Virulence of Pyrenophora tritici-repentis: a minireview

Pyrenophora tritici-repentis is a major wheat pathogen in all wheat (Triticum spp.) growing areas worldwide. Up to date, eight P. tritici-repentis races have been described based on chlorosis, necrosis, or both symptoms caused on race differential wheat genotypes: ‘Glenlea’, 6B662, 6B365, and ‘Salamouni’. Symptom development on differential genotypes depends on the interaction of the pathogen’s necrotrophic effectors named Ptr ToxA, Ptr ToxB, and Ptr ToxC with host susceptibility genes. Ptr ToxA is encoded by the single copy gene ToxA and induces necrosis on sensitive wheat cultivars. Ptr ToxB causes chlorosis and is encoded by the multicopy gene ToxB. The Ptr ToxC is the non-proteinaceous, polar, low molecular mass molecule that also induces chlorosis, but up to date, the gene encoding this toxin is unknown. Races producing Ptr ToxA are predominant in the global Ptr population. There are several reports about new putative races of P. tritici-repentis that do not conform with the current race system, so further research is required. This study aims to collect and systematise available information about the virulence and races of P. tritici-repentis.

Pyrenophora tritici-repentis population structure in the Republic of Kazakhstan and identification of wheat germplasm resistant to tan spot

Pyrenophora tritici-repentis is a causative agent of tan spot in wheat. In recent years, there has been an increasing spread and harmfulness of wheat tan spot. The aim of the research was to study the racial composition of the P. tritici-repentis population in the Republic of Kazakhstan. A collection of 30 common wheat accessions, including promising lines and cultivars from Kazakhstan and CIMMYT–ICARDA, was assessed for resistance to P. triticirepentis in a greenhouse and characterized using the Xfcp623 molecular marker, diagnostic for the Tsn1 gene. Monosporic isolates of P. tritici-repentis isolated from the southeastern region were assigned to certain races based on the manifestation of symptoms of necrosis/chlorosis on standard differentials (Glenlea, 6B662, 6B365). Five races of P. tritici-repentis have been identified, including races 1, 2, 3, 7 and 8. It has been shown that races 1 and 8 of P. tritici-repentis are dominant. As a result of the analysis of the frequency of occurrence of the P. tritici-repentis races, it was found that race 1 (50 %) producing Ptr ToxA and Ptr ToxB and race 8 (35 %) producing Ptr ToxA, Ptr ToxB and Ptr ToxC turned out to be dominant. From a practical point of view, of greatest interest are 16 wheat samples, which demonstrated resistance to race 1 and confirmed insensitivity to Ptr ToxA in a molecular screening. These include eight Kazakhstani (4_PSI, 10204_2_KSI, 10204_3_KSI, 10205_2_KSI, 10205_3_KSI, 605_SP2, 632_SP2, Dana) and seven foreign lines (KR11-20, KR11-03, KR11-9014, 11KR-13, KR12-9025, KR12-07, GN-68/2003). The results of this study are of interest in wheat breeding programs for tan spot resistance.

PacBio genome sequencing reveals new insights into the genomic organisation of the multi-copy ToxB gene of the wheat fungal pathogen Pyrenophora tritici-repentis

Background Necrotrophic effector proteins secreted by fungal pathogens are important virulence factors that mediate the development of disease in wheat. Pyrenophora tritici-repentis (Ptr), the causal agent of wheat tan spot, has a race structure dependent on the combination of effectors. In Ptr, ToxA and ToxB are known proteinaceous effectors responsible for necrosis and chlorosis respectively. While Ptr ToxA is encoded by the single gene ToxA, ToxB has multiple loci in the Ptr genome, which is postulated to be directly related to the level of ToxB production and leaf chlorosis. Although previous analysis has indicated that the majority of the ToxB loci lie on a single chromosome, the exact number and chromosomal locations for all the ToxB loci have not been fully identified. Results In this study, we have sequenced the genome of a race 5 ToxB-producing isolate (DW5), using PacBio long read technology, and found that ToxB duplications are nested in the complex subtelomeric chromosomal regions. A total of ten identical ToxB gene copies were identified and based on flanking sequence identity, nine loci appeared associated with chromosome 10 and a single copy with chromosome 5. Chromosome 10 multiple ToxB gene loci were separated by large sequence regions between 31 and 66 kb within larger segmental duplications in an alternating pattern related to loci strand, and flanked by transposable elements. Conclusion This work provides for the first time the full accompaniment of ToxB loci and surrounding regions, and identifies the organization and distribution of ten ToxB loci to subtelomeric regions. To our knowledge, this is the first report of an interwoven strand-related duplication pattern event. This study further highlights the importance of resolving the highly complex distal chromosomal regions, that remain difficult to assemble, and can harbour important effectors and virulence factors.

IDENTIFICATION WHEAT GENOTYPES RESISTANT TO TAN SPOT PYRENOPHORA TRITICI-REPENTIS

Intensified wheat production, changes in cultural practices including shifts from conventional tillage to reduced tillage practices, and wheat monoculture involving cultivation of susceptible cultivars has resulted in development of tan spot to epidemic proportions in Kazakhstan. Pyrenophora tritici-repentis, causal agent of tan spot on wheat. In recent years, there has been increasing distribution and harmfulness of P. tritici-repentis on wheat. The aim of the study was to identify and select wheat germplasm resistant to tan spot P. tritici-repentis using molecular markers. The results of field evaluation showed resistant reaction to tan spot in 76 wheat varieties (68%). Molecular screening of wheat germplasm was carried out based on the reaction to the fungal inoculum and to host-specific toxins (HST) produced by the P. tritici-repentis. The wheat germplasm insensitive to the toxins HST Ptr ToxA and Ptr ToxB was selected. As a result of molecular screening of 111 wheat genotypes using SSR marker Xfcp623 linked to insensitivity gene to the selective toxin Ptr ToxA of tan spot, 31 carriers of effective tsn1 gene were identified, which accounted for 27,9% of the genotypes studied. Ten samples of wheat (Jubileynaya 60, TOO11/TOOOO7, F3.71/TRM/VORONA/3/OC14, NANJTNG 82149 KAUZ, ECHA/LI115, Akmola 2, Kazakh-stanskaya rannespelaya, Kazakhstanskaya 25, 428g/MK-122A and 190-Naz/GF55) are characterized with complex resistance to the races Ptr 1 and 5, as well as to 2 toxins (ToxA and ToxB) and to the Septoria nodorum blotch isolate SNB7k. 20 promising wheat lines resistant to tan spot were selected. These genotypes also showed a moderate and high level of field resistance and recommended to use in the breeding programs for resistance to tan spot.

Phytopathological screening and molecular marker analysis of wheat germplasm from Kazakhstan and CIMMYT for resistance to tan spot

Tan spot caused by the fungus Pyrenophora tritici-repentis is an important leaf spot disease in wheat growing areas throughout the world. The study aims to identify wheat germplasm resistant to tan spot based on phytopathological screening and molecular marker analysis. A collection of 64 common wheat germplasms, including cultivars and breeding lines from Kazakhstan and CIMMYT, was assessed for tan spot resistance in greenhouse conditions and characterized using the Xfcp623 molecular marker, diagnostic for the Tsn1 gene. All wheat cultivars/lines varied in their reaction to tan spot isolate race 1, ranging from susceptible to resistant. Most accessions studied (53 %) were susceptible to Ptr race 1. Spring wheat cultivars were more susceptible to race 1 than winter wheat cultivars. As a result of genotyping, an insensitive reaction to Ptr ToxA was predicted in 41 wheat cultivars (64 %). The tsn1 gene carriers identified included 27 Kazakhstani and 14 CIMMYT cultivars/lines, demonstrating insensitivity to Ptr ToxA. The majority of the Tsn1 genotype were sensitive to race 1 and showed susceptibility to the pathogen in the field. Disease scores from seedling stage positively correlated with field disease ratings. Of particular interest are 27 wheat accessions that demonstrated resistance to spore inoculation by Ptr race 1, were characterized by insensitivity to ToxA and showed field resistance to the pathogen. The results of this study will contribute to wheat breeding programs for tan spot resistance with Marker Assisted Selection using the closely flanking markers.

Identifcation of genotypescarriers of resistance to tan spot Ptr ToxA and Ptr ToxB of Pyrenophora tritici-repentis in common wheat collection

Pyrenophora tritici-repentis(Ptr) is the causative agent of tan spot, one of the yield limiting diseases of wheat, rapidly increasing in wheat growing countries including Kazakhstan. The aim of this study was the identifcation of wheat genotypes with resistance to Ptr race 1 and race 5 and their host­selective effectors (toxins) Ptr ToxA and Ptr ToxB. A common wheat collection of 41 accessions (38 experimental and 3 controls) was characterized using the molecular markersXfcp623andXBE444541, diagnostic for theTsn1andTsc2genes conferring sensitivity to fungal toxins. The coincidence of the markerXBE444541with resistance to race 5 was 92.11 %, and with Ptr ToxB, 97.37 %. Genotyping results using the markerXfcp623confrmed the expected response to Ptr ToxA; the presence/absence of the markerXfcp623completely (100 %) coincided with sensitivity/resistance to race 1 and Ptr ToxA. This demonstrates the reliability of the diagnostic markerXfcp623for identifying wheat genotypes with resistance to the fungus and insensitivity to Ptr ToxA. The study of the reaction of wheat germplasm to the fungal inoculation and toxin infltration showed that out of 38 genotypes analyzed 30 (78 %) exhibited resistance to both race 1 and race 5, and insensitivity to toxins Ptr ToxA and ToxB. Of most signifcant interest are eight wheat genotypes that showed resistance/insensitivity both to the two races and two toxins. The results of phenotyping were reconfrmed by the molecular markers used in this study. Sensitivity to Ptr ToxB is not always correlated with susceptibility to race 5 and is dependent on the host’s genetic background of the wheat genotype, i. e. on a specifc wheat genotype. The results of the study are of interest for increasing the efciency of breeding based on the elimination of the genotypes with the dominant allelesTsn1andTsc2sensitive to the toxins Ptr ToxA and ToxB. The genotypes identifed will be used in wheat breeding for resistance to tan spot.