Morphological and molecular confirmation of Albugo resedae (Albuginales; Oomycota) as a distinct species from A. candida

The genus Albugo s.str. causes white blister rust on four families of the Brassicales, Brassicaceae, Capparaceae, Cleomaceae, and Resedaceae. Recent phylogenetic studies have revealed that several host specific lineages are present within Albugo on Brassicales, while it was also confirmed that Albugo candida has an exceptionally wide host range which extends from Brassicaceae to Cleomaceae and Capparaceae. The Albugo species infecting the Resedaceae was attributed in monographic studies as well as local floras to either A. resedae or, applying a broader species concept, to A. candida. In the present study, A. resedae specimens were morphologically and molecularly compared to the five Albugo species so far confirmed from Brassicales, A. candida, A. koreana, A. laibachii, A. lepidii, and A. voglmayrii. Both morphological differences of oospore ornamentation and phylogenetic analysis of cox2 mtDNA sequences provided evidence that A. resedae is distinct from A. candida and from the additional four species so far described from Brassicaceae. It thus seems possible that so far unknown factors restrict Albugo candida to Brassicaceae and its sister families, Cleomaceae and Capparaceae.     

A new presumably widespread species of Albugo parasitic to Strigosella spp. (Brassicaceae)

White blister rust is one of the most common diseases in Brassicaceae. Recently, molecular approaches revealed that apart from Albugo candida, several other more specialized species of the genus are causing this disease on Brassicaceae and the diversity of this group still remains largely unexplored. All newly described species have so far been sampled only from a limited geographic range, except for Albugo species which followed their invasive host from Europe to other continents. In this study we show that a previously unknown species of Albugo is causing white blister rust disease on two species of Strigosella. This species can be distinguished from other species of the genus Albugo both by its phylogenetic position and its unique oospore ornamentation that might be an adaptation to the harsh environment of the host plants. As a consequence, Albugo arenosa is described and illustrated as a new species, so far known from Strigosella africana in Iran and Spain, and Strigosella brevipes in Iran. Apart from Albugo candida and Albugo lepidii, this is the third hitherto known species of Albugo s. str. with a confirmed native distribution range of several thousand kilometres.     

Phylogeny of an Albugo sp. infecting Barbarea vulgaris in Denmark and its frequency of symptom development in natural populations of two evolutionary divergent plant types

The oomycete Albugo candida has long been considered a broad spectrum generalist pathogen, but recent studies suggest that it is diverged into several more specialized species in addition to the generalist Albugo candida sensu stricto. Whereas these species cause the disease white blister rust in many crucifer plants, asymptomatic endophytic infections may be important in the epidemiology of others. One of the plant species attacked by Albugo sp. is the wild crucifer Barbarea vulgaris ssp. arcuata, which is diverged into two phytochemically and genetically different types with different geographical distributions in Europe. These were previously shown to differ strongly in propensity to develop white rust upon controlled infections in the greenhouse. Here, we analyse the phylogenetic relatedness of this local Albugo sp. field isolate to other species and lines of Albugo spp., including others collected on B. vulgaris. We further ask whether the difference in incidence of white rust between the two types of B. vulgaris are also expressed in natural populations.     

A new species of <Emphasis Type="Italic">Pustula</Emphasis> (Oomycetes,Albuginales) is the causal agent of sunflower white rust

White blister rust of sunflower is an emerging disease that is among the most important diseases in this crop in South Africa and has recently spread to Europe. For the genus Albugo, it has been demonstrated that species are mostly at least host genus specific and that several previously overlooked species are present on Brassicaceae. It thus seems likely that previously unrecognised species are also present in the genus Pustula. Based on previous phylogenetic reconstruction in combination with differences in oospore ornamentation, it is revealed that Pustula on sunflower, previously attributed to Pustula tragopogonis (syn. Albugo tragopogonis), is distinct from Pustula on Tragopogon. Therefore, this pathogen is described as a new species, Pustula helianthicola. In addition, taxonomic observations revealed that Pustula tragopogonis is an incorrect name and is replaced by the new combination, Pustula obtusata.     

A new perspective on the evolution of white blister rusts: <Emphasis Type="Italic">Albugo</Emphasis> s.str. (Albuginales; Oomycota) is not restricted to Brassicales but also present on Fabales

For almost all groups of pathogens, unusual and rare host species have been reported. Often, such associations are based on single or few collections only, which are frequently hard to access. Many of them later prove to be due to misidentification of the host, the pathogen, or both. Therefore, such reports are often disregarded, or treated anecdotally in taxonomic and phylogenetic studies, regardless of their potential importance to unravelling the evolution of the entire group. Concerning oomycete biotrophs there are several reports of unusual and rare hosts for hardly known pathogens. In the order Fabales, for example, a single species of Albugo, A. mauginii, was described as parasitic to Onobrychis crista-galli about 80 years ago, but not recorded again. All other confirmed members of Albugo s.str. are parasitic to representatives of the families Brassicaceae, Capparaceae, Cleomaceae, and Resedaceae in the order Brassicales. In the present study, molecular phylogenetic analysis of cox2 mtDNA sequences and morphological investigations on an original specimen confirmed the occurrence of a member of Albugo on Fabaceae hosts, with the characteristic thin wall of the secondary sporangia, which is almost uniform in thickness. In phylogenetic analyses the species results as embedded within Albugo s.str. Therefore, it is concluded that the natural host range of Albugo s.str. extends from Brassicales to Fabales via host jumping. Our results underscore that unrevised reports of pathogens from unusual hosts should be reconsidered carefully to obtain a more complete picture of pathogen diversity and evolution.     

Three new phylogenetic lineages are the closest relatives of the widespread species Albugo candida

White blister rust caused by the obligate biotroph Albugo candida (Albuginaceae; Oomycota) is one of the most notorious and common diseases of Brassicaceae. During the past 5 y, A. candida specimens collected from about 30 host genera were phylogenetically and morphologically investigated in several studies. These not only revealed that A. candida s.str. has a broad host range, encompassing a large number of host plants belonging to Brassicales, but also the presence of previously overlooked species of Albugo with hosts in this order. In this study, we examined specimens from Alyssum, Barbarea, and Rorippa, of which many species were commonly recorded as host plants of A. candida but could not be included in previous works due to the paucity of specimens available. It was revealed that Albugo specimens from Alyssum montanum, Barbarea vulgaris, and various Rorippa species, were placed in three phylogenetically distinct clades, but closer to A. candida s.str. than any previously reported species. Oospores were observed from Albugo specimens parasitic to Rorippa and could be distinguished morphologically from A. candida. Therefore, Albugo rorippae sp. nov. is described and illustrated here. In addition, a key of Albugo species described previously from Brassicales is given. The present study reveals that a large number of Albugo species remain still undiscovered, and that species close to A. candida exist. This could help elucidating the basis of the broad host range of A. candida as opposed to the narrow specialisation that is seemingly present in other species of Albugo on the Brassicaceae.     

The only known white blister rust on a basal angiosperm is a member of the genus <Emphasis Type="Italic">Albugo</Emphasis>

Rare pathogens on unusual hosts are often providing valuable insight into the evolution of the pathogen group concerned, but it is often challenging to obtain sequence data for these, as because only very few, often decades-old specimens are available. One such example is Albugo tropica, the white blister pathogen of a basal angiosperm in the genus Peperomia (Piperaceae). For this species, only two, more than 70 and over 120-year-old collections available. Here, sequence data for A. tropica are reported and phylogenetic reconstructions reveal it as the sister group to all other white blister rusts of the genus Albugo. Its isolated position is also reflected by several morphological differences to the other species of the genus, such as very thin-walled sporangia and almost smooth oospores. The isolated phylogenetic position of the pathogen and its host might indicate that it is a relict species trapped on its host. The sister-group relationship to all members of the genus Albugo s.str., which have been investigated using molecular phylogenetics, hints at the possibility, that Albugo might have originated in South America or Gondwana and has later radiated in the holarctic on members of the Brassicales.     

Association of Peronospora parasitica with Albugo candida on Brassica juncea Leaves

Associated infections between Peronospora parasitica and Albugo candida were observed on Brassica juncea leaves. A. Candida appeared first and was followed by P. parasitica. A. candida predisposes the host tissues towards susceptibility to P. parasitica.     

Obligate biotrophic pathogens of the genus Albugo are widespread as asymptomatic endophytes in natural populations of Brassicaceae

Mutualistic interactions of plants with true fungi are a well‐known and widespread phenomenon, which includes mycorrhiza and non‐mycorrhizal endophytes like species of Epichloë. Despite the fact that these organisms intrude into plants, neither strong defence reactions nor the onset of symptoms of disease can be observed in most or even all infested plants, in contrast to endophytic pathogens. Oomycetes are fungal‐like organisms belonging to the kingdom Straminipila, which includes diatoms and seaweeds. Although having evolved many convergent traits with true fungi and occupying similar evolutionary niches, widespread oomycete endophytes are not known to date, although more than 500 endophytic pathogens, including species of the obligate biotrophic genus Albugo, have been described. Here, we report that oomycetes of the genus Albugo are widespread in siliques of natural host populations. A total of 759 plants, encompassing four genera with rare reports of white blister incidents and one with common incidents, were collected from 25 sites in Germany. Nested PCR with species‐specific primers revealed that 5–27% of the hosts with rare disease incidence carried asymptomatic Albugo in their siliques, although only on a single plant of 583 individuals, an isolated pustule on a single leaf could be observed. Control experiments confirmed that these results were not because of attached spores, but because of endophytic mycelium. Vertical inheritance of oomycete infections has been reported for several plant pathogens, and it seems likely that in nature this way of transmission plays an important role in the persistence of asymptomatic endophytic Albugo species.     

The effect of mating system on growth of Arabidopsis lyrata in response to inoculation with the biotrophic parasite Albugo candida

The effects of variation in host reproductive systems on response to pathogens are not well understood. We inoculated individuals from outcrossing and inbreeding populations of North American Arabidopsis lyrata with Albugo candida (white blister rust) to test the effect of mating system and heterozygosity on disease response. We observed three host infection phenotypes, classified as fully resistant, partially resistant and fully susceptible. Overall, inbreeding populations had more susceptible and fewer partially resistant individuals than outcrossing populations, but the highest proportion of resistant individuals was found in two of the inbreeding populations. Mating system did not affect relative growth rate of inoculated plants, but there were strong effects of population and infection phenotype. We conclude that mating system per se does not determine the resistance of natural A. lyrata populations to infection by Albugo, but that the increased variability in responses among inbreeding populations may be due to reduced effective population size.     

Antifungal activity of berberine iodide, a constituent ofFumaria indica

Berberine iodide, an isoquinoline alkaloid was isolated fromFumaria indica and its efficacy was tested against spore germination of some plant parasitic as well as saprophytic fungi. The alkaloid significantly curbed spore germination ofCurvularia lunata, Erysiphe cichoracearum, E. pisi, Fusarium udum andPenicillium species. Complete inhibition (100%) of spore germination was observed inE. cichoracearum andPenicillium species at 1.5 g/L.Albugo candida andAlternaria species were not affected by the chemical.     

Wolbachia endosymbiont is essential for egg hatching in a parthenogenetic arthropod

Wolbachia pipientis can induce a range of sex ratio distortions including parthenogenesis. Recently Wolbachia has been detected in the diploid, parthenogenetic, collembolan species Folsomia candida. In this paper we address the effect of Wolbachia on reproduction in F. candida. Wolbachia infection was removed by antibiotic and heat treatment, and quantitative PCR techniques confirmed the success of the treatments. Complete loss of Wolbachia-infection led to the production of normal clutch sizes, but was associated with full egg hatching failure. Our results demonstrate that F. candida is strictly dependent on Wolbachia to produce viable offspring. This is one of the few cases of obligatory Wolbachia infection in arthropods. Our data suggest a unique mechanism underlying Wolbachia-dependence of egg development. One of our more salient results is that hatching success increased in consecutive egg clutches of antibiotic-treated individuals, probably due to restoration of bacterial densities over time. These observations suggest that reproduction in F. candida is a threshold effect requiring a critical Wolbachia density as is hypothesized by the bacterial dosage model. Quantitative PCR analysis showed that heat or antibiotic treated individuals with egg hatching failure had low average bacterial densities, but bacterial densities were not significantly different from those of treated individuals with successfully eclosing eggs. Additional experiments with partially cured F. candida are needed to prove the dosage model.     

A study of host specificity in the entomopathogenic fungus Beauveria bassiana (Hypocreales,Clavicipitaceae)

Beauveria bassiana (Balsamo – Crivelli) Vuillemin based mycoinsecticides are used against agricultural, veterinary and medical insect pests. The fungus has a very diverse and extensive host range. Variation in virulence among isolates of B. bassiana to different insect species has been abundantly documented. Given the effect of multiple factors on virulence, it is not certain whether the observed difference in virulence can be labelled as host specificity. Environmental conditions and susceptibility of the insect population are two main factors that affect successful fungal infection. Keeping the environmental factors constant, if virulence of an isolate to different insect species and different populations within an insect species is compared, the scale of difference between the two responses can be estimated. If differences in virulence of an isolate to different insect species are greater than the difference in virulence to different insect populations within an insect species, then, the isolate can be considered as exhibiting specific preference to those insect species towards which it exhibits high virulence. To examine this feature, a worldwide sample of B. bassiana was bioassayed on nine insect species and two different populations within two insect species. Laboratory bioassays were done on: Bombyx mori (Lepidoptera), Spodoptera litura (Lepidoptera), Chilo partellus (Lepidoptera), Helicoverpa armigera (Lepidoptera), Epilachna vigintioctopunctata (Coleoptera), Mylabris pustulata (Coleoptera), Aphis craccivora (Homoptera), Maconellicoccus hirsutus (Hemiptera) and Oecophylla smaragdina (Hymenoptera). The range of variation in virulence of a B. bassiana isolate to different insect species was not more than that observed with different populations within a single insect species. B. bassiana is thus a generalist with no strict host preference. B. bassiana based biopesticide can be used as a broad spectrum insecticide against a myriad of insect pests.     

Antibiotic treatment leads to the elimination of Wolbachia endosymbionts and sterility in the diplodiploid collembolan Folsomia candida

Background  

Wolbachia is an extremely widespread bacterial endosymbiont of arthropods and nematodes that causes a variety of reproductive peculiarities. Parthenogenesis is one such peculiarity but it has been hypothesised that this phenomenon may be functionally restricted to organisms that employ haplodiploid sex determination. Using two antibiotics, tetracycline and rifampicin, we attempted to eliminate Wolbachia from the diplodiploid host Folsomia candida, a species of springtail which is a widely used study organism.     

Evolutionary trade-offs at the Arabidopsis WRR4A resistance locus underpin alternate Albugo candida race recognition specificities

The oomycete Albugo candida causes white rust of Brassicaceae, including vegetable and oilseed crops, and wild relatives such as Arabidopsis thaliana. Novel White Rust Resistance (WRR) genes from Arabidopsis enable new insights into plant/parasite co-evolution. WRR4A from Arabidopsis accession Columbia (Col-0) provides resistance to many but not all white rust races, and encodes a nucleotide-binding, leucine-rich repeat immune receptor. Col-0 WRR4A resistance is broken by AcEx1, an isolate of A. candida. We identified an allele of WRR4A in Arabidopsis accession Øystese-0 (Oy-0) and other accessions that confers full resistance to AcEx1. WRR4A^Oy-0 carries a C-terminal extension required for recognition of AcEx1, but reduces recognition of several effectors recognized by the WRR4A^Col-0 allele. WRR4A^Oy-0 confers full resistance to AcEx1 when expressed in the oilseed crop Camelina sativa.     

Recent outbreaks of downy mildew on grape ivy (Parthenocissus tricuspidata, Vitaceae) in Germany are caused by a new species of Plasmopara

Plasmopara viticola is one of the most destructive pathogens in grape production and was introduced from North America to Europe more than a century ago. The pathogen is thought to affect a broad range of hosts in the Vitaceae, including the genera Vitis and Parthenocissus. However, the latter has not previously been reported as a host in Germany. Here, it is shown on the basis of phylogenetic and morphological data that recent outbreaks of downy mildew on Parthenocissus tricuspidata in the Stuttgart area of Southern Germany are not caused by Plasmopara viticola, but by an undescribed species introduced here as Plasmopara muralis. The new species has rapidly spread in the Stuttgart area throughout the past 2 years and poses a potential threat to ornamental vines grown as wall cover should its spread not be controlled.     

Cryptic speciation in the ectocommensal Bdelloura candida (Platyhelminthes,Tricladida, Maricola) follows habitat specialization of the American horseshoe crab,Limulus polyphemus

In strict symbiotic associations, the genetic structure of the symbiont often mirrors that of its host, with interesting implications for population dynamics and phylogeography. An unresolved case of symbiotic specificity and phylogeographic consequence is the relationship between the marine triclad Bdelloura candida and its host, the American horseshoe crab, Limulus polyphemus. A recent study by Riesgo et al. (2017, Marine Biology, 164, 111) identified a strong genetic break between populations of B. candida in the Gulf of Mexico and the Atlantic Ocean but had minimal sampling around the Florida peninsula such that the exact location of the boundary zone was not specified. To solve this, a comprehensive analysis of 16S rRNA and ITS2 genetic markers was conducted from new collections around the Florida peninsula. A clear and significant genetic break was identified between populations of supposed B. candida between Cumberland Island, Georgia, and Mosquito Lagoon, Florida. This genetic break establishes two cryptic lineages, an Atlantic population as far south as Georgia and a Floridian population inclusive of the entire peninsula and Gulf of Mexico, potentially due to niche partitioning of the unique intertidal habitats of its horseshoe crab hosts in Florida. This result directly refutes the previous hypothesis that a population break exists between the coasts of the Atlantic Ocean and Gulf of Mexico, and instead matches the genetic break of its host. Furthermore, a third cryptic lineage was identified in Key West. Overall, this work demonstrates the challenges in maintaining genetic connections between populations of both B. candida and L. polyphemus across their distributions, and poses meaningful implications for both species in the larger context of marine conservation and biodiversity.     

Biology and host specificity of Ceutorhynchus scrobicollis (Curculionidae; Coleoptera), a root-crown mining weevil proposed as biological control agent against Alliaria petiolata in North America

Ceutorhynchus scrobicollis is a root-crown mining weevil proposed for release as biological control agent of Alliaria petiolata (Brassicaceae, Thlaspideae), a European biennial herb, currently invading temperate North America. Using a combination of laboratory, common garden and fieldwork we studied biology, ecology and host range of C. scrobicollis, a univoltine species that oviposits and develops in A. petiolata rosettes in fall and spring. Individual C. scrobicollis can be long-lived (>2 years) and females show a second oviposition period. Weevils did not attack any of 31 test plant species outside the Brassicaceae. Within the Brassicaceae, five species allowed complete larval development under no-choice conditions. In subsequent choice tests, three of these five species (Nasturtium officinale, Peltaria alliacea and Thlaspi arvense; which are of European origin) were attacked. North American Rorippa sinuata was the only native species to be attacked by C. scrobicollis and only under no-choice conditions. Results of subsequent impact experiments showed that C. scrobicollis attack changed plant architecture but had no effect on overall plant vigour and reproductive output of R. sinuata, suggesting lack of impact on demography or population dynamics. A petition for field release of C. scrobicollis in North America has been submitted.     

Ultrastructural characterisation of the host–pathogen interface in white blister-infected Arabidopsis leaves

In this study transmission electron microscopy (TEM) was used to examine details of the host–pathogen interface in Arabidopsis thaliana cotyledons infected by Albugo candida, causal agent of white blister. After successful entry through stomatal pores, the pathogen developed a substomatal vesicle and subsequently produced intercellular hyphae. TEM observations revealed that coenocytic intercellular hyphae ramified and spread intercellularly throughout the host tissue forming several haustoria in host mesophyll cells. Intracellular haustoria were spherical and 4.5 μm in diameter. Each haustorium was connected to intercellular hyphae by a narrow, slender haustorium neck. The cytoplasm of the haustorium included the organelles characteristic of the pathogen. No obvious response was observed in host cells following formation of haustoria. Most of the mesophyll cells contained normal haustoria and the host cytoplasm displayed a high degree of structural integrity. Absence of host cell wall alteration and cell death in penetrated host cells suggest that the pathogen exerts considerable control over basic cellular processes and in this respect, response to this biotrophic Oomycete differs considerably from responses to other pathogens such as necrotrophs. Modification of the host plasma membrane (PM) along the cell wall and around the haustoria, was detected by applying the periodic acid-chromic acid-phosphotungstic acid (PACP) staining technique. After staining with PACP, the host PM was found to be intensely electron dense where it was adjacent to the host cell wall and the distal region of the haustorial neck. By contrast, the extrahaustorial membrane, where the host PM surrounded the haustorium, was consistently very lightly stained.     

Morphological variability of the water lily (<Emphasis Type="Italic">Nymphaea</Emphasis>) in the Polesie Zachodnie region,Eastern Poland

Morphological analysis of the water lily (Nymphaea) was carried out to verify distribution of two closely related species Nymphaea alba and N. candida in the large swampy area in Eastern Poland, Polesie Zachodnie. A total of 25 lakes were investigated across the region. Data were collected from 20 lakes of different origin. Based on morphological characters of flowers and leaves the specimens varied continuously from “pure” N. candida to N. alba with high number of plants with mixed morphology, commonly known as N. × borealis. The intermediate morphotypes were common in all over the study area, whereas plants with exact morphology of N. candida were found mostly in humic lakes, in the eastern part of the region. The results provide evidences that N. candida occurs beyond formerly recognized southern border of its range in the country.