Virulence and Molecular Polymorphism in International Collections of the Wheat Leaf Rust Fungus Puccinia triticina

ABSTRACT Collections of Puccinia triticina, the wheat leaf rust fungus, were obtained from Great Britain, Slovakia, Israel, Germany, Australia, Italy, Spain, Hungary, South Africa, Uruguay, New Zealand, Brazil, Pakistan, Nepal, and eastern and western Canada. All single-uredinial isolates derived from the collections were tested for virulence polymorphism on 22 Thatcher wheat lines that are near-isogenic for leaf rust resistance genes. Based on virulence phenotype, selected isolates were also tested for randomly amplified polymorphic DNA (RAPD) using 11 primers. The national collections were placed into 11 groups based on previously established epidemiological zones. Among the 131 single-uredinial isolates, 105 virulence phenotypes and 82 RAPD phenotypes were described. In a modified analysis of variance, 26% of the virulence variation was due to differences in isolates between groups, with the remainder attributable to differences within groups. Of the RAPD variation, 36% was due to differences in isolates between groups. Clustering based on the average virulence distance (simple distance coefficient) within and between groups resulted in eight groups that differed significantly. Collections from Australia-New Zealand, Spain, Italy, and Britain did not differ significantly for virulence. Clustering of RAPD marker differences (1 - Dice coefficient) distinguished nine groups that differed significantly. Collections from Spain and Italy did not differ significantly for RAPD variation, neither did collections from western Canada and South America. Groups of isolates distinguished by avirulent/virulent infection types to wheat lines with resistance genes Lr1, Lr2a, Lr2c, and Lr3 also differed significantly for RAPD distance, showing a general relationship between virulence and RAPD phenotype. The results indicated that on a worldwide level collections of P. triticina differ for virulence and molecular backgrounds.     

43个中国小麦品种(系)抗叶锈性研究

 选用12个墨西哥叶锈菌生理小种对43个中国小麦品种(系)所携带的抗叶锈病基因进行了推导,在25个品种(系)中推导出6个抗叶锈基因Lr1,Lr10,Lr13,Lr14a,Lr16和Lr26,9个品种(系)对本试验所使有的12个叶锈菌生理小种都表现感病反应,另有9个品种(系)携带未知的抗叶锈基因。在墨西哥2个地点进行的田间成株期抗叶锈性试验表明,12个品种(系)表现慢叶锈性,在将来的抗病育种中有一定的应用价值。     

Evaluation of Wheat Leaf Rust Resistance Genes in 10 Wheat Genotypes

Journal of Plant Diseases and Protection - Wheat leaf rust (Puccinia triticina) is one of the most important plant diseases in the world, and growing resistant cultivars is the most economical,...     

PCR-based Assays to Assess Wheat Varietal Resistance to Blotch (Septoria Tritici and Stagonospora Nodorum) and Rust (Puccinia Striiformis and Puccinia Recondita) Diseases

A multiplex Polymerase Chain Reaction (PCR) assay was developed to detect and quantify four fungal foliar pathogens in wheat. For Septoria tritici (leaf blotch) and Stagonospora nodorum (leaf and glume blotch), the -tubulin gene was used as the target region. Diagnostic targets for Puccinia striiformis (stripe or yellow rust) and P. recondita (brown rust) were obtained from PCR products amplified with -tubulin primer sequences. Final primer sets were designed and selected after being tested against several fungi, and against DNA of infected and healthy wheat leaves. For detection of the four pathogens, PCR products of different sizes were amplified simultaneously, whereas no products were generated from wheat DNA or other non-target fungi tested. The presence of each of the diseases was wheat tissue- and cultivar specific. Using real-time PCR measurements with the fluorescent dye SYBR Green I, PCR-amplified products could be quantified individually, by reference to a standard curve generated by adding known amounts of target DNA. Infection levels for each of the diseases were measured in the flag leaf of 19 cultivars at Growth Stage (GS) 60–64 in both 1998 and 1999. The infection levels for the cultivars were ranked, and showed, with a few exceptions, a good correlation with the NIAB Recommended List for winter wheat, which is based on visual assessment of symptoms. With PCR, the presence of the different pathogens was accurately diagnosed and quantification of pre-symptomatic infection levels was possible. Although sampling and DNA detection methods need further optimisation, the results show that multiplex PCR and quantitative real-time PCR assays can be used in resistance screening to measure the interaction between different pathogens and their hosts at different growth stages, and in specific tissues. This should enable an earlier identification of specific resistance mechanisms in both early-stage breeding material and field trials.     

Performance and Mapping of Leaf Rust Resistance Transferred to Wheat from Triticum timopheevii subsp. armeniacum

ABSTRACT Host plant resistance is an economical and environmentally sound method of control of leaf rust caused by the fungus Puccinia triticina, which is one of the most serious diseases of wheat (Triticum aestivum) worldwide. Wild relatives of wheat, including the tetraploid T. timopheevii subsp. armeniacum, represent an important source of genes for resistance to leaf rust. The objectives of this study were to (i) evaluate the performance of leaf rust resistance genes previously transferred to wheat from three accessions of T. timopheevii subsp. armeniacum, (ii) determine inheritance and allelic relationship of the new leaf rust resistance genes, and (iii) determine the genetic map location of one of the T. timopheevii subsp. armeniacum-derived genes using microsatellite markers. The leaf rust resistance gene transferred to hexaploid wheat from accession TA 28 of T. timopheevii subsp. armeniacum exhibited slightly different infection types (ITs) to diverse races of leaf rust in inoculated tests of seedlings compared with the gene transferred from TA 870 and TA 874. High ITs were exhibited when seedlings of all the germ plasm lines were inoculated with P. triticina races MBRL and PNMQ. However, low ITs were observed on adult plants of all lines having the T. timopheevii subsp. armeniacum-derived genes for resistance in the field at locations in Kansas and Texas. Analysis of crosses between resistant germ plasm lines showed that accessions TA 870 and TA 874 donated the same gene for resistance to leaf rust and TA 28 donated an independent resistance gene. The gene donated to germ plasm line KS96WGRC36 from TA 870 of T. timopheevii subsp. armeniacum was linked to microsatellite markers Xgwm382 (6.7 cM) and Xgdm87 (9.4 cM) on wheat chromosome arm 2B long. This new leaf rust resistance gene is designated Lr50. It is the first named gene for leaf rust resistance transferred from wild timopheevi wheat and is the only Lr gene located on the long arm of wheat homoeologous group 2 chromosomes.     

Specificity of Prehaustorial Resistance to Puccinia hordei and to Two Inappropriate Rust Fungi in Barley

ABSTRACT To elucidate the specificity of prehaustorial resistance to inappropriate rust fungi, we studied two populations of recombinant inbred lines of barley that segregated for partial resistance (PR) to Puccinia hordei and for the resistance to the inappropriate rust species P. recondita f. sp. tritici and P. hordei-murini. PR to P. hordei is prehaustorial and nonhypersensitive, and its level can be assessed accurately by measuring the latent period of the fungus. The resistance to the inappropriate rust species is a combination of prehaustorial (nonhypersensitive) and posthaustorial (hypersensitive) mechanisms. The amount of nonhypersensitive, early abortion of P. recondita f. sp. tritici and P. hordei-murini sporelings reflects the degree of prehaustorial defense to the two inappropriate rust species. All lines showing a long latent period of P. hordei also had a relatively high level of early abortion of the growth of P. recondita f. sp. tritici and P. hordei-murini. This indicates that genes for PR to P. hordei are also effective against these two inappropriate rust species. The reverse was not necessarily true; some lines showing a high level of early abortion of P. recondita f. sp. tritici and P. hordei-murini had a low level of PR to P. hordei. Moreover, lines with a similar level of prehaustorial resistance to P. recondita f. sp. tritici could differ considerably in their prehaustorial resistance to P. hordei-murini. This indicates that genes for prehaustorial resistance may exhibit rust species specificity.     

Gene Action and Linkage of Avirulence Genes to DNA Markers in the Rust Fungus Puccinia graminis

ABSTRACT Two strains of the wheat stem rust fungus, Puccinia graminis f. sp. tritici, were crossed on barberry, and a single F(1) progeny strain was selfed. The parents, F(1), and 81 F(2) progeny were examined for virulence phenotypes on wheat differential cultivars carrying stem rust resistance (Sr) genes. For eight Sr differentials, phenotypic ratios are suggestive of single dominant avirulence genes AvrT6, AvrT8a, AvrT9a, AvrT10, AvrT21, AvrT28, AvrT30, and AvrTU. Avirulence on the Sr; (Sr 'fleck') differential showed phenotypic ratios of approximately 15:1, indicating epistatic interaction of two genes dominant for avirulence. Avirulence on Sr9d favored a 3:13 over a 1:3 ratio, possibly indicating two segregating genes-one dominant for avirulence and one dominant for avirulence inhibition. Linkage analysis of eight single dominant avirulence genes and 970 DNA markers identified DNA markers linked to each of these avirulence genes. The closest linkages between AvrT genes and DNA markers were between AvrT6 and the random amplified polymorphic DNA marker crl34-155 (6 centimorgans [cM]) AvrT8a and the amplified fragment length polymorphism marker eAC/mCT-197 (6 cM) and between AvrT9a and the amplified fragment length polymorphism marker eAC/mCT-184 (6 cM). AvrT10 and AvrTU are linked at distance of 9 cM.     

Identification and Molecular Mapping of a Gene in Wheat Conferring Resistance to Mycosphaerella graminicola

ABSTRACT Septoria tritici leaf blotch (STB), caused by the ascomycete Mycosphaerella graminicola (anamorph Septoria tritici), is an economically important disease of wheat. Breeding for resistance to STB is the most effective means to control this disease and can be facilitated through the use of molecular markers. However, molecular markers linked to most genes for resistance to STB are not yet available. This study was conducted to test for resistance in the parents of a standard wheat mapping population and to map any resistance genes identified. The population consisted of 130 F(10) recombinant-inbred lines (RILs) from a cross between the synthetic hexaploid wheat W7984 and cv. Opata 85. Genetic analysis indicated that a single major gene controls resistance to M. graminicola in this population. This putative resistance gene is now designated Stb8 and was mapped with respect to amplified fragment length polymorphism (AFLP) and microsatellite markers. An AFLP marker, EcoRI-ACG/MseI-CAG5, was linked in repulsion with the resistance gene at a distance of approximately 5.3 centimorgans (cM). Two flanking microsatellite markers, Xgwm146 and Xgwm577, were linked to the Stb8 gene on the long arm of wheat chromosome 7B at distances of 3.5 and 5.3 cM, respectively. The microsatellite markers identified in this study have potential for use in marker-assisted selection in breeding programs and for pyramiding of Stb8 with other genes for resistance to M. graminicola in wheat.     

Partial Resistance to Septoria Tritici Blotch (Mycosphaerella graminicola) in Wheat Cultivars Arina and Riband

ABSTRACT Partial resistance to Septoria tritici blotch (STB) and its inheritance were investigated in a doubled-haploid population of a cross between cvs. Arina and Riband. The former has good partial resistance whereas the latter is susceptible. In adult plant trials in polytunnels, STB disease scores were negatively correlated with heading date. Resistance was not specific to any of the three fungal isolates used in these tests. A quantitative trait locus (QTL) for partial resistance to STB was identified in Riband on chromosome 6B and is named QStb.psr-6B-1. No QTL controlling a major part of the Arina resistance was identified, suggesting that its resistance may be dispersed and polygenic. There was no correlation between the lines' mean disease scores at the seedling and adult stages, implying that partial resistance to STB is developmentally regulated. Seedling resistance to the isolate IPO323 was isolate-specific and controlled by a single gene in Arina, probably allelic with the Stb6 gene in cv. Flame that confers resistance to the same isolate. The implications of these results for wheat breeding programs are discussed.     

Quantitative Trait Loci in Sweet Corn Associated with Partial Resistance to Stewart's Wilt, Northern Corn Leaf Blight, and Common Rust

ABSTRACT Partial resistance to Stewart's wilt (Erwina stewartii, syn. Pantoea stewartii), northern corn leaf blight (NCLB) (Exserohilum turcicum), and common rust (Puccinia sorghi) was observed in an F(2:3) population developed from a cross between the inbred sweet corn lines IL731a and W6786. The objective of this study was to identify quantitative trait loci (QTL) associated with partial resistance using restriction fragment length polymorphic markers. Phenotypic data were collected for 2 years for Stewart's wilt, NCLB, and common rust but, due to significant family-environment interaction, analysis was conducted individually on data from each year. In 2 years of evaluation for the three diseases, a total of 33 regions in the maize genome were associated with partial resistance describing from 5.9 to 18% of the total phenotypic variability. Of six regions common in both years, three were associated with partial resistance to Stewart's wilt (chromosomes 4:07, 5:03, and 6:04), one was associated with NCLB (chromosome 9:05), and two were associated with common rust (chromosomes 2:04 and 3:04). The rust QTL on 3S mapped to within 20 cM of the rp3 locus and explained 17.7% of the phenotypic variability. Some of the QTL associated with partial resistance to the three diseases have been reported previously, and some are described here for the first time. Results suggest it may be possible to consolidate QTL from various elite backgrounds in a manner analogous to the pyramiding of major resistance genes. We also report here on two QTL associated with anthocyanin production on chromosomes 10:6 and 5:03 in the general location of the a2 gene.     

Roles of peroxidases and NADPH oxidases in the oxidative response of wheat (Triticum aestivum) to brown rust (Puccinia triticina) infection

The goal of this work was to establish which enzymes – peroxidases or NADPH oxidases – play the most important role in the resistance‐related oxidative burst response of wheat to infection by brown rust (Puccinia triticina). The expression of four peroxidases and two NADPH oxidases was analysed in the susceptible wheat cv. Thatcher and isogenic lines with different Lr resistance genes after pathogen inoculation. Of the peroxidases, TaPrx118 and TaPrx112 were induced several times more strongly than TaPrx103 and TaPrx107. The induction of peroxidases was more pronounced than that of NADPH oxidases. The patterns of peroxidase expression clearly differentiated moderately resistant from highly resistant lines and corresponded to oxidative response profiles. The possible involvement of peroxidases or NADPH oxidases was verified with enzyme‐specific inhibitors. The oxidative burst in the susceptible cv. Thatcher and in the lines TcLr24, TcLr25, TcLr9 was peroxidase‐dependent, while the response in line TcLr26 was NADPH‐oxidase‐dependent. It is postulated that class III peroxidases play a leading role in the formation of reactive oxygen species molecules during the response of wheat to pathogen infection. The results suggest a high level of redundancy of some peroxidase genes induced in biotic stress. The role of both enzyme systems in wheat response/resistance to brown rust is discussed in relation to the oxidative response, the efficiency of resistance, and the presence and origin of particular Lr resistance genes.     

Effect of host genotype on leaf rust (Puccinia triticina) lesion development and urediniospore production in wheat seedlings

Experiments were performed in controlled conditions to compare several aggressiveness components of an isolate of Puccinia triticina on different wheat ( Triticum aestivum ) genotypes. Latency period, urediniospore production per lesion and per unit of sporulating surface, lesion size, and nitrogen and carbon content of the spores were measured on four host genotypes. Three of the host cultivars (Soissons, Altria and Isengrain) were susceptible and the fourth (Trémie) was resistant to the isolate used in the experiments, producing a mesothetic infection type. In all the analyses, lesion density was used as a covariable. The latent period was identical on the susceptible host cultivars, and there were no marked differences in the urediniospore production capacity when related to the sporulating surface area. In cv. Isengrain, differences in lesion size relative to the other susceptible cultivars resulted in lower urediniospore production per lesion. Urediniospore production per lesion and lesion size were density dependent and decreased exponentially with increasing lesion density. Urediniospore production per unit of sporulating surface was either independent of lesion density or marginally dependent (decreased linearly with a limited rate). On the resistant cultivar, Trémie, the latent period was longer and of a much greater variance compared with the susceptible cultivars. Urediniospore production per lesion was considerably reduced, but this reduction was fully accounted for by a reduction in lesion size, the urediniospore production capacity of the diseased tissue remaining equivalent to that observed in the susceptible cultivars. In all cultivars the C and N contents of the urediniospores were considered unchanged, even if minor differences were detected.     

Chromosomal Location of Genes for Resistance to Puccinia striiformis in the Wheat Line TP1295 Selected from the Cross of Soissonais-Desprez with Lemhi

Crosses of a wheat line TP1295 with the cultivar Chinese Spring monosomic series were used to locate, on chromosome 1D, a major gene for resistance to isolate WYR 85-22 of race 6E0 of Puccinia striiformis. The gene is designated as Yr25 and is probably present in several of the cultivars currently widely used for differentiating races of this pathogen. The expression of the gene was modified by the environment and by at least one minor gene which may be located on chromosome 6A. In F2 and F3 generations from a cross between TP1295 and euploid Chinese Spring, a wide range of variation in infection type (IT) was observed. This precluded the classification of the plants as either resistant or susceptible, so they were assigned to 6 classes and analyzed by factorial correspondence analysis and non-hierarchical classification. When all F3 plants in a family were fully resistant, like TP1295 itself (IT ;), both Yr25 and the modifying gene were assumed to be present and homozygous. In environments favourable to expression of the gene, families thought to carry Yr25 alone had a distribution of ITs from fully resistant (IT ;) to intermediate (IT 2, rarely 3 or 3+). This F3 analysis indicated that use of IT data alone, in the monosomic analysis, would not reveal the chromosomal location of the genes and that chromosome counting of numerous plants was necessary. As well as indicating the chromosomes carrying the genes for resistance to isolate WYR 85-22, the data showed that plants monosomic for chromosomes 5B and 5D were more resistant than the corresponding disomics, indicating that these chromosomes promote susceptibility and supporting other evidence of the effects of these chromosomes on yellow rust resistance.     

Identification and Molecular Mapping of a Gene Conferring Resistance to Pyrenophora tritici-repentis Race 3 in Tetraploid Wheat

ABSTRACT Race 3 of the fungus Pyrenophora tritici-repentis, causal agent of tan spot, induces differential symptoms in tetraploid and hexaploid wheat, causing necrosis and chlorosis, respectively. This study was conducted to examine the genetic control of resistance to necrosis induced by P. tritici-repentis race 3 and to map resistance genes identified in tetraploid wheat (Triticum turgidum). A mapping population of recombinant inbred lines (RILs) was developed from a cross between the resistant genotype T. tur-gidum no. 283 (PI 352519) and the susceptible durum cv. Coulter. Based on the reactions of the Langdon-T. dicoccoides (LDN[DIC]) disomic substitution lines, chromosomal location of the resistance genes was determined and further molecular mapping of the resistance genes for race 3 was conducted in 80 RILs of the cross T. turgidum no. 283/Coulter. Plants were inoculated at the two-leaf stage and disease reaction was assessed 8 days after inoculation based on lesion type. Disease reaction of the LDN(DIC) lines and molecular mapping on the T. turgidum no. 283/Coulter population indicated that the gene, designated tsn2, conditioning resistance to race 3 is located on the long arm of chromosome 3B. Genetic analysis of the F(2) generation and of the F(4:5) and F(6:7) families indicated that a single recessive gene controlled resistance to necrosis induced by race 3 in the cross studied.     

Genes for race-specific resistance to yellow rust (Puccinia striiformis) in Indian wheat cultivars

Seedlings of Indian wheat cultivars (Kalyansona, Sonalika, WL711 and eight others released commercially) were tested with 13 British and four alien races of Puccinia striiformis. The data indicated the probable presence of the resistance gene Yr2 in the three cultivars named above and in six of the others. Reactions of the remaining two cultivars, PWB12 and WL2265, were consistent with the presence of the gene 177. The presence of Yr2 in Kalyansona, Sonalika and WL711 was supported by evidence from crosses between them and with Heines VII, which is known to carry Yr2. In crosses of Sonalika with a susceptible cultivar, Kharchia Local and also with WL711, tests of F1, F2 and F3 generations indicated that, in addition to Yr2 , Sonalika possesses at least two other genes. Both these genes were difficult to detect but the F3 data supported the hypothesis that there is a single partially recessive gene giving resistance to alien race 6E16 and a different, possibly complementary, gene system effective against another alien race, 39E134. The presence of resistance in addition to Yr2 was also detected in WL711 and HD2329.     

Comparison of Resistance to Tomato Leaf Curl Virus (India) and Tomato Yellow Leaf Curl Virus (Israel) among Lycopersicon Wild Species, Breeding Lines and Hybrids

The objective of this study was to screen wild and domesticated tomatoes for resistance to Tomato yellow leaf curl virus, Israel (TYLCV-Is) and Tomato leaf curl virus from Bangalore isolate 4, India (ToLCV-[Ban4]) to find sources of resistance to both viruses. A total of 34 tomato genotypes resistant/tolerant to TYLCV-Is were screened for resistance to ToLCV-[Ban4] under glasshouse and field conditions at the University of Agricultural Sciences, Bangalore, India. Resistance was assessed by criteria like disease incidence, symptom severity and squash-blot hybridization. All the tomato genotypes inoculated with ToLCV-[Ban4] by the whitefly vector Bemisia tabaci (Gennadius) produced disease symptoms. In some plants of the lines 902 and 910, however, the virus was not detected by hybridization. The tomato genotypes susceptible to ToLCV-[Ban4] by whitefly-mediated inoculation were also found susceptible to the virus under field conditions. However, there were substantial differences between genotypes in disease incidence, spread, symptom severity and crop yield. Despite early disease incidence, many genotypes produced substantially higher yields than the local hybrid, Avinash-2. Sixteen tomato genotypes from India resistant/tolerant to ToLCV-[Ban4] were also tested for TYLCV-Is resistance at the Hebrew University of Jerusalem, Rehovot, Israel. Accessions of wild species, Lycopersicon hirsutum LA 1777 and PI 390659 were the best sources of resistance to both viruses. Lines 902 and 910, which were, resistant to TYLCV-Is were only tolerant to ToLCV-[Ban4] and accession Lycopersicon peruvianum CMV Sel. INRA, resistant to ToLCV-[Ban4], was only tolerant to TYLCV-Is. Implications of using the resistant lines in breeding programme is discussed.     

Identification, Chromosome Location, and Diagnostic Markers for a New Gene (YrCN19) for Resistance to Wheat Stripe Rust

ABSTRACT Several wheat lines and cultivars of wheat (Triticum aestivum) originating from the southwestern region of China were found to be highly resistant to stripe rust by inoculation with the prevalent races (CYR30, CYR31, and CYR32) and newly emerged races (H46-4, SY11-4 and SY11-14) of the pathogen. An inheritance study of the resistance to stripe rust was carried out by crossing resistant AIM6 with susceptible BeiZ76. Results indicated that the resistance to stripe rust was controlled by a single dominant gene. The 112 F(2) plants chosen from the cross BeiZ76/ AIM6 were analyzed with 218 pairs of microsatellite primers to determine the map location of the resistance gene. A simple sequence repeat marker on chromosome arm 2BS, Xgwm410, showed polymorphism and co-segregation between stripe rust resistant and susceptible plants. From the pedigree, inheritance, molecular marker, and resistance response, it is concluded that the stripe rust resistance gene in wheat cv. Chuan-nong19 (CN19) and wheat lines AIM5 and AIM6 is a novel gene, designated YrCN19. The microsatellite primer Xgwm410 is a diagnostic marker of the resistance gene YrCN19, which has potential for application in the marker-assisted breeding of wheat.