Maize Head Smut Research Articles

Transcriptome Analysis Reveals Contrasting Plant Responses of Sorghum bicolor upon Colonization by Two Formae Speciales of Sporisorium reilianum

The biotrophic fungus Sporisorium reilianum exists in two host-adapted formae speciales that cause head smut in maize (S. reilianum f. sp. zeae; SRZ) and sorghum (S. reilianum f. sp. reilianum; SRS). In sorghum, the spread of SRZ is limited to the leaves. To understand the plant responses to each forma specialis, we determined the transcriptome of sorghum leaves inoculated either with SRS or SRZ. Fungal inoculation led to gene expression rather than suppression in sorghum. SRZ induced a much greater number of genes than SRS. Each forma specialis induced a distinct set of plant genes. The SRZ-induced genes were involved in plant defense mainly at the plasma membrane and were associated with the Molecular Function Gene Ontology terms chitin binding, abscisic acid binding, protein phosphatase inhibitor activity, terpene synthase activity, chitinase activity, transmembrane transporter activity and signaling receptor activity. Specifically, we found an upregulation of the genes involved in phospholipid degradation and sphingolipid biosynthesis, suggesting that the lipid content of the plant plasma membrane may contribute to preventing the systemic spread of SRZ. In contrast, the colonization of sorghum with SRS increased the expression of the genes involved in the detoxification of cellular oxidants and in the unfolded protein response at the endoplasmic reticulum, as well as of the genes modifying the cuticle wax and lipid composition through the generation of alkanes and phytosterols. These results identified plant compartments that may have a function in resistance against SRZ (plasma membrane) and susceptibility towards SRS (endoplasmic reticulum) that need more attention in the future.

Identification of QTL for resistance to head smut in maize (Zea mays. L)

Head smut (HS) is one of the most devastating diseases of maize in spring production regions in China. Quantitative trait loci (QTL) for HS resistance were identified in this study to provide theoretical and applied tools for breeding HS resistance in maize. QTL associated with HS resistance were identified in a F2:3 population derived from a T32 (highly resistant genotype) × HC (highly susceptible genotype) cross. Analysis in each of three environments and a collective analysis across all three environments were used to identify QTL in the F2:3 population. A significant difference in HS resistance was found between the inbred lines, ‘T32’ and ‘HC’. Large genetic variation and transgressive segregation in the F2:3 population were observed between the three different sites, Guian (GA), Huaxi (HX), and Pingba (PB). Two stable and novel QTL for resistance to HS were detected in the different environments that were located within the bnlg1014 to umc2224 (qHS1) interval on chromosome 1 and in the umc1006 to umc1857 (qHS6) interval on chromosome 6. Both QTL can be used for further fine mapping, marker-assisted selection breeding, and theoretical studies on HS resistance in maize.

Gains in recurrent selection cycles for grain yield and resistance to head smut in maize

El basidiomiceto <em>Sporisorium reilianum</em> f. sp. <em>zeae</em> infecta al cultivo de maíz y causa importantes pérdidas en regiones donde existen condiciones propicias para el desarrollo de la enfermedad. En este trabajo se evaluó la respuesta a la selección de seis ciclos de selección recurrente de maíces de endospermo blanco y cinco ciclos de maíces de endospermo amarillo para caracteres agronómicos deseables, rendimiento de grano y resistencia al carbón de la espiga. Los diferentes ciclos en las dos poblaciones se obtuvieron siguiendo el método de selección recurrente de familias S1, los cuales fueron evaluados para determinar el avance genético. En un ensayo anexo, las poblaciones fueron inoculadas artificialmente para medir el progreso de la resistencia a la enfermedad y evaluar un nuevo método de inoculación. Se utilizó un diseño experimental de bloques completos al azar con tres repeticiones para el ensayo de rendimiento y dos repeticiones para el ensayo inoculado. Los ciclos evaluados en las dos poblaciones mostraron un incremento en la resistencia de 0.94 y 1.2% ciclo-1, una ganancia genética para rendimiento de grano de 272.9 y 620 kg ha-1 ciclo-1 para la población blanca y amarilla, respectivamente. Los resultados mostraron que la selección recurrente de familias S1 fue eficiente para mejorar el rendimiento de grano y resistencia al carbón de la espiga en ambas poblaciones.

Maize Head Smut: Pathogenesis, Epidemiology, and Management Options

Maize (Zea mays) head smut caused by Sphacelotheca reiliana, a basidiomycete with worldwide distribution, can cause devastating crop losses that pose a food security threat. While Kenya has experienced high incidences of the disease in the recent years, the seed certification regulation has a zero tolerance on S. reiliana. The spores of S. reiliana remain viable in soil for many years and serve as the main inoculum source as they germinate when the conditions are favourable and infect the host in the early stages of growth after germination. After penetration, the fungus grows systemically as the plant matures eventually transforming part or all of the inflorescence (ears and tassels) tissues into smut galls. The symptoms develop because the inflorescences have increased levels of reactive oxygen species, auxin, and misregulation of floral regulatory transcription factors. The most practical control strategy for maize head smut encompasses the use of resistant/tolerant cultivars, fungicide treatment (of seed or drenching of rows immediately after seeding), and field hygiene/ sanitation. Crop rotation may help when host crops are not planted for between 2-3 years or even more. Resistance genes, including ZmWAK, found in the major quantitative trait locus qHSR1/qHS2.09 regulate resistance of maize to head smut. The objective of this review paper is to provide an understanding of the head smut disease pathogenesis, epidemiology, and effective management options. Key words: Sphacelotheca reiliana, basidiomycete, seed certification, smut galls, resistance gene

Resistance to Sporisorium reilianum f. sp. zeae in native maize germplasm

El carbón de la espiga del maíz (<em>Sporisorium reilianum</em> f. sp. <em>zeae</em>) es una enfermedad que se caracteriza porque el patógeno remplaza las inflorescencias por soros llenos de teliosporas. El objetivo de este estudio fue investigar la respuesta en campo de 55 poblaciones de maíz nativo a la infección por <em>S. reilianum</em>. Las poblaciones de maíz se colectaron en los estados de Guerrero (13), Oaxaca (13), Puebla (seis), Tlaxcala (12) y Estado de México (11). La semilla fue inoculada con teliosporas del patógeno, usando grenetina como adherente. El híbrido Az 41801 se utilizó como testigo. Las poblaciones fueron evaluadas en Mixquiahuala, Hidalgo, en los ciclos 2015 y 2016. La incidencia de la enfermedad se registró mediante la observación directa de signos y síntomas en las inflorescencias. La incidencia máxima de la enfermedad en las poblaciones de maíz fue de 28.8% y 22.2% en la primera y segunda evaluación, respectivamente, mientras que el testigo (Az 41801) presentó una incidencia de 70.7% y 42.3%. Considerando el origen geográfico, los maíces del Estado de México y Tlaxcala presentaron una menor incidencia de la enfermedad respecto al resto de las poblaciones, lo que indica la presencia de genes de resistencia a la enfermedad.

Molecular Variation of the Phytopathogenic Fungus Sporisorium reilianum in Valle del Mezquital, Hidalgo

Head smut in maize (Zea mays) is a systemic disease caused by the phytopathogenic fungus Sporisorium reilianum. One of the most severely-affected regions in Mexico is Valle del Mezquital in the state of Hidalgo, a high production zone with irrigated agriculture that produces over 50% of maize supplies in the state. This study was designed to analyze the genetic diversity of this phytopathogen isolated from different zones of Valle del Mezquital using the following molecular markers: ITS, aspartyl protease, β-tubulin and the mitochondrial endonuclease LAGLIDADG. Our objective was to estimate the level of genetic variability in a regional space and understand the evolutionary processes to which non-model organisms are subjected. A total of 53 strains of S. reilianum were isolated from the sampling sites and then purified. A Bayesian Analysis of Population Structure (BAPS) analysis of the genetic structuring allowed us to recover three genetic groupings (K = 3), while molecular variance analysis showed a very low fixation index (FST = 0.0146), which was not statistically significant and indicated little between-group genetic differentiation. Also, the value determined for the coefficient of genetic differentiation (Gst) showed little variation among the genetic groups, while the gene flow number (Nm) was 8.25. The Tajima´s D estimator and haplotype network recently indicated a rapid population expansion. Together, these results suggest that reproduction and migration are key factors in the evolution and virulence of this fungus.

Screening of Secreted Proteins of Sporisorium reilianum f. sp. zeae for Cell Death Suppression in Nicotiana benthamiana.

Sporisorium reilianum f. sp. zeae (SRZ) is a biotrophic fungus causing head smut in maize. Maize infection with SRZ leads to very little cell death suggesting the presence of cell-death suppressinpg effectors. Several hundred effector proteins have been predicted based on genome annotation, genome comparison, and bioinformatic analysis. For only very few of these effectors, an involvement in virulence has been shown. In this work, we started to test a considerable subset of these predicted effector proteins for a possible function in suppressing cell death. We generated an expression library of 62 proteins of SRZ under the control of a strong constitutive plant promoter for delivery into plant cells via Agrobacterium tumefaciens-mediated transient transformation. Potential apoplastic effectors with high cysteine content were cloned with signal peptide while potential intracellular effectors were also cloned without signal peptide to ensure proper localization after expression in plant cells. After infiltration of Nicotiana benthamiana leaves, infiltration sites were evaluated for apparent signs of hypersensitive cell death in absence or presence of the elicitin INF1 of Phytophthora infestans. None of the tested candidates was able to induce cell death, and most were unable to suppress INF1-induced cell death. However, the screen revealed one predicted cytoplasmic effector (sr16441) of SRZ that was able to reliably suppress INF1-induced cell death when transiently expressed in N. benthamiana lacking its predicted secretion signal peptide. This way, we discovered a putative function for one new effector of SRZ.

Sporisorium reilianum possesses a pool of effector proteins that modulate virulence on maize.

The biotrophic maize head smut fungus Sporisorium reilianum is a close relative of the tumour-inducing maize smut fungus Ustilago maydis with a distinct disease aetiology. Maize infection with S.reilianum occurs at the seedling stage, but spores first form in inflorescences after a long endophytic growth phase. To identify S.reilianum-specific virulence effectors, we defined two gene sets by genome comparison with U.maydis and with the barley smut fungus Ustilago hordei. We tested virulence function by individual and cluster deletion analysis of 66 genes and by using a sensitive assay for virulence evaluation that considers both disease incidence (number of plants with a particular symptom) and disease severity (number and strength of symptoms displayed on any individual plant). Multiple deletion strains of S.reilianum lacking genes of either of the two sets (sr10057, sr10059, sr10079, sr10703, sr11815, sr14797 and clusters uni5-1, uni6-1, A1A2, A1, A2) were affected in virulence on the maize cultivar 'Gaspe Flint', but each of the individual gene deletions had only a modest impact on virulence. This indicates that the virulence of S.reilianum is determined by a complex repertoire of different effectors which each contribute incrementally to the aggressiveness of the pathogen.

SUPPRESSOR OF APICAL DOMINANCE1 of Sporisorium reilianum Modulates Inflorescence Branching Architecture in Maize and Arabidopsis.

The biotrophic fungus Sporisorium reilianum causes head smut of maize (Zea mays) after systemic plant colonization. Symptoms include the formation of multiple female inflorescences at subapical nodes of the stalk because of loss of apical dominance. By deletion analysis of cluster 19-1, the largest genomic divergence cluster in S. reilianum, we identified a secreted fungal effector responsible for S. reilianum-induced loss of apical dominance, which we named SUPPRESSOR OF APICAL DOMINANCE1 (SAD1). SAD1 transcript levels were highly up-regulated during biotrophic fungal growth in all infected plant tissues. SAD1-green fluorescent protein fusion proteins expressed by recombinant S. reilianum localized to the extracellular hyphal space. Transgenic Arabidopsis (Arabidopsis thaliana)-expressing green fluorescent protein-SAD1 displayed an increased number of secondary rosette-leaf branches. This suggests that SAD1 manipulates inflorescence branching architecture in maize and Arabidopsis through a conserved pathway. Using a yeast (Saccharomyces cerevisiae) two-hybrid library of S. reilianum-infected maize tissues, we identified potential plant interaction partners that had a predicted function in ubiquitination, signaling, and nuclear processes. Presence of SAD1 led to an increase of the transcript levels of the auxin transporter PIN-FORMED1 in the root and a reduction of the branching regulator TEOSINTE BRANCHED1 in the stalk. This indicates a role of SAD1 in regulation of apical dominance by modulation of branching through increasing transcript levels of the auxin transporter PIN1 and derepression of bud outgrowth.

Real-Time PCR Assay for Detection of Sphacelotheca reiliana Infection in Maize (Zea mays) Seedlings and Evaluation of Seed Treatment Efficacy.

Head smut of maize, caused by the fungus Sphacelotheca reiliana, is an economically important disease in all major maize-producing countries. Although seed treatments are commonly used for management purposes, evaluating these treatments for efficacy is both time consuming and inefficient. Therefore, in order to improve the capacity for evaluating seed treatment fungicides, we developed a real-time PCR-based seedling assay for S. reiliana infection. We optimized growth chamber conditions and inoculation methods to achieve infection incidence of 60 to 80% in inoculated, nontreated controls. The effects of five commercially available fungicidal seed treatment formulations on seedling infection incidence were compared by PCR analysis of root and mesocotyl tissues. Tebuconazole, fludioxonil, sedaxane, and Maxim Quattro (fludioxonil+mefenoxam+azoxystrobin+thiabendazole) reduced the incidence of infection (P < 0.05) compared with the control, but no difference was found between the azoxystrobin treatment and the control. All rates tested for both sedaxane and tebuconazole were equally effective for seeds coated with 106 teliospores∙seed-1. Sedaxane, at a rate of 0.1 mg/kernel, eliminated seedling infection if seeds were infested with a low inoculum concentration (101 teliospores∙seed-1). The assay developed here is a valuable tool not only for the detection of fungal infection at the seedling stage, but also for testing the relative efficacies of seed treatments for reducing incidence of infection.

ZmWAK, a quantitative resistance gene to head smut in maize, improves yield performance by reducing the endophytic pathogen Sporisorium reiliana

Head smut is a soil-borne systemic maize disease caused by the causal pathogen Sporisorium reiliana. ZmWAK is the gene within the major QTL qHSR1 conferring resistance to head smut disease. ZmWAK has been transferred by marker-assisted backcrossing into elite inbred lines with high-yield potential but susceptible to head smut. The converted inbred lines, together with their hybrids, were grown in 2012 and 2013 to investigate resistance performance, yield per plot, and yield-related traits. ZmWAK-mediated resistance to maize head smut follows a dominant genetic model, and a single ZmWAK allele could achieve high disease resistance to prevent yield loss from S. reiliana infection. Genetically, ZmWAK itself had no detectable negative impacts on yield-related traits. Instead, ZmWAK could indirectly improve most yield components, particularly ear-related traits, in symptomless plants by reducing the amount of endophytic S. reiliana. Overall, ZmWAK is beneficial to both head smut resistance and yield-related traits in maize.

A maize wall-associated kinase confers quantitative resistance to head smut.

Head smut is a systemic disease in maize caused by the soil-borne fungus Sporisorium reilianum that poses a grave threat to maize production worldwide. A major head smut quantitative resistance locus, qHSR1, has been detected on maize chromosome bin2.09. Here we report the map-based cloning of qHSR1 and the molecular mechanism of qHSR1-mediated resistance. Sequential fine mapping and transgenic complementation demonstrated that ZmWAK is the gene within qHSR1 conferring quantitative resistance to maize head smut. ZmWAK spans the plasma membrane, potentially serving as a receptor-like kinase to perceive and transduce extracellular signals. ZmWAK was highly expressed in the mesocotyl of seedlings where it arrested biotrophic growth of the endophytic S. reilianum. Impaired expression in the mesocotyl compromised ZmWAK-mediated resistance. Deletion of the ZmWAK locus appears to have occurred after domestication and spread among maize germplasm, and the ZmWAK kinase domain underwent functional constraints during maize evolution.

Analysis of gene expression profiles in response to Sporisorium reilianum f. sp. zeae in maize (Zea mays L.)

article i nfo Background: Head smut of maize, which is caused by Sporisorium reilianum f. sp. zeae (Kuhn), is a serious disease in maize. In order to reveal the molecular mechanism of the resistance to head smut in maize, a microarray containing ~14,850 probes was used to monitor the gene expression profiles between a disease resistant near isogenic line (NIL) and a highly susceptible inbred line after S. reilianum was injected with an artificial inoculation method. Results: Levels of expression for 3,532 genes accounting for 23.8% of the total probes changed after inoculation. Gene Ontology analysis revealed that the differentially expressed genes participated in physiological and biochemical pathways. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that plant-pathogen interaction, natural killer cell mediated cytotoxicity and benzoxazinoid biosynthesis pathways play important roles in resistance to head smut. Three head smut resistance-related candidate genes, CLAVATA1, bassinosteroid insensitive 1-associated receptor kinase 1 and LOC100217307 with leucine-rich repeat (LRR) conserved domains were identified, each of which is in maize mapping bin 2.09, a region previously shown to include a major QTL for head smut resistance. Furthermore, LOC100217307 was validated by quantitative real-time (qRT)-PCR inferring that this gene may be involved in the resistance to head smut of maize. Conclusions: This study provided valuable information for cloning, functional analysis and marker assisted breeding of head smut resistance genes.

Microencapsulation of seed-coating tebuconazole and its effects on physiology and biochemistry of maize seedlings

Tebuconazole is a triazole systemic fungicide that is commonly used to treat fungal pathogens of crops, but at high doses can reduce seed germination. Seeds with microcapsulated tebuconazole were investigated to determine effects of this method on maize seedlings and the bioefficacy against maize head smut (Sphacelotheca reiliana). The ethyl cellulose (EC)-based microcapsules had encapsulation efficiency of 90.6%, and average size of 1.6μm. A release kinetic study revealed that tebuconazole release from EC-based microcapsules fits the model (Mt/Mz=ktn+C). Glasshouse studies indicated that maize seedling emergence and growth were negatively affected in an exponential manner as predicted by model Y=A+B×e(−x/k). However, microencapsulation could induce tebuconazole's growth promoting effects by increasing emergence, shoot fresh weight, root fresh weight, carotenoid and chlorophyll content. Phytohormone analysis indicated the beneficial effects of microencapsulated tebuconazole were due to the sustained release of tebuconazole that appeared to influence the balance of phytohormones in maize seedlings. Contrary to conventional tebuconazole, microencapsulated seed-coated tebuconazole can lead to slightly increased gibberellins (GA) level and disappearance of abscisic acid (ABA) accumulation in maize. In addition, microcapsule formulation of tebuconazole was found to provide better protection against maize head smut when compared to conventional formulation.

Purification and Characterization of Xylanase SRXL1 from Sporisorium reilianum Grown in Submerged and Solid-State Fermentation

Sporisorium reilianum is a phytopathogenic fungus that produces the maize head smut. Hydrolytic enzymes such as xylanases have not been studied in this basidiomycete, although these enzymes are widely used within the industry. In this study, the characterization of a xylanase produced by S. reilianum grown in submerged and solid-state fermentation using different culture media was performed. Submerged fermentation with a medium containing birch xylan and yeast extract showed the highest xylanase activity (12.6 U/mL). The enzyme, purified through ion exchange chromatography, had a molecular weight of 42 kDa, as determined by SDS-PAGE, and a Km and Vmax of 1.72 mg/mL and 2.48 μmol/mL/min, respectively. The optimal values of pH and temperature were 5.0 and 70 °C, respectively. The enzyme showed activity over a broad range of temperatures and pHs. Zn2+, Fe3+, and Mn2+ ions increased xylanase activity. Sequence protein analysis showed 100% similarity with the theoretical protein encoded by the sr14403 gene of S. reilianum, encoding a putative endo-Beta-1,4-xylanase. This is the first report on the production and purification of a xylanase from this fungus, which has interesting biochemical characteristics for application in biotechnological processes.

Genome-wide association study (GWAS) of resistance to head smut in maize

Head smut, caused by the fungus Sphacelotheca reiliana (Kühn) Clint, is a devastating global disease in maize, leading to severe quality and yield loss each year. The present study is the first to conduct a genome-wide association study (GWAS) of head smut resistance using the Illumina MaizeSNP50 array. Out of 45,868 single nucleotide polymorphisms in a panel of 144 inbred lines, 18 novel candidate genes were associated with head smut resistance in maize. These candidate genes were classified into three groups, namely, resistance genes, disease response genes, and other genes with possible plant disease resistance functions. The data suggested a complicated molecular mechanism of maize resistance against S. reiliana. This study also suggested that GWAS is a useful approach for identifying causal genetic factors for head smut resistance in maize.

Transcriptomic analysis of Sporisorium reilianum in response to the strigolactone analogue GR24

A suppression subtractive hybridization (SSH) approach was used to generate cDNA libraries representing genes differentially expressed in the haploid cells of the maize head smut pathogen Sporisorium reilianum exposed to GR24, a strigolactone analogue. Strigolactones are present in root exudates and have been known to trigger germination of parasitic plant seeds and signal mycorrhizal fungi to connect to root systems forming mutualistic relationships. Cell respiration increased within 1 h after GR24 addition, but decreased after 5 and 8 h. All induced cells were used to construct a cDNA library, which contained 1440 clones. The cDNA ESTs were deposited on macro-array membranes and hybridized with P32 cDNA probes obtained from mRNA isolated from S. reilianum yeast cultures exposed or not to 100 nM GR24 for three time intervals. A total of 678 ESTs were identified as differentially expressed during three time-courses in response to GR24. A set of 36 candidate genes were analyzed by qRT-PCR that presented an induction of the genes at 1 h, confirming the hybridization data. Induced genes mostly affect catalysis functions (45%) like cell respiration (27%), and cell signaling (26%). Although the biological significance of this perception remains hypothetical, these results indicate that strigolactones could have a wider biological influence in the rhizosphere than previously recognized.

Characterization and molecular mapping of RsrR, a resistant gene to maize head smut

AbstractMaize head smut (MHS) caused by the fungi Sporisorium reilianum (Kühn) Landon and Fullerton (S. reilianum) is the most serious disease occurred in China recently. There are only a few reports concerning the genetics of this disease and the resistant gene of maize. In this paper a new dominant resistant gene to MHS caused by the pathogen Shenyang-1 of S. reilianum was discovered from a newly developed resistant inbred line ‘R24’ and was named RsrR. RsrR gene was molecular tagged and mapped on the long arm of maize chromosome 1 via simple sequence repeat-bulked segregant analysis (SSR-BSA) and sequence related amplified polymorphism (SRAP)-BSA analysis, the nearest linkage marker is 2.5 cM apart from the RsrR gene. The gene RsrR has been transferred into two elite maize inbred lines, ‘Huangzao4’ and ‘Qi319’, through traditional hybridization and marker assisted selection. The converted lines can be used in maize MHS-resistance breeding.

Effect of GR24, a synthetic analogue of strigolactones, on gene expression of solopathogenic strain of Sporisorium reilianum

Sporisorium reilianum f.sp zeae, a basidiomycetous fungus belonging to Ustilaginaceae, is the causal agent of maize head smut. Its pathogenicity is initiated by fusion of two compatible sporidia which give rise to the formation of a dikaryotic pathogenic hyphe. In addition, pathogenic dimorphic diploid strains called solopathogens can be formed where no mating occurs. Strigolactones are the trace molecules in plant root exudates perceived by some fungi at subnanomolar concentrations that have been implicated in inducing spore germination in fungi. Cell respiration in fungal cells was measured through polarography and fluorescence assay. GR24, a synthetic analogue of strigolactones, induced a burst of cell respiration 1 h after adding GR24 (10 -7 M) that gradually decreased at 5 and 8 h after addition of GR24. Quantitative polymerase chain reaction (qPCR) analysis showed that transcription levels of genes involved in cell respiration and a 12 kDa heat shock protein were up-regulated 1 h after the addition of GR24 to a culture of the solopathogenic strain but no influence was observed on the other pathogenicity genes and on the culture morphology. These results suggest that strigolactones influence the rhizosphere and play a role in plant-microbe interactions Key word: GR24, strigolactones, Sporisorium reilianum, solopathogen root exudates.

Development of Sequence Characterized Amplified Region (SCAR) Primers for the Detection of Resistance to Sporisorium reiliana in Maize

Head smut of maize ( Zea mays L.), which was caused by Sporisorium reiliana, occurred in most of the maize growing areas of the world. The purpose of this study was to develop SCAR markers for map-based cloning of resistance genes and MAS. Two sets of BC3 progenies, one (BC3Q) derived from the cross Qi319 (resistance) × Huangzao 4 (susceptible), the other (BC3M) from Mo17 (resistance) × Huangzao 4 (susceptible), were generated. Huangzao 4 was the recurrent parent in both progenies. A combination of BSA (bulked segregant analysis) with AFLP (amplified fragment length polymorphism) method was applied to map the genes involving the resistance to S. reiliana, and corresponding resistant and susceptible bulks and their parental lines were used for screening polymorphic AFLP primer pairs. One fragment of P13M61–152 was converted into SCAR (sequence charactered amplified fragment) marker S130. The marker was mapped at chromosome bin 2.09, the interval of a major QTL region previously reported to contribute to S. reiliana resistance. Furthermore, S130 was highly associated with resistance to S. reiliana, and could be useful for marker-assisted selection and facilitate map-based cloning of resistance genes.