Susceptibility of European bread and durum wheat cultivars to Tilletia indica

Representative European wheat cultivars were tested under quarantine containment for their susceptibility to Tilletia indica, the cause of Karnal bunt of wheat. Fifteen winter and 15 spring wheat ( Triticum aestivum ) and 11 durum wheat ( Triticum durum ) cultivars were inoculated by boot injection just prior to ear emergence to test their physiological susceptibility. Selected cultivars were then re-tested by spray inoculation after ear emergence to determine their morphological susceptibility, which is a better predictor of field susceptibility. At maturity, the ears and seeds were assessed for incidence and severity of disease. For the physiological susceptibility tests, 13/15 winter wheat cultivars were infected and the percentage of infected seeds ranged from 1 to 32%. For spring cultivars, 13/15 cultivars were infected and the percentage of infected seeds ranged from 1 to 48%. For the durum cultivars, 9/11 were infected and the percentage of infected seeds ranged from 2 to 95%. Across all cultivars, 35/41 were infected. Based on historical Karnal bunt susceptibility categories using coefficients of infection, one cultivar was classed as highly susceptible, three as susceptible, 11 as moderately susceptible, 20 as resistant and only six as highly resistant. The spray-inoculation morphological susceptibility tests broadly confirmed the physiological susceptibility results, although lower levels of infection were observed. Overall, the range of susceptibility was similar to that found in cultivars grown in Karnal bunt affected countries. The results demonstrate that European wheat cultivars are susceptible to T. indica and thus could potentially support the establishment of T. indica if introduced into Europe.     

Susceptibility of Wheat to Tilletia indica During Stages of Spike Development

ABSTRACT Karnal bunt of wheat is caused by the fungus Tilletia indica, which partially converts kernels into sori filled with teliospores. Despite minor overall yield and quality losses, the disease is of considerable international quarantine concern. Plant development stages reported susceptible to infection vary considerably. A study was designed to better define the susceptibility period by inoculating wheat spikes at different growth stages with naturally liberated secondary sporidia under optimal conditions for disease development. Spikes of a resistant and susceptible cultivar were inoculated at eight growth stages from awns emerging to soft dough. Spikes became susceptible only after emerging from the boot and continued to be susceptible up to soft dough stage at which low levels of disease occurred. Disease severity in both cultivars peaked when spikes were inoculated after complete emergence, but before the onset of anthesis. Disease levels tapered off gradually in spikes inoculated after anthesis. The results broaden the known susceptibility period of wheat to T. indica to include stages long after anthesis, and indicate that infection from airborne inoculum is not possible during boot or awns emerging stages, which are commonly referred to as the most susceptible stages.     

Towards a more reasoned assessment of the threat to wheat crops from <Emphasis Type="Italic">Tilletia indica</Emphasis>, the cause of Karnal bunt disease

The interpretation of information used to defend an assessment that T. indica, the cause of Karnal bunt of wheat, has a high risk of establishing in Europe and of causing significant yield/quality losses is questioned. Karnal bunt has only established in locations that are arid or semi-arid with hot summers and cool/mild winters. There is very strong circumstantial evidence that substantial amounts of seed contaminated with teliospores of T. indica were sown in Europe in the past without the appearance of Karnal bunt. It is unlikely that sufficient numbers of teliospores would survive long enough on the soil surface under European conditions and then synchronise germination during the period at heading when wheat is vulnerable to infection to guarantee disease expression. Karnal bunt is regarded as a minor disease everywhere it occurs. Almost two thirds of European wheat cultivars inoculated by a severe boot injection method have been categorised as either resistant or highly resistant to T. indica. Yield/quality losses would be expected to be low even if the pathogen were capable of establishing in Europe. The status of T. indica as an important quarantine pest is based on the indirect economic consequences of the appearance of the pathogen and not on the direct damage it causes to wheat crops. Arguments in this and previous reviews advocating a more reasoned and comprehensive assessment of the threat to Europe, North America and other locations from T. indica need to be taken into consideration in any new pest risk analyses. David Jones is a retired employee of the Central Science Laboratory (CSL). Statements and opinions expressed in this paper are those of the author and are not the views of CSL.     

Emmer wheat, a potential new host of Tilletia indica

Two representative Italian emmer wheat (Triticum dicoccum) landraces, two selected lines and three improved emmer wheat cultivars, derived from crosses with durum wheat (Molisano landrace × ‘Simeto’), were tested for their susceptibility to Tilletia indica, the cause of Karnal bunt of wheat. Plants of emmer wheat were inoculated by injecting allantoid sporidial suspensions into the boot cavity of plants, just prior to ear emergence. A highly susceptible Indian spring wheat cultivar (Triticum aestivum) was used as a comparative control. At maturity of the plants, the seeds were harvested and assessed for incidence and severity of disease. All emmer wheat genotypes tested were infected but showed differing levels of susceptibility. The percentage of infected seeds for individual genotypes ranged from 5.4 to 75.0% compared with 99.1% for WL-711. The severity of infection was less in the old landraces, but it was higher in all the improved emmer wheat cultivars. In conclusion, Italian cultivars of emmer wheat were found to be highly susceptible to T. indica, and are potentially able to support the establishment of the pathogen.Authors L. Riccioni and M. Valvassori contributed equally to this work and should both be considered as first author.     

Identification and Differentiation of Tilletia indica and T. walkeri Using the Polymerase Chain Reaction

ABSTRACT Karnal bunt of wheat, caused by Tilletia indica, was found in regions of the southwestern United States in 1996. Yield losses due to Karnal bunt are slight, and the greatest threat of Karnal bunt to the U.S. wheat industry is the loss of its export market. Many countries either prohibit or restrict wheat imports from countries with Karnal bunt. In 1997, teliospores morphologically resembling T. indica were isolated from bunted ryegrass seeds and wheat seed washes. Previously developed PCR assays failed to differentiate T. indica from the recently discovered ryegrass pathogen, T. walkeri. The nucleotide sequence of a 2.3 kb region of mitochondrial DNA, previously amplified by PCR only from T. indica, was determined for three isolates of T. indica and three isolates of T. walkeri. There was greater than 99% identity within either the T. indica group or the T. walkeri group of isolates, whereas there was =3% divergence between isolates of these two Tilletia species. Five sets of PCR primers were made specific to T. indica, and three sets were designed specifically for T. walkeri based upon nucleotide differences within the mitochondrial DNA region. In addition, a 212 bp amplicon was developed as a target sequence in a fluorogenic 5' nuclease PCR assay using the TaqMan system for the detection and discrimination of T. indica and T. walkeri.     

基于MaxEnt模型的小麦印度腥黑穗病在中国的适生性分析

本文用MaxEnt软件对小麦印度腥黑穗病(Karnalbunt)在我国的潜在分布进行了预测。结果表明,小麦印度腥黑穗病在中国适生范围广。根据适生值的大小,中风险区和高风险区主要分布在我国华北、黄淮海、西南、西北、长江中下游和东北地区的小麦主产区,加强这些区域的调查监测和检疫对保护我国的小麦生产有重要意义。     

Rainfall and temperature distinguish between Karnal bunt positive and negative years in wheat fields in Texas

Karnal bunt of wheat, caused by the fungus Tilletia indica, is an internationally regulated disease. Since its first detection in central Texas in 1997, regions in which the disease was detected have been under strict federal quarantine regulations resulting in significant economic losses. A study was conducted to determine the effect of weather factors on incidence of the disease since its first detection in Texas. Weather variables (temperature and rainfall amount and frequency) were collected and used as predictors in discriminant analysis for classifying bunt-positive and -negative fields using incidence data for 1997 and 2000 to 2003 in San Saba County. Rainfall amount and frequency were obtained from radar (Doppler radar) measurements. The three weather variables correctly classified 100% of the cases into bunt-positive or -negative fields during the specific period overlapping the stage of wheat susceptibility (boot to soft dough) in the region. A linear discriminant-function model then was developed for use in classification of new weather variables into the bunt occurrence groups (+ or -). The model was evaluated using weather data for 2004 to 2006 for San Saba area (central Texas), and data for 2001 and 2002 for Olney area (north-central Texas). The model correctly predicted bunt occurrence in all cases except for the year 2004. The model was also evaluated for site-specific prediction of the disease using radar rainfall data and in most cases provided similar results as the regional level evaluation. The humid thermal index (HTI) model (widely used for assessing risk of Karnal bunt) agreed with our model in all cases in the regional level evaluation, including the year 2004 for the San Saba area, except for the Olney area where it incorrectly predicted weather conditions in 2001 as unfavorable. The current model has a potential to be used in a spray advisory program in regulated wheat fields.     

DArTseq physical mapping of QTLs linked to Karnal bunt (Tilletia indica) resistance in two historical wheat populations

Karnal bunt, a disease of wheat, durum, rye, and triticale, is subject to strict quarantine restrictions worldwide. The disease is considered a major threat to food security, due to its use as a non-tariff trade barrier by several wheat-importing countries. In this paper, we analysed seven years of phenotypic data to search for quantitative trait loci (QTLs) associated with resistance in common wheat, validated the QTLs using an independent population, and assessed the potential of genomic selection as a tool for pre-emptive breeding. The QTL study used phenotypic data collected from artificially inoculated field experiments involving two historical Karnal bunt resistance populations: WH542 × HD29 and WH542 × W485. QTL analyses detected four significant (p < 0.001) QTLs on chromosomes 1A, 3A, 4B, and 6B, which explained between 13.7% and 15.7% of the phenotypic variation. A panel of 130 cultivars was used to validate QTL effects. These were genotyped with the same DArTseq protocol, and two of the four QTLs were significantly (p < 0.001) associated with Karnal bunt resistance in the validation panel. The potential of genomic selection was investigated by comparing accuracies of a model trained with all available markers and a model based solely on validated QTL information from the biparental populations. Genomic prediction accuracy, based on the two scenarios, averaged 0.43 and 0.33, respectively, suggesting that even in situations where phenotyping is difficult due to quarantine restrictions, the prospects for pre-emptive breeding against Karnal bunt are encouraging, and resources are now available that will reduce the cost burden.     

Association study of resistance to Soilborne wheat mosaic virus in U.S. winter wheat

Soilborne wheat mosaic virus (SBWMV) is one of the most important winter wheat pathogens worldwide. To identify genes for resistance to the virus in U.S. winter wheat, association study was conducted using a selected panel of 205 elite experimental lines and cultivars from U.S. hard and soft winter wheat breeding programs. Virus symptoms were evaluated twice in virus-infected fields for the panel at Manhattan, KS in spring 2010 and 2011 and for a subpanel of 137 hard winter wheat accessions at Stillwater, OK in spring 2008. At the two locations, 69.8 and 79.5% of cultivars were resistant or moderately resistant to the disease, respectively. After 282 simple-sequence repeat markers covering all wheat chromosome arms were scanned for association in the panel, marker Xgwm469 on the long arm of chromosome 5D (5DL) showed a significant association with the disease rating. Three alleles (Xgwm469-165bp, -167bp, and -169bp) were associated with resistance and the null allele was associated with susceptibility. Correlations between the marker and the disease rating were highly significant (0.80 in Manhattan at P < 0.0001 and 0.63 in Stillwater at P < 0.0001). The alleles Xgwm469-165bp and Xgwm469-169bp were present mainly in the hard winter wheat group, whereas allele Xgwm469-167bp was predominant in the soft winter wheat. The 169 bp allele can be traced back to 'Newton', and the 165 bp allele to Aegilops tauschii. In addition, a novel locus on the short arm of chromosome 4D (4DS) was also identified to associate with the disease rating. Marker Xgwm469-5DL is closely linked to SBWMV resistance and highly polymorphic across the winter wheat accessions sampled in the study and, thus, should be useful in marker-assisted selection in U.S. winter wheat.     

Meta-analysis of regression coefficients for the relationship between fusarium head blight and deoxynivalenol content of wheat

ABSTRACT A total of 126 field studies reporting deoxynivalenol (DON; ppm) content of harvested wheat grain and Fusarium head blight index (IND; field or plot-level disease severity) were analyzed to determine the overall mean regression slope and intercept for the relationship between DON and IND, and the influence of study-specific variables on the slope and intercept. A separate linear regression analysis was performed to determine the slope and intercept for each study followed by a meta-analysis of the regression coefficients from all studies. Between-study variances were significantly (P < 0.05) greater than 0, indicating substantial variation in the relationship between the variables. Regression slopes and intercepts were between -0.27 and 1.48 ppm per unit IND and -10.55 to 32.75 ppm, respectively. The overall mean regression slope and intercept, 0.22 ppm per unit IND and 2.94 ppm, respectively, were significantly different from zero (P < 0.001), and the width of the 95% confidence interval was 0.07 ppm per unit IND for slope and 1.44 ppm for intercept. Both slope and intercept were significantly affected by wheat type (P < 0.05); the overall mean intercept was significantly higher in studies conducted using winter wheat cultivars than in studies conducted using spring wheat cultivars, whereas the overall mean slope was significantly higher in studies conducted using spring wheat cultivars than in winter wheat cultivars. Study location had a significant effect on the intercept (P < 0.05), with studies from U.S. winter wheat-growing region having the highest overall mean intercept followed by studies from Canadian wheat-growing regions and U.S. spring wheat-growing regions. The study-wide magnitude of DON and IND had significant effects on one or both of the regression coefficients, resulting in considerable reduction in between-study variances. This indicates that, at least indirectly, environment affected the relationship between DON and IND.     

Electron Microscopy in Biology

Abstract

Farmer surveys were conducted in the Yaqui Valley, Sonora, Mexico during 1983 and 1985 to investigate the relationship between agronomic practices used by the farmers and the incidence of Karnal bunt (Tilletia indica) disease of wheat. The variables investigated included soil type, land preparation, origin of seed, irrigation, nitrogen fertilizer, weed control and crop rotations. None of these variables were found to have a significant influence on the disease. This indicates that although management practices may have some effect on the incidence of the disease they are not the primary factor. The coincidence of rainfall and/or high humidity at flowering time appears to be of more importance than agronomic practices.     

Differentiation of citrus bacterial canker strains in Korea by host range,rep-PCR fingerprinting and 16S rDNA analysis

Soil-borne cereal mosaic virus (SBCMV) causes a severe disease in susceptible cultivars of winter wheat. The virus is vectored by the soil-borne protist Polymyxa graminis. Experiments were conducted to investigate whether SBCMV RNA2 could persist in seed from SBCMV-infected susceptible cultivars of winter wheat. Over 7,000 seedlings were generated from seed collected from two cultivars of SBCMV-infected winter wheat. Seedlings were grown in a glasshouse compartment and batch tested for the presence of SBCMV using real-time RT-PCR. The majority of batches tested positive for SBCMV, indicating an RNA2 transmission rate of 1.8–9.4% in wheat. The presence of the virus was confirmed by amplifying and sequencing a larger (400 bp) fragment of viral RNA2 in a sub-set of the seedlings testing positive by real-time RT-PCR. Root extracts from this sub-set tested negative for P. graminis using real-time PCR. The implications for disease epidemiology of this virus are discussed. The authors are British Civil Servants and as such their work is subject to British Crown Copyright. This means the exclusive copyright for the article cannot be transferred.     

河南省主要推广品种对小麦黄花叶病毒抗性的评价

为了评价河南省主要推广品种对小麦黄花叶病毒(Wheat yellow mosaic virus, WYMV)的抗性,于2006—2010年在河南省西平县病圃进行了田间抗性鉴定试验和室内间接ELISA检测,并分析了病害严重程度对产量的影响。结果表明,在供试的62个品种中,仅有新麦208表现为免疫;豫麦70-36、泛麦5号、阜麦936、山东95519、豫麦70、高优503、豫麦9676、郑麦366和陕麦229等9个品种表现为抗病,占供试品种的14.5%;濮优938、兰考矮早8、新原958、花培2号、温优1号、豫麦18、郑麦9023、豫麦47、豫农201、偃展4110、豫麦36、百农878和豫麦49-198等13个品种表现为中抗,占供试品种的21.0%;另外39个品种表现为感病,占供试品种的62.9%。对48个品种进行了产量与病害严重度分析,发现随着病害的严重度增加,小麦的穗数、千粒重以及产量都有明显下降,严重度为1级时,平均减产9.6%;严重度达到2级和3级时,平均减产分别为30.3%和33.5%。     

Disease levels in winter wheat,rye and triticale grown on soil artificially inoculated withCephalosporium gramineum

Field experiments with winter cereals grown on soil inoculated withC. gramineum showed that wheat and rye cultivars possess some resistance to the pathogen, while the triticale cultivars were the most susceptible. Higher tolerance of the tested wheat cultivars was connected mainly with slow development of disease symptoms; rye cultivars had, on average, lower percentages of plants infected byC. gramineum. The greatest variation in susceptibility toC. gramineum occurred among the selected cultivars of triticale.     

Organic Seed-treatment as a Substitute for Chemical Seed-treatment to Control Common Bunt of Wheat

Common bunt of wheat, caused by Tilletia laevis and T. tritici, is a major seed and soil-borne disease in West Asia and North Africa. The use of resistant cultivars and chemical seed-treatments are the current control measures used to combat this disease. The aim of this study was to investigate alternative control measures to chemical seed-treatments that are environmentally friendly to support cultivar resistance. Several organic nutrients [skimmed milk powder, hucket (local skimmed milk) and wheat flour] at a concentration of 160g per kg of seeds were used as seed-treatments on two susceptible bread and durum wheat cultivars (Bau and Sebou, respectively) to examine their effectiveness in controlling the disease. Field trials over four years showed that skimmed milk powder, hucket, and wheat flour reduced common bunt infection levels by 96%, 93% and 62%, respectively. In most cases, the effectiveness of the skimmed milk powder and hucket was equal to the chemical seed-treatment; thus, these organic nutrients offer an effective and environmentally safe alternative to chemical treatments. However, a study of their economic value as well as of their effect on seed germination, and field emergence is needed.     

Effect of seed treatment in preventing transmission of dwarf bunt of winter wheat to new areas

The effectiveness of various fungicide formulations containing carboxin and thiabendazole in eradicating teliospores of Tilletia controversa (dwarf bunt) from winter wheat seed was evaluated in the laboratory and the field. In the laboratory tests, the viability of teliospores washed from treated infested seed was greatly reduced as compared to the viability of teliospores washed from untreated infested seed, suggesting that carboxin and thiabendazole were primarily fungicidal. Field tests in two different years were established to determine if teliospores from treated infested seed could infect nearby untreated noninfested seed. Formulations containing carboxin were highly effective in reducing infection of healthy seed. No dwarf bunt infected plants developed in either year from seed treated with ‘Vitaflo 250’. Furthermore, seedborne common bunt caused by Tilletia foetida was nearly eradicated by carboxin-containing formulations in one field test. Carboxin treatment of winter wheat seed infested with dwarf bunt teliospores will greatly reduce the danger of introducing this pathogen into new areas.     

Abscisic acid is a potent inhibitor of growth and sporidial formation inneovossia indica cultures: Dual mode of actionvia loss of polyamines and cellular turgidity

The inhibition of polyamine biosynthesis inNeovossia indica (Mitra) Mundkur by D,L— α-difluoromethylornithine (DFMO) caused an effective reduction of mycelial growth and sporidial production underin vitro conditions, which was reversed by ornithine application. Abscisic acid proved to be similarly effective, not only in inhibiting mycelial growth and sporidial formation but also the germination of teliospores, which constitute the primary inoculum of the pathogen. ABA-mediated inhibition resulted in decreased polyamine levels and loss of cellular turgidity of mycelial cultures. Scanning electron microscopy of ABA-treated cultures revealed extremely shrunken hyphae, in marked contrast to the turgid controls. It is suggested that the manipulation of ABA levels and/or tissue sensitivity in wheat could be a strategy to combat ‘Karnal’ bunt, the disease caused byN. indica.     

Teliospore Survival and Plant Quarantine Significance of Tilletia Indies (causal agent of Karnal bunt) Particularly in Relation to China

The influence of temperature, light and soil conditions on teliospore germination of Tilletia (Neovossia) Indica was investigated. The teliospores were germinated by incubation in an aqueous suspension in a concavity on a water agar surface and the germination percentage was scored directly under low magnification of the compound microscope. The optimal temperature range for germination was found to be 15 to 22°C. Germination was extremely low at 2°C, but total inhibition was only observed after prolonged exposure to 35®C (in darkness) or to - 18°C under dry conditions. Light was shown to have a stimulating effect on germination (artificial daylight was found to be better that near ultraviolet light). No stimulation was achieved by prolonging the illumination from 12 to 24 h. Teliospores from 5-year-old wheat samples were shown to be viable. The weather conditions of the various wheat- growing areas of China were compared with the results obtained for the survival of teliospores of T. indica. It is concluded that a considerable risk exists for establishment and spread of the Karnal bunt disease if first accidentally introduced into China. Strict quarantine precautions, and policies can minimize or even eliminate this risk.     

Seed-borne Pathogens on Wheat (Particularly Smuts) in Turkey1

The important seed-borne diseases of wheat in Turkey are common bunt ( Tilletia caries and T. foetida ), loose smut ( Ustilago nuda tritici ), dwarf bunt ( T. contraversa ) and flag smut ( Urocystis tritici ). Seed treatment against common bunt is widely used in Turkey. The damage is about 10–15 %, sometimes 90 % in some fields where seeds are sown without treatment. About 10 % of wheat fields were infected with bunt during 1972–1974. Seed treatment with carboxin (Vitavax) is recommended against loose smut of wheat in Turkey, but has not yet been used. Some wheat varieties, especially Siirak (1593/51) wheat, are highly susceptible to loose smut. The proportion of smut-infected heads reached 30–40 % in this variety and about 32 % of wheat fields were infected with loose smut during 1972–1974. The damage was about 6 % in infected fields. Yield losses were at least 35,000 tons. Dwarf bunt is distributed at high altitude (1300–2000 m) in eastern Turkey; 2500 ha of wheat fields are infected with dwarf bunt. The disease causes 1–80 % losses in some fields. The wheat variety Yayla 305 is recommended for infected areas. Flag smut is present in some areas of Turkey, but yield losses are not important. Seed treatment is effective against seed-borne spores.     

Ultrastructural and immunocytochemical investigation of pathogen development and host responses in resistant and susceptible wheat spikes infected by Fusarium culmorum

Pathogen development and host responses in wheat spikes of resistant and susceptible cultivars infected by Fusarium culmorum causing Fusarium head blight (FHB), were investigated by means of electron microscopy as well as immunogold labelling techniques. The studies revealed similarities in the infection process and the initial spreading of the pathogen in wheat spikes between resistant and susceptible cultivars. However, the pathogen’s development was obviously more slow in the resistant cultivars as in comparison to a susceptible one. The structural defence reactions such as the formation of thick layered appositions and large papillae were essentially more pronounced in the infected host tissues of the resistant cultivars, than in the susceptible one. β -1,3-glucan was detected in the appositions and papillae. Furthermore, immunogold labelling of lignin demonstrated that there were no differences in the lignin contents of the wheat spikes between susceptible and resistant cultivars regarding the uninoculated healthy tissue, but densities of lignin in host cell walls of the infected wheat spikes differed distinctly between resistant and susceptible cultivars. The lignin content in the cell walls of the infected tissues of the susceptible wheat cultivar increased slightly, while the lignin accumulated intensely in the host cell walls of the infected wheat spikes of the resistant cultivars. These findings indicate that lignin accumulation in the infected wheat spikes may play an important role in resistance to the spreading of the pathogen in the host tissues. Immunogold labelling of the Fusarium toxin DON in the infected lemma showed the same labelling patterns in the host tissues of resistant and susceptible cultivars. However, there were distinct differences in the toxin concentration between the tissues of the susceptible and resistant cultivars. At the early stage of infection, the labelling densities for DON in resistant cultivars were significantly lower than those in the susceptible one. The present study indicates that the FHB resistant cultivars are able to develop active defence reactions during infection and spreading of the pathogen in the host tissues. The lower accumulation of the toxin DON in the tissues of the infected spikes of resistant cultivars which results from the host’s defence mechanisms may allow more intensive defence responses to the pathogen by the host.