Phragmidium satoanum,a new rust pathogen of Rosa hirtula in Japan

The causal fungus of a rust disease of Rosa hirtula, endemic to mountainous areas of Fuji-Hakone-Izu National Park, Japan, was thought to be a common species Phragmidium rosae-multiflorae. Continued field observations, morphological examination, and experimental inoculations proved that the fungus produced laterally three-angled aeciospores and urediniospores together with multi-cellular teliospores on the same R. hirtula trees. These morphological features were different from those of P. rosae-multiflorae. The fungus parasitized only R. hirtula. Experimental inoculations and field observations did not prove that R. banksiae, R. laevigata, and R. multiflora supported infection and sporulation of the fungus. Under the field observations, R. multiflora, the most common host of P. rosae-multiflorae, was not proven to harbor the R. hirtula fungus. Therefore, the fungus was concluded to be a species distinct from P. rosae-multiflorae; and a new name, P. satoanum, was proposed for it.     

Expression of apoplast-targeted plant defensin <Emphasis Type="Italic">MtDef4.2</Emphasis> confers resistance to leaf rust pathogen <Emphasis Type="Italic">Puccinia triticina</Emphasis> but does not affect mycorrhizal symbiosis in transgenic wheat

Rust fungi of the order Pucciniales are destructive pathogens of wheat worldwide. Leaf rust caused by the obligate, biotrophic basidiomycete fungus Puccinia triticina (Pt) is an economically important disease capable of causing up to 50 % yield losses. Historically, resistant wheat cultivars have been used to control leaf rust, but genetic resistance is ephemeral and breaks down with the emergence of new virulent Pt races. There is a need to develop alternative measures for control of leaf rust in wheat. Development of transgenic wheat expressing an antifungal defensin offers a promising approach to complement the endogenous resistance genes within the wheat germplasm for durable resistance to Pt. To that end, two different wheat genotypes, Bobwhite and Xin Chun 9 were transformed with a chimeric gene encoding an apoplast-targeted antifungal plant defensin MtDEF4.2 from Medicago truncatula. Transgenic lines from four independent events were further characterized. Homozygous transgenic wheat lines expressing MtDEF4.2 displayed resistance to Pt race MCPSS relative to the non-transgenic controls in growth chamber bioassays. Histopathological analysis suggested the presence of both pre- and posthaustorial resistance to leaf rust in these transgenic lines. MtDEF4.2 did not, however, affect the root colonization of a beneficial arbuscular mycorrhizal fungus Rhizophagus irregularis. This study demonstrates that the expression of apoplast-targeted plant defensin MtDEF4.2 can provide substantial resistance to an economically important leaf rust disease in transgenic wheat without negatively impacting its symbiotic relationship with the beneficial mycorrhizal fungus.     

Life cycle of Melampsora coleosporioides,a leaf rust of Salix babylonica in Japan

Melampsora coleosporioides produces uredinia and telia on the leaves of Salix babylonica. Since the life cycle of this fungus is largely unknown, inoculation experiments were conducted to find alternate host plants. Results showed that M. coleosporioides can use Corydalis incisa as a spermogonial and aecial host. The morphological characteristics of the spermogonial and aecial states of the fungus are first described. Furthermore, field observations and histological studies demonstrated that the fungus was able to overwinter in twigs of S. babylonica and produced urediniospores in early spring. Thus, the leaf rust occurs on S. babylonica every year and alternation with the spermogonial and aecial host is not necessary.     

A Novel Fungal Hyperparasite of Puccinia striiformis f. sp. tritici,the Causal Agent of Wheat Stripe Rust

Puccinia striiformis f. sp. tritici (Pst), the causal fungus of wheat stripe rust, was previously reported to be infected by Lecanicillium lecanii, Microdochium nivale and Typhula idahoensis. Here, we report a novel hyperparasite on Pst. This hyperparasitic fungus was identified as Cladosporium cladosporioides (Fresen.) GA de Vries based on morphological characteristics observed by light and scanning electron microscopy together with molecular data. The hyperparasite reduced the production and viability of urediniospores and, therefore, could potentially be used for biological control of wheat stripe rust.     

Putative natural hybrid between Puccinia lagenophorae and an unknown rust fungus on Senecio madagascariensis in KwaZulu-Natal,South Africa

Several specimens of an aecial rust fungus were collected on Senecio madagascariensis during a field survey carried out in KwaZulu-Natal, South Africa. As telia were not present in the specimens collected, DNA sequence analyses were undertaken to determine the identity of the rust species. ITS and β-tub1 sequencing confirmed that one of the isolates recovered is Puccinia lagenophorae sensu lato. On the other hand, sequencing and RFLP analysis revealed the presence of two divergent copies of ITS and β-tub1 in all the other six isolates investigated. In both phylogenetic trees, one copy of the gene region grouped within a well supported clade with sequences of P. lagenophorae accessions from different geographical origins and hosts, and the Australian rusts Puccinia saccardoi and Puccinia stylidii. The other copy of these gene regions grouped within a separate clade comprising European accessions of Puccinia dioicae (ITS) and Uromyces sommerfeltii (β-tub1) that occur on Asteraceae hosts. Multiple copies of these gene regions were not observed in Australian isolates of P. lagenophorae. Our study provides some evidence that an interspecific hybrid rust fungus, with P. lagenophorae as one of its parents, may occur on S. madagascariensis in South Africa. The identity of the other parent remains unknown.     

Cystomyces antheropori,a new rust fungus on Antheroporum glaucum from Thailand

A rust fungus was found on leaves of Antheroporum glaucum (Fabaceae, Faboideae) in Provinces of Petchaburi and Ratchaburi, western Thailand. No rust fungus was previously reported on this legume tree species. The new rust fungus was apparently microcyclic, producing telia surrounding dense aggregates of spermogonia, on the abaxial leaf surface. Teliospores were produced by sympodial proliferation from a basal sporogenous cell and mostly composed of three fertile cells subtended by two hygroscopic cysts. A germ slit was present near the septum in two of three fertile cells. A simple persistent pedicel was attached to the teliospore cysts. Because of resemblance in the teliospore morphology, this fungus was compared to Bibulocystis, Caetea, Cystomyces, and Spumula. Radially symmetric teliospores in the horizontal plane with three more or less globose fertile cells subtended by two hygroscopic cysts and the presence of a germ slit near the septum in fertile cells are the characteristics of the genus Cystomyces. Therefore, it was concluded that the fungus should be classified in Cystomyces, and a new name, C. antheropori was proposed.     

<Emphasis Type="Italic">Melampsora salicis-bakko</Emphasis>, a new species on willows in Japan evidenced by morphological and molecular phylogenetic analyses

Through the morphological and molecular examinations of Melampsora species on willows, we clarified the taxonomic identity of the rust specimens on Salix bakko, S. hultenii and S. leucopithecia from Japan and described the following rust fungus as a new species, Melampsora salicis-bakko. This rust fungus resembled M. caprearum in morphology of teliospores, but it differed from M. caprearum mainly in the density of spines on the urediniospores. Molecular phylogenetic analyses using the rDNA ITS region (complete ITS1, 5.8S rRNA gene and ITS2) revealed that M. salicis-bakko was monophyletic, and that this rust fungus was distinct from other Melampsora species, including M. caprearum.     

Predicted potential distribution of Sydowia japonica in Japan

Sydowia japonica, a dothidealean fungus, is a parasite that is specific to the male strobili of Japanese cedar. The fungus is a candidate for the control of Japanese cedar pollen dispersal, which is a cause of pollinosis. To evaluate S. japonica for bioherbicidal applications, it is necessary to characterise its potential distribution and environmental niche. Here, we predicted the distribution of S. japonica in Japan using field surveys and a maximum entropy model, and identified the environmental variables that influence its distribution. We identified S. japonica from a total of 87 localities in Japan through field surveys. Based on presence data and associated environmental variables, our model predicted that S. japonica is widely distributed in Japan, but concentrated in the Hokuriku and Kinki areas along the Sea of Japan. The model also predicted that the most important environmental variables influencing fungal distribution were sunshine duration in the winter and precipitation in the summer. This new information will contribute to the development of bioherbicidal applications for S. japonica.     

<Emphasis Type="Italic">Melampsora pakistanica</Emphasis> sp. nov., a new rust fungus on <Emphasis Type="Italic">Euphorbia helioscopia</Emphasis> (Sun spurge) from Pakistan

A rust fungus was found on the leaves of Euphorbia helioscopia during a field study in Pakistan. Previously, Melampsora euphorbiae, M. euphorbiae-gerardianae and M. helioscopiae have been reported on E. helioscopia, the first two of which are also known from Pakistan. Morphological observations of the newly collected rust samples detected some differences from the previously described Melampsora species on E. helioscopia. The molecular analysis of the ITS and LSU sequences also detected that the rust is different from the previously reported rusts described from E. helioscopia. Based on both morphological comparisons and sequence analysis, the rust is described here as M. pakistanica sp. nov. This species could have potential as a bio-control agent against its host plant—E. helioscopia—which is a weed of wheat fields in Pakistan and elsewhere in the world.     

Pathogenicity and taxonomy of Tenuignomonia styracis gen. et sp. nov., a new monotypic genus of Gnomoniaceae on Styrax obassia in Japan

Since 2010, an unknown fungus in the Gnomoniaceae has been found on overwintered leaves and petioles of Styrax obassia (Styracaceae) in Japan. This fungus is characterized by dark brown immersed or partially erumpent ascomata with long necks and fusiform to obovoid asci each with an acute or long tapering stipe. Each ascus bears eight fusiform to filiform ascospores. Our morphological observation and phylogenetic analyses based on the markers LSU, rpb2, and tef-1α indicated that this is a new monotypic genus in the Gnomoniaceae (Diaporthales), and Tenuignomonia styracis gen. et sp. nov. was descried herein. Members of the Gnomoniaceae are commonly isolated as endophytes, saprobes, and plant pathogens from a broad diversity of herbaceous, shade tree, and agriculturally significant plants. We thus carried out a pathogenicity test to determine if T. styracis is the causative agent of leaf blotch on S. obassia. One week after inoculation, this fungus produced small necrotic spots on the leaves and petioles, and all leaves having necrotic spots were abscised in a short time. We thus confirmed that this fungus has weak pathogenicity on S. obassia. This new species may promote early defoliation of S. obassia during the fall.     

An Interspecies Comparative Analysis of the Predicted Secretomes of the Necrotrophic Plant Pathogens Sclerotinia sclerotiorum and Botrytis cinerea

Phytopathogenic fungi form intimate associations with host plant species and cause disease. To be successful, fungal pathogens communicate with a susceptible host through the secretion of proteinaceous effectors, hydrolytic enzymes and metabolites. Sclerotinia sclerotiorum and Botrytis cinerea are economically important necrotrophic fungal pathogens that cause disease on numerous crop species. Here, a powerful bioinformatics pipeline was used to predict the refined S. sclerotiorum and B. cinerea secretomes, identifying 432 and 499 proteins respectively. Analyses focusing on S. sclerotiorum revealed that 16% of the secretome encoding genes resided in small, sequence heterogeneous, gene clusters that were distributed over 13 of the 16 predicted chromosomes. Functional analyses highlighted the importance of plant cell hydrolysis, oxidation-reduction processes and the redox state to the S. sclerotiorum and B. cinerea secretomes and potentially host infection. Only 8% of the predicted proteins were distinct between the two secretomes. In contrast to S. sclerotiorum, the B. cinerea secretome lacked CFEM- or LysM-containing proteins. The 115 fungal and oomycete genome comparison identified 30 proteins specific to S. sclerotiorum and B. cinerea, plus 11 proteins specific to S. sclerotiorum and 32 proteins specific to B. cinerea. Expressed sequence tag (EST) and proteomic analyses showed that 246 S. sclerotiorum secretome encoding genes had EST support, including 101 which were only expressed in vitro and 49 which were only expressed in planta, whilst 42 predicted proteins were experimentally proven to be secreted. These detailed in silico analyses of two important necrotrophic pathogens will permit informed choices to be made when candidate effector proteins are selected for function analyses in planta.     

Signatures of Adaptation in Human Invasive Salmonella Typhimurium ST313 Populations from Sub-Saharan Africa

Two lineages of Salmonella enterica serovar Typhimurium (S. Typhimurium) of multi-locus sequence type ST313 have been linked with the emergence of invasive Salmonella disease across sub-Saharan Africa. The expansion of these lineages has a temporal association with the HIV pandemic and antibiotic usage. We analysed the whole genome sequence of 129 ST313 isolates representative of the two lineages and found evidence of lineage-specific genome degradation, with some similarities to that observed in S. Typhi. Individual ST313 S. Typhimurium isolates exhibit a distinct metabolic signature and modified enteropathogenesis in both a murine and cattle model of colitis, compared to S. Typhimurium outside of the ST313 lineages. These data define phenotypes that distinguish ST313 isolates from other S. Typhimurium and may represent adaptation to a distinct pathogenesis and lifestyle linked to an-immuno-compromised human population.     

<Emphasis Type="Italic">Sordaria fimicola</Emphasis>-like ascomycete isolated from <Emphasis Type="Italic">Pinus coulteri</Emphasis> needles in Slovakia

This is the first report of Sordaria fimicola-like ascomycete which was encountered during a diversity study of injured tissues of coulter pine in Slovakia. The fungus was identified as Sordaria fimicola by morphological analyses. Sequence analysis of internal transcribed spacer region (ITS) showed that the fungus is highly related to the ITS sequences of several S. fimicola isolates documenting wide ecological valence and geographical distribution of S. fimicola-like ascomycetes.     

Ochropsora staphyleae,a new rust pathogen of Japanese bladdernut,found in central Japan

Japanese bladdernut shrub, Staphylea bumalda (Staphyleaceae), was found infected by a fungus in Prefectures of Ibaraki and Tochigi, central Japan. The fungus produced telia beneath the host epidermis and basidiospores on metabasidia emerging from the sori on the abaxial leaf surface. Probasidia were single-celled, sessile, short-cylindrical, thin-walled, and laterally free. A four-celled metabasidium arose from a probasidium by apical elongation. Apparent biotrophic nature and the basidium morphology of the Staphylea fungus suggested its taxonomic affinity to the rust genus Ochropsora. Molecular phylogenetic analyses of D1/D2 region of LSU rDNA showed that two collections of the fungus were grouped with Ochropsora ariae (type species of the genus) and O. nambuana but formed an independent clade. The fungus under consideration was, therefore, classified in Ochropsora, and a new name, O. staphyleae, was proposed for it.     

The Complete Chloroplast Genome of Guadua angustifolia and Comparative Analyses of Neotropical-Paleotropical Bamboos

To elucidate chloroplast genome evolution within neotropical-paleotropical bamboos, we fully characterized the chloroplast genome of the woody bamboo Guadua angustifolia. This genome is 135,331 bp long and comprises of an 82,839-bp large single-copy (LSC) region, a 12,898-bp small single-copy (SSC) region, and a pair of 19,797-bp inverted repeats (IRs). Comparative analyses revealed marked conservation of gene content and sequence evolutionary rates between neotropical and paleotropical woody bamboos. The neotropical herbaceous bamboo Cryptochloa strictiflora differs from woody bamboos in IR/SSC boundaries in that it exhibits slightly contracted IRs and a faster substitution rate. The G. angustifolia chloroplast genome is similar in size to that of neotropical herbaceous bamboos but is ~3 kb smaller than that of paleotropical woody bamboos. Dissimilarities in genome size are correlated with differences in the lengths of intergenic spacers, which are caused by large-fragment insertion and deletion. Phylogenomic analyses of 62 taxa yielded a tree topology identical to that found in preceding studies. Divergence time estimation suggested that most bamboo genera diverged after the Miocene and that speciation events of extant species occurred during or after the Pliocene.     

Cloning and Functional Characterization of SAD Genes in Potato

Stearoyl-acyl carrier protein desaturase (SAD), locating in the plastid stroma, is an important fatty acid biosynthetic enzyme in higher plants. SAD catalyzes desaturation of stearoyl-ACP to oleyl-ACP and plays a key role in determining the homeostasis between saturated fatty acids and unsaturated fatty acids, which is an important player in cold acclimation in plants. Here, four new full-length cDNA of SADs (ScoSAD, SaSAD, ScaSAD and StSAD) were cloned from four Solanum species, Solanum commersonii, S. acaule, S. cardiophyllum and S. tuberosum, respectively. The ORF of the four SADs were 1182 bp in length, encoding 393 amino acids. A sequence alignment indicated 13 amino acids varied among the SADs of three wild species. Further analysis showed that the freezing tolerance and cold acclimation capacity of S. commersonii are similar to S. acaule and their SAD amino acid sequences were identical but differed from that of S. cardiophyllum, which is sensitive to freezing. Furthermore, the sequence alignments between StSAD and ScoSAD indicated that only 7 different amino acids at residues were found in SAD of S. tuberosum (Zhongshu8) against the protein sequence of ScoSAD. A phylogenetic analysis showed the three wild potato species had the closest genetic relationship with the SAD of S. lycopersicum and Nicotiana tomentosiformis but not S. tuberosum. The SAD gene from S. commersonii (ScoSAD) was cloned into multiple sites of the pBI121 plant binary vector and transformed into the cultivated potato variety Zhongshu 8. A freeze tolerance analysis showed overexpression of the ScoSAD gene in transgenic plants significantly enhanced freeze tolerance in cv. Zhongshu 8 and increased their linoleic acid content, suggesting that linoleic acid likely plays a key role in improving freeze tolerance in potato plants. This study provided some new insights into how SAD regulates in the freezing tolerance and cold acclimation in potato.     

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     

Endogone corticioides sp. nov. from subalpine conifer forests in Japan and China,and its multi-locus phylogeny

Endogone corticioides sp. nov. was described from subalpine conifer forests in Japan and China. This species is similar to E. pisiformis in its yellowish sporocarps and zygosporangia on the equal empty gametangia. Endogone corticioides was clearly distinguished from E. pisiformis based on its resupinate sporocarp, the presence of hyphal peg-like structures on the peridium, and abundant thick-walled hyphae in the gleba. Phylogenetic analysis of the small and large subunit nuclear ribosomal RNA and the elongation factor-1α genes suggested that E. corticioides is closely related to E. pisiformis. Limited vegetative growth from the surface-sterilized sporocarps on agar media was shown.     

Ricoseius loxocheles,a phytoseiid mite that feeds on coffee leaf rust

One of the most important diseases of coffee plants is the coffee leaf rust fungus Hemileia vastatrix Berkeley and Broome (Uredinales). It can cause 30 % yield loss in some varieties of Coffea arabica (L.). Besides fungus, the coffee plants are attacked by phytophagous mites. The most common species is the coffee red mite, Oligonychus ilicis McGregor (Acari: Tetranychidae). Predatory mites of the Phytoseiidae family are well-known for their potential to control herbivorous mites and insects, but they can also develop and reproduce on various other food sources, such as plant pathogenic fungi. In a field survey, we found Ricoseius loxocheles (De Leon) (Acari: Phytoseiidae) on the necrotic areas caused by the coffee leaf rust fungus during the reproductive phase of the pathogen. We therefore assessed the development, survivorship and reproduction of R. loxocheles feeding on coffee leaf rust fungus and measured predation and oviposition of this phytoseiid having coffee red mite as prey under laboratory conditions. The mite fed, survived, developed and reproduced successfully on this pathogen but it was not able to prey on O. ilicis. Survival and oviposition with only prey were the same as without food. This phytoseiid mite does not really use O. ilicis as food. It is suggested that R. loxocheles is one phytoseiid that uses fungi as a main food source.