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Stripe rust, caused by Puccinia striiformis f. sp. tritici ( Pst ), is one of the main diseases of wheat worldwide. Mianyang of Sichuan province in Southwest China is one of main regions for winter Pst inoculum production and spring epidemic and provides urediniospores for infecting wheat in the surrounding regions. Understanding the urediniospore dynamics is important to predict and manage stripe rust. In this study, spore trapping coupled with a TaqMan real-time quantitative PCR method was used to monitor airborne Pst urediniospores from December 2019 to December 2022 in Mianyang. Weather conditions (temperature, relative humidity, daily sunshine duration, and precipitation) were collected for the same period. These data were used to study the relationship of airborne urediniospore density with climatic conditions. The results showed that Pst urediniospores were captured all year round, and the annual peak of urediniospore densities occurred in the period from March to April in which the urediniospores accounted for the largest proportion of the annual total urediniospores. The density of urediniospores in the period of March to April was linearly related to the average sunshine duration of 20 days and average temperature of 15 days prior to the final day of a 7-day trapping period. This relationship needs to be tested in other regions where Pst can sporulate during the winter before it can be integrated with Pst infection conditions to predict rust development., Competing Interests: The author(s) declare no conflict of interest.

Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici Erikss. ( Pst ), is a devastating disease resulting in yield reduction. Because the temperature limits the overwintering and oversummering of Pst , it cannot complete the whole year cycle in most areas of China. Longnan, located in the southeast of Gansu Province, is one of the annual cycle areas of Pst , which can supply urediniospores in autumn to eastern wheat-growing areas in China. In this study, a TaqMan real-time quantitative PCR (TaqMan-qPCR) detection system for Pst urediniospores was established, and the detection limit was a single urediniospore. The dynamics of Pst urediniospores in Longnan were monitored by spore trapping and TaqMan-qPCR for 3 years. Meanwhile, the meteorological conditions including air temperature, relative humidity, and precipitation were recorded. Results showed that Pst urediniospores can be captured from March to December, and two peaks of urediniospore density appeared in May and June, respectively. The density of urediniospore is closely related to temperature and precipitation from March to June. In addition, we found that the density of Pst urediniospore had the peak value when the average air temperature was 10 to 21°C, and the relative humidity was 60 to 85% from May to June. The exponential model could describe the variation of Pst urediniospore density based on average temperature and precipitation from March to June. It is worth mentioning that the exponential model based on average temperature 7 days before spore capture has a great advantage in predicting the urediniospore density in the air. This study laid a foundation for establishment of a prediction model for wheat stripe rust based on the density of urediniospores and meteorological factors., Competing Interests: The author(s) declare no conflict of interest.

The binary power law (BPL) is often used to characterize spatial heterogeneity of disease incidence. A hierarchical mixed model, coupled with multiple imputation to randomly generate any missing standard errors, was used to conduct a meta-analysis of >200 published values of the estimated aggregation ( b ) parameter of the BPL. Approximately 50% of estimated b values ranged from 1.1 to 1.3. Moderator variable analysis showed that the number of individuals per sampling unit ( n ) had a strong positive effect on b , with a linear relation between estimated b and ln( n ). Estimated expected value of b for the population of published regressions at a reference n of 15 was 1.22. The increase in the variance due to the imputations was only 0.03, and the efficiency exceeded 0.98. Results were confirmed with an alternative mixed model that considered a range of possible within-trial correlations of the estimated b values and with a random-coefficient mixed model fitted to the subset of the data. Cropping system, dispersal mode, and pathogen type all had significant effects on b , with annuals having larger expected value than woody perennials, soilborne and rain-splashed dispersed pathogens having the largest expected values for dispersal mode, and bacteria and oomycetes having the largest expected values for pathogen type. However, there was considerable variation within each of the levels of the moderators, and the differences of expected values from smallest to largest were small, ≤0.16. Results are discussed in relation to previously published findings from stochastic simulations.

In the main wheat production area of China (the Huang Huai Plain [HHP]), both Fusarium graminearum and Fusarium asiaticum , the causal agents of Fusarium head blight (FHB), are present. We investigated whether the relative prevalence of F . graminearum and F . asiaticum is related to cropping systems and/or climate factors. A total of 1,844 Fusarium isolates were obtained from 103 fields of two cropping systems: maize-wheat and rice-wheat rotations. To maximize the differences in climatic conditions, isolates were sampled from the north and south HHP regions. Based on the phylogenetic analysis of EF - 1 α and Tri101 sequences, 1,207 of the 1,844 isolates belonged to F . graminearum , and the remaining 637 isolates belonged to F . asiaticum . The former was predominant in the northern region: 1,022 of the 1,078 Fusarium isolates in the north were F . graminearum . The latter was predominant in the southern region: 581 of the 766 Fusarium isolates belonged to F . asiaticum . Using an analysis based on generalized linear modeling, the relative prevalence of the two species was associated more with climatic conditions than with the cropping system. F . graminearum was associated with drier conditions and cooler conditions during the winter but also with warmer conditions in the infection and grain-colonization period as well as with maize-wheat rotation. The opposite was true for F . asiaticum . Except for the 15-acetyldeoxynvalenol genotype, the trichothecene chemotype composition of F . asiaticum differed between the two cropping systems. The 3-acetyldeoxynivalenol genotype was more prevalent in the maize-wheat rotation, whereas the nivalenol genotype was more prevalent in the rice-wheat rotation. The results also suggested that environmental conditions in the overwintering period appeared to be more important than those in the infection, grain-colonization, and preanthesis sporulation periods in affecting the relative prevalence of F . graminearum and F . asiaticum . More research is needed to study the effect of overwintering conditions on subsequent epidemic in the following spring.

Xiaoyan6 (XY6) is a wheat ( Triticum aestivum ) cultivar possessing nonrace-specific high-temperature seedling plant (HTSP) resistance against stripe rust, caused by Puccinia striiformis f. sp. tritici . Previously, we identified one particular gene, TaRPS2 , for its involvement in the HTSP resistance. To elucidate the role of TaRPS2 in the HTSP resistance, we cloned the full length of TaRPS2 from XY6. The transcriptional expression of TaRPS2 was rapidly upregulated (19.11-fold) under the normal-high-normal temperature treatment that induces the HTSP resistance. The expression level of TaRPS2 in leaves was higher than that in the stems and roots. Quantification of the endogenous hormones in wheat leaves after P. striiformis f. sp. tritici inoculation showed that 1-aminocyclopropane-1-carboxylic acid, salicylic acid (SA), and jasmonic acid were involved in the HTSP resistance. In addition, detection of hydrogen peroxide (H 2 O 2 ) accumulation indicated that reactive oxygen species burst was also associated with the HTSP resistance. Two hours after exogenous H 2 O 2 treatment or 0.5 h after SA treatment, the expression level of TaRPS2 was increased by 2.66 and 2.35 times, respectively. The subcellular localization of enhanced green fluorescent protein-TaRPS2 fusion protein was in the nuclei and plasma membranes. Virus-induced gene silencing of TaRPS2 reduced the level of HTSP resistance in XY6. Compared with the nonsilenced leaves, the TaRPS2 -silenced leaves had the reduction of necrotic cells but a greater number of uredinia. These results indicated that TaRPS2 positively regulates the HTSP resistance of XY6 against P. striiformis f. sp. tritici and is related to the SA and H 2 O 2 signaling pathways.

Stripe rust, caused by Puccinia striiformis f. sp. tritici Erikss., is an important disease of wheat worldwide. Identification of pathogen survival hot spots is important for predicting disease onset and subsequent spread to other regions, and such understanding is essential for developing integrated management strategies. We conducted field studies to determine the relationship of P. striiformis f. sp. tritici overwintering potential to winter temperatures in 10 sites in the northwest of China in three growing seasons (2011-12, 2012-13, and 2013-14). The fungus survived better in wheat cultivars with stronger winter hardiness than in those with weaker winter hardiness. Windowpane analysis suggested that P. striiformis f. sp. tritici winter survival is related to temperatures in the coldest period from mid-December to late January. β Regression showed that P. striiformis f. sp. tritici winter survival was negatively related to the number of days with daily average temperature <-2°C for cultivars with weak winter hardiness and <-4°C for cultivars with both moderate and strong winter hardiness. Models developed under constant temperature experiments greatly underestimate the P. striiformis f. sp. tritici overwinter potential under field conditions. This underestimation probably results from the possibility that P. striiformis f. sp. tritici in wheat tissues may have increased tolerance to low temperatures because of acclimation to gradual temperature changes under field conditions. The present models can be used to predict quantitative risks of P. striiformis f. sp. tritici overwintering in winter-wheat-growing regions for timely implementation of regional disease management strategies.

High-resolution aerial imaging with an unmanned aerial vehicle (UAV) was used to quantify wheat powdery mildew and estimate grain yield. Aerial digital images were acquired at Feekes growth stage (GS) 10.5.4 from flight altitudes of 200, 300, and 400 m during the 2009-10 and 2010-11 seasons; and 50, 100, 200, and 300 m during the 2011-12, 2012-13, and 2013-14 seasons. The image parameter lgR was consistently correlated positively with wheat powdery mildew severity and negatively with wheat grain yield for all combinations of flight altitude and year. Fitting the data with random coefficient regression models showed that the exact relationship of lgR with disease severity and grain yield varied considerably from year to year and to a lesser extent with flight altitude within the same year. The present results raise an important question about the consistency of using remote imaging information to estimate disease severity and grain yield. Further research is needed to understand the nature of interyear variability in the relationship of remote imaging data with disease or grain yield. Only then can we determine how the remote imaging tool can be used in commercial agriculture.

WRKY transcription factors (TFs) play crucial roles in plant resistance responses to pathogens. Wheat stripe rust, caused by the fungal pathogen Puccinia striiformis f. sp. tritici (Pst), is a destructive disease of wheat (Triticum aestivum) worldwide. In this study, the two WRKY genes TaWRKY49 and TaWRKY62 were originally identified in association with high-temperature seedling-plant resistance to Pst (HTSP) resistance in wheat cultivar Xiaoyan 6 by RNA-seq. Interestingly, the expression levels of TaWRKY49 and TaWRKY62 were down- and up-regulated, respectively, during HTSP resistance in response to Pst. Silencing of TaWRKY49 enhanced whereas silencing TaWRKY62 reduced HTSP resistance. The enhanced resistance observed on leaves following the silencing of TaWRKY49 was coupled with increased expression of salicylic acid (SA)- and jasmonic acid (JA)-responsive genes TaPR1.1 and TaAOS, as well as reactive oxygen species (ROS)-associated genes TaCAT and TaPOD; whereas the ethylene (ET)-responsive gene TaPIE1 was suppressed. The decreased resistance observed on leaves following TaWRKY62 silencing was associated with increased expression of TaPR1.1 and TaPOD, and suppression of TaAOS and TaPIE1. Furthermore, SA, ET, MeJA (methyl jasmonate), hydrogen peroxide (H2O2) and abscisic acid (ABA) treatments increased TaWRKY62 expression. On the other hand, MeJA did not affect the expression of TaWRKY49, and H2O2 reduced TaWRKY49 expression. In conclusion, TaWRKY49 negatively regulates while TaWRKY62 positively regulates wheat HTSP resistance to Pst by differential regulation of SA-, JA-, ET and ROS-mediated signaling.

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is a devastating disease of wheat (Triticum aestivum) worldwide. Wheat high-temperature seedling plant (HTSP) resistance to Pst is non-race-specific and durable. WRKY transcription factors have been proven to play important roles in plant defence responses to attacks by several pathogens. However, there is no direct evidence as to whether WRKY transcription factors play a role in HTSP resistance to Pst. We isolated a WRKY gene, named TaWRKY70, from wheat cultivar Xiaoyan 6. The expression level of TaWRKY70 was increased significantly when exposed to high temperatures (HTs) during the initial symptom expression stage of Pst infection. The expression of this gene increased in plants treated with ethylene (ET), salicylic acid (SA) and cold (4°C) stresses, but decreased in plants treated with methyl jasmonate (MeJA) and heat (40°C) stresses. Silencing of TaWRKY70 led to greater susceptibility to Pst (in terms of the increase in length of uredinial pustules and the decrease in the number of necrotic cells) compared with non-silenced plants when exposed to HT during the initial symptom expression stage of Pst infection, coinciding with expression changes of the ET- and SA-responsive genes TaPIE1 and TaPR1.1. In contrast, the expression level of the jasmonic acid (JA)-responsive gene TaAOS was not affected by TaWRKY70. These results indicate that TaWRKY70 is positively involved in HTSP resistance, during which SA and ET signalling are probably activated., (© 2016 BSPP AND JOHN WILEY & SONS LTD.)

In agricultural research, it is often difficult to construct a single "best" predictive model based on data collected under field conditions. We studied the relative prediction performance of combining empirical linear models over the single best model in relation to number of models to be combined, number of variates in the models, magnitude of residual errors, and weighting schemes. Two scenarios were simulated: the modeler did or did not know the relative of performance of the models to be combined. For the former case, model averaging is achieved either through weights based on the Akaike Information Criterion (AIC) statistic or with arithmetic averaging; for the latter case, only the arithmetic averaging is possible (because the relative model predictive performance is not known for a common dataset). In addition to two experimental datasets on oat mycotoxins in relation to environmental variables, two datasets were generated assuming a consistent correlation structure among explanatory variates with two magnitudes of residual errors. For the majority of cases, model averaging resulted in improved prediction performance over the single-model predictions, especially when a modeler does not have the information of relative model performance. The fewer variates in the models to be combined, the greater is improvement of model averaging over the single-model predictions. Combining models led to very little improvement over individual models when there were many variates in individual models. Overall, simple arithmetic averaging resulted in slightly better performance than the AIC-based weighted averaging. The advantage in model averaging is also noticeable for larger residual errors. This study suggests that model averaging generally performs better than single-model predictions, especially when a modeler does not have information on the relative performance of the candidate models.

Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most important diseases of wheat worldwide. Understanding the survival of Pst during the overwintering period is critical for predicting Pst epidemics in the spring. Real-time quantitative PCR (qPCR) methods quantifying Pst DNA and RNA (cDNA) were developed and compared for the ability to quantify viable Pst in leaf tissues. Both qPCR of DNA and RNA can provide reliable measurement of viable Pst in plant tissues prior to the late sporulation stage for which qPCR of DNA gave a much higher estimate of fungal biomass than qPCR of RNA. The percentage of Pst biomass that was viable in detached and attached leaves under low temperatures decreased over time. Pst survived longer on attached leaves than on detached leaves. The survival of Pst in cultivars with strong winter-hardiness at 0°C and -5°C was greater than those with weak winter-hardiness. However, such differences in Pst survival among cultivars were negligible at -10, -15 and -20°C. Results indicated that Pst mycelia inside green leaves can also be killed by low temperatures rather than through death of green leaves under low temperatures. The relationship of Pst survival in attached leaves with temperature and winter-hardiness was well described by logistic models. Further field evaluation is necessary to assess whether inclusion of other factors such as moisture and snow cover could improve the model performance in predicting Pst overwintering potential, and hence the epidemic in spring.

To determine the influence of plant density and powdery mildew infection of winter wheat and to predict grain yield, hyperspectral canopy reflectance of winter wheat was measured for two plant densities at Feekes growth stage (GS) 10.5.3, 10.5.4, and 11.1 in the 2009-2010 and 2010-2011 seasons. Reflectance in near infrared (NIR) regions was significantly correlated with disease index at GS 10.5.3, 10.5.4, and 11.1 at two plant densities in both seasons. For the two plant densities, the area of the red edge peak (Σdr680-760 nm), difference vegetation index (DVI), and triangular vegetation index (TVI) were significantly correlated negatively with disease index at three GSs in two seasons. Compared with other parameters Σdr680-760 nm was the most sensitive parameter for detecting powdery mildew. Linear regression models relating mildew severity to Σdr680-760 nm were constructed at three GSs in two seasons for the two plant densities, demonstrating no significant difference in the slope estimates between the two plant densities at three GSs. Σdr680-760 nm was correlated with grain yield at three GSs in two seasons. The accuracies of partial least square regression (PLSR) models were consistently higher than those of models based on Σdr680760 nm for disease index and grain yield. PLSR can, therefore, provide more accurate estimation of disease index of wheat powdery mildew and grain yield using canopy reflectance.

Quantification of Verticillium dahliae microsclerotia is an important component of wilt management on a range of crops. Estimation of microsclerotia by dry or wet sieving and plating of soil samples on semiselective medium is a commonly used technique but this method is resource-intensive. We developed a new molecular quantification method based on Synergy Brands (SYBR) Green real-time quantitative polymerase chain reaction of wet-sieving samples (wet-sieving qPCR). This method can detect V. dahliae microsclerotia as low as 0.5 CFU g(-1) of soil. There was a high correlation (r=0.98) between the estimates of conventional plating analysis and the new wet-sieving qPCR method for 40 soil samples. To estimate the inoculum threshold for cotton wilt, >400 soil samples were taken from the rhizosphere of individual plants with or without visual wilt symptoms in experimental and commercial cotton fields at the boll-forming stage. Wilt inoculum was estimated using the wet-sieving qPCR method and related to wilt development. The estimated inoculum threshold varied with cultivar, ranging from 4.0 and 7.0 CFU g(-1) of soil for susceptible and resistant cultivars, respectively. In addition, there was an overall relationship of wilt incidence with inoculum density across 31 commercial fields where a single composite soil sample was taken at each field, with an estimated inoculum threshold of 11 CFU g(-1) of soil. These results suggest that wilt risk can be predicted from the estimated soil inoculum density using the new wet-sieving qPCR method. We recommend the use of 4.0 and 7.0 CFU g(-1) as an inoculum threshold on susceptible and resistant cultivars, respectively, in practical risk prediction schemes.

Fusarium head blight (FHB) of wheat and barley has been extensively researched worldwide; in contrast, there is limited information on the effects of environmental conditions on Fusarium toxin accumulation in oat grains. More than 300 samples of oat grain from various regions of the United Kingdom from 2006 to 2008 were analyzed for mycotoxin contamination due to infection by Fusarium spp. HT2 and T2 toxins were the two most commonly detected, and their concentrations in individual samples were highly correlated. Hourly weather data were obtained from meteorological recording stations near most of the sampling sites. Statistical modeling was applied to both the original toxin (HT2 plus T2) data and the toxin data adjusted for oat cultivars and number of cereal crops in the previous four seasons. Accumulation of HT2 and T2 toxin was positively correlated with warm and wet conditions during early May and dry conditions thereafter. Using a collection of 51 environmental variables summarized over three lengths (10, 15, and 20 days) of time periods encompassing early May, late May, and early July, all-subsets regression showed that many models, consisting of three to six predictor variables, could be identified with similar explanatory strength for the effect of environmental conditions on toxin accumulation. Most important predictor variables were related to wet conditions during the early-May period, which was before anthesis. These results suggest that the predominant period for Fusarium langsethiae infection of oat is likely to be before rather than during anthesis, as for other head blight pathogens. These empirical models may be further improved by using quantified pathogen biomass within the grains and weather predictor variables summarized in relation to plant growth stages (instead of calendar times).

Background: Deployment of cultivars with different resistance in mixtures is one means to manage plant diseases and prolong the life of resistance genes. One major concern in adopting mixtures is the development of 'super-races' that can overcome many resistance genes present in the mixture. A stochastic simulation model was developed to study the dynamics of virulence alleles in two-cultivar mixtures of perennial crops, focusing on the effects of cost of virulence and pathogen reproduction mechanism. The simulated mechanism of virulence has characteristics of both major and minor genes., Results: Random genetic drift due to repeated population crashes during the overwintering phase led to fixation of a single fungal genotype (in terms of its virulence), often within 100 seasons. Overall, cost of virulence is most important in determining the virulence dynamics under the present model formulation. With cost of virulence incorporated, nearly all simulation runs ended up with a single fungal genotype that can infect only one of the two cultivars. In absence of cost of virulence, most of the simulation runs ended up with fungal genotypes that can infect both host cultivars but in many cases do not contain the maximum possible number of virulence alleles due to random drift. A minimum of 20% sexual reproduction between strains from different cultivars is necessary to ensure that the final fixed strains are able to infect both cultivars. Although the number of virulence alleles in the final genotype and the time to fixation are affected by simulation factors, most of the variability was among replicate simulation runs (i.e. stochastic in nature). The time to fixation is generally long relative to cropping cycles., Conclusions: A single fungal genotype will dominate a population due to the bottleneck in overwintering with cost of virulence primarily determining whether the dominant genotype can infect both cultivars. However, the dominant genotype is unlikely to accumulate all the virulence alleles due to genetic drift. The risk of emergence and spread of super-races is insufficiently great to prevent the use of cultivar mixtures of perennial crops as a means to reduce disease development provided that host resistance structure in mixtures is altered every cropping cycle.

There is increasing use of networks in ecology and epidemiology, but still relatively little application in phytopathology. Networks are sets of elements (nodes) connected in various ways by links (edges). Network analysis aims to understand system dynamics and outcomes in relation to network characteristics. Many existing natural, social, and technological networks have been shown to have small-world (local connectivity with short-cuts) and scale-free (presence of super-connected nodes) properties. In this review, we discuss how network concepts can be applied in plant pathology from the molecular to the landscape and global level. Wherever disease spread occurs not just because of passive/natural dispersion but also due to artificial movements, it makes sense to superimpose realistic models of the trade in plants on spatially explicit models of epidemic development. We provide an example of an emerging pathosystem (Phytophthora ramorum) where a theoretical network approach has proven particularly fruitful in analyzing the spread of disease in the UK plant trade. These studies can help in assessing the future threat posed by similar emerging pathogens. Networks have much potential in plant epidemiology and should become part of the standard curriculum.

Understanding the mechanisms responsible for divergence and specialization of pathogens on different hosts is of fundamental importance, especially in the context of the emergence of new diseases via host shifts. Temporal isolation has been reported in a few plants and parasites, but is probably one of the least studied speciation processes. We studied whether temporal isolation could be responsible for the maintenance of genetic differentiation among sympatric populations of Ampelomyces, widespread intracellular mycoparasites of powdery mildew fungi, themselves plant pathogens. The timing of transmission of Ampelomyces depends on the life cycles of the powdery mildew species they parasitize. Internal transcribed spacer sequences and microsatellite markers showed that Ampelomyces populations found in apple powdery mildew (Podosphaera leucotricha) were genetically highly differentiated from other Ampelomyces populations sampled from several other powdery mildew species across Europe, infecting plant hosts other than apple. While P. leucotricha starts its life cycle early in spring, and the main apple powdery mildew epidemics occur before summer, the fungal hosts of the other Ampelomyces cause epidemics mainly in summer and autumn. When two powdery mildew species were experimentally exposed to Ampelomyces strains naturally occurring in P. leucotricha in spring, and to strains naturally present in other mycohost species in autumn, cross-infections always occurred. Thus, the host-related genetic differentiation in Ampelomyces cannot be explained by narrow physiological specialization, because Ampelomyces were able to infect powdery mildew species they were unlikely to have encountered in nature, but instead appears to result from temporal isolation., (© 2011 Blackwell Publishing Ltd.)

Background: Myclobutanil, a demethylation inhibitor (DMI) fungicide, is an important fungicide for controlling apple scab and powdery mildew. Overuse of this fungicide has led to establishment of scab isolates with reduced sensitivity to this fungicide in several countries. Experiments were conducted to determine the sensitivity of the causal agent of apple scab, Venturia inaequalis (Cooke) Winter, to myclobutanil in the UK, in order to assess whether there is a relationship between fungal insensitivity and the number of DMI applications, and establishing whether fungal sensitivity varied greatly within an orchard., Results: Reduced sensitivity of V. inaequalis to myclobutanil was positively related linearly to the number of DMI applications. ED(50) values ranged from 0.028 to 1.017 mg L(-1) (average = 0.292) for the baseline population, whereas isolates from two other orchards had much greater ED(50) values, ranging from 0.085 to 5.213 mg L(-1) (average = 1.852). There was significant variation in fungal sensitivity to myclobutanil among fungal isolates from different locations within a single orchard., Conclusions: Spatial spread of insensitive isolates of V. inaequalis to myclobutanil is likely to be limited in distance. Conidia may be an important source of primary inoculum. Myclobutanil should still be effective for most field isolates, but its use should be strategically integrated with other groups of fungicides., ((c) 2009 Society of Chemical Industry.)

Research on the pathogen components involved in Fusarium head blight (FHB) along with the effects of their interactions on disease development and mycotoxin accumulation is reviewed. The fungal components within the FHB complex differ significantly in different environments. Individual species may respond differently to, and be differentially influenced by, particular disease control measures. Almost all published co-inoculation studies on wheat spikes or grains show that competitive interactions among FHB pathogens are the rule when fungal/disease development is considered. However, the fungi with the competitive advantage do not usually gain any advantage from the presence of other weaker competing fungi. Total mycotoxin production in mixed inoculations may decrease, increase, or remain unchanged compared with single-isolate inoculations, depending on the fungal species concerned and environmental conditions. A few recent studies, where each individual fungal component was quantified using molecular methods, suggest that mycotoxin productivity in mixed inoculations generally increases.

Published research on the effects of fungal interaction on disease development and subsequent mycotoxin accumulation was reviewed, focusing on pathogens related to Fusarium Head Blight (FHB). Almost all published studies showed that competitive interactions are the rule when fungal/disease development is considered. The fungi with the competitive advantage did not usually colonise significantly more than when inoculated alone, i.e. there was no advantage gained by the dominant pathogen from the presence of other weaker competing fungi. However, the effects of fungal interactions on mycotoxin accumulation were generally more complicated. Total mycotoxin production in mixed inoculation may decrease, increase or remain at a similar level compared with single-isolate inoculation, depending on the fungal species concerned and environmental conditions. However, the lack of accurate quantification of each competing fungal component in mixed inoculations in many studies prevented an accurate estimation of mycotoxin productivity per unit fungal biomass. A few recent studies, where each individual fungal component was quantified using molecular methods, suggested that mycotoxin productivity in mixed inoculations generally increased.

Transcription factors (TFs) bind to the promoters of target genes to regulate gene expression in response to different stimuli. The functions and regulatory mechanisms of transcription factors (TFs) in Verticillium dahliae are, however, still largely unclear. This study showed that a C2H2-type zinc finger TF, VdCf2 (V. dahliae chorion transcription factor 2), plays key roles in V. dahliae growth, melanin production, and virulence. Transcriptome sequencing analysis showed that VdCf2 was involved in the regulation of expression of genes encoding secreted proteins, pathogen-host interaction (PHI) homologs, TFs, and G protein-coupled receptors (GPCRs). Furthermore, VdCf2 positively regulated the expression of VdPevD1 (VDAG_02735), a previously reported virulence factor. VdCf2 thus regulates the expression of several pathogenicity-related genes that also contribute to virulence in V. dahliae. VdCf2 also inhibited the transcription of the Vd276-280 gene cluster and interacted with two members encoding proteins (VDAG_07276 and VDAG_07278) in the gene cluster. IMPORTANCE Verticillium dahliae is an important soilborne phytopathogen which can ruinously attack numerous host plants and cause significant economic losses. Transcription factors (TFs) were reported to be involved in various biological processes, such as hyphal growth and virulence of pathogenic fungi. However, the functions and regulatory mechanisms of TFs in V. dahliae remain largely unclear. In this study, we identified a new transcription factor, VdCf2 (V. dahliae chorion transcription factor 2), based on previous transcriptome data, which participates in growth, melanin production, and virulence of V. dahliae. We provide evidence that VdCf2 regulates the expression of the pathogenicity-related gene VdPevD1 (VDAG_02735) and Vd276-280 gene cluster. VdCf2 also interacts with VDAG_07276 and VDAG_07278 in this gene cluster based on a yeast two-hybrid and bimolecular fluorescence complementation assay. These results revealed the regulatory mechanisms of a pivotal pathogenicity-related transcription factor, VdCf2 in V. dahliae.

Rhizosphere microbial communities are known to be related to plant health; using such an association for crop management requires a better understanding of this relationship. We investigated rhizosphere microbiomes associated with Verticillium wilt symptoms in two cotton cultivars. Microbial communities were profiled by amplicon sequencing, with the total bacterial and fungal DNA quantified by quantitative polymerase chain reaction based on the respective 16S and internal transcribed spacer primers. Although the level of V. dahliae inoculum was higher in the rhizosphere of diseased plants than in the healthy plants, such a difference explained only a small proportion of variation in wilt severities. Compared to healthy plants, the diseased plants had much higher total fungal/bacterial biomass ratio, as represented by quantified total fungal or bacterial DNA. The variability in the fungal/bacterial biomass ratio was much smaller than variability in either fungal or bacterial total biomass among samples within diseased or healthy plants. Diseased plants generally had lower bacterial alpha diversity in their rhizosphere, but such differences in the fungal alpha diversity depended on cultivars. There were large differences in both fungal and bacterial communities between diseased and healthy plants. Many rhizosphere microbial groups differed in their abundance between healthy and diseased plants. There was a decrease in arbuscular mycorrhizal fungi and an increase in several plant pathogen and saprophyte guilds in diseased plants. These findings suggested that V. dahliae infection of roots led to considerable changes in rhizosphere microbial communities, with large increases in saprophytic fungi and reduction in bacterial community. [ABSTRACT FROM AUTHOR]

Verticillium wilt caused by Verticillium dahliae is a common soil-borne disease worldwide, affecting many economically important crop species. Soil microbes can influence plant disease development. We investigated rhizosphere and endosphere microbiomes in relation to cotton cultivars with differential susceptibility to Verticillium wilt. Soil samples from nine cotton cultivars were assessed for the density of V. dahliae microsclerotia; plants were assessed for disease development. We used amplicon sequencing to profile both bacterial and fungal communities. Unlike wilt severity, wilt inoculum density did not differ significantly among resistant and susceptible cultivars. Overall, there were no significant association of alpha diversity indices with wilt susceptibility. In contrast, there were clear differences in the overall rhizosphere and endosphere microbial communities, particularly bacteria, between resistant and susceptible cultivars. Many rhizosphere and endosphere microbial groups differed in their relative abundance between resistant and susceptible cultivars. These operational taxonomic units included several well-known taxonomy groups containing beneficial microbes, such as Bacillales, Pseudomonadales, Rhizobiales, and Trichoderma , which were higher in their relative abundance in resistant cultivars. Greenhouse studies with sterilized soil supported that beneficial microbes in the rhizosphere contribute to reduced wilt development. These findings suggested that specific rhizosphere and endosphere microbes may contribute to cotton resistance to V. dahliae. [ABSTRACT FROM AUTHOR]