Ribosomal DNA-ITS sequence polymorphism in the sugarcane rust,Puccinia kuehnii

The two highly divergent ITS types previously observed in isolates ofPuccinia kuehnii were amplified from the same isolates through the use of ITS type-specific primers for PCR and are therefore considered as polymorphisms of ITS regions within the species. Although homology of sequences of one of the ITS types with otherPuccinia species was of expected levels, significantly high homology of sequences of the other type with those ofCronartium members indicates that abnormal genetic events led to the occurrence of these polymorphic regions. These results indicate that ITS gene tree phylogeny may not reflect true species phylogeny in this group of rust fungi. Rather, D1/D2 region tree phylogeny, which was concordant with the differences in morphology with and among related rusts, more correctly reflects phylogenetic relationships of sugarcane and related grass rusts. contribution No. 159, Laboratory of Plant Parasitic Mycology, Institute of Agriculture and Forestry, University of Tsukuba.     

Phylogenetic relationships of sugarcane rust fungi

The phylogenetic positions of Puccinia spp. infecting sugarcane (a complex hybrid of Saccharum spp.) were determined using 38 newly generated rust sequences and 26 sequences from GenBank. Rust specimens on sugarcane were collected from 164 locations in 23 countries and identified based on light microscopy. The morphology for all samples matched that of Puccinia kuehnii or P. melanocephala, the orange and brown rust pathogens of sugarcane, respectively. Nuclear ribosomal DNA sequences (rDNA) including portions of the 5.8S rDNA, the complete internal transcribed spacer 2 (ITS2) and 5′ region of the large subunit (nLSU) rDNA were obtained for each species along with 36 additional rust taxa. Despite a shared host, the two Puccinia spp. on sugarcane are not closely related within the Pucciniales. Phylogenetic analyses place P. melanocephala most closely to P. miscanthi, P. nakanishikii, and P. rufipes infecting Miscanthus sinensis, Cymbopogon citratus, and Imperata cylindrica, respectively. Puccinia kuehnii is basal to a clade of Poaceae-infecting rusts including P. agrophila, P. polysora, P. substriata, and Uromyces setariae-italicae infecting Schizachyrium spp., Zea mays, Digitaria spp., and Urochloa mosambicensis, respectively. Light and scanning electron microscopy images highlight morphological differences distinguishing the two sugarcane-infecting species. This study confirms the separation of rust species infecting Poaceae from Cyperaceae- and Juncaceae-infecting rusts and also provides support for the presence of an additional group that includes P. kuehnii and other grass-infecting relatives.     

<Emphasis Type="Italic">Puccinia calystegiae-soldanellae</Emphasis>, a new rust species on <Emphasis Type="Italic">Calystegia soldanella</Emphasis> from Japan

A rust species on Calystegia soldanella in Japan has been treated as Puccinia convolvuli to date. However, morphological characteristics of specimens on C. soldanella collected from Japan are significantly different from those of specimens on other Calystegia and Convolvulus species from different areas of the world. It is proved by inoculation experiment that the rust on C. soldanella is specific to C. soldanella. Based on these results, Puccinia rust on C. soldanella from Japan is described as a new species, Puccinia calystegiae-soldanellae.     

Puccinia carici-adenocauli, a new rust fungus onCarex, and its anamorph,Aecidium adenocauli

Field observations and inoculation experiments showed thatAecidium adenocauli onAdenocaulon himalaicum was an anamorph of aPuccinia onCarex hakonensis andC. uda. From comparative morphology with caricicolous puccinias, the rust on theseCarex species was considered as a new species and was named asPuccinia carici-adenocauli. Contribution No. 142, Laboratory of Plant Pathology and Mycology, Institute of Agriculture and Forestry, University of Tsukuba     

牛紏吴茱萸柄锈菌新种

本文描述了采自海南省琼中县牛紏吴茱萸Evodia trichotoma (Lour.) Pierre(芸香科)上的柄锈菌一新种——Puccinia evodiae-trichotomae J. -y. Zhuang。本种与巴布亚一新几内亚产的吴茱萸Evodia sp.上的另一种柄锈菌Puccinia evodiae Cumm.略近似,但冬孢子堆裸露呈垫状,而后者则深埋于病瘿中。冬孢子形状与大小和后者相仿,然而少数孢子可长达60μm,顶壁有时略增厚,颜色很浅或近几色。镜下可见一些孢子已萌发,推测可能属无眠冬孢型。芸香科植物上的柄锈菌目前仅知10种,本种特征与它们均不相同,故鉴定为新种。本新种模式标本保藏在中国科学院微生物研究所真菌标本室。     

Aecidium dispori is the aecial anamorph ofPuccinia albispora, sp. nov. (Uredinales)

Aecidium dispori forms spermogonium and aecium onDisporum sessile andD. smilacinum, which are distributed in East Asia. TheAecidium species is found to be an aecial anamorph of aPuccinia fungus, with its uredinial-telial stage being formed onCarex conica, C. dolichostachya subsp.multifolia, C. pisiformis subsp.alterniflora andC. rugata. Urediniospores of this fungus are large, colorless, thick-walled with 4–5 equatorial germ pores. The morphological characteristics of urediniospores and the spermogonial-aecial host do not fit to any set of circumscribing characters of previously described species. We consider the fungus to be a new species and propose a new name,Puccinia albispora, for the fungus.     

Genetic diversity of Puccinia kuehnii,the causal agent of orange rust of sugarcane,from Brazil

The use of resistant genotypes is the preferred method to control orange rust of sugarcane (Saccharum spp) caused by Puccinia kuehnii. This approach has been adopted in Brazil but outbreaks of the disease on previously resistant varieties showed that the efficacy of this method is limited and requires a better understanding of pathogen diversity. Nevertheless, adequate molecular markers for examining pathogen diversity at population level are not available, which limits the success of orange rust control by genetic resistance. Therefore, two independent investigations were conducted to examine genetic diversity of P. kuehnii from São Paulo state, the most important sugarcane growing state of Brazil. First, simple-sequence repeat (SSR) markers were developed in the present work and genotypic diversity of orange rust isolates from different locations investigated. Second, phenotypic diversity was examined by the single-pustule inoculation technique on P. kuehnii isolates retrieved from three susceptible commercial sugarcane cultivars. A total of 96 SSR markers were generated and tested for this species. Subsequently, 29 isolates of P. kuehnii were fingerprinted with nine SSR markers to estimate the genotypic diversity by neighbour-joining and 3D principal coordinates. The 29 isolates of the pathogen clustered into four main groups, which were identified by three SSR markers (NPRL_PK_108a, NPRL_PK_162_spka and NPRL_PK_221_spka). Phenotypic data at 21 days after the single-pustule inoculation showed that P. kuehnii from highly susceptible commercial cultivars harboured a small proportion of variants capable of causing disease on resistant cultivars. A differential reaction was demonstrated for the most virulent variant in a repeated experiment confirming the existence of races within P. kuehnii in Brazil.     

Relatedness Among Agronomically Important Rusts Based on Mitochondrial Cytochrome b Gene and Ribosomal ITS Sequences

A fragment of the mitochondrial cytochrome b (cyt b) gene of 13 agronomically important plant pathogenic Basidiomycetes was sequenced, including several Puccinia spp., Uromyces appendiculatus, Phakopsora pachyrhizi, Hemileia vastatrix and Rhizoctonia solani. The deduced amino acid sequences (residues 142–266) were used to study the relatedness of these pathogens as compared to other species of the Basidiomycetes, Ascomycetes and Oomycetes. The relatedness was also studied at nuclear level using the Internal Transcribed Spacers (ITS) in the ribosomal DNA. Phylogenic trees were constructed with the maximum parsimony (MP) and the neighbour‐joining (NJ) methods. On the basis of both cytochrome b and ITS sequences, the Puccinia species pathogenic to graminaceous crop plants, such as Puccinia recondita f. sp. tritici, Puccinia graminis f. sp. tritici, Puccinia striiformis f. sp. tritici, Puccinia coronata f. sp. avenae, Puccinia hordei, P. recondita f. sp. secalis and Puccinia sorghi, together with Puccinia horiana from Chrysanthemum, were very closely related to each other, whereas Puccinia arachidis (from peanut) was closely related to U. appendiculatus (from beans) but more distant from the other Puccinia species. Both rusts on soybean (P. pachyrhizi) and coffee (H. vastatrix) were outside the Puccinia cluster. All rusts were separated from other Basidiomycetes such as R. solani and the strobilurin producing species Strobilurus tenacellus and Mycena galopoda. Our results demonstrate that the amino acid sequence of the mitochondrial cytochrome b is a valid tool to study phylogenic relatedness among plant pathogenic Basidiomycetes and supports taxonomic grouping based on morphological structures and host specificity. Because of their high variability, ITS sequences were able to discriminate Puccinia species, which were identical on the basis of the cytochrome b amino acid sequence. Thus, ITS sequences could better show differences among species or within a species, whereas cytochrome b is more suitable than ITS for phyologenic inference at family or genus level. In addition, the sequence data obtained during this study represent essential information for easy isolation of the cyt b gene and detection of point mutations conferring resistance to Qo Inhibitor fungicides that eventually may evolve.     

Three species of rust fungi (Uredinales) from Okinawa,Japan

Three species of rust fungi were reported based on the specimens collected in the islands of the Yaeyama group and Okinawa, Japan.Puccinia tarennicola onTarenna gracilipes andUredo daphniphylli onDaphniphyllum teijsmannii were described as new species.Villebrunea frutescens was added to the host plants ofUredo pipturi. Contribution No. 115, Laboratories of Plant Pathology and Mycology, Institute of Agriculture and Forestry, University of Tsukuba.     

Observations on the ultrastructure of the uredinial stage ofPuccinia polypogonis onPolypogon monspeliensis

The ultrastructure of the uredinial stage of the rust fungus,Puccinia polypogonis onPolypogon monspeliensis is described, using scanning and transmission electron microscopy. This study examined the urediniospores, intercellular hyphae, and haustoria of the fungus. The formation and structure of urediniospores is similar to those of otherPuccinia species. The ultrastructure of intercellular hyphae and haustoria is similar to those of other rust fungi, but with some differences. No modifications are observed in the wall of the haustorial mother cells during penetration. A collar is found only around old haustoria. In most cases, one nucleus is detected inside the haustorial body and no nucleoli are seen in the nuclei of intercellular hyphae and haustoria. The host-parasite interface, including extrahaustorial matrix and extrahaustorial membrane, is also discussed and compared with those of other rust fungi.     

Accumulation of genes for susceptibility to rust fungi for which barley is nearly a nonhost results in two barley lines with extreme multiple susceptibility

Nonhost resistance is the most common type of resistance in plants. Understanding the factors that make plants susceptible or resistant may help to achieve durably effective resistance in crop plants. Screening of 109 barley (Hordeum vulgare L.) accessions in the seedling stage indicated that barley is a complete nonhost to most of the heterologous rust fungi studied, while it showed an intermediate status with respect to Puccinia triticina, P. hordei-murini, P. hordei-secalini, P. graminis f. sp. lolii and P. coronata ff. spp. avenae and holci. Accessions that were susceptible to a heterologous rust in the seedling stage were much more or completely resistant at adult plant stage. Differential interaction between barley accessions and heterologous rust fungi was found, suggesting the existence of rust-species-specific resistance. In particular, many landrace accessions from Ethiopia and Asia, and naked-seeded accessions, tended to be susceptible to several heterologous rusts, suggesting that some resistance genes in barley are effective against more than one heterologous rust fungal species. Some barley accessions had race-specific resistance against P. hordei-murini. We accumulated genes for susceptibility to P. triticina and P. hordei-murini in two genotypes called SusPtrit and SusPmur, respectively. In the seedling stage, these accessions were as susceptible as the host species to the target rusts. They also showed unusual susceptibility to other heterologous rusts. These two lines are a valuable asset to further experimental work on the genetics of resistance to heterologous rust fungi.Electronic Supplementary Material Supplementary material is available in the online version of this article at http://dx.doi.org/10.1007/s00425-004-1319-1Abbreviations ff. spp Formae speciales - RIL Recombinant inbred line - DC Double cross - DC-S Progeny produced by selfing of double-cross plants     

Tissue culture of the alternate hosts of wheat,rye and oat rusts and their response to in vitro inoculation with the rust pathogens

Alternate host plants of cereal rust fungi are necessary for studying the rust sexual cycle and pathogenicity. These plants are usually difficult to propagate through cloning, while seed-propagated plants may have variable responses to the pathogen. To overcome these obstacles, tissue culture, under controlled and aseptic conditions, was utilized for clonal propagation and in vitro inoculation of the following species: Rhamnus palaestinus Boiss., the alternate host of oat (Avena spp.) crown rust (Puccinia coronata Corda); Thalictrum speciosissimum L., the alternate host of brown leaf rust of wheat (Puccinia recondita f. sp. tritici Eriks. & Henn.); and Lycopsis arvensis L., the alternate host of rye (Secala spp.) leaf rust (Puccinia recondita f. sp. recondita Rob. & Desm.). Shoot culture procedures for initial establishment and proliferation were developed for all three alternate host species. Shoot cultures were multiplied at rates ranging from 0.3 to 1.7 shoots/week. Successful infection following inoculation with teliospores of the corresponding rust fungi was obtained for R. palaestinus and T. speciosissimum but not for L. arvensis. The hardening and acclimatization efficiency of rooted T. speciosissimum and L. arvensis was of 80–90%. The propagation efficiency for R. palaestinus was not successful because of the low rate and poor quality of its rooting. It is concluded that the in vitro system might be used as an alternative method for inoculation and multiplication of alternate hosts of cereal rusts, although more experimentation is needed to define accurately the appropriate conditions for the proper infection response. This revised version was published online in June 2006 with corrections to the Cover Date.     

Comparative morphology of uredinia and urediniospores of six Puccinia species parasitic on Poaceae in Saudi Arabia

Uredinia and urediniospores of six Puccinia species growing on Poaceae in southwestern Saudi Arabia were morphologically compared by light and scanning electron microscopy (SEM). Puccinia cenchri, P. fragosoana and P. isiacae were recorded for the first time in Saudi Arabia. Many differences between uredinia and urediniospores of studied Puccinia species were recorded. These differences are not related to host plant but may be due to the species of Puccinia itself. Observations by SEM led to more information in distinguishing between these Puccinia species particularly the presence of paraphyses and density and length of spines.     

准噶尔柄锈菌:新疆荒漠的雀麦锈菌——新种

本文描述了南疆雀麦Bromus gedrosianus Penz.上的锈菌一新种:准噶尔柄锈菌Puccinia junggarensis S.-X. Wei & J.-Y. Zhuang sp. nov.。模式标本采自新疆古尔班通古特沙漠克拉玛依附近,保藏在中国科学院微生物研究所真菌标本室。此菌曾被本文第二作者暂定为Puccinia burnettii Griff.。原标本经第一作者复查发现夏孢子表面纹饰为细庆而非细刺,夏孢子和冬孢子的综合特征不同于禾本科植物上迄今已知的所有柄锈菌属的种,故鉴定为新种。     

Life cycle and life strategy features of Puccinia glechomatis (Uredinales) favorable for extending the natural range of distribution

The objective of this study was to find features in microcyclic rust fungi (Uredinales) on wild host plants favorable for extension of the natural range of distribution. Puccinia glechomatis, a leptosporic rust fungus and its herbal host Glechoma hederacea (Lamiaceae), both natives to Eurasia and introduced in North America, were used for this study. Although the host has been known from North America since the beginning of the nineteenth century, the rust fungus was first observed there only in recent years. Favorable features were identified by studying the life cycle of the rust, including nuclear conditions and seasonal characteristics as well as its spread in North America. The life cycle was studied macroscopically by inoculation experiments, by various light microscope techniques, and by scanning electron microscopy. The spread of the pathogen and its host were reconstructed by evaluating host plant herbarium specimens and databases, literature, and field study data. The studies on P. glechomatis show that, generally for microcyclic rust fungi, establishment and potential for spread are based on several favorable features of both the host (e.g., synanthropic occurence and dispersal, genetic stability, regeneration of vegetative plant parts) and the rust fungus (asexual reproduction/genetic stability, homothallism, propagation with host plant, formation of both leptospores and thick-walled teliospores).     

Epidemic Development and Virulence in 1995–1998 of Puccinia coronata, a Potential Biocontrol Agent of Wild Oats on San Clemente Island

The biology of islands differs from that of large land masses in having less complex ecosystems. Introduced exotic weeds are often a major threat to fragile island ecosystems because of their expansion into habitats previously occupied by endemic species. San Clemente Island, 120 km off the California coastline, is an example of this process in which numerous exotic weed species have been introduced and some are endangering the native flora. Crown rust of oats caused by Puccinia coronata f.sp. avenae was investigated as a potential biocontrol agent against two wild oat species, Avena barbata and Avena fatua, introduced on San Clemente Island. Epidemiology and virulence of this rust were studied. The island was surveyed from 1995 to 1998 for occurrence of P. coronata on wild oats. Wild oats were found sprouting in the northern part of the island shortly after autumn rainfall and subsequently covered the main grasslands of the island. The rust also appeared first on the northern part of the island and progressively spread south. However, disease severities in the south were considerably lower than those in the north. Diverse virulence types, although related to Californian and Mexican forms, were detected among the isolates. The potential use of P. coronata as an augmentative biocontrol agent for wild oat species on San Clemente Island is discussed.     

Taxonomic identity of caricicolousPuccinia host-alternating onPetasites in Japan

Uredinial-telialPuccinia fungi onCarex shimizuensis in Nagano Pref. and onC. dimorpholepis in Ibaraki Pref. were proven to host-alternate onPetasites japonicus by field observations and inoculation experiments. These fungi from the two localities were morphologically similar and were compared with three describedPuccinia species that host-alternate betwenCarex species andP. japonicus in Japan, i.e.,P. caricis-petasitidis, P. caricis-flabellatae andP. caricis-podogynae. The three previously described species and the newly foundPuccinia fungi were morphologically indistinguishable at all stages of the life cycle; therefore, it was concluded that three species and the two newly found fungi are taxonomically identical, in whichP. caricis-petasitidis has nomenclatural priority.     

竹上一柄锈新种

本文描述了采自湖南龙山的竹锈一新种:竹生柄锈Puccinia bambusicola S.-x. Wei & J.-y. Zhuang。模式标本保藏在中国科学院微生物研究所真菌标本室.此菌与其它已知的寄生在竹上的柄锈比较,冬孢子很小(25-38 × 14-21 µm),顶壁也不厚(仅3-6 gym),近似于南美产的竹柄锈P. bambusarum Arth.,但后者冬孢子隔膜多为斜生且颜色较淡。附我国竹生柄锈属已知种的检索表。     

中国蓼科植物柄锈菌的分类研究

蓼科植物((Polygonaceae)具有重要的经济意义.其中有粮食作物,著名中药,蜜源植物,观赏植物以及其它经济植物.柄锈菌(Puccinia)是蓼科植物的重要的病原真菌.本文对我国寄生于蓼科植物的柄锈菌进行了分类研究. 在中国首先报告蓼科植物柄锈的是日本人三宅市郎(1914).我国真菌学家从30年代起分别描述或记载过各种蓼科植物的柄锈,并发表过一些中国特有的新种. 本文作者重新研究了中国科学院微生物研究所真菌标本室收藏的全部蓼科植物柄锈的标本以及近年来采集的未定名标本.共鉴定标本262号,得柄锈30种,包括5个新种:汶川柄锈(Pucinia wencuanensis),雾灵柄锈(P. wulingensis),南迩巴瓦柄锈(P.namjagbarwana),太白柄锈(P. taibaiana),掌叶大黄柄锈(P. rhei palmati),和五个国内新记录种:装饰柄锈(P, ornata),岩手山柄锈(P. iwateyamensis),山蓼柄锈(P. oxyriae),赫尔顿柄锈(P.hultenii),箭叶蓼柄锈(P.polygoni-sieboldii).寄主植物包括蓼科8属47种.