Diversity among Finnish net blotch isolates and resistance in barley

Summary Seedlings of a differential barley (Hordeum vulgare L.) series (21 genotypes) and of six check genotypes were used in the greenhouse to assess variation in virulence among 20 single-spore isolates of the net blotch pathogen. Pyrenophora teres Drechs. f.teres Smedeg., collected from various sites in Finland. The experiment was run twice and symptom expression was recorded on the first three leaves. Analysis of second leaf symptom scores from Run 1 indicated differences in virulence between isolates, all of which were pathogenic, and differential resistance among the barleys. The virulence of P. teres isolates appeared to be conditioned by the host barley from which the isolate derived; the average virulence of isolates collected from a susceptible host was greater than that of isolates collected from a less susceptible host. Results from Run 2 were similar regarding resistance within the barleys, but variation in virulence among the P. teres isolates was not consistent with that from Run 1. CI 9819 caries duplicate genes for resistance to P. teres (Rpt1b and Rpt1c), and CI 7548 possesses Rpt3d. Both genotypes were highly resistant to all isolates. The Rpt1a gene of Tifang (CI4407) conferred resistance to all isolates in Run 2, but only to about half of the isolates in Run 1. The checks, including two of the symptomatically most resistant Nordic barley genotypes, were universally susceptible during these stringent tests. No selective pressure has been placed on Finnish isolates of P. teres through previous deployment of major resistance genes, and it is speculated that any variation in virulence among isolates is likely to be due to a combination of evolutionary forces including, natural selection, random genetic drift and gene flow.     

Generation Mean Analysis of Inheritance of Resistance to Pyrenophora teres in Barley

F1, F2, Fl × parentl (BC1), and Fl × parent2 (BC2) generations resulting from four crosses among seven cultivars of barley used in national and international breeding programmes were tested at the seedling stage for their resistance to a mixture of five isolates of Pyrenophora teres. Four methods were used to assess disease resistance: infection type (IT), average lesion size (ALS), number of lesions per unit leaf area (NL) and percent leaf area infected (PLAI). Gene actions were estimated by generation mean analysis on each of the four crosses and on each of the evaluation methods. Significant additive and additive × additive epistatic effects were found. Infection type and percent leaf area infected were found to be highly correlated in all four crosses. These results suggest that barley breeders could improve the level of resistance to P. teres by making appropriate crosses between highly susceptible barley cultivars.     

Barley mutants with enhanced level of resistance to Swedish isolates of Bipolaris sorokiniana,casual agent of spot blotch

Spot blotch, caused by Bipolaris sorokiniana, has become an increasingly important disease on barley in Sweden. In order to enhance our knowledge of this disease, eight Swedish B. sorokiniana isolates were assessed for their virulence pathotype on 30 barley genotypes including eight lines of mutant origin, designated B. sorokiniana tolerant (bst). A leaf screen assay revealed the presence of virulence groups 1 and 7 in Sweden. Several mutant genotypes demonstrated enhanced resistance responses (P = 0.0001), of which bst1:3 had the lowest leaf scoring value to the most virulent isolate, Swe2 (virulence group 7). To evaluate the host root responses, two contrasting isolates Swe2 and Swe4 (virulence group 7 and 1) were used to inoculate a set of eight differential lines, including four promising mutant lines from the leaf screen. Mutant bst1:6 was significantly less susceptible (P = 0.001) compared with the other barley genotypes tested, independent of the two isolates used. Mutants bst1, bst1:3 and bst1:4, which displayed less or no spots, are also considered promising genotypes for further breeding attempts.     

Inheritance of resistance to Pyrenophora teres f. teres in spring barley

The inheritance of seedling resistance to a Swedish isolate of Pyrenophora teres f. teres was investigated in four resistance sources of spring barley. Accessions CI 2330, CI 5791, CI 5822 and CI 9779 were used as resistance sources, and the cultivar ‘Alexis’ was used as a susceptible parent in different crosses. From the disease reaction in the F₁, F₂ and F₃ generations it was concluded that the resistance was governed by the same two complementary genes in CI 5791, CI 822 and CI 9776. One of these genes was present in CI 2330. The first three cultivars were highly resistant to the isolate used in this investigation. These results, when combined with earlier studies, suggest that CI 5791, CI 5822 and CI 9776 may be of great value as sources of resistance to barley net blotch. Spearman's rank correlation between the disease reaction of F₂ plants and their F₃ progeny was highly significant (r = 0.75; P ≥ 0.001) It is suggested that selection in the F₂ generation is effective. In a backcross breeding scheme, single plant reactions in F₁ or F₂ need to be confirmed in later generations.     

Adaptation of poplar leaf rust to Eastern cottonwood

Summary Genotypes of eastern cottonwood, Populus deltoides, from Kentucky or Mississippi were inoculated using a leaf disk assay with isolates of poplar leaf rust, Melampsora medusae f. sp. deltoidae (Mmd), from both locations to determine if pathogenic races differ at these locations. Disease severity measurements, including infection probability (number of uredia produced per uredospore), sporulation (number of uredospores/cm²), and the progeny/parent ratio (number of daughter uredia produced per mother uredium) were significantly higher when trees of southern origin were inoculated with isolates from southern locations as compared to isolates from northern locations. In addition, larger variation in disease severity measurements attributable to specific interactions, i.e. the variance due to isolates adjusted for virulence on a universal suscept, were found in northern than in southern isolates. These results suggest that isolates of southern origin have broader adaptation and are more virulent on local southern host genotypes than isolates of northern origin. However, the larger variance attributable to specific interactions in northern isolates indicates the potential for pathogen adaptation to resistant host genotypes in Kentucky.Journal series paper 92-8-81 of the University of Kentucky Agricultural Experiment Station     

Genes for resistance in barley to Finnish isolates of Rhynchosporium secalis

Summary Twenty Finnish isolates of Rhynchosporium secalis (Oud.) J.J. Davis, the causal agent of scald, were taken from infected barley (Hordeum vulgare L.) plants and inoculated on to seedlings of a differential series of barley containing a range of major genes for resistance to the fungus, as well as on to six Nordic 6-row spring barleys and three winter ryes (Secale cereale L.). These fungal isolates derived from four sites and three host varieties. Disease development was monitored on two leaves of seedlings in the greenhouse employing a standard scale, and on adult plants in the field by assessing the diseased area on the three uppermost leaves. A comparison was also made between the pathogenicity and virulence of ten Finnish and ten Canadian R. secalis isolates. The Finnish isolates varied in virulence, but with the exception of Algerian (CI 1179) seedlings and adult La Mesita (CI 7565) all seedlings and adult plants of the entire differential series were resistant to all isolates. Canadian isolates were, on average, less virulent than Finnish isolates. All the Nordic checks were susceptible to all Finnish and seven Canadian isolates, but differences in the degree of susceptibility were evident. Isolates of R. secalis from barley were non-pathogenic on rye, isolates from Elymus repens L. were non-pathogenic on barley and rye, and isolates from rye were only pathogenic on rye. Finnish R. secalis isolates contain no redundant pathogenic diversity. The differential series represents a useful, but as yet untapped, source of resistance to R. secalis for Finnish barley breeders.     

Virulence studies of Swedish net blotch isolates (Drechslera teres) and identification of resistant barley lines

Six Swedish and one Canadian single spore isolate of Drechslera teres f. teres were used to screen 109 barley lines for disease resistance and to select a differential set of barley lines for use in assessing pathogen virulence. A large variation for net blotch resistance was found among the 109 barley lines which were classified into four groups, those showing: 1) only resistant reactions; 2) differential reactions; 3) only intermediate reactions and 4) only susceptible reactions. The European commercial varieties included, showed susceptibility to all Swedish isolates, but a few were resistant to the Canadian isolate. The 18- member differential set separated 25 Swedish and two Canadian isolates of D. teres into 14 pathotypes, three of which made up 59% of the isolates. Only one barley differential (CI 9776) was resistant to all net form isolates. Host selection on the pathogen seems to be present as all six isolates obtained from cv. Golf belonged to the same pathotype and 4 of 5 isolates from cv. Karin shared the same virulence pattern. The net form of net blotch (D. teres f. teres) predominated in the sampled regions and only one of 26 Swedish isolates was of the spot form (D. teres f. maculata). This revised version was published online in July 2006 with corrections to the Cover Date.     

Pathogenic variation among isolates of Rhynchosporium secalis from cultivated barley growing in Victoria,Australia

Summary Three hundred and nineteen Rhynchosporium secalis isolates from cultivated barley were divided into five groups on the basis of their virulence on 15 differential barley varieties. Pathogenic variation was also demonstrated for isolates from different scald lesions within the same crop and amongst different spores from the same lesions.     

Genetic diversity of Phaeoisariopsis griseola in the State of Minas Gerais, Brazil

Due to the importance of common bean angular leaf spotin the state of Minas Gerais-Brazil and to the greatvariability of the pathogen, Phaeoisariopsisgriseola, monitoring races becomes an important toolfor breeding programs aiming at genetic resistance.The pathogenic variability of 30 isolates of the P. griseola, collected from various locations in thestate of Minas Gerais, was studied using the followingcommon bean differential series (Don Timóteo,Bolón Bayo, Montcalm, G 5686, Amendoin, G 11796,BAT 332, PAN 72, Cornell 49-242, México 54, Florde Mayo and G 2858). The first trifoliate leaf wasinoculated with a 2 × 10⁴ conidia/mL. Plants weremaintained at 20–22 ^°C and 95% relativehumidity for 48 hours. Symptom evaluation wasperformed 15 days after inoculation. Thirteen raceswere identified demonstrating the wide geneticvariability of the pathogen in the state of MinasGerais. Race 63.63 was the most virulent, whereas race63.23 was the most frequent (10 of 30 isolates), beingwidely distributed among the regions studied. Thevirulence phenotype indicated that the races studiedbelonged to the Mesoamerican group, which wasconfirmed when the 30 isolates were compared to Andeanand Mesoamerican standards using RAPD markers.     

Analysis of durable resistance to stem rust in barley

Summary Since the mid-1940's, barley cultivars grown in the northern Great Plains of the USA and Canada have been resistant to stem rust caused byPuccinia graminis f. sp.tritici. This durable resistance is largely conferred by a single gene,Rpg1, derived from a single plant selection of the cultivar Wisconsin 37 and an unimproved Swiss cultivar. At the seedling stage, barley genotypes withRpg1 generally exhibit low mesothetic reactions at 16–20° C and slightly higher mesothetic reactions at 24–28° C to many stem rust pathotypes. This resistance is manifested by a low level of rust infection and mostly incompatible type uredia on adult plants.Rpg1 reacts in a pathotype-specific manner since some genotypes ofP. g. f. sp.tritici are virulent on cultivars carrying this gene in the field. Several factors may have contributed to the longevity of stem rust resistance in barley, a) since barley is planted early and matures early, it can sometimes escape damage from stem rust inoculum carried from the south; b) one or more minor genes may augment the level of resistance already provided byRpg1; c) the cultivation of resistant wheat cultivars and eradication of barberry have reduced the effective population size and number of potential new pathotypes ofP. g. f. sp.tritici, respectively; and d) virulent pathotypes ofP. g. f. sp.tritici andP. g. f. sp.secalis have not become established. This situation changed in 1989 when a virulent pathotype (Pgt-QCC) ofP. g. f. sp.tritici became widely distributed over the Great Plains. However,Rpg1 may still confer some degree of resistance to pathotype QCC because stem rust severities have been low to moderate and yield losses light on barley cultivars carrying the gene during the last four seasons (1989–1992). Several sources of incomplete resistance to pathotype QCC have been identified in barley. To facilitate the transfer of resistance genes from these sources into advanced breeding lines, molecular marker assisted selection is being employed.     

Evaluation of greenhouse inoculation techniques to screen sorghum for resistance to downy mildew

Summary Six inoculation techniques were compared for the artificial promotion of downy mildew (Peronosclerospora sorghi) in sorghum. These were (1) sprouted seeds incubated between sporulating infected leaves, (2) sprouted seeds depped in conidial suspension, (3) sprouted seeds sprayed with conidial suspension, (4) seedlings at plumule stage inoculated with drops of a conidial suspension, (5) seedlings at plumule stage sprayed with a conidial suspension, and (6) seedling showered with conidia falling from infected leaves. Seedlings at the one-leaf stage sprayed with a conidial suspension (6 × 10⁵ ml^-1) showed the highest systemic infection (100%) in the susceptible lines IS 643 and IS 18433. This technique is effective, repeatable, and allows the deposition of a conidial suspension as a fine mist on the entire seedling surface. In the greenhouse, the technique was used to test the downy mildew reaction of genotypes previously reported as resistant (< 5% incidence) in 3–4 years of field screenings. Of the 61 genotypes tested, 21 were free from downy mildew, 14 had less than 5% incidence, and the rest showed variable susceptible reactions. Therefore, the technique can be reliably and effectively used in the greenhouse to detect disease escapes and to indentify resistance.     

Pathotypes and NIP1 gene sequences of Finnish Rhynchosporium secalis isolates from barley, couch grass and rye

Rhynchosporium secalis (Oud.) J.J. Davis, causal agent of scald of barley (Hordeum vulgare L.), was isolated from barley, couch grass (Elymus repens L.) and rye (Secale cereale L.). Isolates were used to inoculate seedlings of a differential barley series containing several sources of major gene resistance to the disease. The series included Atlas46, (resistance gene Rrs1) and the isogenic line Atlas, which lacks the gene (rrs1). The necrosis-inducing peptide NIP1 has been suggested to be the product of the avirulence gene AvrRrs1 (NIP1) that with the barley resistance gene Rrs1 determines the incompatibility of the pathogen-host interaction. All R. secalis isolates were virulent only on the susceptible barley cultivars Arve and Chan, irrespective of spore concentration or original host species. There were no indications of redundant virulence among the R. secalis isolates. The NIP1 gene was sequenced from each isolate and there was no correlation with source of the isolate or sequence modification and virulence. Four isolates, from barley and couch grass, were characterised by a basic Type I NIP1sequence. The recorded NIP1 sequence changes are consistent withR. secalis populations not receiving selection through deployment of the Rrs1 gene in commercial barley cultivars in Finland. This revised version was published online in July 2006 with corrections to the Cover Date.     

Resistance spectra of wheat genotypes and virulence patterns of <Emphasis Type="Italic">Mycosphaerella graminicola</Emphasis> isolates in Iran

Co-evolution of wheat and its devastating pathogen Mycosphaerella graminicola (anamorph Septoria tritici), the causal agent of septoria tritici blotch, a foliar disease of wheat, is suggested to occur in Fertile Crescent as their center of origin and, thus, interaction between pathogen virulence and host resistance is important subject to be addressed. We have investigated resistance spectra of 54 wheat genotypes including a set of differentials carrying known resistance genes and virulence patterns of 14 M. graminicola isolates at seedling stage under controlled environmental conditions. The isolates were collected in Iran from five provinces. Diversity in virulence and aggressiveness was observed among the isolates from four provinces. Isolates collected from Golestan province were virulent to all wheat genotypes from germplasm of Iran, while specific resistances were identified to the isolates from other provinces. Among wheat genotypes, cvs. Chamran, Morvarid and Hirmand had the greatest number of specific resistances as well as partial resistance. Wheat genotypes of the differential set also differed in their reactions to the isolates. Arina, Flame and TE 9111 were specifically resistant to the greatest number of isolates from different provinces. Most isolates were virulent to the other differentials such as cvs. Shafir, Estanzuela federal and Courtot indicating that extensive adaption of virulence to most of the known resistance genes (Stb) has occurred in these regions. The new sources of resistance to highly virulent isolates from Iran may also be utilized in wheat breeding programs to develop resistant cultivars against pathogen populations in other countries.     

The importance of abortive stoma penetration in the partial resistance and nonhost reaction of adult barley and wheat plants to leaf rust fungi

Summary Early stages of the infection process of Puccinia hordei isolate 1.2.1 and of a P. recondita f.sp. tritici isolate were studied on adult plants of four barley lines and one wheat line. Two of the barley lines are extremely susceptible to P. hordei, the other two have a very high level of partial resistance.A histological study based on a trypan blue staining indicated that stoma penetration by P. hordei isolate 1.2.1 was equally successful on the susceptible as on the partially resistant adult barley plants. Abortion of substomatal vesicles was rare in all lines. These results do not support a hypothesis that mechanisms of partial resistance in adult plants differ from those in seedlings by a substantial abortive stoma penetration.Also in the nonhost combinations wheat-P. hordei and barley-P. recondita f.sp. tritici inhibition of stoma penetration and of substomatal vesicle development appears to play a biologically insignificant role in adult plants.The proportion of stoma penetration on the leaf sheaths of two of the barley lines was as high as on the leaf blades of the flag leaf and the leaf below the flag leaf. There was no evidence for stomatal exclusion as a crucial factor in the relatively low infectibility of leaf sheaths to leaf-blade specialized rust species.     

Mapping seedling resistance to net form of net blotch (Pyrenophora teres f. teres) in barley using detached leaf assay

To develop molecular markers against Pyrenophora teres f. teres in barley, a detached leaf assay was conducted on two DH populations with a set of 11 single conidial lines (SCLs). Out of these, three showed different reactions in the DH population ‘Uschi × HHOR3073’ and two in ‘(P x V) × HHOR9484’. For SCL ‘QLB’, a 1r : 1s (χ² = 2.78) segregation was observed in the population ‘Uschi × HHOR3073’. In contrast to this, a continuous variation was observed for the SCL ‘WvB’ and ‘d8_4’ in the DH population ‘Uschi × HHOR3073’ and for ‘AR’ and ‘net1840’ in ‘(P × V) × HHOR9484’. With respect to resistance to the SCL ‘QLB’, a single major gene was located on chromosome 7H, and for resistance against SCL ‘WvB’, two QTLs were detected on chromosome 3H and 7H, and against SCL ‘d8_4’, two loci were mapped on chromosome 3HS in ‘Uschi × HHOR3073’. In the DH population ‘(P x V) × HHOR9484’, one locus conferring resistance to the SCL ‘AR’ was located on chromosome 3H. For resistance to SCL ‘net1840’, two QTLs were mapped on chromosome 4H and 5H.     

Effect of leaf age on components of partial resistance in rice to leaf blast

Summary Ten rice genotypes were inoculated with a virulent isolate of Magnaporthe grisea to study the effect of leaf age on components of partial resistance, and evaluate associations between these components. The number of sporulating lesions per cm² leaf declined with increase of leaf age in all genotypes. The number of lesions per cm² leaf area in one week old leaves was about 25% that of the number in very young leaves in the susceptible cultivar CO39, but less than 2% in the more resistant cultivars IR36, IR60 and IR62. Large differences between genotypes were found for the number of sporulating lesions that developed, and this factor was closely related to the period that leaves remained susceptible after appearance. The number of lesions in the most susceptible cultivar CO39 was about 7 times that in the cultivars IR60 and IR64. Differences between genotypes were also found for lesion size. The effect of aging on average lesion size was less pronounced than on lesion density. Lesion size and lesion density were positively correlated, but a rapid decline of density was not necessarily accompanied by a rapid decline of size. No apparent differences between genotypes were observed for latent period. Genotypes with leaves that became highly resistant soon after appearance expressed higher levels of partial resistance in the field.     

Identification and characterization of seedling and adult plant resistance to Puccinia hordei in Chinese barley germplasm

Leaf rust of barley, caused by Puccinia hordei, occurs in all barley‐growing regions of Australia causing significant yield losses under epidemic conditions. The development and use of resistant cultivars are the most economical and environmentally sustainable method to control leaf rust which in turn relies on ongoing efforts to identify and characterize new sources of resistance. The aim of this study was to postulate known genes and/or identify new sources of resistance to P. hordei. Fifty‐two genotypes were assessed at the seedling and adult plant growth stages. On the basis of multipathotype tests, 39 genotypes lacked detectable seedling resistance, and nine were postulated to carry the genes Rph2, Rph4, Rph12 and Rph19 singly. Four genotypes carried uncharacterized seedling resistance; however, the gene(s) present in each were ineffective to at least one of the pathotypes used. Field tests at the adult plant growth stage revealed the presence of adult plant resistance (APR) in 12 genotypes. Tests of allelism and marker analysis indicated that resistance genes present in these genotypes were independent of the APR gene Rph20.     

Resistance to powdery mildew in selections from Moroccan barley landraces

Seventy-five barley landraces from Morocco were tested for resistance to powdery mildew and a number of different resistance genes were detected. Thirty-five isolates of Blumeria graminis f. sp. hordei and the Pallas isoline differential set were used. Isolates used in the experiment had virulences corresponding to all major resistance genes used in Europe. Forty-four of the tested landraces showed resistant reactions. From each of these landraces, one to five resistant plants were selected and 92 single plant lines were created. Six lines selected from 3landraces were assumed to carry the mlo gene but they were discarded after microscopic investigation. Seventeen lines were tested in the seedling stage with 17isolates and another 69 lines were tested with 23 differential isolates. These lines showed 71 reaction spectra to isolates of powdery mildew. Eight lines (9%), 255-3-3, 282-3-4, 286-1-1, 294-2-3,294-2-4, 295-1-2, 308-1-2 and 327-2-1, selected from 7 landraces showed resistance to all isolates. Seventy-eight lines (90%) showed a resistant infection type 2with more than 50% of the isolates used. In most of the selected lines (86%) unknown genes, alone or in combination with known specific resistance genes, were detected. Four different resistance alleles (Mlat, Mla6, Mla14 and Mla1) were postulated to be present in the tested lines. The most common was Mlat, which was postulated in 35 (41%) lines. The use of newly identified sources of powdery mildew resistance in barley breeding is discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Pathogenic variation among isolates of Rhynchosporium secalis from barley grass growing in South Eastern Australia

Summary The pathogenicity of 182 single spore isolates of Rhynchosporium secalis from Hordeum leporinum and 94 isolates from H. vulgare collected from throughout southeastern Australia was tested on 15 barley varieties, each having different combinations of resistance genes. Forty five percent of the barley grass isolates were pathogenic on 5 or more varieties but only 6% of the cultivated barley isolates attacked this range of varieties. On the basis of reaction type 20 different pathogenicity groups were recognised, with barley grass isolates being classified into 19 and the cultivated barley isolates into 5, four of which were the same as the barley grass isolates. Numerical analysis of data on leaf area damage inferred 33 groups, 24 of which were unique to barley grass isolates, two to cultivated barley isolates and 9 common to both groups. There was as much variation in pathogenicity among single spore isolates from the same lesion as between isolates from different lesions collected from the same or different locations.     

Variation in virulence and resistance in the bean-bean rust pathosystem in Ecuador

Twenty two Uromyces appendiculatus isolates were tested on 20 differential and 25 Ecuadorian Phaseolus vulgaris cultivars in the seedling stage. Based on the infection types 20 races could be discerned. The Ecuadorian cultivars differed greatly in their reaction to the isolates, from resistant to only one isolate (`Red Small Garden') to resistant to all isolates (’G2333‘). The isolates showed a wide range of virulence to the Ecuadorian cultivars, from virulent to only two cultivars (isolate 13) to virulent to 21 cultivars (isolates 5 and 23). Seven cultivars with a basically susceptible infection type appeared to differ greatly in quantitative resistance when tested in three consecutive crop cycles. The disease severities in percentage leaf area affected averaged over the three cycles ranged between 83.9% for ‘Red Small Garden’ and 13.1% for ‘INIAP-414’. Race-specific resistance does not seem an advisable breeding strategy, but the quantitative resistance offers a good alternative.