Phylogenetic relationships among Puccinia hemerocallidis, P. funkiae, and P. patriniae (Uredinales) inferred from ITS sequence data

Puccinia hemerocallidis and P. funkiae resemble each other morphologically; however, they are biologically and taxonomically distinct, with telia of the former being restricted to species of Hemerocallis and the latter to Hosta species. However, both fungi share a macrocyclic and heteroecious life cycle with Patrinia villosa as the spermogonial and aecial host. An additional microcyclic rust fungus, P. patriniae, is also known on P. villosa. This microcyclic fungus is similar to the two macrocyclic fungi in its telial structure and teliospore morphology. These similarities in morphology and host relationships suggest the three fungi may also share a close evolutionary relationship. To determine the phylogenetic relationships of the three species, a portion of the nuclear ribosomal DNA repeat encoding the ITS and 5.8S subunit regions was amplified by PCR, sequenced, and analyzed. The resulting phylogenetic trees showed that P. hemerocallidis and P. funkiae share a recent common ancestor and that P. patriniae is closely allied with P. hemerocallidis. The results suggest a possible evolutionary derivation of microcyclic P. patriniae from macrocyclic heteroecious P. hemerocallidis, which fits the evolutionary interpretation of correlated species known as Tranzschel's law.     

The diversity of nuclear cycle in microcyclic rust fungi (Uredinales) and its ecological and evolutionary implications

 Nine types with 11 variations of nuclear cycle and associated metabasidium development were distinguished in microcyclic rust fungi. An additional type was recognized in rust fungi with an expanded life cycle. A significant proportion of rust fungi with a reduced life cycle is assumed to have lost a sexual genetic recombination process, being either apomictic or asexual in reproduction. Most species that retain a sexual process in the microcyclic life cycle seem to have become homothallic. During life cycle evolution by the omission of spore stages, these traits might have had a selective advantage for those species that had less opportunity to encounter a genetically different but sexually compatible mate because of isolated patchy distribution or a short growing season. The findings that different populations of a morphologically identifiable species exhibit two or more distinct patterns of nuclear cycle and different metabasidium development indicate that microcyclic lineages might have evolved independently and repeatedly from a macrocyclic parental species. Those lineages are morphologically the same but would differ from each other in their genetics and biology. Received: July 5, 2002 / Accepted: August 5, 2002 Acknowledgment This work was supported in part by a Grant-in-Aid for Scientific Research (no. 09640744) from the Ministry of Education, Science and Culture (now the Ministry of Education, Culture, Sports, Science and Technology), Japan. Correspondence to:Y. Ono     

Genetic evidence for sexual reproduction and multiple infections of Norway spruce cones by the rust fungus Thekopsora areolata

Rust fungi are obligate parasites, of plants, with complex and in many cases poorly known life cycles which may include host alteration and up to five spore types with haploid, diploid, and dikaryotic nuclear stages. This study supports that Thekopasora areolata, the causal agent of cherry‐spruce rust in Norway spruce, is a macrocyclic heteroecious fungus with all five spore stages which uses two host plants Prunus padus and Picea abies to complete its life cycle. High genotypic diversity without population structure was found, which suggests predominantly sexual reproduction, random mating and a high gene flow within and between the populations in Fennoscandia. There was no evidence for an autoecious life cycle resulting from aeciospore infection of pistillate cones that would explain the previously reported rust epidemics without the alternate host. However, within cones and scales identical multilocus genotypes were repeatedly sampled which can be explained by vegetative growth of the fertilized mycelia or repeated mating of mycelium by spermatia of the same genotype. The high genotypic diversity within cones and haplotype inference show that each pistillate cone is infected by several basidiospores. This study provides genetic evidence for high gene flow, sexual reproduction, and multiple infections of Norway spruce cone by the rust fungus T. areolata which expands the general understanding of the biology of rust fungi.     

Volatiles modulate the development of plant pathogenic rust fungi

Rust fungi are obligate biotrophic pathogens that differentiate a series of specialized cells to establish infection. One of these cells, the haustorium, which serves to absorb nutrients from living host cells, normally develops only in planta. Here, we show that the rust fungus Uromyces fabae (Pers.) Schroet. stimulates volatile emission of its host, broad bean (Vicia faba L.). Volatiles were identified and shown to be perceived by the fungus in in vitro assays that excluded the host. Three of them, nonanal, decanal, and hexenyl acetate promoted the development of haustoria on artificial membranes. In contrast, the terpenoid farnesyl acetate suppressed this differentiation. In assays using whole plants, farnesyl acetate reduced rust disease not only on broad bean but also on several cereals and legumes including soybean. This natural substance was effective against all rusts tested when directly applied to the host. This demonstrated that farnesyl acetate may serve as a powerful novel tool to combat rust fungi including Phakopsora pachyrhizi that currently threatens the production of soybeans world-wide.     

Does Rust Infection of Willow Affect Feeding and Oviposition Behavior of Willow Leaf Beetles?

Willows are often attacked by both herbivorous insects and rust fungi. Little is known about interactions between these two willow enemies. We studied whether feeding and oviposition behavior of the willow leaf beetle Plagiodera versicolora upon the willow hybrid Salix x cuspidata is affected when the rust fungus Melampsora allii-fragilis has attacked the plant. Laboratory bioassays revealed that adult willow leaf beetles significantly avoided feeding and oviposition on rust-infected leaves when compared to healthy leaves. Further bioassays aimed to elucidate the temporal and spatial scale of effects of rust infection on feeding behavior of adults. While infected parts of leaves were avoided at all times past infection tested (8, 12, and 16 days), symptom-free parts of infected leaves were only avoided 16 days past infection. Systemic effects extended only one leaf position up and two leaf positions down from the infection site.     

Tests for Parasite-mediated Frequency-dependent Selection in Natural Populations of an Asexual Plant Species

Genetic variation in plant populations for resistance to pathogens and herbivores might be maintained by parasite-mediated negative frequency-dependent selection (FDS). But it is difficult to observe the time-lagged oscillations between host and parasite genotypes that should result from FDS. To evaluate the potential for FDS, we tested for local adaptation of parasites to common clones, the role of host genetic diversity in resistance to parasites, and genetic correlations among fitness, parasitism, and the frequency of host clones. We studied three populations of Arabis holboellii, a short-lived apomictic (asexual by seed) plant attacked by rust fungi and insect herbivores. To estimate clone frequency, we used polymorphic allozyme markers on 200 individuals in each population in 1990 and in 2000. We also recorded levels of parasitism and host fitness (fruit production). Only the rust fungi showed evidence for local host adaptation; they usually increased in incidence as a function of clone frequency, and they tracked temporal change in clone frequency. In further support of FDS, parasitism was lower in populations with higher genetic diversity. However, total parasitism (herbivory and disease combined) decreased as host clone frequency and fitness increased. Thus, although the highly virulent rust pathogen showed potential for driving the cycles that result from FDS, this apparently does not occur in the populations studied because the host clones were also attacked by herbivores.Co-ordinating editor: J.F. Stuefer     

Aplopsora corni sp. nov. onCornus controversa from Hokkaido,Japan

Aplopsora corni sp. nov. is proposed for a rust fungus whose uredinial and basidial stage occurs onCornus controversa (Cornaceae) in Hokkaido. This new species is separated by its larger urediniospores and probasidia from the morphologically closely relatedA. nyssae onNyssa aquatica andN. sylvatica (Cornaceae) distributed in southern North America.     

Life cycle and nuclear behavior in three rust fungi (Uredinales)

Kuehneola japonica has a microcyclic life cycle with a regular alternation of generations. Single basidiospore inoculations onto Rosa wichuraiana resulted in teliospore production, indicating its homothallic nature. Dikaryotization in a vegetative mycelium in the host seemed to occur through nuclear division that was not followed by septum formation. Karyogamy and meiosis took place through teliospore and metabasidium development; this fungus was considered to reproduce genetically homogeneous progenies. Puccinia lantanae and P. patriniae were also microcyclic in their life cycle; however, these fungi differed from K. japonica in the mode of nuclear behavior. In the former two fungi, both vegetative and reproductive cells were uninucleate. No karyogamy was observed, and nuclear division in the metabasidium development was thought to be mitotic. In P. lantanae, a basidiospore was formed on a sterigma, whereas a whiplike hypha emerged from each metabasidium cell in P. patriniae. Inoculations of Justicia procumbens with a single basidiospore of P. lantanae resulted in teliospore production. The fungus seemed to remain uninucleate, either haploid or diploid, throughout the life cycle. Thus, reproduction was considered to be apomictic. Received: August 16, 2001 / Accepted: October 1, 2001     

Experiments on competitiveness of three endomycorrhizal fungi

Summary This paper examines competition between three VA endophytes places respectively as “inoculant” fungus in an inner compartment (300 ml) below the germinating seed, and as “indigenous” fungus in an outer compartment (900 ml). Previously prepared infested soil was used as source of the endophytes. Inner and outer compartment roots were harvested separately and percentage infection determined for each fungus, using diagnostic anatomical features of each fungus. Changes in colonisation patterns were followed up to 4 and 5 months. Treatments with non-infested soil in the outer compartment were used to compare infectivity and spread of the different endophytes. All three endophytes were able to invade soil already colonised by either of the other two, butGlomus fasciculatum (E₃) was more invasive thanGlomus tenue. This pattern appeared to be related to rates of spread of the fungi in the non-infested soil. The pattern was not affected by host, clover or sorghum, or by position, immediately below the seed or in the outer compartment. It seemed that the endophyte in the inner compartment had overriding and persistent effects on plant growth, unrelated to its dominance pattern or share of overall root infection at the later harvests.     

Fungal–plant signalling in the Ustilago maydis–maize pathosystem

The smut fungus Ustilago maydis needs the host plant maize for completion of its sexual life cycle. Recent experiments highlight the importance of cAMP and mitogen-activated protein (MAP) kinase signalling for cell fusion as well as for subsequent stages of plant colonisation and induction of disease symptoms. During these distinct developmental stages the same signalling cascades must be differentially regulated and accommodate multiple inputs and outputs.     

Frequency-dependent selection in a wild plant-pathogen system

This study investigated the occurrence of the rust fungus Puccinia chondrillina in clonal populations of its host plant, Chondrilla juncea. In 16 populations spread across eastern Turkey, 48 different multilocus isozyme phenotypes were identified in the host. Of these clones, 88% were restricted to single localities, while the remaining 12% were found in 2–11 populations. For 13 of the 16 plant populations the commonest host clone was always infected. Indeed, at ten sites this clone was the only one found to carry disease. In the remaining three populations the rusted plants were all of the second commonest isozyme type. The possibility of such a tight association of rust incidence with host clone frequency simultaneously across a wide geographic area is very low (P0.023), supporting the contention that the pathogen P. chondrillina may be imposing negative frequency-dependent selection on these C. juncea populations.     

Prospects for Biocontrol of Invasive Rosa rugosa

The biota of herbivorous arthropods and pathogenic microorganisms associated with Rosa rugosa in its native and exotic ranges is reviewed. This is done as an initial step towards the identification of potential agents for biological control of this plant species invasive in Europe and North America. It is shown that more insect (but apparently not fungal) species attack R. rugosa in its native range than in its exotic range, and that most of the specialized insect and fungal enemies are confined to its native range. Among the close relatives of R. rugosa in its exotic ranges are many native species, as well as economically important crop plants. Few organisms appear to be narrowly specialized to R. rugosa, but true host specificity can only be identified through experimental testing. Based on the literature, the most promising candidates for biocontrol seem to be the aphids Myzus japonensis and Amphorophora amurensis, the leaf hopper Empoasca ussurica, the tortricid moth Notocelia longispina, the cynipid gall-wasp Diplolepis fukudae, and the rust fungi Phragmidium rosae-rugosae and P. yezoense. A screening programme is suggested, investigating the impact of these organisms on R. rugosa performance, their host specificity and the risk of undesired indirect effects in the ecosystem where agents are released. In addition, demographic studies of the target plant should be integrated to provide guidance for the stage in the life cycle most sensitive to control and, thus, enable selection of the most efficient and safe biocontrol agents.     

Phylogenetic analyses of <Emphasis Type="Italic">Uromyces viciae-fabae</Emphasis> and its varieties on <Emphasis Type="Italic">Vicia</Emphasis>, <Emphasis Type="Italic">Lathyrus</Emphasis>, and <Emphasis Type="Italic">Pisum</Emphasis> in Japan

A pea rust fungus, Uromyces viciae-fabae, has been classified into two varieties, var. viciae-fabae and var. orobi, based on differences in urediniospore wall thickness and putative host specificity in Japan. In principal component analyses, morphological features of urediniospores and teliospores of 94 rust specimens from Vicia, Lathyrus, and Pisum did not show definite host-specific morphological groups. In molecular analyses, 23 Uromyces specimens from Vicia, Lathyrus, and Pisum formed a single genetic clade based on D1/D2 and ITS regions. Four isolates of U. viciae-fabae from V. cracca and V. unijuga could infect and sporulate on P. sativum. These results suggest that U. viciae-fabae populations on different host plants are not biologically differentiated into groups that can be recognized as varieties.Contribution no. 184, Laboratory of Plant Parasitic Mycology, Institute of Agriculture and Forestry, University of Tsukuba, Japan     

Cronartium orientale, sp. nov., segregation of the pine gall rust in eastern Asia fromCronartium quercuum

The Asian pine gall rust, which has hitherto been assigned toCronartium quercuum distributed in North America, is described asC. orientale, sp. nov. The spermogonial and aecial states occur on hard pines, and the uredinial and telial states occur on various oaks.Cronartium orientale has globose, almost hyaline basidiospores in contrast to the ellipsoid, yellow-orange ones of North AmericanC. quercuum s.l. Characteristics of the new rust species in morphology, DNA analysis, and host alternation are discussed.     

Biology and life cycle ofAtelocauda koae,an unusual demicyclic rust

Atelocauda koae, a rust of the native HawaiianAcacia koa, is considered as a demicyclic species, having spermogonial, aecial, and telial states, but is unusual in production of aeciospores simultaneously with teliospores rather than consecutively. Host inoculation with spores of each state separately confirmed that the life cycle was perpetuated by the telial state, but the aeciospores, while capable of germination and stomal penetration, did not produce detectable infection. This rust therefore behaves as a microcyclic species, and appears to be in evolutionary transition toward this reduced state. Teliospores produced vestigial, permanently attached basidiosopores which germinated to produce infective hyphae. The hyphae entered the host either through stomata or penetrated the epidermis directly, with the latter method being more common. Unusual nuclear associated with teliospore germination, in which meiosis occurs in more than one diploid nucleus was observed, in confirmation of an earlier study.     

Pre-release impact assessment of two stem-boring weevils proposed as biological control agents for Alliaria petiolata

Experimental studies can be useful tools to test plant responses to herbivory and to quantify the impact of potential biological control agents prior to their release. We evaluated the per-capita effect of Ceutorhynchus alliariae and C. roberti, two stem-boring weevils currently being investigated as potential biological control agents for garlic mustard, Alliaria petiolata, in North America. Weevils were released at three different densities in individual and mixed-species treatments onto potted plants of A. petiolata. Damage by C. roberti alone and by both weevils combined caused an increase in the numbers of inflorescences produced per plant. Although plants could compensate for low levels of damage, moderate to high levels of damage by both C. alliariae and C. roberti, individually and in combination, caused a decrease in plant height and a reduction in seed output per plant. The damage inflicted by both weevil species is similar so the overall impact of both species combined can be predicted by summing the impact of each species alone. Provided they are sufficiently host specific, both weevils could be released as biocontrol agents. Because reduced seed production is necessary to suppress A. petiolata populations, both species have the potential to contribute to control of A. petiolata in North America.     

Caeoma tsukubaense n. sp., a rhododendron rust fungus of Japan and southern Asia, and its relationship to Chrysomyxa rhododendri

A rust fungus found in Japan on Rhododendron kaempferi, R. kiusianum, and R. dauricum has previously been identified as Chrysomyxa rhododendri. Light and scanning electron microscopy of fresh and herbarium materials of the rust fungus, however, show that the spore surface morphology differs from the urediniospores of C. rhododendri, and the spores are slightly smaller. Furthermore, the DNA sequence of the 5′-end of the large subunit of ribosomal DNA differs from that of C. rhododendri by 3%. Telia have not been found; therefore, it is redescribed as a new anamorphic species, Caeoma tsukubaense. Several specimens from North Korea, Tibet, and Nepal bearing a similar rust fungus are also included in the species.Contribution no.193 from the Laboratory of Plant Parasitic Mycology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Japan     

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     

Host range of Ceutorhynchus assimilis (Coleoptera: Curculionidae), a candidate for biological control of Lepidium draba (Brassicaceae) in the USA

Ceutorhynchus assimilis has been selected as a potential biological control agent of Lepidium draba, which is a Eurasian invasive weed in North America. Preliminary studies indicated specificity of this weevil collected in southern France on L. draba. This result was in discord with the pest status of C. assimilis found in the literature. Host-specificity tests based both on field and laboratory experiments showed heterogeneity in the host spectrum of the weevils reared from different host-plants as determined by larval development. However, no distinguishable morphological differences could be visually detected between the populations feeding on different host-plants. All sampled populations of weevils were polyphagous as adults. Weevils reared from L. draba were specific to this plant for their complete larval development. Conversely, populations living on other wild and cultivated Brassicaceae species were not able to use L. draba as a host plant. Such differentiation is further highlighted by other biological aspects such as plant infestation rates, sex-ratio, duration of larval development, and differences in the timing of their life cycles. These results demonstrate that C. assimilis, an insect species formerly considered as a pest of Brassicaceae, is characterized by its host-range variability, with one population being potentially useful in the biological control of L. draba. Moreover, this example points to the need to test multiple populations of biological control agents in assessing risk.     

Impact of the belowground herbivore and potential biological control agent, Ceutorhynchus scrobicollis, on Alliaria petiolata performance

We studied the influence of the root-crown weevil Ceutorhynchus scrobicollis on its host plant Alliaria petiolata, a European biennial herb that is currently invading much of temperate North America. Varying timing of attack and herbivore densities in a common garden allowed to assess seasonality of plant response, density-dependence of impact, and the effect of intraspecific competition on C. scrobicollis recruitment (number of F₁ generation adults emerged). Data collected in the common garden were compared with data collected at field sites. C. scrobicollis is a common weevil in Europe, frequently attaining high attack levels on its host plant. In the common garden, weevil attack decreased plant survival by up to 43%, reduced plant height by 54%, increased the number of shoots by up to four–fold and delayed seed ripening, but had no significant negative effect on seed production. Plants infested in spring allocated less biomass to aboveground plant parts, and remained smaller than plants attacked in autumn, indicating that the latter were able to partly compensate for weevil attack. Increasing weevil density rarely had an effect on A. petiolata performance, and did not increase F₁ recruitment, suggesting strong intraspecific competition. At field sites, C. scrobicollis attack is spread over a long time period, which probably alleviates intraspecific competition. In summary, attack by the root-crown feeding weevil, C. scrobicollis, can substantially reduce growth and survival of A. petiolata. If introduced as a biological control agent into North America, C. scrobicollis is likely to decrease the fitness and competitive superiority of A. petiolata.