Characteristics of strains of Septoria nodorum adapted to wheat or to barley

Sixty-four strains of Septoria which had been isolated from a range of hosts at different locations and in different years were characterized for their adaptation to wheat or barley, growth at near-maximum temperature, fluorescence, colony morphology, conidial (pycnidiospore) length and hexokinase and alkaline phosphatase isozymes. For each character except conidial length, the strains could be divided into two or three discrete groups. The variation in these six characters was strongly associated, such that 60 strains could be classified into two groups, designated W-type and B-type. W-type strains are adapted to wheat, produce large colonies at 31 C, fluoresce, produce brown-pigmented colonies, and have fast isozymes. B-type strains are adapted to barley, produce small colonies at 31 C, do not fluoresce, produce pink-pigmented colonies, and have slow isozymes. A few strains differed from these norms in one of the six characters, but only one showed an atypical host adaptation. The four unclassified strains differed from W- or B-type in two or more characters. The many differences between the W- and B-types suggest they are genetically distinct populations within Septoria nodorum.     

Adaptation of Septoria nodorum to wheat or barley on detached leaves

Single-spore cultures of wheat- and barley-adapted isolates of Septoria nodorum were serially passaged through detached leaves of wheat or barley. Rare instances of change in host adaptation were observed after passaging but associated changes in other characters, including heterokaryon compatibility, identified the changed isolates as contaminants. Inoculation of progressively more dilute spore suspensions to detached leaves revealed that some isolates were highly infective to the host to which they were adapted, with a high probability of a single spore initiating infection. Inocula deliberately contaminated with a small proportion of spores of opposite host adaptation revealed changes when passaged on wheat that mimicked the rare changes observed on passaging pure wheat- or barley-adapted isolates. These results suggest that adaptation to wheat or barley is a stable property of pure cultures of S. nodorum and that the occasional changes in host adaptation observed on passaging result from selection of contaminants.     

Response of wheat and barley isolates of Septoria nodorum to passage through barley and wheat cultivars

A single-spore wheat isolate of S. nodorum was passaged once through barley and a barley isolate passaged once through wheat. Reisolations from barley leaves were designated as either 'original-type wheat' (OTW) and resembled the wheat isolate used for inoculations in morphology, or 'sector-type barley' (STB) resembling the known morphology of barley isolates. Reisolations from wheat were similarly designated either OTB or STW. Sub-cultures of STB were significantly more pathogenic to barley in a glasshouse experiment than OTW, and vice versa. Similarly STW was significantly more pathogenic to wheat than OTB, and vice versa.
A single-spore wheat isolate was also passaged 0, 1, 2, 3 or 4 times through barley and a barley isolate passaged similarly through wheat. Disease assessment and yield data from a field experiment showed that the pathogenicity of the passaged wheat isolate remained largely unaltered from that of the original isolate. However, the barley isolate responded after the third wheat passage by causing significantly less disease and significantly higher yields in barley than the unpassaged isolate; conversely, on wheat, disease levels significantly increased and yield slgnficantly decreased after the second wheat passage.
Data obtained are discussed in the context of cultural observations made on colony morphology during host passage sequences and possible genetic mechanisms responsible.     

A method to test wheat leaves for their reactions to inoculation with Septoria species

The construction and application is described of a polystyrol humidity box in which wheat leaves, while continuing to function as parts of living plants, can be tested for their reactions toSeptoria spp. in an atmosphere nearly saturated with water, as is required for successful infection. The method is simple, accurate ans inexpensive.Samenvatting Voor dit doel is een z.g. vochtdoos geconstrueerd (Fig. 1A). Het te toetsen blad wordt daar doorheen geleid en in de doos geïnoculeerd met een druppel conidiën-suspensie van de schimmel. Onder het blad staat wat water in de doos (Fig. 1B). Na afsluiting ontstaat in de doos de hoge luchtvochtigheid van bijna 100% die nodig is voor de infectie. Dit wordt op deze wijze eenvoudig en goedkoop gerealiseerd. De bladeren die zo worden getoetst blijven onbeschadigd functioneren aan de plant. Na enkele dagen kan de doos worden geopend en kan de symptoom-ontwikkeling worden afgewacht en gevolgd (Fig. 3A, 3B en 4). De methode leent zich voor nauwkeurig werk en vereist zeer weinig infectiemateriaal. De Technische en Fysische Dienst voor de Landbouw (TFDL), Wageningen, ontwierp en construeerde een statief voor het gebruik van de vochtdozen in serie (Fig. 2).     

Yield reduction in wheat in relation to leaf necrosis caused by Septoria tritici

In 1980, field inoculations withSeptoria tritici were performed on winter wheat cv. Okapi. Leaf necrosis progress curves were established; they were sigmoid and could be transformed into logit lines. There was a great difference in the slope of the logit lines between inoculated plots and non-inoculated controls. The mid-time values varied according to inoculum dosage and post-inoculation humidity treatment. Yields (kg ha^?1) and average grain weights (mg grain^?1) differed significantly between inoculated and non-inoculated plots, but seldom among inoculation treatments. Decrease of average grain weight completely explained yield loss, which was 878 kg ha^?1 or 12% of control (7045 kg ha^?1). This crop loss is considerable for a cultivar which is not particularly susceptible. A good correlation was found between the amount of necrosis at development stages DC=75 and 77 and yield depression. The integral of necrosis over time gave a high correlation with loss of kernel weight.     

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.     

Inoculum density — host response relationships of spring wheat cultivars to infection by Septoria tritici

Eightinoculumdensities (ID), increasing in geometric progression from 0.1×10⁵ to 12.8×10⁵ conidia ml^–1 ofSeptoria tritici were sprayed onto the leaves of five cultivars ofTriticum aestivum, differing in resistance toS. tritici. Host response to infection byS. tritici, measured as either thelatencyperiod (LPD) orpercentage ofleafarea covered bylesions with pycnidia (PLAL), was significantly linearly correlated with log₁₀ID (r=0.84 to 0.96, with 6 degrees of freedom). As the ID increased from 0.2×10⁵ to 12.8×10⁵ conidia ml^–1, LPD decreased by 1 to 2 days for the susceptible and resistant cultivars and by 3 to 5 days for the moderately susceptible cultivars. Within the same ID range, PLAL increased from 73% to 90% for the susceptible cultivar and 2 to 12% for the resistant cultivar. The ID₅₀ value (the number of conidia required for 50% PLAL) calculated from the regression equations relating log₁₀ID to PLAL for each cultivar, ranged from 6.42×10³ conidia ml^–1 for the susceptible cultivar to 7.13×10¹³ conidia ml^–1 for the resistant cultivar. Estimates of the number of conidia ml^–1 required to initiate a lesion (obtained by substituting 1% PLAL into the regression equations relating log₁₀ ID to PLAL) ranged from 16 conidia ml^–1 for the susceptible cultivar to 1.0×10⁴ conidia ml^–1 for the resistant cultivar. Using the change of PLAL with ID, the restriction of lesion development for each cultivar could be partitioned into that due to host resistance and that due to interaction between lesions. Log₁₀LPD was significantly correlated negatively with PLAL. As a general rule when screening cultivars for resistance toS. tritici, it is advisable to inoculate them with several inoculum densities and to determine the latency period.Samenvatting Sporensuspensies vanSeptoria tritici die in dichtheid (ID) varieerden volgens een geometrische reeks van 0,1 tot 12,8×10⁵ sporen.ml^–1, werden gebruikt bij de inoculatie van kiemplanten van vijf cultivars vanTriticum aestivum die verschilden in resistentie tegenS. tritici.De reactie van de waardplanten werd gemeten aan de latentie periode (LPD) en aan het percentage bladoppervlak dat ingenomen werd door vlekken met pycniden (PLAL). Beide grootheden toonden een significante lineaire correlatie met log₁₀ID (r=0,84 tot 0,96 bij 6 vrijheidsgraden). Bij een toename van ID van 0,2×10⁵ tot 12,8×10⁵ sporen.ml^–1 nam de lengte van de latentie periode af met 1 tot 2 dagen bij de vatbare en de resistente cultivars, en met 3 tot 5 dagen bij de matig vatbare cultivars (Fig. 2).In hetzelfde traject van inoculumdichtheid nam de PLAL toe van 73 tot 90% bij vatbare cultivars, en van 2 tot 12% bij de resistente cultivars (Fig. 3). De ID₅₀-waarde, het aantal sporen dat nodig is om 50% aantasting te verkrijgen, werd berekend uit de regressievergelijkingen die voor iedere cultivar het verband aangeven tussen log₁₀ ID en het aantastingspercentage (PLAL). Deze waarde varieerde van 6,42×10³ sporen per ml voor vatbare tot 7,13×10³ sporen per ml voor de resistente cultivars (Tabel 1). Schattingen voor het aantal sporen dat nodig is om een vlek te doen ontstaan werden verkregen door de 1% aantastingswaarde te substitueren in de eerder genoemde regressievergelijkingen. Deze waarden varieerden van 16 sporen.ml^–1 voor de vatbare cultivar tot 10000 sporen.ml^–1 voor de resistente cultivar (Tabel 1). Via de toename van de aantastingsgraad (PLAL) per eenheid stijging in de inoculumdichtheid kon de remming op de vlekontwikkeling opgesplitst worden in een gedeelte dat toegeschreven werd aan de resistentie van de plant en een gedeelte dat te wijten was aan de interactie tussen de vlekken (Fig. 4 en 5; Tabel 2).Log₁₀ LPD was negatief gecorreleerd met PLAL (Fig. 6). Bij het toetsen van cultivars op resistentie tegenS. tritici zouden sporensuspensies van verschillende dichtheden gebruikt moeten worden en zou de latentie periode, evenals de aantastingsgraad bepaald moeten worden.     

Observations on the host range of an isolate of Septoria nodorum from wheat

An isolate ofS. nodorum from wheat was inoculated onto grasses in the field. Re-isolates from these grasses were tested in a cross-inoculation experiment, performed in a growth chamber. The wheat isolate was pathogenic to each element of a set consisting ofElytrigia repens, Hordeum vulgare, Lolium perenne, Poa annua, andTriticum aestivum. Re-isolates from any of the elements of this set were pathogenic to all other elements. The effects of hosts and inoculum-density treatments were statistically significant. A significant isolate x host interaction suggests a form of specialization, which is possibly due to a passage effect. These observations may contribute to a better understanding of the epidemiology ofS. nodorum in the Netherlands.Samenvatting Grassen en granen te velde werden geïnoculeerd met eenS. nodorum isolaat van tarwe. Symptomen van aantasting door de schimmel werden gevonden opElytrigia repens, Hordeum vulgare, Lolium perenne, Poa annua enTriticum aestivum. Van deze vijf waardplanten werden her-isolaten gewonnen, die vervolgens in een klimaatkamer geinoculeerd werden op ieder van de vijf waardplanten. Aldus ontstond een kruisinoculatiematrix van bladaantastingspercentages (Tabel 2), die aan een variantieanalyse (Tabel 3) kon worden onderworpen. De volgende effecten waren significant: waardplanten, inoculumdichtheden, interactie herisolaat x waardplant, en interactie herisolaat x waardplant x inoculumdichtheid. De interacties suggereren beïnvloeding door de waardplant, dus een passage-effect. Bij passage over een waardplant zou enige mate van fysiologische specialisatie kunnen optreden. Dit wordt bevestigd door gegevens van Weber (Tabel 5). Deze waarnemingen over de waardplantreeks kunnen van belang zijn voor een beter begrip van de epidemiologie vanS. nodorum in Nederland.     

Genetic variability within <Emphasis Type="Italic">Septoria carvi</Emphasis> Syd.- a pathogen of caraway <Emphasis Type="Italic">Carum carvi</Emphasis> L

Genetic variability within Septoria carvi isolates obtained from various organs of caraway cultivated in south-eastern and central Poland was studied using the RAPD-PCR technique. The tests were performed using randomly selected primers. The DNA profiles obtained using four primers proved useful in determining genetic variability among the genotypes of Septoria carvi isolates. The present study characterized the differences in the nucleotide sequence within the internal transcribed spacer region of rDNA (ITS1, 5.8S, ITS2) of selected S. carvi isolates and reference strains of Septoria spp. Moreover, eight isolates were sequenced for three loci: actin, calmodulin and translation elongation factor 1-alpha, and the obtained sequences were compared with the sequences of Septoria reference strains affecting other plants of the family Apiaceae. Phylogenetic analysis showed distinct differences of the tested isolates, which allowed to treat them Septoria carvi species affecting the above-ground organs of caraway Carum carvi L. This study is the first report on the genetic characteristics of the species S. carvi.     

A detached seedling leaf technique to study resistance to Mycosphaerella graminicola (anamorph Septoria tritici) in wheat

A detached seedling leaf technique was developed to screen for resistance to septoria tritici blotch of wheat and to detect specific interactions between cultivars and isolates. Wheat seedlings were inoculated with spore suspensions of Mycosphaerella graminicola . Detached primary leaves were then placed in a clear plastic box such that their cut ends were sandwiched between layers of agar containing benzimidazole, with a gap below the middle of the leaves. Mean levels of disease were affected by light and temperature, and also by the concentration of benzimidazole, such that higher concentrations resulted in less disease. Second leaves were more susceptible than seedling primary leaves. However, none of these factors affected ranking of disease among cultivars or cultivar-by-isolate interactions. Kavkaz–K4500 1.6.a.4, Synthetic 6x and Triticum macha showed specific susceptibility and resistance to different isolates. The detached leaf technique could be a useful complement to field trials and an alternative to whole seedling assays in assessing cultivar resistance and investigating the genetics of the host–pathogen interaction.     

Evaluation of the susceptibility of modern,wild, ancestral,and mutational wheat lines to Septoria tritici blotch disease

Globally, bread wheat production is threatened by fungal diseases, including the devastating disease Septoria tritici blotch (STB). Given the global importance of STB, and the difficulty in identifying novel sources of resistance to this disease, we screened a variety of wheat genotypes, including wild, ancestral, and mutagenized lines, for their STB response. This delineated a panel of wild wheat relatives and Watkins collection lines with exceptional resistance to a range of Zymoseptoria tritici isolates, some of which are highly virulent on modern, elite wheat varieties. Additionally, we characterized the STB susceptibility of 500 lines of the wheat cultivar Cadenza TILLING population and developed backcross derivatives of two TILLING lines that show dominant partial resistance to STB. These backcross lines are partially resistant to multiple isolates of Z. tritici, and, with the wild and ancestral lines identified, provide a useful reservoir of STB-resistant germplasm for use in wheat breeding programmes.     

A genetic study of the relationship between height, time of ear emergence and resistance to Septoria nodorum in wheat

Winter wheat cultivars of varying height were intercrossed and the progeny were studied in field trials to determine whether the greater resistance to S. nodorum of the taller cultivars was fortuitous or the result of genetic association.
Variation in height, in resistance and in time of ear emergence was continuous, providing no evidence of the existence of individual genes of major effect. In F₂ and backcross generations, the inheritance of all three characters was principally additive. Among random F₃ and F₄ families there was a clear tendency for tall straw and resistance to segregate together, proving that the association between these characters was not fortuitous and providing evidence of linkage or pleiotropy in their genetic control. There was a similar though less consistent tendency for late ear emergence and resistance to segregate together. There was substantial residual variation in resistance, not associated with variation in height or time of ear emergence, but its heritability was difficult to prove by comparisons between F₃ and F₄ generations, perhaps partly because of genotype x environment interaction.
It is proposed that resistance is under polygenic control and pleiotropy rather than linkage is suggested as the explanation of its genetic association with height and with time of ear emergence. The existence of genes affecting resistance independently of height and time of ear emergence is also inferred. It is suggested that they permit the breeding of resistant cultivars of any height, though with increasing difficulty as height is reduced.     

Effects of treatments to perennial ryegrass on the development of Septoria spp. in a subsequent crop of winter wheat

An experiment started in spring 1979 tested the effects of different treatments to perennial ryegrass, established as pure stands or undersown in spring wheat, on the severity of septoria diseases on the grass and in a following crop of winter wheat, sown in autumn 1980. Other plots were fallowed in 1979 and 1980 before sowing to winter wheat, also in autumn 1980.
Septoria tritici was most severe in winter wheat after fallow and least severe in wheat that had been direct-drilled after ryegrass. These effects were attributed to differences in amounts of available nitrogen, and consequent differences in crop growth, rather than to any differences in primary inoculum.
Symptoms attributed to S. nodorum on the ryegrass were more common where the grass had been established under spring wheat than where it had been sown as a pure stand. They also tended to be more common where ryegrass had been inoculated with spores of S. nodorum than where it had been sprayed with captafol. Similar effects on symptoms attributed to Septoria spp. (mostly S. tritici ) on the wheat were apparent in July. The results support the conclusion that the greater severity of septoria diseases that often occurs on wheat after grass than after non-graminaceous breaks, is probably due in part to survival of the pathogens on grass and, in some circumstances, on debris remaining from a previous wheat crop. However, other factors are also likely to be involved.     

Occurrence of Mycosphaerella graminicola, teleomorph of Septoria tritici, on wheat debris in the UK

Ascocarps of Mycosphaerella graminicola regularly developed in debris of wheat crops from eastern England after natural weathering. Identity with the Septoria tritici anamorph and pathogenicity to wheat were demonstrated by isolation and inoculation. Ascospores were present from autumn onwards and were most frequent in December and January. They may contribute to the primary inoculum for epidemics on autumn-sown wheat.     

The histology of processes associated with the infection of resistant and susceptible wheat cultivars with Septoria tritici

The development of Septoria tritici blotch on seedling leaves of the wheat cultivars Shafir (susceptible) and KVZ-K4500 L.6.A.4 (resistant) was studied by light and fluorescence microscopy. Appressoria were usually located above a stomatal opening. Percentage germination, formation of appressoria and the pattern of invasion of the substomatal chamber were the same in the susceptible and resistant cultivars. Growth of hyphae was observed in the intercellular spaces of the leaf tissue of both cultivars.
Autofluorescence was more commonly observed in the resistant accession, as was the affinity for trypan blue dye. Some lignification of cell walls and sparse deposition of callosic materials occurred in the resistant line. Leaves of KVZ-K4500 L.6.A.4, which macroscopically appeared pycnidia-free, revealed microscopically the presence of arrested, immature, pycnidia-like aggregations that did not progress further under prolonged environmental conditions conducive for pycnidial induction.