Sources and Origin of Resistance to Xanthomonas campestris pv. campestris in Brassica Genomes

ABSTRACT Two hundred and seventy-six accessions of mainly Brassica spp. were screened for resistance to Xanthomonas campestris pv. campestris races. In Brassica oleracea (C genome), the majority of accessions were susceptible to all races, but 43% showed resistance to one or more of the rare races (2, 3, 5, and 6) and a single accession showed partial resistance to races 1, 3, 5, and 6. Further searches for resistance to races 1 and 4, currently the most important races worldwide, and race 6, the race with the widest host range, were made in accessions representing the A and B genomes. Strong resistance to race 4 was frequent in B. rapa (A genome) and B. napus (AC genome), indicating an A genome origin. Resistance to races 1 and 4 was present in a high proportion of B. nigra (B genome) and B. carinata (BC genome) accessions, indicating a B genome origin. B. juncea (AB genome) was the most resistant species, showing either strong resistance to races 1 and 4 or quantitative resistance to all races. Potentially race-nonspecific resistance was also found, but at a lower frequency, in B. rapa, B. nigra, and B. carinata. The combination of race-specific and race-nonspecific resistance could provide durable control of black rot of crucifers.     

Differential Responses to Pea Bacterial Blight in Stems, Leaves and Pods Under Glasshouse and Field Conditions

Resistance to pea bacterial blight (Pseudomonas syringae pv. pisi) in different plant parts was assessed in 19 Pisum sativum cultivars and landraces, carrying race-specific resistance genes (R-genes) and two Pisum abyssinicum accessions carrying race-nonspecific resistance. Stems, leaves and pods were inoculated with seven races of P. s. pv. pisi under glasshouse conditions. For both race-specific and nonspecific resistance, a resistant response in the stem was not always associated with resistance in leaf and pod. Race-specific genes conferred stem resistance consistently, however, there was variability in the responses of leaves and pods which depended on the matching R-gene and A-gene (avirulence gene in the pathogen) combination. R2 generally conferred resistance in all plant parts. R3 or R4 singly did not confer complete resistance in leaf and pod, however, R3 in combination with R2 or R4 enhanced leaf and pod resistance. Race-nonspecific resistance conferred stem resistance to all races, leaf and pod resistance to races 2, 5 and 7 and variable reactions in leaves and pods to races 1, 3, 4 and 6.Disease expression was also studied in the field under autumn/winter conditions. P. sativum cultivar, Kelvedon Wonder (with no R genes), and two P. abyssinicum accessions, were inoculated with the most frequent races in Europe under field conditions (2, 4 and 6). Kelvedon Wonder was very susceptible to all three races, whereas P. abyssinicum was much less affected. The combination of disease resistance with frost tolerance in P. abyssinicum enabled plants to survive through the winter. A breeding strategy combining race-nonspecific resistance derived from P. abyssinicum with race-specific R-genes should provide durable resistance under severe disease pressure.     

Race-specific reaction of resistance to black rot in Brassica oleracea

Several black rot-resistant varieties of Brassica oleracea showed a race-specific hypersensitive response (HR) to inoculation with Xanthomonas campestris pv. campestris isolates of different races. In progenies of cabbage line PI436606, Portuguese kale ISA454 and Chinese kale SR1 the HR to race 1 of the pathogen was controlled by a dominant gene named R1, when a recessive gene r5 was responsible for the HR to race 5. Genes with a similar race-specific reaction were assumed on the basis of gene-for-gene interaction in black rot-resistant Japanese cabbage cultivars and double haploid lines obtained from them. Homology of gene r5 in cabbage lines PI436606, Fujiwase 01 and kale ISA454 was postulated in crosses between those lines or their progenies. In a cross between SR1 and PI436606, interaction between resistance to race 1 and non-specific resistance localized in the stem vascular system was found. On the basis of pedigree information and homology of resistance genes in the cultivars of East-Asian cabbage and Portuguese kales, the probable origin of race-specific resistance to black rot of cole crops was suggested to be in heading Mediterranean kale. Some evidence was found for a gene conferring resistance to race 4 in B. oleracea.     

Pathogenic variation in Xanthomonas campestris pv. phaseoli, the causal agent of common bacterial blight in Phaseolus beans

A total of 93 isolates of Xanthomonas campestris pv. phaseoli was collected from five countries over a wide range of altitude (500–2320 m above sea level) in eastern Africa. Collections were made from a range of cultivars of the common bean ( Phaseolus vulgaris ), including known sources of resistance, as well as from wild alternative but symptomless hosts including Senna ( Cassia ) hirsuta and Digitaria scalarum . From these, 30 isolates were selected for detailed characterization. Although variation was found in parameters including phage type, the production of brown pigment in culture media and in-vitro growth rate, there was little consistent relationship between these characters and pathogenic variation.
Studies of pathogenic variation among the 30 isolates on 20 genotypes of P. vulgaris revealed quantitative host non-specific differences (aggressiveness). On tepary bean ( P. acutifolius ), however, the 30 isolates interacted differentially with the genotypes and eight distinct physiologic races of X . c . pv. phaseoli were defined, suggestive of an underlying gene-for-gene relationship.
Despite this apparent gene-for-gene interaction, resistance to common bacterial blight in P. vulgaris , derived from earlier use of P. acutifolius , has apparently remained non-specific and essentially durable. It is suggested that, since levels of resistance already available within P. vulgaris provide adequate protection, bean breeders should resist the temptation to incorporate new complete resistance from P. acutifolius as it would risk destabilizing the host–bacterium relationship by introducing race-specific resistance that is more likely to prove transient in agriculture.     

Resistance to Pseudomonas syringae in a collection of pea germplasm under field and controlled conditions

A total of 242 Pisum accessions were screened for resistance to Pseudomonas syringae pv. pisi under controlled conditions. Resistance was found to all races, including race 6 and the recently described race 8. Fifty‐eight accessions were further tested for resistance to P. syringae pv. syringae under controlled conditions, with some highly resistant accessions identified. Finally, a set of 41 accessions were evaluated for resistance to P. syringae pv. pisi and pv. syringae under spring‐ and winter‐sowing field conditions. R2, R3 and R4 race‐specific resistance genes to P. syringae pv. pisi protected pea plants in the field. Resistance sources to race 6 identified under controlled conditions were ineffective in the field. Frost effects were also evaluated in relation to disease response. Results strongly suggest that frost tolerance is effective in lowering the disease effects caused by P. syringae pv. pisi and pv. syringae under frost‐stress conditions, even in the absence of disease resistance genes, although the highest degree of this protection is reached when frost tolerance and disease‐resistance genes are combined in the same genetic background.     

Identification of race-specific resistance in North American Vitis spp. limiting Erysiphe necator hyphal growth

Race-specific resistance against powdery mildews is well documented in small grains but, in other crops such as grapevine, controlled analysis of host-pathogen interactions on resistant plants is uncommon. In the current study, we attempted to confirm powdery mildew resistance phenotypes through vineyard, greenhouse, and in vitro inoculations for test cross-mapping populations for two resistance sources: (i) a complex hybrid breeding line, 'Bloodworth 81-107-11', of at least Vitis rotundifolia, V. vinifera, V. berlandieri, V. rupestris, V. labrusca, and V. aestivalis background; and (ii) Vitis hybrid 'Tamiami' of V. aestivalis and V. vinifera origin. Statistical analysis of vineyard resistance data suggested the segregation of two and three race-specific resistance genes from the two sources, respectively. However, in each population, some resistant progeny were susceptible in greenhouse or in vitro screens, which suggested the presence of Erysiphe necator isolates virulent on progeny segregating for one or more resistance genes. Controlled inoculation of resistant and susceptible progeny with a diverse set of E. necator isolates clearly demonstrated the presence of fungal races differentially interacting with race-specific resistance genes, providing proof of race specificity in the grape powdery mildew pathosystem. Consistent with known race-specific resistance mechanisms, both resistance sources were characterized by programmed cell death of host epidermal cells under appressoria, which arrested or slowed hyphal growth; this response was also accompanied by collapse of conidia, germ tubes, appressoria, and secondary hyphae. The observation of prevalent isolates virulent on progeny with multiple race-specific resistance genes before resistance gene deployment has implications for grape breeding strategies. We suggest that grape breeders should characterize the mechanisms of resistance and pyramid multiple resistance genes with different mechanisms for improved durability.     

小麦品种抗条锈性分化的初步研究

 以小麦条锈菌野生菌系(CYR29-1)及其7个毒性突变菌株测定小麦品种的抗锈性分化,发现水源11、抗引655和无芒中四群体对某些菌系有抗病性分化,一些抗源品种和后备品种也出现比例不同的抗感植株和不同低反应型的植株,证实这些品种群体对不同菌系具有抗病异质性。讨论了品种抗锈性分化的原因及在抗病品种丧失抗锈性中的作用,对改进小种鉴定、应用抗源等提出了建议。     

New genes for resistance to downy mildew (Peronospora parasitica) in oilseed rape (Brassica napus ssp. oleifera)

The response of cotyledons of oilseed rape ( Brassica napus ssp. oleifera ) accessions to infection by isolates of Peronospora parasitica under controlled conditions was assessed on a 0–7 scale (disease reaction). In interactions scored 0–3, 4–5 and 6–7, the host was considered resistant, partially resistant and susceptible, respectively. Accession RES-26, selected from the spring oilseed rape cultivar Janetzkis, was partially resistant to isolate R1 and resistant to isolate P003 of P. parasitica , which distinguishes it from three previously described differential response groups ('A', 'B' and 'C') of accessions in B. napus . The resistance of RES-26 to isolate P003 seemed to be conditioned by a single, partially dominant gene and the resistance of RES-02, which belongs to group 'A' (resistant to R1 and P003), by two independent partially dominant genes. The gene for resistance to P003 in RES-26 is either closely linked, allelic or identical to one of the two genes for resistance in RES-02. Resistance of RES-02 to R1 is conditioned by a single, incompletely dominant gene. The genes for resistance to isolates R1 and P003 in RES-02 are either closely linked, allelic or identical. The cotyledonary leaves of each seedling responded independently when inoculated simultaneously each with a different isolate of the pathogen.     

Histopathology of durable adult plant resistance to leaf rust in the Brazilian wheat variety Toropi

Leaf rust, caused by the fungus Puccinia triticina is a major disease of wheat (Triticum aestivum) worldwide. This disease is prevalent in southern South America where the environmental conditions and high genetic variability of P. triticina favour epidemics. The primary means of controlling pathogenic P. triticina races has been through using wheat varieties containing race-specific resistance genes. The defence mechanisms involved in durable race non-specific resistance to P. triticina are probably distinct from those involved in non-durable race-specific resistance. We investigated the histological components of resistance to P. triticina present in three wheat genotypes: the race non-specific resistant Brazilian variety Toropi; the race-specific resistant line RL6010 Lr9; and the susceptible Brazilian variety BRS 194. Plants of these three genotypes were inoculated with P. triticina race MFP and tissue samples excised from flag leaves at various times after inoculation to assess the number of infective structures, frequency of cell death and the accumulation of autofluorescent cells and hydrogen peroxide. The genotypes showed different resistance mechanisms active at different times during the infection process. Our results for Toropi indicate that there was a reduction in the extent of formation of stomatal appressoria and all subsequent structures. During attempted penetration we also observed the production of autofluorescent compounds and late cell death, but not peroxide formation. This non-specific resistance to P. triticina involves both pre-haustorial and post-haustorial mechanisms which may be responsible for maintaining the low disease severity observed in this variety even under high inoculum pressure.     

Resistance to Taiwanese race 1 strains of Ralstonia solanacearum in wild tomato germplasm

A total of 252 wild Solanum accessions and one population of 49 introgression lines of LA716 were screened for resistance to a race 1/biovar 4/phylotype I strain Pss186 of Ralstonia solanacearum. Most wild tomato accessions were highly susceptible. However, five accessions of S. pennellii, i.e. LA1943, LA716, LA1656, LA1732 and TL01845 were resistant to strain Pss186. These accessions were then challenged against two other race 1/phylotpye I strains Pss4 and Pss190, which were more aggressive. All the five S. pennellii accessions were susceptible to Pss4, but displayed high to moderate resistance to Pss190 with a percentage of wilted plants ranging from 0% to 60%. Pss190 is an aggressive strain that made a resistant tomato line Hawaii 7996 susceptible. Thus, the results found in this study provide evidence of the presence of strain-specific resistance. LA3501, which has an introgression segment on chromosome 6, was found to be resistant to Pss186 among the screened introgression lines. This confirms the importance of resistance trait loci on chromosome 6 that have been identified by other studies. This is the first report of S. pennellii being resistant to bacterial wilt. These new resistant sources will provide breeders with more resources to breed for stable resistance to bacterial wilt of tomato.     

A Cluster of Major Specific Resistance Genes to Leptosphaeria maculans in Brassica napus

ABSTRACT Two types of genetic resistance to Leptosphaeria maculans usually are distinguished in Brassica napus: qualitative, total resistance expressed at the seedling stage and quantitative, partial resistance expressed at the adult plant stage. The latter is under the control of many genetic factors that have been mapped through quantitative trait loci (QTL) studies using 'Darmor' resistance. The former usually is ascribed to race-specific resistance controlled by single resistance to L. maculans (Rlm) genes. Three B. napus-originating specific Rlm genes (Rlm1, Rlm2, and Rlm4) previously were characterized. Here, we report on the genetic identification of two novel resistance genes, Rlm3 and Rlm7, corresponding to the avirulence genes AvrLm3 and AvrLm7. The identification of a novel L. maculans- B. napus specific interaction allowed the detection of another putative new specific resistance gene, Rlm9. The resistance genes were mapped in two genomic regions on LG10 and LG16 linkage groups. A cluster of five resistance genes (Rlm1, Rlm3, Rlm4, Rlm7, and Rlm9) was strongly suggested on LG10. The relation between all these specific resistance genes and their potential role in adult-plant field resistance is discussed. These two Rlm-carrying regions do not correspond to major QTL for Darmor quantitative resistance.     

Expression of biotic stress response genes to Phytophthora infestans inoculation in White Lady,a potato cultivar with race-specific resistance to late blight

Expression changes of biotic stress response genes were analyzed during a 65 h period post inoculation with Phytophthora infestans in potato cultivar White Lady that possesses race-specific resistance to this pathogen. All analyzed respiratory burst oxidase homologs, the PR proteins, the serine-, cysteine- and aspartic protease inhibitors, as well as the Rpi-bt1 gene homolog were up-regulated in the biotrophic phase. The R1 and R2 gene homologs showed up-regulation only at 65 hpi, and interestingly, the R3a gene showed only a very slight expressional increase. It is concluded, that beside the constitutively expressed R genes a number of non-specific stress response genes contribute to the successful resistance response in race-specific defense.     

A detailed evaluation method to identify sources of quantitative resistance to Fusarium oxysporum f. sp. pisi race 2 within a Pisum spp. germplasm collection

Fusarium oxysporum f. sp. pisi (Fop) is an important pathogen of field pea (Pisum sativum) worldwide. The constant evolution of the pathogen drives the necessity to broaden the genetic basis of resistance to Fop. To achieve this, it is important to have a large germplasm collection available and an accurate and efficient method for disease assessment. Here, a detailed evaluation method coupling disease incidence, disease rating over time and its related area under the disease progression curve (AUDPC) was established and used to screen a Pisum spp. germplasm collection against one isolate of Fop race 2. A large variation in the disease response of specific pea accessions ranging from highly resistant to susceptible was observed within the collection, indicating the quantitative expression of the resistance. The repetition of the inoculation experiments on a subset of 19 accessions, including two susceptible accessions, indicated that the scoring method was robust and reproducible and confirmed the highly resistant phenotypes of 11 accessions. To initiate the characterization of resistance mechanisms within these accessions, the external and internal stem symptoms were compared between these selected pea accessions, together with the extent of fungal colonization within plants. All these tests indicated that, in all resistant accessions, the resistance mechanisms efficiently stopped pathogen progression at the crown. Incorporation of these sources of resistance to breeding programmes will contribute to improved Fop resistance in pea cultivars.     

Identification of the rice blast resistance gene Pib in the National Small Grains Collection

The Pib gene in rice confers resistance to a wide range of races of the rice blast pathogen, Magnaporthe oryzae, including race IE1k that overcomes Pita, another broad-spectrum resistance gene. In this study, the presence of Pib was determined in 164 rice germplasm accessions from a core subset of the National Small Grains Collection utilizing DNA markers and pathogenicity assays. The presence of Pib was evaluated with two simple sequence repeat (SSR) markers and a dominant marker (Pib-dom) derived from the Pib gene sequence. Pathogenicity assays using two avirulent races (IE1k and IB1) and a virulent race (IB54) were performed to verify the resistance responses of accessions. Of the 164 accessions evaluated, 109 contained the Pib gene as determined using both SSR markers and pathogenicity assays, albeit different haplotypes were detected. The remaining 52 germplasm accessions were different in their responses to the blast races IB54, IE1k, and IB1, thus indicating the presence of R gene(s) other than Pib. The accessions characterized in this study could be used for marker-assisted breeding to improve blast resistance in indica and japonica cultivars worldwide.     

Inheritance of Race-Specific Resistance to Xanthomonas campestris pv. campestris in Brassica Genomes

ABSTRACT The inheritance of resistance to three Xanthomonas campestris pv. campestris races was studied in crosses between resistant and susceptible lines of Brassica oleracea (C genome), B. carinata (BC genome), and B. napus (AC genome). Resistance to race 3 in the B. oleracea doubled haploid line BOH 85c and in PI 436606 was controlled by a single dominant locus (Xca3). Resistance to races 1 and 3 in the B. oleracea line Badger Inbred-16 was quantitative and recessive. Strong resistance to races 1 and 4 was controlled by a single dominant locus (Xca1) in the B. carinata line PI 199947. This resistance probably originates from the B genome. Resistance to race 4 in three B. napus lines, cv. Cobra, the rapid cycling line CrGC5, and the doubled haploid line N-o-1, was controlled by a single dominant locus (Xca4). A set of doubled haploid lines, selected from a population used previously to develop a restriction fragment length polymorphism map, was used to map this locus. Xca4 was positioned on linkage group N5 of the B. napus A genome, indicating that this resistance originated from B. rapa. Xca4 is the first major locus to be mapped that controls race-specific resistance to X. campestris pv. campestris in Brassica spp.     

A Single Gene Cluster Controls Incompatibility and Partial Resistance to Various Melampsora larici-populina Races in Hybrid Poplars

ABSTRACT Complete cosegregation for race-specific incompatibility with three Melampsora larici-populina rust races was observed in five F(1) hybrid progenies of Populus, with different patterns among the various progenies. A single gene cluster could explain these segregations: one locus with multiple alleles or two tightly linked loci controlling complete resistance to E1 and E3, and two tightly linked loci for E2. The random amplified polymorphic DNA marker OPM03/04_480 was linked to that cluster in all families (<1 cM). This marker accounted for more than 70% of the genetic variation for field resistance in each family (heritability approximately 0.40). The same marker accounted for up to 64% of the clonal variation for growth in the nursery under natural inoculum pressure; the weak tolerance to rust of F(1) interspecific hybrids was attributed to a genetic background effect. Partial resistance was split into epidemiological components (heritability ranged from 0.35 to 0.87). Genotypic correlations among resistance traits for the different races were high (0.73 to 0.90). However, correlations among different resistance components for a single race were not all significant. A major quantitative trait locus for all components of partial resistance to E2 was associated to the cluster controlling incompatibility to E1 and E3 and marked by OPM03/04_480 (R(2)from 48 to 68%).     

Analysis of quantitative trait Loci for resistance to southern leaf blight in juvenile maize

ABSTRACT A set of 192 maize recombinant inbred lines (RILs), derived from a cross between the inbred lines Mo17 and B73, were evaluated as 3-week-old seedlings in the greenhouse for resistance to southern leaf blight, caused by Cochliobolus heterostrophus race O. Six significant (LOD >3.1) quantitative trait loci (QTL) were identified for disease resistance, located on chromosomes 1, 2, 3, 6, 7, and 8. Results were compared with a previous study that had used the same RIL population and pathogen isolate, but had examined resistance in mature rather than juvenile plants. There was a very weak but significant correlation between the overall resistance phenotypes of the RILs scored as mature and juvenile plants. Two QTL were found in similar positions on chromosomes 1 and 3 at both growth stages. Other QTL were specific to one growth stage or the other. Twenty-three of these RILs, together with the parental lines, were inoculated in the greenhouse with four C. heterostrophus isolates. Results indicated that the quantitative resistance observed was largely isolate non-specific.     

Genetics of resistance to race TTKSK of Puccinia graminis f. sp. tritici in Triticum monococcum

Race TTKSK (or Ug99) of Puccinia graminis f. sp. tritici possesses virulence to several stem rust resistance genes commonly present in wheat cultivars grown worldwide. New variants detected in the race TTKSK lineage further broadened the virulence spectrum. The identification of sources of genetic resistance to race TTKSK and its relatives is necessary to enable the development and deployment of resistant varieties. Accessions of Triticum monococcum, an A-genome diploid wild and cultivated wheat, have previously been characterized as resistant to stem rust. Three resistance genes were identified and introgressed into hexaploid wheat: Sr21, Sr22, and Sr35. The objective of this study was to determine the genetic control and allelic relationships of resistance to race TTKSK in T. monococcum accessions identified through evaluations at the seedling stage. Generation F(2) progeny of 8 crosses between resistant and susceptible accessions and 13 crosses between resistant accessions of T. monococcum were evaluated with race TTKSK and often with North American races, including races QFCSC, TTTTF, and MCCFC. For a selected population segregating for three genes conferring resistance to race TTKSK, F(2:3) progeny were evaluated with races TTKSK, QFCSC, and TTTTF. In that population, we detected two genes conferring resistance to race TTKSK that are different from Sr21, Sr22, and Sr35. One of the new genes was effective to all races tested. The identification of these genes will facilitate the development of varieties with new resistance to race TTKSK.     

Genetic analysis of the resistance to eight anthracnose races in the common bean differential cultivar Kaboon

Resistance to the eight races (3, 7, 19, 31, 81, 449, 453, and 1545) of the pathogenic fungus Colletotrichum lindemuthianum (anthracnose) was evaluated in F(3) families derived from the cross between the anthracnose differential bean cultivars Kaboon and Michelite. Molecular marker analyses were carried out in the F(2) individuals in order to map and characterize the anthracnose resistance genes or gene clusters present in Kaboon. The analysis of the combined segregations indicates that the resistance present in Kaboon against these eight anthracnose races is determined by 13 different race-specific genes grouped in three clusters. One of these clusters, corresponding to locus Co-1 in linkage group (LG) 1, carries two dominant genes conferring specific resistance to races 81 and 1545, respectively, and a gene necessary (dominant complementary gene) for the specific resistance to race 31. A second cluster, corresponding to locus Co-3/9 in LG 4, carries six dominant genes conferring specific resistance to races 3, 7, 19, 449, 453, and 1545, respectively, and the second dominant complementary gene for the specific resistance to race 31. A third cluster of unknown location carries three dominant genes conferring specific resistance to races 449, 453, and 1545, respectively. This is the first time that two anthracnose resistance genes with a complementary mode of action have been mapped in common bean and their relationship with previously known Co- resistance genes established.     

Identification of Lentil Germ Plasm Resistant to Colletotrichum truncatum and Characterization of Two Pathogen Races

ABSTRACT A total of 1,771 lentil accessions from the U.S. lentil collection (U.S. Department of Agriculture-Agricultural Research Service, Pullman, WA) and the Institut für Pflanzengenetik und Kulturpflanzenforschung (Gatersleben, Germany) were screened for resistance to Colletotrichum truncatum, the cause of anthracnose. About 95% of the accessions were susceptible when inoculated with a single isolate in the field. Retesting, under controlled conditions, of accessions rated as resistant or moderately resistant in the field resulted in identification of anthracnose resistance in four accessions from the U.S. collection (PI 320937, PI 320952 [cv. Indianhead], PI 345629, and 468901), and 12 accessions from the German collection (Lens 3, 102, 104, 106, 107, 119, 122, 134, 135, 177, 195, and 209). Seven of the accessions were used as host differentials to characterize pathogenic variability of 50 single-spore isolates collected in Manitoba and Saskatchewan, Canada. The presence of two distinct races was demonstrated. Isolates of C. truncatum avirulent on cv. Indianhead, PI 320937, PI 345629, PI 468901, Lens 102, Lens 104, and Lens 195 were designated race Ct1. Isolates that were virulent on these seven entries were designated race Ct0, indicating their lack of avirulence genes. Race Ct0 was isolated more frequently from commercial seed samples than race Ct1, but the two races were isolated with similar frequency from plants in commercial fields planted to susceptible cultivars. Race Ct0, to which no resistance has yet been identified, presents a high risk to lentil production in Canada and potentially worldwide.