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.     

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.     

Temperature-sensitive wheat stem rust resistance gene Sr15 is effective against Puccinia graminis f. sp. tritici race TTKSK

The wheat stem rust fungus, Puccinia graminis f. sp. tritici (Pgt), race TTKSK and related races pose a serious threat to world wheat production. Knowing the effectiveness of wheat stem rust resistance (Sr) genes against Pgt race TTKSK is fundamental in mitigating this threat through resistance breeding. Sr15 was previously identified as being ineffective against Pgt race TTKSK. Here, multirace disease phenotyping data, linkage analyses, allelism testing and haplotype analyses are presented to support the conclusion that Sr15 is effective against Pgt race TTKSK. Resistance to race TTKSK was mapped to Sr15 in a biparental population. Thirty-two accessions with Sr15 displayed seedling resistance phenotypes against race TTKSK. However, these accessions were susceptible as seedlings at high temperatures (22–25 °C), consistent with previous reports that the interaction between avirulent Pgt isolates and Sr15 is temperature-sensitive. Markers STS638, wri4 and KASP_IWB30995 were found to predict the presence of Sr15, suggesting the utility of these assays for marker-assisted selection in breeding programmes. The effectiveness of Sr15 to specific Pgt races and temperatures makes it a less-desirable TTKSK-effective gene. Wheat lines assayed as resistant to race TTKSK at the seedling stage may possess Sr15 and breeders should be aware of the limitations of Sr15 for conferring stem rust resistance.     

<Emphasis Type="Italic">Berberis</Emphasis> phyllody is a phytoplasma-associated disease

Berberis thunbergii atropurpurea, also known as Red barberry, is a small ornamental shrub in the family Berberidaceae. In recent years, a phyllody symptom has been observed frequently, spreading in the shrubs in northwestern China. A phytoplasma 16S rDNA specific fragment was amplified by PCR from Berberis plants with the phyllody symptoms. DNA sequencing and restriction fragment length polymorphism analysis revealed the phytoplasma belongs to 16SrV-B subgroup. This is the first report that Berbegis thunbergii atropurpurea is a host for 16SrV-B phytoplasma. The disease was named Berberis phyllody.     

African wheat germplasm – a valuable resource for resistance to rust diseases

Known and unknown genes conferring seedling and adult plant resistance (APR) to leaf rust, stem rust and stripe rust were detected either singly or in combination in a set of 136 African wheat genotypes using multi-pathotype tests with characterized Australian Puccinia triticina (Pt), P. graminis f. sp. tritici (Pgt) and P. striiformis f. sp. tritici (Pst) pathotypes. Lines Beladi 132, IYN 68/9.44, Kenya Kifaru and Kenya Mbweha were postulated to carry resistance against multiple pathotypes of Pt, Pgt and Pst, whereas IAR/W/163-3, Grano Di Moggio Tipo 44 and Trigo 48 had resistance against all pathotypes tested in the current study. Field evaluation with the three rust pathogens detected low to high APR in more than 50% of lines, and while most tested positive with markers linked to known APR genes (csLV34, csLV46G22, TM10KASPAR, csGS, Cfb5006 and csSr2), many carried unidentified and useful resistance to all three rusts. Genetic analysis of F₃ mapping populations based on seven genotypes showed either monogenic or digenic inheritance of APR to leaf rust, stem rust and stripe rust. The lines postulated to carry effective uncharacterized seedling genes and APR genes are of great potential value in diversifying resistance to help achieve durable control of all three rust diseases of wheat.     

Genetic Diversity in Relation to Sexual and Asexual Reproduction in Populations of Melampsora larici-epitea

Genetic diversity of Melampsora larici-epitea leaf rust from three cultivated stands of the willow Salix viminalis was studied using AFLP polymorphisms at 60 loci. One population was located in Northern Ireland and two in Sweden. Analysis of molecular variance (AMOVA) showed that most of the genetic variation was distributed on a fine scale within the field in all populations. Both Swedish populations displayed a very high genotypic diversity (normalized Shannon's indices of 0.95 and 1.00) and random association among loci. These results suggested that sexual reproduction had an important influence on the Swedish populations. The occurence of the alternate host (larch) adjacent to one of the Swedish rust populations did not affect the genetic diversity. However, severe rust attacks started earlier in the season in this population. The M. larici-epitea population in Northern Ireland was characterized by a low genotypic diversity (normalized Shannon's index = 0.54) and non-random association among loci was indicated by test of multilocus association and by pairwise tests among loci. These results suggested that asexual reproduction had a major effect on the genetic structure of this population, probably because of the overwintering of asexual spores and/or a population bottleneck associated with the annual sexual phase.     

The wheat stem rust pathogen in the central region of the Russian Federation

A total of 387 isolates of Puccinia graminis f. sp. tritici (Pgt) collected in the central region of the Russian Federation from 2000 to 2009 was analysed with North American differential sets comprising 16 genotypes for samples of 2000–2006 and 20 genotypes for samples from 2007–2009. Forty‐five races were identified. The race composition of the local population underwent changes during this period. Race MKBT was the predominant race in the earlier years, but TKNT and TKNTF were in the majority later. During 2000–2009 there were no stem rust epidemics in the region. It was assumed that the local pathogen population cycled on wild grasses (including Elytrigia, Agropyron, Festuca, Dactylis, Phleum and Lolium spp.) and not only on wheat. The existence of host communities of wheat stem rust was supported by random amplified polymorphic DNA (RAPD) markers produced with high‐GC primers. The local population of Pgt was considered to be sexual based on the relatively high diversity of races isolated from various hosts and the absence of correlation between virulence attributes and molecular markers.     

Characterization of Puccinia graminis f. sp. tritici isolates derived from an unusual wheat stem rust outbreak in Germany in 2013

An unusual stem rust infestation occurred in German wheat fields in summer 2013. This study analysed 48 isolates derived from 17 Puccinia graminis f. sp. tritici (Pgt) samples and six races were identified: TKTTF, TKKTF, TKPTF, TKKTP, PKPTF and MMMTF. Infection type and genotypic data confirmed that none of these races belonged to the TTKS (Ug99) race group. German isolates of race TKTTF are phenotypically different to the ones responsible for the stem rust epidemic in Ethiopia in 2013–2014. Forty isolates were genotyped using a custom SNP array. Phylogenetic analysis showed that these 40 isolates represented two distinct lineages (clade IV and clade V). Thirty‐eight isolates clustered into clade IV, which previously was defined by Ethiopian isolates of race TKTTF. Race TKKTP is of special concern due to its combined virulence to stem rust resistance genes Sr24, SrTmp and Sr1RS^Amigo. The vulnerability to race TKKTP in US and international winter wheat was confirmed as 55% of North American and international cultivars and breeding lines resistant to race TTKSK (Ug99) became susceptible to TKKTP. Races identified in Germany in 2013 confirmed the presence of virulence to important resistance genes that are effective against race TTKSK. This information should be useful for breeders to select diverse and effective resistance genes in order to provide more durable stem rust resistance and reduce the use of fungicides.     

甘肃陇南感病小檗在小麦条锈病发生中起提供(初始)菌源作用的直接证据

 陇南是中国小麦条锈菌易变区、小麦条锈病的常发流行区和防治的关键地区。明确陇南小麦条锈菌转主寄主小檗在小麦条锈病发生中的作用,对阐释该地区小麦条锈菌新小种产生的来源和指导小麦条锈病的综合防控具有重要意义。本研究从陇南春季自然受锈菌侵染的堆花小檗及其邻近的小麦上分离获得小麦条锈菌菌系,19个来自发病小檗的单夏孢子堆菌系在中国小麦条锈菌鉴别寄主上产生5个不同毒力类型VP1~VP5,均为新小种;在近等基因系和载体品种组成的鉴别寄主上产生17个不同毒力类型。29个来自邻近发病小檗的小麦上条锈菌菌系在中国小麦条锈菌鉴别寄主上产生了10个不同的毒力类型VP1~VP10,均为新小种;在近等基因系和载体品种组成的鉴别寄主上产生了24个不同的毒力类型。两个小麦条锈菌群体享有共同的毒力类型VP1~VP5,同时来自发病小檗上菌系的一些毒力类型不同于小麦上的毒力类型。主坐标分析(PCoA)和毒性表型聚类分析表明来自于自然受锈菌侵染小檗及其邻近小麦上的小麦条锈菌存在菌源交流。综合研究表明在自然条件下,陇南小麦条锈菌在野生感病小檗上进行有性生殖是常年发生的,感病小檗在新小种产生和在陇南小麦条锈病的发生中起提供菌源的作用。因此,在陇南小麦条锈病的综合治理策略上加强对小檗的处理,降低新小种产生速率和品种抗性丧失速率,从而延长品种使用年限,同时减少新小种向东部广大麦区传播,保障小麦生产安全。     

小檗公式在小麦条锈菌侵染过程中的表达特征

 小檗是小麦条锈菌的转主寄主,自然条件下在小檗上进行有性生殖不仅是条锈菌毒性变异的重要途径,也为病害的流行提供了初侵染源。目前条锈菌与小麦互作已经有较为细致深入的研究,而条锈菌与小檗分子互作的研究未见报道。NPR1是植物调控防卫反应的一个关键基因,本研究克隆得到小檗NPR1基因(BhNPR1),并对BhNPR1及其下游病程相关基因的表达模式进行了分析。与小麦NPR1基因受条锈菌夏孢子侵染诱导不同,BhNPR1的表达水平在条锈菌担孢子侵染小檗过程中保持稳定。另外,条锈菌侵染小檗过程中NPR1下游的病程相关基因BhPR5和BvPDF1.2a的表达量变化也不明显,而BhPR1和BhPR2则显著上调表达。这些结果表明BhNPR1及多个病程相关基因不参与对条锈菌的防卫反应,这可能是导致小檗先天免疫缺陷而成为多种锈菌共同的转主寄主。     

The elasticity of the epidemic growth rate to observed weather patterns with an application to yellow rust

ABSTRACT We extend a previously developed method that quantifies the sensitivity of the exponential epidemic growth rate, r, to weather changes, through a pathogen's life cycle components (basic reproduction number, latent period, and mean and standard deviation of the spore production curve). Here a method is developed to study the elasticities of the system and subsequently the model is linked to observed weather patterns. This enables a direct comparison between the effects of different weather variables (temperature, surface wetness duration, and light quantity) under realistic weather scenarios. The three sites studied represent areas within the United Kingdom with contrasting climates. Yellow rust, caused by Puccinia striiformis, on winter wheat is studied as a key application. Our results show that temperature and more importantly changes in temperature through their effect on pathogen reproduction have the largest effect on r. The long latent period at low winter temperatures is not a key component in the epidemic development, which is contrary to general beliefs. The results combined with long term average yellow rust severity patterns show that it is winter survival and not summer survival that controls the eventual disease severity. The results also show that within the current United Kingdom spraying regime on wheat crops against yellow rust, the first spray should mainly affect the basic reproduction number, i.e., should be a protectant spray, whereas the second spray should also affect the latent period, i.e., should also have curative action.     

A unique race of the wheat stem rust pathogen with virulence on Sr31 identified in Spain and reaction of wheat and durum cultivars to this race

Wheat stem rust, caused by Puccinia graminis f. sp. tritici, is a re-emerging disease, posing a threat to wheat production. In Spain, stem rust has been rarely detected since the 1970s, but infection was observed in wheat fields in 2018. We analysed six stem rust samples collected in Rota, Cádiz province and one from Monteagudo del Castillo, Teruel province. All the samples from Rota were typed as race TKTTF, whereas the sample from Monteagudo del Castillo, collected in a wheat field adjacent to barberry bushes, was typed as race TKHBK. This race has a unique and significant virulence combination that includes virulence to Sr31, Sr33, Sr53 and Sr59, and is avirulent to Rusty, a durum line developed for universal susceptibility to the wheat stem rust pathogen. TKHBK is the first race outside the Ug99 race group with virulence to Sr31 and the first known race with virulence to Sr59. Genotyping studies indicate that race TKHBK does not belong to the Ug99 or TKTTF race groups and constitutes a previously unknown lineage. Two hundred bread and durum wheat cultivars and breeding lines from Spain were evaluated against TKHBK, TKTTF, and six additional races. Resistance was observed to all the races evaluated. Molecular markers confirmed the presence of Sr7a, Sr24, Sr31, Sr38 and Sr57 in bread wheat, and Sr13 in durum wheat. The re-emergence of wheat stem rust in Spain and the occurrence of unique virulences underscore the need to continue surveying and monitoring this disease.     

青海东部小麦条锈菌转主寄主小檗资源调查与鉴定

 青海省是中国小麦条锈病的重要流行区。研究证实小檗是小麦条锈菌的转主寄主。然而,关于青海省分布的小檗能否作为小麦条锈菌转主寄主鲜有报道。本研究对青海东部的小檗资源进行调查,并对该地区小檗能否作为小麦条锈菌转主寄主进行了室内人工鉴定。结果表明:有10种小檗在青海东部常见且分布广泛,其中匙叶小檗、置疑小檗、松潘小檗、甘肃小檗和近似小檗,是小麦条锈菌的转主寄主小檗的新种类。本研究对进一步研究青海东部条锈菌有性生殖与毒性变异和小麦条锈病的流行奠定了基础。     

Evaluation of morphology of infection structures in distinguishing between different Allium rust fungi

Fifteen isolates of rust fungi were collected in the United Kingdom and in the Netherlands from severalAllium species. The samples represented three putative rust species. The morphology of the telia, teliospores and urediniospores was investigated. From urediniospores infection structures were induced on leek, and their morphology was described. Telia and teliospores were not available on every sample. Morphology of the infection structures clearly differentiated between ‘leek’ type and ‘chive’ type isolates. The morphology of infection structures ofPuccinia mixta is very similar to that ofUromyces ambiguus, but clearly distinct fromPuccinia allii sensu Jennings from leek. Quantitative differences in urediniospore characters differentiated between the putative species, but there was overlap between the taxa. We conclude that morphology of infection structures of urediniosporelings is a useful trait for identification of rust isolates fromAllium. This is especially true where more than one rust species may occur on the same host species, as withAllium fistulosum.     

小麦条锈菌有性生殖与毒性变异的研究进展

小麦条锈病是由条形柄锈菌小麦专化型Puccinia striiformis f.sp.tritici Erikss.et Henn.引起的、最具毁灭性的小麦真菌病害之一,在全世界小麦种植的国家或地区均有发生,其流行常常导致小麦严重的产量损失。利用抗病品种是防治小麦条锈病最为经济、有效的措施。然而,由于病原菌毒性变异不断产生新小种,频繁导致品种抗病性"丧失",继而引发病害流行成灾。小麦条锈菌的有性阶段过去一直未被发现,有性生殖与条锈菌变异及其在病害发生中的作用研究一直处于空白状态。近年来,随着小麦条锈菌转主寄主的发现,国内外学者开展了相关的研究工作,取得了一系列重要的研究进展。本文综述了小麦条锈病的发生症状、病菌的生物学特性、转主寄主、有性生殖与条锈菌毒性变异、转主寄主在条锈病发生中的作用、影响有性生殖发生的因素等方面的最新进展,并展望了深入开展小麦条锈菌有性生殖的研究思路。     

Extended survival of Puccinia graminis f. sp. tritici urediniospores: implications for biosecurity and on‐farm management

Puccinia graminis f. sp. tritici (Pgt), the causal organism of stem rust, is of global importance across wheat‐growing countries. However, some epidemics commence without the obvious presence of ‘alternate’ or ‘green bridge’ hosts, suggesting urediniospores can survive in the absence of suitable host plants for many weeks. Testing a range of inert material types, including metals, plastics, fabrics and woods, highlighted a significant effect of material type and temperature on urediniospore viability (P < 0.001), with urediniospores remaining attached and viable on these materials (aluminium, paper, rubber, all fabric and all woods) for up to 365 days at 23/8 °C day/night. At 36/14 °C day/night, urediniospore viability was retained for a maximum of 300 days on denim and jute. Furthermore, at 45/15 °C day/night, urediniospores remained viable for a maximum of 180 days on cotton and jute. The frequency of recovery of attached urediniospores was also dependent upon the material type, with significant differences between materials in their abilities to retain urediniospores after washing (P < 0.001). Urediniospores recovered even after 300 or 365 days from the lower two temperature regimes successfully initiated infections of wheat seedlings. Results confirm the potential importance of inert materials as long‐term carriers of viable Pgt urediniospores, highlighting risks of spread of new pathotypes and strains across wheat‐growing regions, the significant biosecurity implications for contaminated carrier materials, and its likely survival across seasons without a host.     

Effects of Ampropylfos (RS)-1-aminopropylphosphonic acid) on zoospore formation,repeated zoospore emergence and oospore formation in Aphanomyces spp.

The effects on growth, asexual and sexual reproduction of different Aphanomyces species by the new fungicide ampropylfos ((RS)-1-aminopropylphosphonic acid) were tested using highly synchronized cultures. Ampropylfos reduced mycelial growth in Aphanomyces astaci, A. euteiches and A. stellatus. The fungicide was found to reduce zoospore numbers in all Aphanomyces species tested except A. euteiches. In one species, A. stellatus, zoospore production was inhibited at much lower fungicide concentrations than in the other species tested. In the pea pathogen A. euteiches, ampropylfos reduced the number of oospores, whereas the fungicide had little effect on asexual reproduction.     

源自陕、甘野生小檗的小麦条锈菌有性生殖后代的毒性分析

 小麦条锈病是我国最具毁灭性的小麦病害之一,其流行常常造成小麦严重减产。种植抗病品种是防治该病害最经济、有效和环保的措施。但是,由于新毒性菌系的出现,抗病品种在种植短短数年内便“丧失”其抗病性。研究证实病原菌毒性变异产生新菌系是导致小麦品种抗病性“丧失”的根本原因。近年来,随着小麦条锈菌转主寄主小檗的确定,发现自然条件下我国小麦条锈菌在野生小檗上可以完成有性生殖。本研究通过对2015年自然发病小檗小麦条锈菌的分离及其单夏孢子堆纯化,利用中国鉴别寄主进行了毒性测定分析。从陕西、甘肃两省的3种感病小檗共分离获得小麦条锈菌菌系8个,其中有1个菌系与已知小种Su11-126的毒性完全匹配,其余7个为新菌系;93个单夏孢子堆群体可分为47个不同的致病类型,包括14个已知小种类型,33个新小种类型;有56个菌系与已知小种的毒性完全匹配,37个为新小种。本研究再次获得了条锈菌自然条件下存在有性生殖并因此导致新菌系产生的证据,证实了野生小檗在我国小麦条锈菌的生活史和病害循环中具有作用。     

Genetic variation in Puccinia graminis collected from oats, rye, and barberry

Puccinia graminis, the causal agent of stem rust, was collected from its alternate host barberry (Berberis spp.) and two different uredinial hosts, oats (Avena sativa) and rye (Secale cereale). The samples were analyzed using 11 polymorphic simple sequence repeat (SSR) markers. There were large differences between fungal populations on oats (P. graminis f. sp. avenae) and rye (P. graminis f. sp. secalis), and the genetic variation within the different formae speciales was also high. It was possible to distinguish between the two formae speciales on barberry. Additional genotypic groups not present in the field samples from oats and rye were also identified on barberry. Our results confirm the importance of barberry in maintaining the populations of P. graminis in Sweden and the importance of the sexual stage for the survival of the pathogen.