Pathotype-specific Resistance Research Articles

Characterization of a virulence factor in Plasmodiophora brassicae, with molecular markers for identification.

Symptom severity on differential host lines is currently used to characterize and identify pathotypes of Plasmodiophora brassicae, which is an obligate, soil-borne chromist pathogen that causes clubroot disease on canola (Brassica napus) and other brassica crops. This process is slow, variable and resource intensive; development of molecular markers could make identification of important pathotypes faster and more consistent for deployment of cultivars with pathotype-specific resistance. In the current study, a variant of gene 9171 was identified in the whole-genome sequences of only the highly virulent pathotypes of P. brassicae from around the world, including the new cohort of virulent pathotypes in Canada; its presence was confirmed using three KASP marker pairs. The gene was not present in the initial cohort of pathotypes identified in Canada. The putative structure, domains, and gene ontogeny of the protein product of gene 9171 were assessed using on-line software resources. Structural analysis of the putative protein produced by gene 9171 indicated that it was localized in the cytosol, and likely involved in cellular processes and catalytic activity. Identification of gene 9171 represents a potentially useful step toward molecular identification of the pathotypes of P. brassicae.

Screening for pathotype-specific resistance to broad bean wilt virus 2 and cucumber mosaic virus in pepper (Capsicum annuum L.)

AbstractTwo aphid-transmitted RNA viruses, broad bean wilt virus 2 (BBWV2) and cucumber mosaic virus (CMV), are the most prevalent viruses in Korean pepper fields and cause chronic damage in pepper production. In this study, we employed a screening system for pathotype-specific resistance of pepper germplasm to BBWV2 and CMV by utilizing infectious cDNA clones of different pathotypes of the viruses (two BBWV2 strains and three CMV strains). We first examined pathogenic characteristics of the BBWV2 and CMV strains in various plant species and their phylogenetic positions in the virus population structures. We then screened 34 commercial pepper cultivars and seven accessions for resistance. While 21 pepper cultivars were resistant to CMV Fny strain, only two cultivars were resistant to CMV P1 strain. We also found only one cultivar partially resistant to BBWV2 RP1 strain. However, all tested commercial pepper cultivars were susceptible to the resistance-breaking CMV strain GTN (CMV-GTN) and BBWV2 severe strain PAP1 (BBWV2-PAP1), suggesting that breeding new cultivars resistant to these virus strains is necessary. Fortunately, we identified several pepper accessions that were resistant or partially resistant to CMV-GTN and one symptomless accession despite systemic infection with BBWV2-PAP1. These genetic resources will be useful in pepper breeding programs to deploy resistance to BBWV2 and CMV.

Leaf Rust Resistance Genes in Wheat Cultivars Registered in Russia and Their Influence on Adaptation Processes in Pathogen Populations

The main growing regions for winter wheat in the Russian Federation are the North Caucasian, Central Black Earth, and Central agroecological regions. Spring wheat crops dominate in the Urals, Volga region, and Western Siberia. Wheat leaf rust, caused by Puccinia triticina, is an important disease, impacting greatly on wheat production. In Russia, the disease was an annual problem until 2010 but has since been more effectively controlled. However, changes in virulence in pathogen populations may arise from climate change, evolving cropping practices, intense use of chemical protectants, and an increase in the release of resistant cultivars. In the 2000s, the State Register of the Russian Federation included an increase in the number of winter and spring wheat cultivars resistant to leaf rust. However, successful genetic protection requires a diversity of cultivars with different resistance genes (Lr genes). Studies by the All Russian Institute of Plant Protection identified Lr genes in Russian cultivars’ phenotypes and molecular markers. In addition, the prevalence of virulence in pathogen populations was studied and the influence of the cultivar used in wheat production on the changes in these populations was evaluated. This paper reviews research on the genetic diversity of winter and spring wheat cultivars included in the State Register of Russia from 2000 to 2020 and analyzes their impact on the prevalence of virulence in pathogen populations. These data demonstrate the continuous evolution of P. triticina in response to wheat breeding efforts. Populations of the pathogen showed higher variability in regions where pathotype-specific resistance cultivars were commonly grown.

Screening ofBrassicagermplasm for resistance toPlasmodiophora brassicaepathotypes prevalent in Canada for broadening diversity in clubroot resistance

Hasan, M. J., Strelkov, S. E., Howard, R. J. and Rahman, H. 2012. Screening of Brassica germplasm for resistance to Plasmodiophora brassicae pathotypes prevalent in Canada for broadening diversity in clubroot resistance. Can. J. Plant Sci. 92: 501–515. Clubroot disease of crucifers, caused by Plasmodiophora brassicae, poses a threat to the Canadian canola industry, and the development of resistant cultivars is urgently needed. Germplasm resistant to local pathotype(s) is the prime requirement for breeding clubroot-resistant cultivars. The objective of this study was to identify Brassica germplasm possessing resistance to P. brassicae pathotypes prevalent in Alberta. Pathotype-specific resistance was identified in the diploid species Brassica rapa (AA) and B. oleracea (CC), and in the amphidiploid B. napus (AACC). Among B. rapa genotypes, turnip was the most resistant, followed by winter type and spring type oilseed rape. The rutabaga group of B. napus, on the other hand, was homogeneous for resistance to Canadian P. brassicae pathotypes. The diploid species B. nigra (BB) also showed pathotype-specific resistance. However, the two amphidiploids carrying the B. nigra genome, B. juncea (AABB) and B. carinata (BBCC) were completely susceptible to clubroot.

Screening of Brassica germplasm for resistance to Plasmodiophora brassicae pathotypes prevalent in Canada for broadening diversity in clubroot resistance

Hasan, M. J., Strelkov, S. E., Howard, R. J. and Rahman, H. 2012. Screening ofBrassicagermplasm for resistance toPlasmodiophora brassicae pathotypes prevalent in Canada for broadening diversity in clubroot resistance. Can. J. Plant Sci. 92: 501–515. Clubroot disease of crucifers, caused by Plasmodiophora brassicae, poses a threat to the Canadian canola industry, and the development of resistant cultivars is urgently needed. Germplasm resistant to local pathotype(s) is the prime requirement for breeding clubroot-resistant cultivars. The objective of this study was to identify Brassica germplasm possessing resistance to P. brassicae pathotypes prevalent in Alberta. Pathotype-specific resistance was identified in the diploid species Brassica rapa (AA) and B. oleracea (CC), and in the amphidiploid B. napus (AACC). Among B. rapa genotypes, turnip was the most resistant, followed by winter type and spring type oilseed rape. The rutabaga group of B. napus, on the other hand, was homogeneous for resistance to Can...

Construction of a genetic linkage map with SSR, AFLP and morphological markers to locate QTLs controlling pathotype-specific powdery mildew resistance in diploid roses

Disease resistance is a sought-after trait in plant breeding programmes. One strategy to make resistance more durable is to increase the number of resistance genes, thereby increasing the number of pathotypes withstood. One of the most important diseases on roses is powdery mildew (PM) (Podosphaera pannosa). Recent studies show that pathotypes of PM and different types of resistances in roses exist. The results of this study aim to contribute to PM resistance in roses by the development of pathotype-specific markers on a genetic map. A diploid rose population (90 genotypes) derived from a cross between Rosa wichurana and Rosa ‘Yesterday’ was used to construct a genetic linkage map encompassing 20 AFLP primer combinations, 43 SSR, and 2 morphological markers. By applying the F1 pseudo test cross population strategy, two parental linkage maps were constructed (parent ‘Yesterday’ 536 cM; parent R. wichurana 526 cM). Both parental maps consisted of seven linkage groups with an average length of 70 cM (Kosambi) corresponding to the seven haploid rose chromosomes. These new maps were used to identify QTLs controlling disease resistance. The offspring population was screened for resistance to two PM pathotypes, R–E and R–P. QTLs for controlling pathotype-specific disease resistance were mapped by applying Kruskal–Wallis rank-sum tests and simple interval mapping. With two pathotypes analysed, nine QTL loci were detected on linkage groups 2, 3, 5 and 6, explaining 15–73% of the phenotypic variance for pathotype-specific disease response. The genetic maps developed here will be useful for future rose breeding, pathotype-specific resistance research and development of a consensus map for roses.

Interaction of soybean and Phakopsora pachyrhizi , the cause of soybean rust.

Abstract Soybean rust, caused by Phakopsora pachyrhizi Syd., is a major disease limiting soybean [ Glycine max (L.) Merr.] production in many areas of the world. Yield losses of up to 80% have been reported in experimental plots in Taiwan. Although the disease is not always yield limiting, it has the potential under conducive conditions to cause major losses in most soybean-producing countries in the world. The threat of P. pachyrhizi to soybean production in the Western Hemisphere began in Paraguay in 2001, where the disease was found in a limited number of soybean fields in the Parana River basin bordering Brazil. Later, in 2004, P. pachyrhizi was found in the continental USA. There are approximately 150 species in 53 genera the family Fabaceae known to be hosts of P. pachyrhizi . Approximately 120 of the known hosts occur in North and Central America and could become important in the epidemiology of the disease, serving as a bridge or over-wintering host between soybean crops. The interaction between soybean genotypes and P. pachyrhizi isolates appears to follow gene-for-gene theory, with resistance genes controlling resistance to specific pathotypes. In addition to soybean, pathotype-specific resistance has also been found in a number of perennial Glycine species. Dominant genes controlling pathotype-specific resistance to soybean rust have been identified and mapped to five loci in the soybean genome, with multiple alleles at four loci. Many of the resistance genes have become ineffective over time in different geographic areas following their deployment into soybean production. The search for more durable soybean rust resistance will include research on non-host and/or engineered resistance.

PATHOTYPE-SPECIFIC POWDERY MILDEW RESISTANCE IN ROSES

Differences in resistance to two powdery mildew isolates (R-P and R-E) were evaluated on two rose genotypes, Rosa wichurana and 'Yesterday'. Pathotype-specific resistance reactions to the developing fungus were observed by microscopy. Both rose genotypes expressed a comparable reaction, i.e., cell reactions to mycelium development of R-P. On 'Yesterday', pathotype R-E stopped growing after germination, whereas R. wichurana showed cell reactions upon inoculation with R-E. Crosses between the genotypes 'Yesterday' and R. wichurana resulted in some offspring with transgressive segregation for powdery mildew resistance. Most of the F 1 hybrids showed partial resistance by cell reactions, while others segregated for the arrested development of the germ tube of R-E as seen in 'Yesterday'. These reactions suggest that pathotype-specific resistance, based on different resistance mechanisms, can be used for resistance pyramiding in rose breeding practice.

Clonal evaluation for fusiform rust disease resistance: effects of pathogen virulence and disease escape

We evaluated the precision of phenotypic classification for fusiform rust resistance of Pinus taeda L. in a clonally propagated population segregating for the pathotype-specific resistance gene Fr1. In all marker-defined Fr1/fr1 clones screened with low complexity or ambient inoculum, marker–trait cosegregation was complete with no exceptions. Uncommon exceptions (4 of 30) in which marker-defined Fr1/fr1 clones screened with high complexity inoculum were diseased were probably due to a low frequency of spores virulent to Fr1 resistance. Marker–trait cosegregation for fr1/fr1 clones was less reliable, as all ramets of a few clones (5 of 29, 3 of 25, and 4 of 16) remained disease-free with low complexity, high complexity, or ambient inoculum, respectively. We termed disease-free fr1/fr1 ramets “escapes”, since the genetics of the host–pathogen interaction predicted them to be diseased. For nonmarker-defined materials, we considered escapes to be disease-free ramets within clones that had at least one diseased ramet. Narrow-sense heritability estimates for escape rate were 29% and 23% for the low and high complexity inocula, respectively, suggesting that genetic variation in the host is an important component of this resistance mechanism.

Ascochyta blight of chickpea: infection and host resistance mechanisms

Ascochyta rabiei (teleomorph Didymella rabiei) is a directly penetrating, necrotrophic fungus that infects all aboveground parts of chickpea (Cicer arietinum). During spore germination and infection, germ tubes secrete a mucilaginous substance to facilitate attachment to the host surface, and the invading fungus produces cell-wall-lytic enzymes to penetrate the host. The pathogen produces several phytotoxins (solanapyrones A, B, and C, cytochalasin D, and a proteinaceous toxin) that seem to be responsible for necrosis and cell death. The pathogen can degrade antimicrobial compounds and suppress their production in chickpea. On the basis of aggressiveness, the population of A. rabiei can be classified into two broad pathotypes: pathotype I (less aggressive) and pathotype II (aggressive). Complete resistance to A. rabiei has not been found in chickpea; the resistance present in superior cultivars used in chickpea production is partial or incomplete. There is a high degree of variation in resistance among chickpea cultivars, and the resistance declines as the plant matures. The symptoms of infection and disease severity follow a quantitative continuum based on aggressiveness of the pathogen, genetic resistance present in the cultivar, and age of the plant. The well-established defense responses in chickpea are cross-linking of cell walls mediated by hydrogen peroxide, production of pathogenesis-related (PR) proteins (chitinase, β-1,3-glucanase, and thaumatin-like proteins), and accumulation of phytoalexins. However, expression of these induced defense responses does not correlate with pathotype-specific resistance, indicating that other constitutive or unknown components may be involved in providing resistance to aggressive pathotypes. Lack of information about the attribute that makes the pathogen aggressive, as well as inadequate knowledge of pathotype-specific defense mechanisms and the causes for decline in resistance, are major constraints in developing cultivars with durable resistance.

Genetic dissection of pathotype-specific resistance to ascochyta blight disease in chickpea (Cicer arietinum L.) using microsatellite markers.

Ascochyta blight is an economically important disease of chickpea caused by the fungus Ascochyta rabiei. The fungus shows considerable variation for pathogenicity in nature. However, studies on the genetics of pathotype-specific resistance are not available for this plant-pathosystem. The chickpea landrace ILC 3279 has resistance to pathotype I and II of the pathogen. In order to understand the inheritance of pathotype-specific resistance in this crop, both Mendelian and quantitative trait loci analyses were performed using a set of intraspecific, recombinant inbred lines derived from a cross between the susceptible accession ILC 1272 and the resistant ILC 3279, and microsatellite markers. We identified and mapped a major locus (ar1, mapped on linkage group 2), which confers resistance to pathotype I, and two independent recessive major loci (ar2a, mapped on linkage group 2 and ar2b, mapped on linkage group 4), with complementary gene action conferring resistance to pathotype II. Out of two pathotype II-specific resistance loci, one (ar2a) linked very closely with the pathotype I-specific resistance locus, indicating a clustering of resistance genes in that region of the chickpea genome.

Different classes of resistance to turnip mosaic virus in Brassica rapa

Pathotype-specific and broad-spectrum resistance to turnip mosaic virus (TuMV) have been identified in the diploid A genome brassica species Brassica rapa. The pathotype-specific resistance is effective against pathotype 1 isolates of TuMV, which are the most common in Europe. It is almost identical in its specificity to that of a mapped resistance gene (TuRB01) present in the A genome of the amphidiploid species Brassica napus. A mutant of a pathotype 1 isolate of TuMV (UK 1M) that is able to overcome TuRB01 also overcame the B. rapa resistance. This, combined with the fact that a single-nucleotide mutation in the cylindrical inclusion gene of TuMV that has been shown to induce a change from avirulence to virulence against TuRB01, had an identical effect on the B. rapa resistance, suggest that the two resistances are conditioned by the same gene. A second source of resistance in B. rapa prevented systemic spread of all TuMV isolates tested. A third source of resistance that appears to provide immunity to, or severely restrict replication of most isolates of TuMV has been characterised. This resistance source also prevented systemic spread of all TuMV isolates tested. Prior to this study, no resistance to pathotype 4 or pathotype 12 isolates of TuMV had ever been identified. For each of these three resistance sources, plant lines that are not segregating for some of the resistance phenotypes and that are presumably homozygous for the genes controlling these phenotypes have been generated. Strategies for further characterising and deploying these resistances in different Brassica species are described.

Histological characterisation of the incompatible interaction between Arabidopsis thaliana and the obligate biotrophic pathogen Plasmodiophora brassicae

The obligate biotrophic pathogen Plasmodiophora brassicae induces cataplastic galls in the roots of various species of the Brassicaceae. The ecotypes Ze-0, Tsu-0, and Ta-0 of Arabidopsis thaliana carry alleles of a dominant single gene ( RPB1 ) for resistance against this pathogen. The pathotype-specific resistance reactions in the roots were accompanied by a hypersensitive response. Infected cells were surrounded by necrotic boundaries and thereby the pathogen growth was restricted. The pathogen could induce hyperplasia and hypertrophy to a slight extent and develop spores within mini-galls surrounded by the necrotic cells, if it was able to colonise meristematic tissue. Ecotype Ze-0 showed the highest level of resistance to P. brassicae isolate ‘e’ as indicated by the very limited number of infected cells, whereas the resistance of the ecotype Ta-0 was rated lower. The results are discussed in relation to the interaction of the pathogen with Brassica crops.

Potyvirus genome-linked protein (VPg) determines pea seed-borne mosaic virus pathotype-specific virulence in Pisum sativum.

The mechanism of Pisum sativum pathotype-specific resistance to pea seed-borne mosaic potyvirus (PSbMV) was investigated and the coding region determinant of PSbMV virulence was defined. Homozygous recessive sbm-1 peas are unable to support replication of PSbMV pathotype 1 (P-1), whereas biochemically and serologically related pathotype 4 (P-4) is fully infectious in the sbm-1/sbm-1 genotype. We were unable to detect viral coat protein or RNA with double antibody sandwich-enzyme-linked immunosorbent assay and reverse transcription-polymerase chain reaction in sbm-1/sbm-1 P-1-inoculated protoplasts and plants. Lack of viral coat protein or RNA in P-1 transfected sbm-1/sbm-1 protoplasts suggests that sbm-1 resistance is occurring at the cellular level and that inhibition of cell-to-cell virus movement is not the operating form of resistance. In addition, because virus products were not detected at any time post-inoculation, resistance must either be constitutive or expressed very early in the virus infection process. P-1-resistant peas challenged with full-length, infectious P-1/P-4 recombinant clones demonstrated that a specific P-4 coding region, the 21-kDa, genome-linked protein (VPg), was capable of overcoming sbm-1 resistance, whereas clones containing the P-1 VPg coding region were noninfectious to sbm-1/sbm-1 peas. VPg is believed to be involved in potyvirus replication and its identification as the PSbMV determinant of infectivity in sbm-1/sbm-1 peas is consistent with disruption of an early P-1 replication event.

Genetics and Utilization of Pathogen Resistance in Plants.

Until relatively recently, knowledge of plant resistance to patho- gens has resulted primarily from research associated with the selective breeding of crop species. Although resistance is well described at the cellular, whole plant, and population levels in terms of genetics, histology, and associated biochemistry, a full mechanistic understanding of how pathogen resistance is mediated in plants is only now becoming feasible as a re- sult of the isolation and sequencing of severa1 putatively interacting plant and pathogen genes (see Alfano and Colher, 1996; Bent, 1996; Dangl et al., 1996; Hammond-Kosack and Jones, 1996, in this issue). Nevertheless, plant resistance genes have been used beneficially in agriculture for decades, even though their effects have not always been durable. This review provides a short and selective overview of the genetics of pathotype-specific resistance in plants, its past utilization in crop improvement, and some indications of how recent ad- vances may impact the future. Reference to data obtained from investigations with a few well-studied host-pathogen combi- nations (Table

Mapping quantitative trait loci for downy mildew resistance in pearl millet

Quantitative trait loci (QTLs) for resistance to pathogen populations of Scelerospora graminicola from India, Nigeria, Niger and Senegal were mapped using a resistant x susceptible pearl millet cross. An RFLP map constructed using F2 plants was used to map QTLs for traits scored on F4 families. QTL analysis was carried out using the interval mapping programme Mapmaker/QTL. Independent inheritance of resistance to pathogen populations from India, Senegal, and populations from Niger and Nigeria was shown. These results demonstrate the existence of differing virulences in the pathogen populations from within Africa and between Africa and India. QTLs of large effect, contributing towards a large porportion of the variation in resistance, were consistently detected in repeated screens. QTLs of smaller and more variable effect were also detected. There was no QTLs that were effective against all four pathogen populations, demonstrating that pathotype-specific resistance is a major mechanism of downy mildew resistance in this cross. For all but one of the QTLs, resistance was inherited from the resistant parent and the inheritance of resistance tended to be the result of dominance or over-dominance. The implications of this research for pearl millet breeding are discussed.