position resulting in an amino acid change from tyrosine (Y) to phenylalanine (F) in the 134th codon (Y134F) of the cytochrome P450 sterol 14a-demethylase enzyme (CYP51), the target gene of DMI fungicide, was identified in the parental isolate. The 87 tested progeny isolates segregated into 19 homozygous wild type (AA), 40 heterozygous (AT), and 28 homozygous mutant (TT) genotypes, fitting a 1:2:1 ratio (χ2 = 2.43; P = 0.30). The mutant isolates had higher EC50 values than the wild type isolates. Significant differences in logEC50 were found between the mutant isolates and the wild type isolates (P = 2.2e-16). However, homozygous and heterozygous mutant isolates were not significantly different (P = 0.21), indicating dominant mutation. Twenty-two progeny isolates were used to inoculate a susceptible wheat variety, and latency period and lesion growth were recorded to compare wild type and mutant isolates for the pathogenicity fitness components. A moderate but significant negative correlation was detected between lesion growth and sensitivity to triadimefon (r = -0.53; P = 0.01). No significant variation in lesion growth was found between homozygous and heterozygous mutant isolates (P = 0.83). In the case of latency period and triadimefon sensitivity, no significant correlation was found (P = 0.17). These results are useful for understanding reduced sensitivity in the pathogen population and improving stripe rust management.
