The infection biology of Fusarium graminearum: defining the pathways of spikelet to spikelet colonisation in wheat ears

Fusarium graminearum is one of the main causal agents of Fusarium Ear Blight on wheat. How the pathogen colonises the entire ear is not known. There is controversy over whether this mycotoxin producing pathogenic fungus invades wheat floral tissue using a necrotrophic or another mode of nutrition. A detailed microscopic investigation has revealed how wild-type fungal hyphae, of the sequenced strain PH-1, colonised susceptible wheat ears and spread from spikelet to spikelet. At the advancing infection front, colonisation of the host cortex occurred ahead of any vascular colonisation and the hyphae adapted to the available intercellular space between host cells. Intercellular hyphae then became abundant and host cells lost their entire cellular contents just prior to intracellular colonisation. No host cells died ahead of the infection. However, while these deep cortex infections progressed, just below the surface the highly photosynthetic chlorenchyma cells were observed to have died prior to colonisation. Behind the infection front, hyphae were abundant in the vasculature and the cortex, often growing through the pit fields of thick walled cells. This high level of inter- and intracellular fungal colonisation resulted in the collapse of the non-lignified cell-types. In this middle zone of infection, hyphal diameters were considerably enlarged. Far behind the infection front inter- and intracellular hyphae were devoid of contents and had often collapsed. At later stages of infection, the pathogen switched from predominately vertical to lateral growth and accumulated below the surface of the rachis. Here the lignified host cell walls became heavily degraded and hyphae ruptured the epidermis and produced an aerial mycelium.     

Colonization of wheat seedling leaves by Fusarium species as observed in growth chambers: a role as inoculum for head blight infection?

Fusarium species involved in the Fusarium head blight complex in Western Europe were investigated for their potential to infect and colonize non-damaged wheat leaves and to produce conidia on senescing wheat leaves incubated at high relative humidity. Fusarium avenaceum, Fusarium culmorum, Fusarium graminearum, Fusarium poae and Fusarium tricinctum did not directly penetrate the leaf tissue after conidia germination on the leaf surface. Germ tubes grew on the host surface for 24–36 hr forming a mycelial network. After invading the host, some species formed runner hyphae between cell wall layers or underneath the cuticular layer. Macroscopic symptoms developed on leaves and stems from 7 d post inoculation. Inside leaf tissues, hyphae thickened in diameter and were both inter- and intra-cellular. Fusarium tricinctum formed sporophores which erupted through the leaf surface releasing numerous conidia. Incubation of senescing leaves at 100 % relative humidity for 48 hr resulted in sporulation of all Fusarium spp.     

The transcriptome of Fusarium graminearum during the infection of wheat

Fusarium graminearum causes head blight disease in wheat and barley. To help understand the infection process on wheat, we studied global gene expression of F. graminearum in a time series from 24 to 196 h after inoculation, compared with a noninoculated control. The infection was rapid and, after 48 h, over 4,000 fungal genes were expressed. The number of genes expressed increased over time up to 96 h (>8,000 genes), and then declined at the 144- and 192-h post-inoculation time points. After subtraction of genes found expressed on complete medium, during carbon or nitrogen starvation, and on barley, only 355 were found exclusively expressed in wheat, mostly genes with unknown function (72.6%). These genes were mainly found in single-nucleotide polymorphism-enriched islands on the chromosomes, suggesting a higher evolutionary selection pressure. The annotated genes were enriched in functional groups predicted to be involved in allantoin and allantoate transport, detoxification, nitrogen, sulfur and selenium metabolism, secondary metabolism, carbohydrate metabolism, and degradation of polysaccharides and ester compounds. Several putative secreted virulence factors were also found expressed in wheat.     

Colonization of roots by arbuscular mycorrhizal fungi using different sources of inoculum

Arbuscular mycorrhizal fungi (AMF) form a number of different infective propagules that are used to form new mycorrhizal associations. These are spores, extraradical hyphae and infected roots. However, not all fungi are equally capable of colonizing roots with all of the above-mentioned propagules and there is conflicting evidence of major differences in colonization strategy between members of the Glomineae and Gigasporineae. In this study, we tested the abilities of eight fungal species from four different genera to colonize roots using three different types of inoculum. Glomus and Acaulospora isolates colonized from all inoculum types, whereas Gigaspora and Scutellospora isolates colonized mainly from spores and to a limited degree from root fragments. Extraradical hyphae were not suitable propagules for the species of Gigaspora and Scutellospora tested. This indicates that AMF have different colonization strategies and that this is largely differentiated at the suborder level. It is unclear why there is such a difference among the fungi in inoculum types. Future research should examine differences in the anatomy and physiology to discern a mechanism for such differences in life-history strategies.     

Microbiological and SYBR green real-time PCR detection of major Fusarium head blight pathogens on wheat ears

Fusarium head blight (FHB) caused by several Fusarium species is one of the most serious diseases affecting wheat throughout the world. The efficiency of microbiological assays and real-time PCRto quantify major FHB pathogens in wheat ears after inoculation with F. graminearum, F. culmorum, F. avenaceum and F. poae undergreenhouse and field conditions were evaluated. The frequency of infected kernel, content of fungal biomass, disease severity and kernel weight were determined. To measure the fungal biomass an improved DNA extraction method and a SYBR Green real-time PCR were developed. The SYBR Green real-time PCR proved to be highly specific for individual detection of the species in a matrix including fungal and plant DNA. The effect of Fusarium infection on visible FHB severity, frequency of infected kernels and thousand-kernel mass (TKM) significantly depended on the Fusarium species/isolate. F. graminearum resulted in highest disease level, frequency of infected kernels, content of fungal biomass, and TKM reduction followed by F. culmorum, EF avenaceum and F. poae, respectively. The comparison of frequency and intensity of kernel colonization proved differences in aggressiveness and development of the fungi in the kernels. Only for F. graminearum, the most aggressive isolate, application of microbiological and real-time PCR assays gave similar results. For the other species, the intensity of kernel colonization was lower than expected from the frequency of infection.     

Composting potential of different inoculum sources in the modified SEBAC system treatment of municipal solid wastes

The aim of the work described here was to analyse the biomethanization process for three types of organic fraction of municipal solid waste (OFMSW) using two different inoculum sources in a sequential leach-bed anaerobic reactor under thermophilic (55 degrees C) and dry conditions (20%TS). The OFMSWs studied were: separately collected food waste (SC_OFMSW) from restaurants, synthetic waste (ST_OFMSW) and mechanically selected municipal waste (MS_OFMSW). The inoculum studied were digested mesophilic sludge (SLUDGE) and digested SC_OFMSW. The results show that SLUDGE gave the best performance and this was therefore selected for the rest of the experiments. Three assays were performed to analyse the biomethanization processes of three types of organic waste. The results suggest that all three wastes give rise to an acclimation stage with acidogenic/acetogenic activity between days 5 and 20-30 and a stabilization phase associated with methanogenic activity. In conclusion, a mixture of OFMSW (regardless of its origin) and SWINE arranged in layers in the reactor, as well as SLUDGE used with an inoculum source, enhanced the fast start up phase of a modified sequential leach-bed system under dry thermophilic conditions.     

Celery leaf spot: sources of inoculum

The relative importance of infected celery seed, infected leaf debris in the soil, and infected wild celery, in the incidence of Septoria leaf spot in cultivated celery has been investigated. Infection can be caused when the sole source of inoculum is viable spores on the seed surface; such spores are considered to be the main cause of disease outbreaks. Of all the seed samples examined, 93% were infected by Septoria spp. In untreated seed samples, 40% carried viable spores which survived for up to 15 months on seed stored in the laboratory, and for longer periods on seed stored at -20d? C. However, ageing of seed is not recommended as a commercial control measure. The fungus was not found in seed embryos or endosperms but mycelium was present in pericarps and testas. Unconfirmed evidence suggests that in favourable circumstances new spores might be produced in old seed-borne pycnidia.     

不同气象条件下小麦冷源的品质变异

小麦冷源是一种冷温供体材料,有较强的降温作用,当受体小麦接受冷温供体的配子后,其后代冠温呈普遍下降态势,且部分受体小麦的温度型可由非冷型转化为后代的冷型,而冷型小麦具有代谢功能好且利于高产、稳产的特征,故小麦冷源不但对冷型小麦的问世有重要诱导作用,且能更好促进小麦的高产、稳产.将小麦冷源材料放置在不同的气象条件之下,以研究其籽粒品质的变异状况.结果表明,小麦冷源和非冷源相比,17项籽粒品质性状变异等级的加权平均值为1.9877,后者为2 8171,显示出小麦冷源的籽粒品质具有明显变异小、较稳定的特征,这和籽粒形成过程中蛋白质、淀粉及其组分的含量,相互间的比例变异小密切相关.由于小麦冷源具有籽粒品质变异小的特性,因而,这种特性就利于在其后代冷型小麦上显现,从而使冷型小麦的一些优良品质性状能在生产上反复重演,这不但进一步赋予了利于高产、稳产的冷型小麦以优质、稳质的性状,并能最终促进小麦生产的较大发展,其意义是十分重要的.     

Effects of long-term storage on Fusarium toxin concentrations in wheat - sources of error of the analytical results

The concentrations of the Fusarium toxins deoxynivalenol (DON) and zearalenone (ZON) of a heavily contaminated wheat grain batch were followed over a period of 1 year by taking samples 15 times every 28 days. The air temperature and relative humidity at the top of the wheat batch ranged between 7 and 22°C and 44 and 55%, respectively, and corresponded to a variation in the moisture content of the wheat grain between 11.5 and 12.3%. None of these fluctuations were related to ZON and DON concentrations, which varied between 0.46 and 0.66 and 15.0 and 19.5 mg/kg DM. Therefore, the data were used to analyse the error sources for the analytical results. It was found that the variance proportions due to sampling and sample preparation plus analysis were not similar for DON and ZON. The variance proportion due to sampling was found to be 0.62 for ZON, which corresponded to a variance proportion of 0.38 due to sample preparation plus analysis. In contrast, the latter variance proportion for DON was estimated to be 1.0 and consequently completely superimposed the sampling error. It is concluded that long-term storage of contaminated wheat grain does not affect the concentrations of DON and ZON considering the measured fluctuations in ambient temperature, relative humidity and moisture content of the grain. Therefore, no degradation of DON and ZON occurred during the storage of wheat for a period of one year under ambient conditions. The effects of sampling and sample preparation plus analysis on the final analytical results are different for DON and ZON and require further consideration.     

Effects of temperature and inoculum concentration on infection of narcissus bulbs by Fusarium oxysporum f.sp. narcissi

Chlamydospores of Fusarium oxysporum germinated, and mycelium grew on agar, at 10 but not 8°C. Numbers of chlamydospores needed to initiate disease suggest that the principal sources of infection are within the stock of bulbs and not the soil.     

Phases of infection and gene expression of Fusarium graminearum during crown rot disease of wheat

Fusarium graminearum causes head blight (FHB) and crown rot (CR) diseases in wheat. Compared with FHB, CR symptom development occurs slowly, usually taking 4 to 8 weeks to become visible. To characterize CR development, we used histological and real-time quantitative polymerase chain reaction analyses to assess fungal colonization during a timecourse of infection. Three distinct phases of infection were identified: i) initial spore germination with formation of a superficial hyphal mat at the inoculation point, ii) colonization of the adaxial epidermis of the outer leaf sheath and mycelial growth from the inoculation point to the crown, concomitant with a drop in fungal biomass, and iii) extensive colonization of the internal crown tissue. Fungal gene expression was examined during each phase using Affymetrix GeneChips. In total, 1,839 F. graminearum genes were significantly upregulated, including some known FHB virulence genes (e.g., TRI5 and TRI14), and 2,649 genes were significantly downregulated in planta compared with axenically cultured mycelia. Global comparisons of fungal gene expression with published data for FHB showed significant similarities between early stages of FHB and CR. These results indicate that CR disease development involves distinct phases of colonization, each of which is associated with a different fungal gene expression program.     

RAPD polymorphisms in spring wheat cultivars and lines with different level of Fusarium resistance

Random amplified polymorphic DNA (RAPD) markers have been used to characterize the genetic diversity among 35 spring wheat cultivars and lines with different levels of Fusarium resistance. The objectives of this study were to determine RAPD-based genetic similarity between accessions and to derive associations between Fusarium head blight (FHB) and RAPD markers. Two bulked DNA from either highly resistant lines or susceptible lines were used to screen polymorphic primers. Out of 160 screened primers, 17 primers generated reproducible and polymorphic fragments. Genetic similarity calculated from the RAPD data ranged from 0.64 to 0.98. A dendrogram was prepared on the basis of a similarity matrix using the UPGMA algorithm, which corresponded well with the results of principal component analysis and separated the 35 genotypes into two groups. Association analysis between RAPD markers and the FHB index detected three RAPD markers, H19(1000), F2(500) and B1(2400), significantly associated with FHB-resistant genotypes. These results suggest that a collection of unrelated genotypes can be used to identify markers linked to an agronomically important quantitative trait like FHB. These markers will be useful for marker-assistant breeding and can be used as candidate markers for further gene mapping and cloning.     

Significance of inoculum size in the lag time of Listeria monocytogenes

The lag time of Listeria monocytogenes growing under suboptimal conditions (low nutrient concentrations, pH 6, and 6.5 degrees C) was extended when the inoculum was severely stressed by starvation and the inoculum size was very small. Predictive microbiology should deal with bacterial stress and stochastic approaches to improve its value for the agro-food industry.     

Effect of wheat cultivars with different resistance to Fusarium head blight on rhizosphere Fusarium graminearum abundance and microbial community composition

Plant and Soil - Wheat (Triticum aestivum L.) cultivars vary in their resistance to Fusarium head blight (FHB), while it is poorly understood how different cultivars influence FHB-causing Fusarium...