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.     

The ubiquity of non-specific eliciting activity in intercellular washing fluids from rust-infected wheat leaves

Intercellular washing fluids (IWF) were harvested from a range of wheat cultivars infected with either the leaf rust fungus ( Puccinia recondite f.sp. tritici ) or the stem rust fungus ( Puccinia graminis f.sp. tritici ). These fluids were then tested for their ability to elicit symptoms of chlorosis and necrosis in 14 wheat cultivars. Eliciting activity was found in all samples tested, irrespective of both the host cultivar infected by the rust fungus and the genotype of the infecting pathogen isolate. Healthy test cultivars either responded to all IWF preparations or were responsive to none of them. Investigations into the nature of this non-specific eliciting activity revealed that active molecules are periodate sensitive, negatively charged at neutral pH, and associated with a high molecular weight fraction.     

Inheritance and molecular mapping of barley genes conferring resistance to wheat stripe rust

ABSTRACT Most barley cultivars are resistant to stripe rust of wheat that is caused by Puccinia striiformis f. sp. tritici. The barley cv. Steptoe is susceptible to all identified races of P. striiformis f. sp. hordei (PSH), the barley stripe rust pathogen, but is resistant to most P. striiformis f. sp. tritici races. To determine inheritance of the Steptoe resistance to P. striiformis f. sp. tritici, a cross was made between Steptoe and Russell, a barley cultivar susceptible to some P. striiformis f. sp. tritici races and all tested P. striiformis f. sp. hordei races. Seedlings of parents and F(1), BC(1), F(2), and F(3) progeny from the barley cross were tested with P. striiformis f. sp. tritici races PST-41 and PST-45 under controlled greenhouse conditions. Genetic analyses of infection type data showed that Steptoe had one dominant gene and one recessive gene (provisionally designated as RpstS1 and rpstS2, respectively) for resistance to races PST-41 and PST-45. Genomic DNA was extracted from the parents and 150 F(2) plants that were tested for rust reaction and grown for seed of F(3) lines. The infection type data and polymorphic markers identified using the resistance gene analog polymorphism (RGAP) technique were analyzed with the Mapmaker computer program to map the resistance genes. The dominant resistance gene in Steptoe for resistance to P. striiformis f. sp. tritici races was mapped on barley chromosome 4H using a linked microsatellite marker, HVM68. A linkage group for the dominant gene was constructed with 12 RGAP markers and the microsatellite marker. The results show that resistance in barley to the wheat stripe rust pathogen is qualitatively inherited. These genes might provide useful resistance against wheat stripe rust when introgressed into wheat from barley.     

Support for a stepwise mutation model for pathogen evolution in Australasian Puccinia striiformis f.sp. tritici by use of molecular markers

Since its initial detection in Australia in 1979, wheat yellow (stripe) rust ( Puccinia striiformis f.sp. tritici ) has evolved in Australia and New Zealand into more than 20 pathotypes with assorted virulence characteristics. This evolution is believed to have occurred in a stepwise fashion from an original single pathotype, with no subsequent new introductions. A combination of random amplified polymorphic DNA (RAPDs) and amplified fragment length polymorphisms (AFLPs) was used to examine the level of molecular variation in Australian and New Zealand isolates, and to compare this with variation amongst other isolates of P. striiformis . Using 60 RAPD primers on seven Australian isolates representing seven different pathotypes collected between 1979 and 1991, more than 300 potentially polymorphic loci were analysed and no polymorphisms were detected. Using the same primers on two UK isolates, 3% of loci showed a polymorphism. A similar level of polymorphism was found between UK isolates using AFLP primers, and between 5 and 15% of fragments were polymorphic between an isolate from the UK, an isolate from Denmark, and one from Colombia. However, no AFLP polymorphisms were found amongst 14 Australian and New Zealand isolates tested, at over 100 potentially polymorphic loci. The lack of molecular variation in the Australian and New Zealand collection is consistent with the stepwise mutation theory of pathotype evolution from a single introduction.     

2013年我国部分麦区小麦白粉菌群体对温度敏感性及其与毒性的关系

 研究小麦白粉病菌群体对温度的敏感性及其与毒性的关系,可为气候变化背景下小麦白粉病的长期发生趋势预测和防治提供依据。本研究对2013年采自云南、四川、河南、北京和陕西五省（市）的小麦白粉菌标样进行分离纯化,获得90个单孢子堆分离物,采用小麦离体叶段法进行温度敏感性测定,并利用32个已知抗病基因品种（系）进行毒性测定,且对两者之间的相关性进行分析。结果表明,供试菌株ET50分布于19.78℃~25.15℃,平均ET50为23.22℃,其中73.34%的供试菌株ET50值介于22℃到24℃之间,小于22℃的菌株占10%,大于24℃的菌株占16.67%。利用Popgen1.32软件对5个省（市）小麦白粉病菌群体的毒性进行多样性分析,结果表明五省市小麦白粉菌群体毒性水平上的多样性指数 H 值为0.237 0,其中四川群体毒性多样性水平最高,Nei’s基因多样性指数 H 值为0.221 2,河南群体最低, H 值为0.197 5。小麦白粉菌群体对温度敏感性与毒性多样性相关性分析结果表明,ET50与毒性多样性（ H 和 I 值）间存在显著的负相关。     

Susceptibility in oats to stem rust induced by co-infection with leaf rust

Induction of susceptibility in oats to a normally avirulent pathotype of Puccinia graminis f.sp. avenae was studied in the presence of different pathotypes of P. coronata f.sp. avenae . Induction occurred on seedlings only in the presence of a virulent culture of P. coronata avenae and was not dependent on time or order of inoculation of either pathogen. This phenomenon was restricted to seedlings of lines possessing the Pg-a source of oat stem rust resistance. The specificity of induced susceptibility can be used as a valuable bioassay for screening and identifying Pg-a . Induced susceptibility occurred only at the seedling stage, and apparently provides no obstacle to the use of Pg-a as a source of stem rust resistance in oats.     

我国小麦条锈菌体细胞遗传重组的分子证据

 小麦条锈病是影响我国小麦生产的重要病害之一,条锈菌毒性变异是引致小麦品种抗病性丧失的主要原因。本文应用SSR分子标记技术,研究了陇南越夏区小麦条锈菌群体遗传结构,以期寻找条锈菌群体遗传重组的分子证据。研究结果表明,陇南地区小麦条锈菌群体遗传多样性比较丰富而遗传分化较小,遗传变异主要存在于群体内部,而在群体之间,遗传多样性有显著的差异。在11对SSR引物中,有4对能够检测到陇南地区小麦条锈菌群体存在遗传重组,而且重组体出现的频率不同,CPS15揭示的条锈菌重组体出现的频率为20.0%,CPS34揭示的条锈菌重组体出现的频率为18.5%,RJ20揭示的条锈菌重组体出现的频率为12.8%,RJ18揭示的条锈菌重组体出现的频率为15.0%,陇南地区小麦条锈菌群体重组体出现频率平均为16.6%。本文揭示的遗传重组现象表明陇南地区条锈菌体细胞结合十分普遍,由此推测我国小麦条锈菌在自然条件下通过遗传重组而导致毒性变异的可能性。     

First detailed report on Puccinia graminis f. sp. avenae virulence structure and Pg resistance genes effective in Poland

European Journal of Plant Pathology - Occurrence of stem rust, caused by Puccinia graminis f. sp. avenae, on oat fields in Europe may lead to significant yield losses. The last P. graminis...     

Virulence analysis of Puccinia graminis f.sp. tritici populations in Ethiopia with special consideration of Ug99

Wheat stem rust samples were collected in 2006 and 2007 in the Arsi, Bale, Shewa and northwest regions of Ethiopia to determine virulence diversity and race distribution in Puccinia graminis f.sp. tritici populations. Stem rust incidence was high in Arsi, Bale and east Shewa. In northwest Ethiopia, and north and west Shewa, stem rust was prevalent at low levels. A total of 152 isolates was analysed and 22 races were identified. Races TTKSR (Ug99), TTHSR and RRTTR were predominant, with frequencies of 26·6, 17·7 and 11·1%, respectively. These races were also detected in all regions. The highly virulent race designated Ug99 was present throughout the country and dominated in all regions except northwest Ethiopia. A variant of Ug99 virulent against the stem rust resistance gene Sr24 was not detected in this study. Four stem rust resistance genes ( Sr13, Sr30, Sr36 and SrTm p) were found to confer resistance to most of the races prevalent in Ethiopia. With the exception of Sr30 , which is not effective against Ug99, these genes could be used in breeding for resistance to stem rust in Ethiopia.     

茉莉酸诱导小麦抗病虫性初步研究

初步研究了茉莉酸诱导对小麦苗抗病虫能力的影响,结果显示,小麦在喷施茉莉酸后能够提高植株对麦长管蚜和小麦白粉病菌、小麦叶锈病菌的抵抗能力,可显著降低小麦白粉病、叶锈病的发病级别和病斑数量,对麦长管蚜则在体重和产仔数量上有显著的抑制作用。     

Analyzing wheat cultivars grown in Czech Republic for eight stem rust resistance genes

European Journal of Plant Pathology - Stem rust is a disease of wheat caused by a basidiomycete Puccinia graminis f. sp. tritici (Pgt). Emergence of new populations of the pathogen and their spread...     

Allele sequencing of the barley stem rust resistance gene Rpg1 identifies regions relevant to disease resistance

The stem rust resistance gene Rpg1 has protected North American barley cultivars from significant yield losses for over 65 years. The remarkable durability of this gene warrants further study as to its possible origin and allelic variation. Eight Swiss barley (Hordeum vulgare) landraces and eight wild barley (H. vulgare subsp. spontaneum) accessions from diverse geographic regions were analyzed to uncover new alleles of Rpg1 and learn about its possible origin. The two germplasm groups included accessions that were resistant and susceptible to Puccinia graminis f. sp. tritici pathotype MCCF. Allele-specific primers were utilized to amplify 1 kbp overlapping fragments spanning the Rpg1 gene and sequenced if a polymerase chain reaction (PCR) fragment was generated. Variation among the PCR products revealed significant polymorphisms among these Hordeum accessions. Landraces and wild barley accessions susceptible to pathotype MCCF exhibited the highest degree of Rpg1 polymorphism. One resistant landrace (Hv672) and one resistant wild barley accession (WBDC040) yielded all seven Rpg1-specific PCR fragments, but only landrace Hv672 coded for an apparently functional Rpg1 as determined by comparison to previously characterized resistant and susceptible alleles and also resistance to HKHJ, a stem rust pathotype that can specifically detect Rpg1 in the presence of other resistance genes. Accessions resistant to stem rust pathotype MCCF, but completely lacking Rpg1-specific PCR amplification and hybridization with an Rpg1-specific probe, suggested the presence of stem rust resistant gene(s) different from Rpg1 in the Hordeum germplasm pool. Some Rpg1 alleles that retained the ability to autophosphorylate did not confer resistance to Puccinia graminis f. sp. tritici pathotype MCCF, confirming our previous observations that autophosphorylation is essential, but not sufficient for disease resistance. Thus, the RPG1 protein plays a complex role in the stem rust disease resistance-signaling pathway.     

A major gene for powdery mildew resistance transferred to common wheat from wild einkorn wheat

ABSTRACT A major gene for resistance to wheat powdery mildew (Blumeria graminis f. sp. tritici = Erysiphe graminis f. sp. tritici) has been successfully transferred into hexaploid common wheat (Triticum aestivum, 2n = 6x = 42, AABBDD) from wild einkorn wheat (Triticum monococcum subsp. aegilopoides, 2n = 2x = 14, AA). NC96BGTA5 is a germ plasm line with the pedigree Saluda x 3/PI427662. The response patterns for powdery mildew resistance in NC96BGTA5 were tested with 30 differential isolates of B. graminis f. sp. tritici, and the line was resistant to all tested isolates. The analyses of P(1), P(2), F(1), F(2), and BC(1)F(1) populations derived from NC96BGTA5 revealed two genes for wheat powdery mildew resistance in the NC96BGTA5 line. One gene, Pm3a, was from its recurrent parent Saluda, and the second was a new gene introgressed from wild einkorn wheat. The gene was determined to be different from Pm1 to Pm21 by gene-for-gene and pedigree analyses. The new gene was identified as linked to the Pm3a gene based on the F(2) and BC(1)F(1) populations derived from a cross between NC96BGTA5 and a susceptible cultivar NK-Coker 68-15, and the data indicated that the gene was located on chromosome 1A. It is proposed that this new gene be designated Pm25 for wheat powdery mildew resistance in NC96BGTA5. Three random amplified polymorphic DNA markers, OPX06(1050), OPAG04(950), and OPAI14(600), were found to be linked to this new gene.     

Development,characterization and application of genomic SSR markers for the oat stem rust pathogen Puccinia graminis f. sp. avenae

Oat stem rust, caused by Puccinia graminis f. sp. avenae (Pga), is one of the most severe diseases of oats worldwide. Population studies are scarce for this pathogen, mainly due to the lack of polymorphic molecular markers suitable for genetic analysis. In this study, an Australian Pga isolate was sequenced, the abundance of simple sequence repeats (SSRs) was determined and PCR‐based polymorphic markers suitable for genetic diversity analysis were developed. The amplification of 194 primer pairs was initially assessed using a set of 12 isolates of different cereal rust species and their formae speciales. A high frequency of cross‐species amplification was observed for most markers; however, 36 SSRs were diagnostic for P. graminis only. A subset of 19 genome‐derived SSRs were deemed useful for genetic diversity analysis of Pga and were assessed on 66 Pga isolates from Australia, Brazil and Sweden. Brazilian and Australian isolates were characterized by one and two predominant clonal lineages, respectively. In contrast, the Swedish isolates, previously shown to undergo sexual recombination, were highly diverse (nine distinct genotypes out of 10 isolates) and divided into two subpopulations. The genome‐derived SSR markers developed in this study were well suited to the population studies undertaken, and have diagnostic capabilities that should aid in the identification of unknown rust pathogen species.     

The classification of isolates of Gaeumannomyces graminis from wheat, rye and oats using restriction fragment length polymorphisms in families of repeated DNA sequences

Isolates of the take-all fungus, Gaeumannomyces graminis var. avenae , which affects oats, wheat and other grasses, and of G.g. var. tritici , which preferentially affects wheat, rye and barley, contain a high proportion of repeated sequences. Total DNA from 57 fungal isolates collected from many locations and different cereal hosts, and scored for virulence on wheat, rye and oats, revealed many restriction fragment length polymorphisms. These RFLP s were observed either by staining the DNA directly, by hybridization to radioactively labelled total fungal DNA , or by hybridization with labelled wheat ribosomal DNA . With only a few exceptions, the isolates with the same preferred cereal hosts showed more similar patterns of restriction fragments than isolates that had different pathogenicity properties on cereal hosts, irrespective of the geographical origins of the isolates. This was even the case for R isolates of G.g. var. tritici that were virulent on wheat and rye compared with N isolates that were virulent only on wheat. Isolates were identified by hybridizing DNA from infected root samples with ³²P-labelled total fungal DNA . The restriction fragment polymorphisms involving families of repeated sequence can therefore be used as a predictive assay for host preference of an isolate, and have probably arisen by host selection of fungal lineages. The variation between isolates in different pathogenicity groups suggests that there is little gene flow between isolates that can infect different hosts, even though they can coexist in the same field.     

2007年我国部分麦区小麦白粉菌对三唑酮的抗药性监测

采用拌种离体叶段法测定了我国9个省（市）的112个小麦白粉菌株对三唑酮的抗性。结果表明,供试菌株的平均EC50为57.25mg/L,平均抗性水平为27.39倍,其中86.61%供试菌株已经产生抗性,抗药性比敏感菌株高出10~40倍的菌株占47.32%,四川、山东、甘肃和贵州等地菌株的抗性高于其他地区。此结果可为三唑类杀菌剂在我国的推广应用和制定合理的抗药性治理措施提供依据。     

小麦白粉病菌越夏,越冬和春季菌源毒性结构比较

 分析了1992年和1993年不同时期采集的小麦白粉病菌越夏、越冬和春季菌源共238个菌株的生理小种、毒性基因结构和对9个推广品种的毒力频率。结果表明三种菌源在生理小种和毒性基因组成及出现频率是相似的,对9个推广品种的毒力频率均较高。这样在白粉病菌毒性和品种抗病性研究中,可以采集应用越夏和越冬菌株标样,避免了夏季高温季节保存菌株的工作;而且从毒性结构方面进一步证实白粉病菌主要以分生孢子在夏季侵染高海拔地区的自生麦苗而越夏,再逐渐扩展至秋播麦苗越冬后导致春季大田发病,病菌的子囊孢子在平原地区侵染循环上作用不大。     

Usefulness of gene pg10 as a source of stem rust resistance in oat breeding

ABSTRACT Infection types produced by Puccinia graminis f. sp. avenae on plants of Avena sativa with the stem rust resistance gene Pg10 are characterized by moderate-sized uredinia surrounded by an area of chlorosis and a larger variable zone of dark brown necrosis. This study was undertaken to assess the effectiveness of gene Pg10 as a source of resistance to stem rust and to determine the interactions of this gene with other common Pg genes. A derived Pg10 line was tested with 58 distinct pathotypes of P. graminis f. sp. avenae and was crossed to substituted single-gene lines carrying the resistance gene Pg1, Pg2, Pg3, Pg4, Pg8, Pg9, Pg13, Pg15, Pg16, or Pga. The Pg10 line showed moderate resistance to all 58 patho-types, and there was no indication of specificity in virulence by any isolate. Gene Pg10 was inherited independently of the other Pg genes and had a complementary effect on the expression of resistance by these genes. An effective level of resistance conferred by Pg10 was demonstrated in a field nursery artificially inoculated with P. graminis f. sp. avenae. It was concluded that Pg10 is a potentially useful source of stem rust resistance in oat breeding, with its main attributes being an apparent broad base of resistance, ease of combining with other Pg genes, and complementary effects on the expression of other Pg genes.     

Mutants in wheat showing multipathogen resistance to biotrophic fungal pathogens

Five fast-neutron-derived mutants were isolated from the wheat line Hobbit 'sib' that show enhanced field resistance towards Puccinia striiformis f.sp. tritici , the causal agent of yellow rust. Subsequent testing showed the yellow rust resistance phenotypes to differ between mutants, to be expressed at different growth stages and, in some cases, to show an isolate interaction. Three mutants, I3-48, I3-49 and I3-54, exhibited an enhanced yellow rust resistance phenotype from the third seedling leaf onwards, while mutants I3-27 and I3-30 did not show an altered yellow rust phenotype until later growth stages. Additional resistance for brown rust (causal agent Puccinia triticina ) was identified in mutants I3-27, I3-30, I3-48 and I3-49, and for powdery mildew caused by Blumeria graminis f.sp. tritici in mutants I3-27, I3-30, I3-48 and I3-54, although in some cases the resistance was isolate-specific.     

小麦锈菌夏孢子经盐酸处理后的形态观察

为了快速区分小麦条锈菌、叶锈菌和秆锈菌,用不同浓度的盐酸处理3种小麦锈菌的新鲜夏孢子和在4 ℃冰箱内保存4个月以上的锈菌夏孢子,在显微镜下观察夏孢子形态以及原生质体变化。结果表明, 24.5%、28.5%、32.5%、36.5%4种浓度的盐酸处理条锈菌夏孢子后其原生质体均浓缩成多个分散的小团。而同样方法处理叶锈菌和秆锈菌新鲜菌种时其原生质体浓缩成一个大团或多个小团,浓缩成一个大团的比例随盐酸浓度的增大而提高;用不同浓度盐酸处理保存4个月以上的2种小麦锈菌时,95%以上的夏孢子原生质体浓缩成多个分散的小团,少数（不超过5%）的夏孢子原生质体浓缩成一个大团。研究还发现叶锈菌和秆锈菌夏孢子活力、盐酸浓度等对夏孢子原生质体浓缩状况有很大影响,且与病菌生理小种有一定的关系。因此,锈菌夏孢子经36.5%浓盐酸处理后的原生质体浓缩状况只能作为小麦锈病田间快速诊断检测的辅助手段,不能作为锈病种类鉴别的唯一标准,必须结合病害症状特征、孢子形态以及分子生物学方法等进行综合判别。