Effect of Trichothecenes Produced by Fusarium graminearum during Fusarium Head Blight Development in Six Cereal Species

Fusarium head blight (FHB) is a complex cereal disease associated with trichothecene production; these mycotoxins are factors of aggressiveness in wheat. Six species (bread and durum wheat, triticale, rye, barley and oats) were submitted to point inoculations with two isogenic strains of Fusarium graminearum; a wild strain (Tri5 +) produced trichothecenes and the mutated strain (Tri5 –) did not. The trichothecene-producing strain was generally more aggressive than the non-producing strain, but this varied according to crop species. The difference in aggressiveness was less pronounced in rye, a very resistant species. High resistance levels were observed in oats due to the large spacing between florets. In six-row barley, despite the existence of a moderate Type II resistance, the fungus was often observed to move externally from one floret to another within the dense spike, without penetrating the rachis. Bread wheat had low resistance to the trichothecene-producing strain and good resistance to the non-producing strain. Triticale responded to the strains in a similar way but was somewhat more resistant to both: symptoms on the spikelets and rachis of the triticales were restricted to below the point of inoculation. Durum wheat was susceptible to the trichothecene-producing strain and only moderately resistant to the non-producing strain, which was able to cause serious damage only to this species. Our study confirmed that the role of trichothecenes in FHB pathogenesis differs among species. The failure of the trichothecene non-producing F. graminearum strain to spread within the inflorescence of wheat, triticale, rye and barley, and the significant reduction of spread in the durum wheat spike strongly suggested that trichothecenes are a major determinant of fungal spread and disease development in Triticeae.     

Effect of temperature on head blight of wheat caused by Fusarium culmorum and F. graminearum

The effect of small temperature differentials (16 vs. 20°C) on the pathogenicity of deoxynivalenol producing single isolates of Fusarium culmorum and F. graminearum and on the fusarium head blight (FHB) response of eight wheat cultivars was examined. Fusarium culmorum inoculation caused greater visual disease symptoms at 20°C than at 16°C, both overall and on an individual cultivar basis (overall AUDPC = 13·5 and 9·6, respectively) ( P  < 0·05). In contrast, F. graminearum inoculation caused greater overall visual disease symptoms at 16°C than at 20°C, both overall and at the individual cultivar level (overall AUDPC = 12·8 and 10·9, respectively) ( P  < 0·05). Results showed both F. culmorum and F. graminearum inoculations caused a greater loss in yield at 20°C (54·3 and 46·9% relative 1000-grain weight, respectively) compared with 16°C (73·3 and 66·9% relative 1000-grain weight, respectively) ( P  < 0·05). Fusarium culmorum -inoculated heads contained similar amounts of fungal DNA at both 16 and 20°C (1·9 and 1·7 ng mg⁻¹ of plant material, respectively) (not significant), while for F. graminearum inoculation, plants contained higher amounts of fungal DNA at 20°C (2·0 and 1·0 ng mg⁻¹ of plant material, respectively) ( P  < 0·05). Overall, there was a significant negative correlation between AUDPC and percentage relative 1000-grain weight at both 16 and 20°C ( r  =−0·693 and −0·794, respectively, P  < 0·01).     

Biological control of southern blight disease of tomato caused by Sclerotium rolfsii with simplified mycelial formulations of Trichoderma koningii

Two simple formulations of an antagonistic strain of Trichoderma koningii were employed against southern blight disease caused by Sclerotium rolfsii in seedling, potted outdoor and field-grown tomatoes (cvs. Ife No. 1 and Ibadan Local). Corn cob germling inoculum and mycelium powder of T. koningii significantly controlled ( P ≤0·05) symptoms of damping off, blight and wilting in both tomato cultivars. The populations of the antagonist increased from an initial 1 × 10⁴ to about 1 × 10⁶ colony-forming units per g of soil in the protected plants. Moreover, sclerotial counts decreased significantly ( P ≤0·05) in these soils and those sclerotia found had been parasitized by T. koningii. Trichoderma -protected plants were more vigorous than those in the other treatment categories. The significance of these results is discussed in relation to the use of Trichoderma in appropriately simplified formulations.     

Genetic variation and covariation for aggressiveness, deoxynivalenol production and fungal colonization among progeny of Gibberella zeae in wheat

Gibberella zeae (anamorph Fusarium graminearum ) causes head blight of cereals and contaminates grains with mycotoxins such as deoxynivalenol (DON). To determine the correlations among aggressiveness traits, fungal colonization and DON production, 50 progeny from a segregating population of G. zeae were inoculated onto a susceptible winter wheat cultivar in three field environments (year–location combinations). Aggressiveness traits were measured as head-blight rating and plot yield relative to noninoculated plots. Fungal colonization, measured as Fusarium exoantigen (ExAg) content, and DON production were analysed with two ELISA formats. Disease severity was moderate to high based on head-blight rating and relative plot yield. Fusarium ExAg content and DON production ranged from 0·26–1·41 units and from 4·18–43·70 mg kg⁻¹, respectively. Significant ( P  = 0·01) genotypic variation was found for all traits. Heritability for Fusarium ExAg content was rather low because of high progeny–environment interaction and error. DON/ Fusarium ExAg ratio did not vary significantly ( P  > 0·1) among progeny. Correlation between DON production and Fusarium ExAg content across environments was high ( r  = 0·8, P  = 0·01), but no covariation existed between aggressiveness traits and DON/ Fusarium ExAg content ratio.     

Management of phytophthora root rot in radiata pine seedlings

Chemical and biological agents were evaluated for their ability to suppress root rot, caused by Phytophthora cactorum , in field-grown radiata pine seedlings in New Zealand. Trials were conducted over two seasons in an area of a forest nursery with a natural infestation of P. cactorum , and a history of root rot. In each season, symptoms of root rot developed during April, one month after root pruning, when seedlings were approximately six months old. In trial one, root rot incidence by mid July 2007 was 9·1% in untreated plots and 8·4% in plots that had been treated with metalaxyl-M/mancozeb (14 kg ha⁻¹) at seedling emergence. Disease incidence was lowest (2·1%) in plots that received seven monthly applications of phosphorous acid (6·5 L ha⁻¹). Other treatments, including seed coating with thiram or Trichoderma spp., and foliar applications of methyl jasmonate, did not control disease. In trial two, effects of treatment timing relative to root pruning were investigated. By late June 2008, three months after root pruning, root rot incidence was 22·2% in the untreated plots. Phosphorous acid was the most effective treatment and almost completely suppressed disease (0·1% incidence) when applied fortnightly from February until May (seven applications). Metalaxyl-M/mancozeb (15 kg ha⁻¹) was not effective (21·4% incidence) when applied five months before root pruning. However, disease incidence was reduced when the chemical was applied one week after root pruning (14·9% incidence) and greater control was achieved (8·2% incidence) when the application rate was increased to 50 kg ha⁻¹.     

Metalaxyl sensitivity of isolates of Phytophthora erythroseptica in Prince Edward Island

Isolates of Phytophthora erythroseptica , causal agent of pink rot of potato, were obtained from diseased tubers collected in Prince Edward Island (PEI), Canada. In an in vitro assay of field and single-zoospore isolates, all isolates of the pathogen from PEI were sensitive to metalaxyl (EC₅₀ < 0·5  µ g mL⁻¹), unlike moderately or highly resistant reference isolates obtained from Maine, USA. Allozyme-banding patterns at the glucose-6-phosphate isomerase ( Gpi ) locus were identical (91/91) for all isolates of P. erythroseptica examined from PEI and Maine, but could be used to distinguish isolates of P. erythroseptica from local isolates of Phytophthora infestans . Inoculation of potato tubers from plants treated with foliar applications of metalaxyl in the field indicated that compounds inhibitory to metalaxyl-sensitive isolates of P. erythroseptica were present in the periderm, even after 4 months' storage. By contrast, moderately or highly resistant isolates of the pathogen caused significantly ( P  ≤ 0·05) more disease than sensitive isolates in tubers from plants treated with metalaxyl. The effectiveness of metalaxyl in controlling pink rot in a particular region will depend on resistance levels of local populations of P. erythroseptica . Preventing the development and spread of resistant strains of P. erythroseptica will be critical in maintaining metalaxyl as an effective chemical for control of this pathogen.     

Comparison of pathogenicity of the Fusarium crown rot (FCR) complex (<Emphasis Type="Italic">F. culmorum</Emphasis>, <Emphasis Type="Italic">F. pseudograminearum</Emphasis> and <Emphasis Type="Italic">F. graminearum)</Emphasis> on hard red spring and durum wheat

Fusarium species involved in the Fusarium crown rot (FCR) complex affect wheat in every stage of development from seedling to grain fill. This study was designed to compare the aggressiveness of the FCR complex members including F. culmorum, F. pseudograminearum and F. graminearum in causing seedling blight, decreased plant vigour and crown rot. To assess their relative pathogenicity, two hard red spring wheat cultivars and two durum wheat cultivars were inoculated in the field with five isolates from each of the three species for two years. Significant differences in patterns of pathogenicity were identified. In particular, F. culmorum caused greater seedling blight while F. pseudograminearum and F. graminearum caused greater crown rot. Greatest yield reductions were caused by F. pseudograminearum. Cultivar differences were identified with respect to seedling disease and late season crown rot. No interactions were identified between cultivar performance and isolates or species with which they were challenged.     

Genetic diversity of Australian Fusarium graminearum and F. pseudograminearum

Genotypic diversity in Fusarium pseudograminearum and F. graminearum from Australia and the relationship between diversity and pathogen aggressiveness for head blight and/or crown rot of wheat were examined. Amplified fragment length polymorphism (AFLP) analysis revealed a high level of genotypic diversity within each species. Sixty-three of the 149 AFLP loci were significantly different between the two species and 70 of 72 F. pseudograminearum and 56 of 59 F. graminearum isolates had distinct haplotypes. When head blight and crown rot severity data from a recently published work on isolates representing the entire range of aggressiveness were used, only the genotypic diversity of F. pseudograminearum was significantly associated with its aggressiveness for the two diseases. Cluster analyses clearly demonstrated the polyphyletic structures that exist in both pathogen populations. The spatial diversity within F. graminearum was high within a single field, while frequent gene flow ( N _m ∼ 14) and a low fixation index ( G _st = 0·03) were recorded among F. pseudograminearum isolates from the adjacent states of New South Wales and Queensland. The differences in population structure between the heterothallic F. pseudograminearum (teleomorph G. coronicola ) and the homothallic F. graminearum (teleomorph G. zeae ) were not as pronounced as expected given their contrasting mating systems. Neither species was panmictic or strictly clonal. This points to sexual recombination in F. pseudograminearum , suggesting that ascospores of G. coronicola may also play a role in its biology and epidemiology.     

Effect of environmental factors and Sclerotium cepivorum isolate on sclerotial degradation and biological control of white rot by Trichoderma

Laboratory assays demonstrated that two isolates of Trichoderma viride and one isolate of Trichoderma pseudokoningii degraded up to 80% of sclerotia of four isolates of Sclerotium cepivorum in a silty clay soil, and also degraded up to 60% of sclerotia in three other soil types. Relationships were defined between the degree of sclerotial degradation by the two T. viride isolates in the silty clay soil and both temperature and soil water potential. Sclerotia were degraded between 10 and 25°C at −0·00012 MPa, but there was little activity of T. viride at 5°C or at −4 MPa. Degradation of S. cepivorum sclerotia also occurred in the absence of Trichoderma at soil water potentials approaching saturation . Experiments using onion seedling bioassays showed that the efficacy of Trichoderma isolates for the control of white rot using the same selection of soils and S. cepivorum isolates was variable, but that there was significant disease control overall. The importance of environmental factors and pathogen isolate in relation to effective biological control of white rot is discussed.     

脱氧雪腐镰刀菌烯醇在小麦赤霉病病程中的作用

 脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)系小麦赤霉病菌——禾谷镰孢(Fusarium graminearum,有性态Gibberell azeae)产生的一种单端孢霉烯族毒素(trichothecenes,以下简称"单族毒素")。作者以病菌正常产毒的野生型菌株GZ3639及其产毒基因Tri5损坏、不能产毒的转化子GZT40(Tri5^-,TRI^-)为材料,对DON在病菌致病过程中的作用进行了研究。结果显示:GZ3639能引起典型的赤霉病症状,病害可由接种小花发展至其它小穗,病菌也可定殖于其它小穗组织;而GZT40仅在接种小花处产生鸟眼状坏死斑点,不能引起典型的赤霉病症状,病菌仅定殖于接种小花处。当病菌与一定浓度(400 μ g·mL^-1)的DON混合处理小花后,GZT40的定殖时间提前2 d (感病品种上)至4 d (抗病品种上),定殖范围可扩大到1~3个小穗,接种20 d后的病小穗率也由病菌单独接种时的2.8%提高到12.5%。DON也使GZ3639的定殖时间提前1 d (感病品种上)至2 d (抗病品种上),定殖范围更广,其所致的病害发展更迅速,接种后20 d的病小穗率由病菌单独接种时的78.0%提高到100%。因此,作者认为,单族毒素在病害发展和病菌在寄主组织内的扩展中起着决定性作用,一旦丧失产毒能力,病菌虽能侵入寄主,但不能在寄主组织中扩展。DON可以缩短病菌成功定殖寄主所需的时间,更有助于病菌在寄主组织内的扩展。     

A high-throughput glasshouse bioassay to detect crown rot resistance in wheat germplasm

A high-throughput and reliable seedling bioassay to screen wheat germplasm for crown rot resistance was developed. Single wheat seedlings were grown in square seedling punnets in a glasshouse and inoculated with a monoconidial Fusarium pseudograminearum isolate 10 days after emergence. The punnets were laid horizontally on their side and a 10- µ L inoculum droplet placed on the stem base. Seedlings were incubated at near-saturated relative humidity, and crown rot severity was assessed 35 days after inoculation. Studies on the duration of incubation period, inoculum concentration and temperature were carried out to optimize these parameters. Seedling growth at 25/15(±5)°C in a glasshouse and 48-h incubation at near-saturated RH in darkness gave the best results. When crown rot resistance rankings of 16 Australian cultivars from the bioassay were compared with their field performance, Spearman's rank correlation coefficient was highly significant. This indicated that the seedling bioassay mimicked field resistance to crown rot in adult plants. A bootstrap resampling analysis showed little or no improvement in the coefficient of variation with an increasing number of replications, indicating a high level of precision and reproducibility. By detecting small but consistent differences in crown rot severity, the bioassay proved effective in large-scale screening for partial resistance: already over 1400 wheat genotypes have been screened. The high degree of precision makes this an invaluable tool in the understanding of pathogen aggressiveness, host specialization and parasitic fitness.     

Seed transmission of maize downy mildew (Peronosclerospora sorghi) in Nigeria

In an area of Nigeria where downy mildew of maize is present, histological assessment of maize seed revealed the presence of mycelium and oospores of Peronosclerospora sorghi in the kernels. Seed transmission of downy mildew of maize was demonstrated when grain purchased at local markets gave mean seedling infection rates of 12·3% (untreated seeds) and 10·0% (in metalaxyl-treated seeds) within 7 days of emergence, after storage in a desiccator for 30 days. When untreated seeds taken from nubbin ears of systemically infected plants from four states in southern Nigeria were planted at 9 days (17–22% moisture content) and 27 days (9–22% moisture content) after harvest, 20·0% infected seedlings resulted in both trials. Seeds from Borno state in northern Nigeria had 26·6% systemic seedling infection after 9 months of storage at 11% moisture content. When seeds harvested from maize plants inoculated with P. sorghi through silks were examined histologically, hyphae of P. sorghi were observed mostly in the scutellum of the embryo. Transmission of disease to seedlings was observed when the silk-inoculated seeds (9% moisture content) were planted in pots in a greenhouse; however, no disease transmission was observed when such seeds were planted in the field. The epidemiological significance of seed transmission is discussed with particular reference to survival of inoculum and development of epidemics. Also noteworthy is the overall significance of seed transmission in Nigeria, where the major source of seed is that saved by farmers from their grain crop, occasionally supplemented by seed bought from the local market.     

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.     

Rapid preparation of Fusarium DNA from cereals for diagnostic PCR u sing sonification and an extraction kit

Several PCR methods have recently been developed for Fusarium analysis in pure cultures. To use these new techniques in mycological studies and in industrial quality control, a protocol was set up for the rapid preparation of fungal DNA from cereals. An ultrasonification probe (sonotrode) and a lysis buffer were used for mechanical lysis of mycelia from infected grains. Following ultrasonification, DNA was isolated using a commercially available kit. DNA preparation was completed within 5 min per sample. The method resulted in DNA of sufficient quality and quantity for diagnostic PCR. Group- and species-specific primers were used to detect DNA of Fusarium graminearum and F. culmorum in species-specific assays as well as trichothecene-producing Fusarium spp. in a group-specific system. A minimum of one F. graminearum -infected grain added to an uninfected 40 g wheat sample was detectable with a species-specific PCR. The PCR signals produced with primers specific for the tri5 gene of trichothecene-producing Fusarium spp. and with primers for the detection of F. graminearum (gaoA) were in accordance with corresponding concentrations of deoxynivalenol (DON) found in samples by HPLC analysis. The speed of the protocol developed may promote the use of PCR in routine applications in an agro-industrial context.     

Studies on the epidemiology and yield losses from rice black-streaked dwarf disease in a recent epidemic in Zhejiang province, China

The spread of rice black-streaked dwarf disease, which has emerged as a major problem on winter wheat and the two summer rice crops (early indica and late japonica ) grown in central and southern Zhejiang province, China, is documented from 1995 to 2007. The late japonica crop suffered the most: up to 64 640 ha were affected with estimated losses of c . 120 000 t grain per year. Peak adult numbers of the small brown planthopper vector, Laodelphax striatellus , coincided with the seedling stages of both rice crops and the proportion of the insect population carrying virus increased during 1998–2005. Seedlings with three to four leaves were the most susceptible, whereas plants inoculated after the end of tillering developed few or no symptoms. Disease levels were strongly correlated with numbers of viruliferous vectors. In sowing-date experiments with both rice crops, the earliest sowings had the most disease and suffered the greatest yield losses. With the last sowing date (25 days after the first), there were almost no losses. There were yield losses of 0·80% for every 1% increase in disease incidence in early indica rice and rather more (0·92%) in the late japonica crop. There were large differences in susceptibility between cultivars, indicating the possibility, within currently available germplasm, of using more resistant cultivars to help contain the disease. Changes in cropping practice and in recent winter weather conditions have probably contributed to the emergence of the virus as a major pathogen in eastern China.     

Estimating the germination dynamics of Plasmopara viticola oospores using hydro-thermal time

The effects of environmental conditions on the variability in germination dynamics of Plasmopara viticola oospores were studied from 1999 to 2003. The germination course was determined indirectly as the relative infection incidence (RII) occurring on grape leaf discs kept in contact with oospores sampled from a vineyard between March and July. The time elapsed between 1 January and the infection occurrence was expressed as physiological time, using four methods: (i) sums of daily temperatures > 8°C; (ii) hourly temperatures > 10°C; (iii) sums of hourly rates from a temperature-dependent function; or (iv) sums of these rates in hours with a rain or vapour pressure deficit ≤ 4·5 hPa (hydro-thermal time, HT). An equation of Gompertz in the form RII = exp[− a  · exp(− b  · HT)] produced an accurate fit for both separate years ( R ² = 0·97 to 0·99) and pooled data ( R ² = 0·89), as well as a good accuracy in cross-estimating new data ( r between observed and cross-estimated data were between 0·93 and 0·99, P  < 0·0001). It also accounted for a great part of the variability in oospore germination between years and both between and within sampling periods. Therefore, the equation of Gompertz (with a  = 15·9 ± 2·63 and b  = 0·653 ± 0·034) calculated over hydro-thermal time, a physiological time accounting for the effects of both temperature and moisture, produced a consistent modelling of the general relationships between the germination dynamics of a population of P. viticola oospores and weather conditions. It represents the relative density of the seasonal oospores that should have produced sporangia when they have experienced favourable conditions for germination.     

Visual and PCR assessment of light leaf spot (Pyrenopeziza brassicae) on winter oilseed rape (Brassica napus) cultivars

Methods to assess light leaf spot ( Pyrenopeziza brassicae ) on winter oilseed rape cultivars were compared in laboratory, controlled-environment and field experiments. In controlled-environment experiments with seedling leaves inoculated at GS 1,4, the greatest differences in percentage area affected by P. brassicae sporulation were observed with inoculum concentrations of 4 × 10³ or 4 × 10⁴ spores mL⁻¹, rather than 4 × 10² or 4 × 10⁵ spores mL⁻¹, but older leaves had begun to senesce before assessment, particularly where they were severely affected by P. brassicae . In winter oilseed rape field experiments, a severe light leaf spot epidemic developed in 2002/03 (inoculated, September/October rainfall 127·2 mm) but not in 2003/04 (uninoculated, September/October rainfall 40·7 mm). In-plot assessments discriminated between cultivars best in February/March in 2003 and June in 2004, but sometimes failed to detect plots with many infected plants (e.g. March/April 2004). Ranking of cultivar resistance differed between seedling experiments done under controlled-environment conditions and field experiments. The sensitivity of detection of P. brassicae DNA extracted from culture was greater using the PCR primer pair PbITSF/PbITSR than using primers Pb1/Pb2. P. brassicae was detected by PCR (PbITS primers) in leaves from controlled-environment experiments immediately and up to 14 days after inoculation, and in leaves sampled from field experiments 2 months before detection by visual assessment.     

Germination, emergence, and dormancy of Mimosa pudica

Mimosa pudica (common sensitive plant) is a problematic weed in many crops in tropical countries. Eight experiments were conducted to determine the effects of light, seed scarification, temperature, salt and osmotic stress, pH, burial depth, and rice residue on the germination, seedling emergence, and dormancy of M. pudica seeds. Scarification released the seeds from dormancy and stimulated germination, though the germination of the scarified seeds was not influenced by light. The scarification results indicate that a hard seed coat is the primary mechanism that restricts germination. The germination increased markedly with the exposure to high temperature "pretreatment" (e.g. 150°C), which was achieved by placing non-scarified seeds in an oven for 5 min followed by incubation at 35/25°C day/night temperatures for 14 days. The germination of the scarified seeds was tolerant of salt and osmotic stress, as some seeds germinated even at 250 mmol L⁻¹ NaCl (23%) and at an osmotic potential of −0.8 MPa (5%). The germination of the scarified seeds was >74% over a pH range of 5–10. The seedling emergence of the scarified seeds was 73–88% at depths of 0–2 cm and it gradually decreased with an increasing depth, with no seedling emergence at the 8 cm depth. The rice residue applied to the soil surface at rates of ≤6 t ha⁻¹ did not influence the seedling emergence and dry weight. The information gained from this study identifies some of the factors that facilitate M. pudica becoming a widespread weed in the humid tropics and might help in developing components of integrated weed management practises to control this weed.     

Binucleate Rhizoctonia sp. AG G causing root rot in yacon (Smallanthus sonchifolius) in Brazil

A new rot caused by a binucleate Rhizoctonia sp. affecting the tuberous root cortex of the domesticated yacon ( Smallanthus sonchifolius ) has been observed in Brazil. Isolates of a binucleate Rhizoctonia sp. were collected from roots with rot symptoms and characterized by the number of nuclei per cell, hyphal anastomosis, RAPD molecular markers, ITS-5·8S rDNA sequence and pathogenicity tests. All isolates had a mean of 1·9–2·2 nuclei per cell and anastomosed with the binucleate Rhizoctonia sp. AG G-tester strain. RAPD analysis was carried out between 11 isolates recovered from yacon and 11 AG (A, Ba, Bb, Bo, C, D, F, G, O, P, Q) standard testers of binucleate Rhizoctonia sp. Genetic similarities of 94·8–100% were observed among isolates of the binucleate Rhizoctonia sp. from yacon and all isolates were genetically more closely related to the AG G tester than other strains according to upgma analysis using RAPD markers. Homologies of complete ITS nucleotide sequences were 100% between binucleate isolates of Rhizoctonia sp. from yacon and the AG G tester. According to pathogenicity tests, the isolates caused typical rot symptoms of yacon tubers 90 days after inoculation     

Effect of pea plant architecture on spatiotemporal epidemic development of ascochyta blight (Mycosphaerella pinodes) in the field

The effect of pea canopy structure on epidemics of Mycosphaerella pinodes was investigated in four spring pea cultivars (Bridge, Obelisque, Solara and Athos) in two field experiments. These cultivars had similar levels of susceptibility to ascochyta blight and presented different architectural features (branching, standing ability and stem height). Stem height ranged from 77 to 95 cm in 1999 and from 72 to 92 cm in 2000, while leaf area index (LAI) ranged from 3·8 to 5·1 in 1999 and from 3·8 to 4·7 in 2000. Internode lengths varied from 4·6 to 6·0 in 1999 and from 3·8 to 4·7 in 2000. Mean distance between nodes in the canopy (MDN) ranged from 12·2 to 15·3 cm in 1999 and from 11·2 to 13·9 cm in 2000. Canopies with different architecture differed in disease progression on stipules and mainly on pods. The levels of disease on stipules and pods were strongly correlated. Moreover, if disease was considered as a function of stipule height up the stem, large differences in vertical disease distribution were observed between cultivars. Three architectural features acted on disease development: cumulative LAI above the node on which disease was assessed (LAIcum) and large MDN favouring disease development, and large internode length reducing disease severity. Modifying LAI distribution and plant organization could be one way to reduce the impact of ascochyta blight, by direct or indirect effects on environmental and dispersal conditions.