Enhanced field control of wheat take-all using cold tolerant isolates of Gaeumannomyces graminis var. graminis and Phialophora sp. (lobed hyphopodia)

Cold tolerant isolates of Gaeumannomyces graminis var. graminis ( Ggg ) and Phialophora sp. (lobed hyphopodia), which produced at least comparable growth rates at 5°C to those of pathogenic G. graminis var. tritici ( Ggt ), were shown to control take-all disease in wheat effectively in 2 years of field experiments in New South Wales, Australia. The addition of oat inoculum of these fungi at the rate of 60 kg/ha to the seeding furrow significantly ( P  ≤ 0.05) reduced disease and increased grain yields by 33–45% compared to the Ggt alone treatment. The use of 30 kg/ha of oat inoculum also significantly ( P  ≤ 0.05) reduced disease and increased grain yields by 21–44%. These high levels of take-all control were obtained consistently from four field experiments on three different soil types with different pHs. A treatment inoculated with Ggg alone showed no disease symptoms and produced grain yields similar to that of untreated wheat. This fungus is, therefore, non-pathogenic to wheat. At high rates of inoculation of Ggg and Phialophora sp. (lobed hyphopodia), 65–80% of tillering wheat plants (GS 32) had root systems colonized by these fungi. In contrast, two Pseudomonas spp. and an isolate each of Ggg and Phialophora sp. (lobed hyphopodia), which did not grow at 5°C, were ineffective in controlling take-all. Take-all assessments during heading (GS 61-83) were highly correlated ( R ²=0.6047, P ≤0.0005) with the relative yield increase or decrease of inoculated treatments compared to the Ggt alone treatment. The use of a Ggg isolate (90/3B) and a Phialophora sp. (lobed hyphopodia) isolate (KY) for take-all control has been patented. These fungi are being developed for commercial use.     

Biological Control of Take-all by Phialophora radicicola Cain1

Biological control of take-all by Phialophora radicicola Cain and similar fungi is reviewed, and new evidence is presented on the possible role of hyper-parasites, like Pythium oligandrum Drechsler, to augment this. Phialophora radicicola var. graminicola Deacon is abundant in British grasslands. Its role in biological control of take-all has been demonstrated in the glasshouse with natural soils and natural population levels of this control agent. Also there is much circumstantial evidence for its beneficial role in agriculture, especially in cereal monoculture following grass crops, and in natural and amenity grasslands. Some other similar fungi control take-all in the glasshouse, but their roles in current agricultural practice are not known. The take-all fungus, itself, can reduce infection of roots by P. radicicola, so there is a dynamic interaction between these fungi. This is affected by host type, relative inoculum potentials of the fungi, and possibly by environmental conditions. By careful manipulation, therefore, it might be possible to control take-all under a wide range of field conditions. The chief role of P. radicicola and similar fungi is to delay establishment of severe take-all early in cereal monoculture, rather than to combat an existing high disease level. It may therefore be desirable to combine different biocontrols, and hyperparasites like Pythium oligandrum may be important in this respect. This fungus was tested against varieties of Gaeumannomyces graminis Arx & Olivier and Phialophora radicicola in the laboratory; the host responses differed greatly, from marked susceptibility (P. radicicola var. radicicola) to almost complete resistance (G. graminis var. graminis). Hence, P. oligandrum might be used to alter relative population levels of these fungi in the field, but first its behaviour in natural soils must be studied.     

The use of rainfall and accumulated mean temperature to indicate fungicide activity in the control of leaf diseases of winter wheat in the UK1

During 1988/1990, a series of 21 experiments was established in commercial crops of winter wheat. Chlorothalonil, fenpropimorph and propiconazole were chosen as protectant, eradicant or curative fungicides active against leaf diseases of winter wheat in the UK. To test their properties each one was applied once only to separate plots during a period of 7–8 consecutive weeks in May and June (GS 32–39). Disease progress was assessed weekly on adjacent unsprayed control and sprayed plots up to GS 85. Septoria tritici leaf blotch (Mycosphaerella graminicola) was the disease that occurred most frequently and severely across the 21 sites. Powdery mildew (Erysiphe graminis), brown rust (Puccinia recondita) and yellow rust (P. striiformis) occurred at fewer sites and were sufficiently severe to distinguish differences between the active ingredients at only two or three sites. Analysis of the disease progress curves for 75% control of each disease at one site only indicated that chlorothalonil possessed very good protectant but shorter-term eradicant activity against M. graminicola and P. striiformis. Fenpropimorph exhibited only short-term eradicant control of M. graminicola, but gave excellent protectant and eradicant control of E. graminis and P. striiformis; against P. recondita only eradicant activity was apparent. Propiconazole showed activity similar to that of fenpropimorph against P. recondita and excellent protectant and eradicant activity against M. graminicola and P. striiformis; against E. graminis, it gave good protectant and eradicant control. From disease progress curves, it was possible to calculate the period of protectant and eradicant activity in thermal time (accumulated degree days above zero) for each of the three active ingredients and to identify the most effective timing(s) for fungicide application in relation to rainfall or imputed infection.     

Investigation of soilborne mosaic virus diseases transmitted by <Emphasis Type="Italic">Polymyxa graminis</Emphasis> in cereal production areas of the Anatolian part of Turkey

Polymyxa graminis is the vector of several important viruses, including Soilborne cereal mosaic virus, Wheat spindle streak mosaic virus, Barley yellow mosaic virus and Barley mild mosaic virus, of winter cereals worldwide. Surveys were carried out to detect these viruses and their vector P. graminis in 300 soil samples from the main wheat and barley production areas of the Anatolian part of Turkey collected in May 2002, June 2004 and May 2005. For these surveys, various susceptible wheat and barley cultivars were pot grown in the collected soil samples in a greenhouse and then analysed using ELISA and RT-PCR to detect the presence of different virus species. In addition, a combination of light microscopy following roots staining with acid fuchsin and PCR was used for detection of P. graminis. All soil samples analysed were found to be free of these soilborne viruses and their vector.     

我国小麦地方品种蚂蚱麦、小白冬麦、游白兰、红卷芒抗白粉病性遗传分析

我国地方品种是小麦白粉病抗性的重要来源之一,为了对地方品种抗源的利用奠定基础,采用常规杂交方法,以感病品种Chancellor分别与我国小麦抗病地方品种蚂蚱麦、小白冬麦、游白兰、红卷芒进行正交和反交,获得F₁、F₂代。根据白粉菌菌株的毒谱选用E09菌株对Chancellor与小白冬麦、游白兰、红卷芒的杂交后代进行苗期抗性鉴定和统计分析,选用E30菌株对Chancellor与蚂蚱麦的杂交后代进行苗期抗性鉴定和统计分析。结果表明4个品种在正、反交情况下均表现出由一对隐性基因控制的抗性,说明这4个地方品种属于核遗传,其抗性是由一对隐性基因控制的。     

Inoculum potential and host range of Pofymyxa graminis1

The inoculum potential of Pofymyxa graminis was studied for 48 samples of Belgian soils and 8 samples from other European countries. After growing barley bait plants in tubes under controlled conditions on increasing dilutions of the samples in sterile sand, P. graminis was detected in 77% of the Belgian soils, the number of infection units calculated per g of soil ranging from 0.004 to 5.6. The inoculum potentials for the other origins were within these limits. The host range of P. graminis was studied by growing various Gramineae in the presence of cystosori extracted from barley roots cultivated on five soils. Barley was infected by all the isolates. Some isolates infected oat and/or rye but none wheat.     

Fungicide sensitivity of populations of wheat powdery mildew (Erysiphe graminis f.sp. Tritici) in Central Europe in 1993

In 1993 we observed the sensitivity of wheat powdery mildew populations (Erysiphe graminis DC f.sp. tritici Marchal) from the Czech Republic, Austria, Hungary and Slovakia to the fungicides triadimenol, tebuconazole, propiconazole, flutriafol and fenpropimorph. The highest resistance value was shown to triadimenol, which attained a mean resistance factor (MRF) of 29 (expressing how many times the population is more resistant than are standard sensitive isolates) in the mildew population from the Czech Republic. The mildew populations from eastern Slovakia and eastern Hungary, populations geographically isolated from the other populations, showed very high sensitivity to all fungicides tested. There was most sensitivity to fenpropimorph (smallest MRF values) compared with the other fungicides. Cross-resistance was established among all triazoles used, but not between triazoles and fenpropimorph. Sensitivity of wheat powdery mildew populations from Central Europe to these fungicides is considered adequate, and the development of resistance has shown a decreasing tendency in recent years.     

Spatial connectedness of plant species: potential links for apparent competition via plant diseases

This study evaluated the reactions of seven common C4 grasses of the tallgrass prairie of the USA Great Plains to the economically important wheat pathogens Pyrenophora tritici‐repentis and Gaeumannomyces graminis var. tritici (Ggt) isolated from wheat. The P. tritici‐repentis isolates (race 1) were pathogenic on all grasses tested, but symptom severity was markedly low. Three of the grass species inoculated with Ggt were highly susceptible, while four species exhibited no symptoms. Because measures of connectedness can provide a proxy for population processes, connectedness was evaluated within and among the seven grass species in representative tallgrass prairie environments for all potential pathogen‐sharing patterns. Andropogon gerardii was ubiquitous, so all plant species were well connected to it. Andropogon scoparius (= Schizachyrium scoparium), Sorghastrum nutans and Panicum virgatum were fairly common but specialized to particular environments. Bouteloua curtipendula was uncommon but occurred in all environments, while Buchloë dactyloides and Bouteloua gracilis were uncommon and only occurred in upland sites. Co‐occurrence of plant species was generally not reciprocal in that, for many species pairs, species A rarely occurred without potential exposure to inoculum from species B, while species B commonly occurred without species A. The three grass species susceptible to Ggt may act as sources of inoculum for each other within tallgrass prairie, with the potential to influence fitness, and tallgrass prairie and commercial wheat ecosystems in the Great Plains also have the potential to share both pathogens.     

Confirmed resistance to aryloxyphenoxypropionate herbicides in Phalaris minor populations in Iran

Phalaris minor (littleseed canary grass) is a major weed in wheat fields in some parts of Iran. Diclofop‐methyl, fenoxaprop‐P‐ethyl, and clodinafop‐propargyl are three acetyl coenzyme A carboxylase (ACCase)‐inhibiting herbicides that are commonly used to control this grass in wheat fields. Thirty‐four P. minor populations with suspected resistance to ACCase‐inhibiting herbicides were sampled from wheat fields in the provinces of Fars and Golestan in Iran. The dose–response assays that were conducted under controlled greenhouse conditions indicated that 14 populations were resistant to fenoxaprop‐P‐ethyl, seven populations were resistant to both fenoxaprop‐P‐ethyl and diclofop‐methyl, and three populations were resistant to fenoxaprop‐P‐ethyl, diclofop‐methyl, and clodinafop‐propargyl. These populations showed different levels of resistance to the applied herbicides, compared to the susceptible population. These results suggest that different mechanisms of resistance could be involved. The enzyme assay revealed that the existence of modified ACCase in the three most‐resistant populations (AR, MR4, and SR3) is responsible for the resistance of these populations.     

Disease development and genotypic diversity of Puccinia graminis f. sp. avenae in Swedish oat fields

The disease development and population structure of Puccinia graminis f. sp. avenae, which causes stem rust on oats, were studied to investigate if sexual reproduction plays an important role in the epidemiology of the disease. The genetic population structure of P. graminis f. sp. avenae in Sweden was investigated by sampling 10 oat fields in July and August 2008 and seven fields during the same period in 2009. Nine single‐pustule isolates were first used to test simple sequence repeat (SSR) markers developed for P. graminis f. sp. tritici. Eleven of the 68 tested SSR markers were useful for genotyping P. graminis f. sp. avenae. For the main study, DNA from single uredinia was extracted and the SSR markers were used to genotype 472 samples. Both allelic and genotypic diversity were high in all fields, indicating that P. graminis f. sp. avenae undergoes regular sexual reproduction in Sweden. No significant relationship between genetic and geographic distances was found. Disease development was studied on two farms during 2008 and 2009. The apparent infection rates ranged between 0·17 and 0·55, indicating the potential for rapid disease development within fields. The incidence of oat stem rust has increased recently in Sweden. One possible explanation is a resurgence of its alternate host, barberry (Berberis spp.), after the repeal of the barberry eradication law in 1994. Barberry is present in several grain‐producing areas in Sweden, which supports the conclusion that P. graminis f. sp. avenae undergoes regular sexual reproduction there.     

Uptake, translocation and phytotoxicity of root-absorbed haloxyfop in soybean, Festuca rubra L. and Festuca arundinacea Schreb

The concentrations of haloxyfop in nutrient solution required to reduce the total plant dry weight of soybean (Glycine max L. Merr. ‘Evans’), red fescue (Festuca rubra L. ‘Pennlawn’), and tall fescue (Festuca arundinacea Schreb. ‘Houndog’) by 50% (GR₅₀) were determined. The GR₅₀) values for soybean, red fescue and tall fescue were 76 μM, 3μM and 0.4 μM, respectively. The reduction in growth in roots and shoots of soybean was similar. In contrast, the relative reduction in root tissue weight was greater than that for foliar tissue in both grass species. The amount of ¹⁴C-haloxyfop in soybean roots or shoots was higher than in red fescue or tall fescue. Red fescue accumulated less haloxyfop in the foliage than in the roots. On the other hand, similar amounts of ¹⁴C-haloxyfop accumulated in both organs in both soybean and tall fescue. ¹⁴C-haloxyfop appeared to be actively absorbed by the roots of all species. Soybean absorbed more nutrient solution, but utilized it less on a per gram dry matter produced basis than the grass species. Differences in the uptake and translocation of haloxyfop by roots do not account for differences in tolerance between species. However, a higher level of retention of haloxyfop in the roots of red fescue than in tall fescue may provide the former with an additional selectivity advantage under conditions where there is significant root exposure to the herbicide.     

小麦锈菌蛋白激酶基因家族的鉴定与生物信息学分析

为深入分析锈菌结构基因组,阐明锈菌毒性变异的分子机制,本研究通过生物信息分析方法,对3种小麦锈菌蛋白激酶(protein kinases,PKs)超家族预测基因进行了系统分析,利用COG(clusters of orthologous groups of proteins)、KEGG(Kyoto encyclopedia of genes and genomes)、GO(gene ontology)和PHI(pathogen-host interactions)数据库进行注释分析,并对小麦锈菌MAPK基因的蛋白质互作网络进行预测。结果表明,条锈菌Puccinia striiformis、叶锈菌P.triticina和秆锈菌P.graminis的PKs基因数量分别为221、159和159个,不同PKs基因家族类型在3种锈菌中的数量分布上表现出很高的保守性。注释分析表明,PKs家族预测功能涉及病原菌生长发育中的调控作用和病原菌-寄主互作机制,包括信号传导和致病因子等。PKs家族核心基因数目在蛋白激酶基因中占比大于1/3。MAPK基因的催化结构域序列呈现高度相似性。以STRING数据库的酿酒酵母蛋白质互作网络信息为参考,对小麦锈菌MAPK基因的蛋白质互作网络进行预测,共鉴定出25个互作关系,包含29个MAPK基因。研究表明这3种锈菌之间MAPK互作蛋白质分布不均衡,这可能反映了锈菌基因组进化的特殊复杂性。     

Genetic Structure of Mycosphaerella graminicola Populations from Iran, Argentina and Australia

Restriction fragment length polymorphism (RFLP) markers were used to assess the genetic structure of populations of Mycosphaerella graminicola collected from wheat fields. A total of 585 isolates representing 10 field populations were sampled from Iran, Argentina and Australia. The genetic structure of M. graminicola populations from Iran and Argentina is described for the first time. Results were compared to previously investigated populations from Israel, Uruguay and Australia. Populations from Iran exhibited high clonality and low gene diversity, suggesting an inoculation event. Populations from uninoculated fields in Argentina had gene and genotype diversities similar to previously described European and North American populations. Genotype diversity was high for populations from Australia and tests for multilocus associations were consistent with sexual recombination in these populations. Gene diversity was low and fixed alleles were found for several loci. These findings are consistent with a relatively small founding population for Australia. These 10 new populations were integrated into a genetic distance comparison with 13 global populations that were characterized earlier.     

Genetic structure of Mycosphaerella graminicola populations in Iran

To provide insight into the genetic structure of Mycosphaerella graminicola populations in Iran, a total of 221 isolates were collected from naturally infected wheat fields of five major wheat‐growing provinces and analysed using AFLP markers and mating‐type loci. All populations showed intermediate to high genotypic diversity. In the Golestan and Ardabil populations two mating types were found at near‐equal frequencies, whilst all populations were in gametic disequilibrium. Moreover, clonal haplotypes were identified in different sampling sites within a field in both the Khuzestan and Fars provinces, demonstrating that pycnidia are probably the primary source of inoculum. All five populations had low levels of gene diversity and had private bands. Low levels of gene flow and high genetic differentiation were observed among populations and different clustering methods revealed five genetically distinct groups in accordance with the sampling areas. The Golestan and East Azarbaijan populations were more genetically differentiated than the others. Random genetic drift, selection and geographic barriers may account for the differentiation of the populations. The results of this study indicate a population structure of M. graminicola in Iran contrasting to that of most other countries studied.     

SnToxA,SnTox1, and SnTox3 originated in Parastagonospora nodorum in the Fertile Crescent

The centre of origin of the globally distributed wheat pathogen Parastagonospora nodorum has remained uncertain because only a small number of isolates from the Fertile Crescent were included in earlier population genetic and phylogeographic studies. We isolated and genetically analysed 193 P. nodorum strains from three naturally infected wheat fields distributed across Iran using 11 neutral microsatellite loci. Compared to previous studies that included populations from North America, Europe, Africa, Australia, and China, the populations from Iran had the highest genetic diversity globally and also exhibited greater population structure over smaller spatial scales, patterns typically associated with the centre of origin of a species. Genes encoding the necrotrophic effectors SnToxA, SnTox1, and SnTox3 were found at a high frequency in the Iranian population. By sequencing 96 randomly chosen Iranian strains, we detected new alleles for all three effector genes. Analysis of allele diversity showed that all three effector genes had higher diversity in Iran than in any population included in previous studies, with Iran acting as a hub for the effector diversity that was found in other global populations. Taken together, these findings support the hypothesis that P. nodorum originated either within or nearby the Fertile Crescent with a genome that already encoded all three necrotrophic effectors during its emergence as a pathogen on wheat. Our findings also suggest that P. nodorum was the original source of the ToxA genes discovered in the wheat pathogens Phaeosphaeria avenaria f. sp. tritici 1, Pyrenophora tritici-repentis, and Bipolaris sorokiniana.     

Dominant Colonisation of Wheat Roots by Pseudomonas fluorescens Pf29A and Selection of the Indigenous Microflora in the Presence of the Take-all Fungus

Increases in populations of fluorescent pseudomonads on wheat roots are usually associated with take-all decline, natural control of take-all, a disease caused by the fungus Gaeumannomyces graminis var. tritici (Ggt). Colonisation by Pseudomonas fluorescens strain Pf29A was assessed on the roots of healthy plants and of plants with take-all, and the effect of this bacterium on indigenous populations of fluorescent pseudomonads was studied. The efficacy of Pf29A as an agent for the biocontrol of take-all on five-week-old wheat seedlings was tested in non-sterile conducive soil in a growth chamber. RAPD (random amplification of polymorphic DNA) fingerprinting with a decamer primer was used to monitor strain Pf29A and culturable indigenous rhizoplane populations of fluorescent pseudomonad. Pf29A decreased disease severity and accounted for 44.6% of the culturable fluorescent pseudomonads on healthy plant rhizoplane and 75.8% on diseased plant rhizoplane. Fewer RAPD patterns were obtained when Pf29A was introduced into the soil with Ggt. In the presence of Ggt and necrotic roots, Pf29A became the dominant root coloniser and dramatically changed the diversity and the structure of indigenous fluorescent pseudomonad populations. The results show that Ggt and reduced lesion size on roots can trigger a specific increase in antagonist populations and that the introduction of a biocontrol agent in soil influences the structure of indigenous bacterial populations.     

Frequency of mutations associated with fungicide resistance and population structure of <Emphasis Type="Italic">Mycosphaerella graminicola</Emphasis> in Tunisia

The occurrence of fungicide resistance in Mycosphaerella graminicola populations from Tunisia was investigated by examining mutations known to be associated with strobilurin and azole resistance. Few mutations associated with fungicide resistance were detected. No evidence for strobilurin resistance was found among 357 Tunisian isolates and only two among 80 sequenced isolates carried mutations associated with azole resistance. A network analysis suggested that these mutations emerged independently from mutations found in previously described European populations. The population genetic structure of M. graminicola in Tunisia was analyzed using variation at 11 microsatellite loci. Populations in Tunisia were characterized by high gene and genotype diversity. All populations were in gametic equilibrium and mating type proportions did not deviate from the 1:1 ratio expected under random mating, consistent with regular cycles of sexual reproduction. In combination with a high degree of gene flow among sampling sites, M. graminicola must be considered a pathogens with high evolutionary potential. Thus, control strategies against Septoria blotch in Tunisia should be optimized to reduce the emergence and spread of resistant isolates.     

Phylogenetic and morphological analyses of Pythium graminicola and related species

Isolates of Pythium graminicola and related species were differentiated using restriction fragment length polymorphism (RFLP) analyses of the internal transcribed spacer (ITS) regions of rDNA and the cytochrome c oxidase subunit II (COX II) gene. These sequences were used in subsequent phylogenetic analyses. Finally, the phylogenetic placement of species was compared to that determined from morphological characteristics. The 62 isolates tested were divided into seven groups, A–G, based on RFLP analysis of the rDNA-ITS region. In the RFLP analysis of the COX II gene, isolates were divided into groups similar to those based on ITS-RFLP. Groups A and B were each separated into two additional subgroups. Grouping of isolates based on RFLP analyses agreed with the morphological differentiation. Groups A, B, D, E, F, and G were identified as P. graminicola, P. arrhenomanes, P. aphanidermatum, P. myriotylum, P. torulosum, and P. vanterpoolii, respectively. Group C was closely related to group B based on phylogenetic analysis of the rDNA-ITS region and the COX II gene and is similar to P. arrhenomanes. Each of the other species occupied their own individual clades. Although P. arrhenomanes is morphologically similar to P. graminicola, our phylogenetic analyses revealed that it was evolutionarily distant from P. graminicola and more closely related to P. vanterpoolii. Our analysis also revealed that P. torulosum with smaller oogonia is more closely related to P. myriotylum with large oogonia than to P. vanterpoolii, which forms smaller oogonia and is morphologically similar to P. torulosum. P. aphanidermatum with large oogonia and aplerotic oospores was not related to the morphologically similar species P. myriotylum. Results suggest that P. graminicola and related species are phylogenetically distinct, and molecular analyses, in addition to morphological analyses, are necessary for the accurate taxonomic placement of species in this complex.     

Barberry plays an active role as an alternate host of Puccinia graminis in Spain

Stem rust, caused by Puccinia graminis, is a destructive group of diseases. The pathogen uses Berberis species as alternate hosts to complete its life cycle. B. vulgaris and the endemic species B. hispanica and B. garciae are present in Spain. The objective of this study was to investigate the functionality of the indigenous barberry as alternate hosts. Field surveys were conducted in 2018 and 2019 in Huesca, Teruel and Albacete provinces of Spain. Aecial samples on barberry were analysed via infection assays and DNA analysis. B. garciae was predominant in Huesca and Teruel provinces, often found in the field margins of cereal crops. Aecial infections on B. garciae were observed in May and uredinial infections on cereal crops in June. Scattered B. hispanica bushes were occasionally found near cereal crops in Albacete, where aecial infections on B. hispanica were observed in June when most cereal crops were mature. Infection assays using aeciospores resulted in stem rust infections on susceptible genotypes of wheat, barley, rye and oat, indicating the presence of the sexual cycle for P. graminis f. sp. tritici, f. sp. secalis and f. sp. avenae. Sequence analyses from aecial samples supported this finding as well as the presence of Puccinia brachypodii. This study provides the first evidence that indigenous Berberis species play an active role in the sexual cycle of P. graminis under natural conditions in Spain.     

Diversity of rDNA-ITS sequences of <Emphasis Type="Italic">Polymyxa graminis</Emphasis> from wheat and barley in Japan

Polymyxa graminis is a soil-borne obligate organism that transmits bymoviruses and furoviruses to barley and wheat. We analyzed rDNA-ITS sequences of P. graminis from roots of wheat and barley in fields in Japan and obtained five kinds of sequences; two sequences were almost the same as known ribotype Ia and IIa, respectively, and three were close to ribotype Ib. When infection of P. graminis was examined using PCR, ribotype Ia was detected only in barley, but ribotypes Ib and IIa were detected in both wheat and barley. Our analysis suggested that Japanese ribotype Ib transmits furoviruses and bymoviruses.