Phylogenomics, taxonomy and morphological characters of the Microdochiaceae (Xylariales, Sordariomycetes).

Species of the family Microdochiaceae (Xylariales, Sordariomycetes) have been reported from worldwide, and collected from different plant hosts. The proposed new genus and two new species, viz., Macroidriella gen. nov., M.bambusae sp. nov. and Microdochiumaustrale sp. nov., are based on multi-locus phylogenies from a combined dataset of ITS rDNA, LSU, RPB2 and TUB2 with morphological characteristics. Microdochiumsinense has been collected from diseased leaves of Phragmitesaustralis and this is the first report of the fungus on this host plant. Simultaneously, we annotated 10,372 to 11,863 genes, identified 4,909 single-copy orthologous genes, and conducted phylogenomic analysis based on genomic data. A gene family analysis was performed and it will expand the understanding of the evolutionary history and biodiversity of the Microdochiaceae. The detailed descriptions and illustrations of species are provided.

Three New Species of Gongronella (Cunninghamellaceae, Mucorales) from Soil in Hainan, China Based on Morphology and Molecular Phylogeny.

The genus Gongronella is important in agriculture and industry by secreting various natural bioactive metabolites such as chitosanases and organic acids. During the most recent 8 years, a total of 14 new species have been described, remarkably enriching the diversity of this genus. In this study, we added three more new species to this valuable genus, based on a combination of morphological traits and phylogenetic information. Six strains of the genus Gongronella were isolated from soil collected in Hainan Province, China. Phylogenetic analyses of ITS and LSU rDNA sequences grouped these strains into three independent clades. According to their unique morphological characteristics, they were classified as G. multiramosa sp. nov., G. qichaensis sp. nov. and G. oleae sp. nov. The G. multiramosa was characterized by multiple branched sporangiophores and was closely related to G. pedratalhadensis. The G. qichaensis was characterized by obscure collars and closely related to G. butleri, G. hydei and G. banzhaoae. The G. oleae was characterized by the presence of oil droplets in the sporangiospores and was closely related to G. chlamydospora and G. multispora. Their descriptions and illustrations were provided, and their differences from morphological allies and phylogenetic-related species are discussed.

Multi-locus phylogenetic analyses reveal eight novel species of Distoseptispora from southern China.

IMPORTANCE: Distoseptispora as a single genus in Distoseptisporaceae was introduced by morphological and phylogenetic analyses. Members of this genus occur mainly as asexual morphs, forming effuse, hairy colonies on decaying wood, plant stems, bamboo culms, and fallen leaves and shafts in terrestrial and freshwater habitats. In the present study, saprobic hyphomycetes from plant debris were investigated, and eight new Distoseptispora species were introduced based on morphology and phylogenetic analyses of LSU, ITS, TEF1, and RPB2 sequence data. This study provides important data on the species diversity, ecological environment, and geographical area of Distoseptispora, greatly updates the classification of Distoseptispora, and improves our understanding of the taxonomy of Distoseptispora.

Mating-type genes of the anamorphic fungus Ulocladium botrytis affect both asexual sporulation and sexual reproduction.

Ulocladium was thought to be a strictly asexual genus of filamentous fungi. However, Ulocladium strains were shown to possess both MAT1-1-1 and MAT1-2-1 genes as observed in homothallic filamentous Ascomycetes. Here, we demonstrate that the U. botrytis MAT genes play essential roles for controlling asexual traits (conidial size and number). Using reciprocal genetic transformation, we demonstrate that MAT genes from the related heterothallic species Cochliobolus heterostrophus can also influence U. botrytis colony growth, conidial number and size, and have a strong effect on the range of the number of septa/conidium. Moreover, U. botrytis MAT genes can also affect similar aspects of asexual reproduction when expressed in C. heterostrophus. Heterologous complementation using C. heterostrophus MAT genes shows that they have lost the ability to regulate sexual reproduction in U. botrytis, under the conditions we employed, while the reciprocal heterologous complementation demonstrates that U. botrytis MAT genes have the ability to partially induce sexual reproduction in C. heterostrophus. Thus, the genetic backgrounds of C. heterostrophus and U. botrytis play significant roles in determining the function of MAT genes on sexual reproduction in these two fungi species. These data further support the role of MAT genes in controlling asexual growth in filamentous Ascomycetes but also confirm that heterothallic and homothallic Dothideomycete fungi can be interconverted by the exchange of MAT genes.

Acrodictys-like wood decay fungi from southern China, with two new families Acrodictyaceae and Junewangiaceae.

During our continuous survey (2012-2016) of saprobic hyphomycetes from dead branches in the forest ecosystems of southern China, we collected several acrodictys-like species. Acrodictys-like species are characterized by darkly pigmented and muriform conidia produced from holoblastic conidiogenous cells on macronematous, mononematous, cylindrical and unbranched or infrequently branched conidiophores. Phylogenetic analyses of ncLSU, ncSSU, ITS and tub2 sequence data lead us to propose two novel families in Sordariomycetes, Acrodictyaceae and Junewangiaceae. In addition, a new species, Acrodictys hainanensis, two new combinations, Junewangia queenslandica and Distoseptispora martinii, three new Chinese records, Acrodictys liputii, A. peruamazonensis and Junewangia sphaerospora are introduced. Two names, Acrodictys globulosa and A. malabarica, are resurrected.

Microwave ablation versus hepatic resection for the treatment of hepatocellular carcinoma and oesophageal variceal bleeding in cirrhotic patients.

PURPOSE: The aim of this study was to compare the results of microwave ablation (MWA) and hepatic resection (HR) when combined with pericardial devascularisation plus splenectomy (PCDV) for the treatment of patients with cirrhosis complicated by small hepatocellular carcinoma (HCC) and oesophageal variceal bleeding (EVB). MATERIALS AND METHODS: Between 2001 and 2013, 73 patients (median age 53.2 years, 67% male) with small HCC and concomitant EVB who underwent MWA or HR for HCC and PCDV for cirrhotic portal hypertension were selected retrospectively for inclusion in this study. The overall survival curves and recurrence-free survival curves were calculated using the Kaplan-Meier method and compared using log-rank tests. Multivariate analysis was performed using the Cox regression model. RESULTS: The 1-, 3- and 5-year overall survival rates were 95.2%, 71.4% and 38.1% and 96.7%, 53.3% and 43.3% for the HR and MWA groups, respectively; these did not differ significantly between the two groups. However, patients in the HR group had more post-operative complications (52.3% vs. 13.7%; p = 0.002). Multivariate analysis identified albumin and bilirubin levels and tumour size to be statistically significant and independent prognostic factors for overall survival, while BCLC stage was associated with poor recurrence-free survival. Furthermore, albumin levels were shown to be an independent predictive factor for post-operative complications. CONCLUSIONS: For patients with small HCC and concomitant EVB, MWA plus PCDV may reduce the incidence of post-operative complications relative to and provide similar therapeutic benefits as HR plus PCDV, especially for patients with low albumin levels.

Characterization of necrosis-inducing NLP proteins in Phytophthora capsici.

BACKGROUND: Effector proteins function not only as toxins to induce plant cell death, but also enable pathogens to suppress or evade plant defense responses. NLP-like proteins are considered to be effector proteins, and they have been isolated from bacteria, fungi, and oomycete plant pathogens. There is increasing evidence that NLPs have the ability to induce cell death and ethylene accumulation in plants. RESULTS: We evaluated the expression patterns of 11 targeted PcNLP genes by qRT-PCR at different time points after infection by P. capsici. Several PcNLP genes were strongly expressed at the early stages in the infection process, but the expression of other PcNLP genes gradually increased to a maximum at late stages of infection. The genes PcNLP2, PcNLP6 and PcNLP14 showed the highest expression levels during infection by P. capsici. The necrosis-inducing activity of all targeted PcNLP genes was evaluated using heterologous expression by PVX agroinfection of Capsicum annuum and Nicotiana benthamiana and by Western blot analysis. The members of the PcNLP family can induce chlorosis or necrosis during infection of pepper and tobacco leaves, but the chlorotic or necrotic response caused by PcNLP genes was stronger in pepper leaves than in tobacco leaves. Moreover, PcNLP2, PcNLP6, and PcNLP14 caused the largest chlorotic or necrotic areas in both host plants, indicating that these three genes contribute to strong virulence during infection by P. capsici. This was confirmed through functional evaluation of their silenced transformants. In addition, we further verified that four conserved residues are putatively active sites in PcNLP1 by site-directed mutagenesis. CONCLUSIONS: Each targeted PcNLP gene affects cells or tissues differently depending upon the stage of infection. Most PcNLP genes could trigger necrotic or chlorotic responses when expressed in the host C. annuum and the non-host N. benthamiana. Individual PcNLP genes have different phytotoxic effects, and PcNLP2, PcNLP6, and PcNLP14 may play important roles in symptom development and may be crucial for virulence, necrosis-inducing activity, or cell death during infection by P. capsici.

Characterization and phylogenetic analysis of the mating-type loci in the asexual ascomycete genus Ulocladium.

The genus Ulocladium is thought to be strictly asexual. Mating-type (MAT) loci regulate sexual reproduction in fungi and their study may help to explain the apparent lack of sexual reproduction in Ulocladium. We sequenced the full length of two MAT genes in 26 Ulocladium species and characterized the entire MAT idiomorphs plus flanking regions of Ulocladium botrytis. The MAT1-1 ORF encodes a protein with an alpha-box motif by the MAT1-1-1 gene and the MAT1-2 ORF encodes a protein with an HMG box motif by the MAT1-2-1 gene. Both MAT1-1-1 and MAT1-2-1 genes were detected in a single strain of every species. Moreover, the results of RT-PCR revealed that both MAT genes are expressed in all 26 Ulocladium species. This demonstrates that MAT genes of Ulocladium species might be functional and that they have the potential for sexual reproduction. Phylogenies based on MAT genes were compared with GAPDH and Alt a 1 phylograms in Ulocladium using maximum parsimony (MP) and Bayesian analysis. The MAT genealogies and the non-MAT trees displayed different topologies, indicating that MAT genes are unsuitable phylogenetic markers at the species level in Ulocladium. Furthermore, the conflicting topologies between MAT1-1-1 and MAT1-2-1 phylogeny indicate separate evolutionary events for the two MAT genes. However, the intergeneric phylogeny of four closely allied genera (Ulocladium, Alternaria, Cochliobolus, Stemphylium) based on MAT alignments demonstrated that MAT genes are suitable for phylogenetic analysis among allied genera.

New insight into the history of domesticated apple: secondary contribution of the European wild apple to the genome of cultivated varieties.

The apple is the most common and culturally important fruit crop of temperate areas. The elucidation of its origin and domestication history is therefore of great interest. The wild Central Asian species Malus sieversii has previously been identified as the main contributor to the genome of the cultivated apple (Malus domestica), on the basis of morphological, molecular, and historical evidence. The possible contribution of other wild species present along the Silk Route running from Asia to Western Europe remains a matter of debate, particularly with respect to the contribution of the European wild apple. We used microsatellite markers and an unprecedented large sampling of five Malus species throughout Eurasia (839 accessions from China to Spain) to show that multiple species have contributed to the genetic makeup of domesticated apples. The wild European crabapple M. sylvestris, in particular, was a major secondary contributor. Bidirectional gene flow between the domesticated apple and the European crabapple resulted in the current M. domestica being genetically more closely related to this species than to its Central Asian progenitor, M. sieversii. We found no evidence of a domestication bottleneck or clonal population structure in apples, despite the use of vegetative propagation by grafting. We show that the evolution of domesticated apples occurred over a long time period and involved more than one wild species. Our results support the view that self-incompatibility, a long lifespan, and cultural practices such as selection from open-pollinated seeds have facilitated introgression from wild relatives and the maintenance of genetic variation during domestication. This combination of processes may account for the diversification of several long-lived perennial crops, yielding domestication patterns different from those observed for annual species.

[Application of portal vein chemoembolization in major hepatectomy for initially unresectable primary and secondary hepatic tumors].

OBJECTIVE: To investigate the surgical treatment of initially unresectable primary and secondary hepatic tumors. METHODS: For the patients with multiple and bilobar colonic hepatic metastases, a first-stage hepatectomy consisted in a radical resection of sigmoid colonic carcinoma and left lateral hepatic segment. Subsequently, under the guidance of ultrasonography and radiography, a right portal vein chemoembolization (PVCE) was performed via a percutaneous approach through left portal branch to induce the atrophy of right hemiliver and hypertrophy of left hemiliver. At Week 5 post-PVCE, a second-stage hepatectomy was planned to resect the right hemiliver. For patients with huge hepatocellular carcinoma (HCC), transcatheter arterial chemoembolization (TACE) were performed and it was followed by PVCE 1 week later. At Week 4 post-PVCE, a right trisegmentectomy was attempted to resect the right liver tumor. The volume of liver was evaluated with three-dimensional CT scan at Weeks 2 and 4 weeks post-PVCE. RESULTS: At Week 4 post-PVCE, the atrophy of right lobe was induced and the left lobe underwent compensatory hypertrophy. The remnant volumes of right lobe and right trisegmentectomy for HCC decreased from 1380.0 cm(3), 1685.4 cm(3) at pre-PVCE to 740.2 cm(3), 1228.1 cm(3) at post-PVCE. The values increased from pre-PVCE 435.1 cm(3), 151.5 cm(3) to post-PVCE 624.4 cm(3), 560.2 cm(3) for left hepatic lobe remnant of colonic liver metastases and left lateral segment for HCC. The ratios of liver remnant to estimated total liver volume increased from 25.6%, 13.6% at pre-PVCE to 50.0%, 43.1% post-PVCE respectively. The postoperative course was uneventful. The liver function, serum CEA and AFP decreased to the normal levels. Two patients were followed up for 18 and 8 months respectively. There was no tumor recurrence. CONCLUSION: PVCE prevents the hepatic function failure after a major hepatectomy. And it may benefit more patients with previously unresectable liver tumors.

Sinomyces: a new genus of anamorphic Pleosporaceae.

Sinomyces gen. nov. is described for Ulocladium alternariae and two new species from China. These three species differ from Ulocladium in producing simple conidiophores with a single, cicatrized apical pore or 1 or 2 short, uniperforate, geniculate sympodial proliferations. Phylogenetic analysis of partial DNA sequences of the glyceraldehyde-3-phosphate dehydrogenase gene (gpd) gene and a fragment of the Alternaria alternata major allergen (Alt a 1) gene, using Maximum-Parsimony (MP), Maximum-Likelihood (ML) and Bayesian approaches, indicates that Sinomyces is distinct within the Alternaria-Stemphylium complex, although its closest relatives could not be determined. Sinomyces alternariae comb. nov. and the new species Sinomyces obovoideus and Sinomyces fusoideus are proposed.

A revision of the genus Lomaantha, with the description of a new species.

Lomaantha phragmitis sp. nov. is described and illustrated from a specimen collected on dead culms of Phragmites communis in southern China. The fungus differs from other described Lomaantha species in its conidiophores, conidiogenous cells and conidial appendages. Conidial morphology and presence or absence of percurrent proliferation of conidiogenous cells are the main characters distinguishing species within this genus. We provided a key and synoptic table of morphological characters of all three Lomaantha species.

Molecular and morphological description of two new species of Stemphylium from China and France.

Two new species of Stemphylium (anamorphic Pleospora) are described on the basis of morphological characters and molecular phylogenetic analyses. Stemphylium phaseolina and S. variabilis were isolated respectively from diseased leaves of Phaseolus vulgaris L. in Hebei Province, China, and from diseased leaves of Allium sativum L. in Angres, France. The two species exhibit characteristic Stemphylium morphology but are distinct from similar species based on the morphology and development of conidia. The internal transcribed spacer (ITS) nuclear rDNA region and glyceraldehyde-3-phosphate dehydrogenase (gpd) genes were sequenced. The results of maximum parsimony (MP) and maximum likelihood (ML) phylogenetic analyses of the combined DNA sequences of these two gene regions supported S. phaseolina and S. variabilis as two distinct phylogenetic species. The taxonomic descriptions of the new species and their comparison with related species are presented, together with the phylogenetic analysis based on combined DNA sequences of ITS and gpd gene regions.

[Left versus right ventricular Tei index for evaluating third-trimester fetal cardiac function in pregnancy-induced hypertension syndrome].

OBJECTIVE: To investigate the value of Tei index and the sensitivity of left versus right ventricular Tei index in evaluating the fetal cardiac function in pregnancy-induced hypertension syndrome in the third trimester. METHODS: Fetal echocardiograms were performed in 30 women with pregnancy-induced hypertension (PIH) syndrome and 55 with normal pregnancy of the third trimester. Tei index was obtained by calculating the ratio of the isovolumic time (isovolumic contraction and relaxation time) to the ejection time of the left and right ventricle. Comparisons of the Tei index were made between the PIH group and control group, and also between the left and right ventricles in each group. RESULTS: Significant difference was found in the left and right ventricular Tei index between PIH group and control group. No difference was noted between the left and right ventricular Tei index in the PIH group. CONCLUSIONS: Tei index is a useful indicator in evaluating fetal global cardiac function, for which purpose the left ventricular Tei index can be as sensitive as the right ventricular Tei index.

Ulocladium cantlous sp. nov. isolated from northwestern China: its morphology and molecular phylogenetic position.

A new species of Ulocladium was isolated from diseased leaves from two Cucumis sp. growing in Sinkiang and Gansu provinces of China. Conidia were isolated from necrotic leaves and used to establish single-spore pure cultures. Conidia were harvested from cultures 7 d after incubation for morphological comparisons. The morphology of this species resembles that of U. botrytis and U. consortiale. However it is distinguished from these two species by the sizes of obovoid to broadly ellipsoidal conidia and longer conidiophores. A taxonomic description of U. cantlous, comparison with related species in this genus, and a species phylogeny based on the partial nucleotide sequence of the glyceraldehyde-3-phosphate dehydrogenase (gpd) gene and the Alternaria alternata major allergen (Alt a 1) gene are provided.

Evolution of the population structure of Venturia inaequalis, the apple scab fungus, associated with the domestication of its host.

Evaluating the impact of plant domestication on the population structure of the associated pathogens provides an opportunity to increase our understanding of how and why diseases emerge. Here, we investigated the evolution of the population structure of the apple scab fungus Venturia inaequalis in response to the domestication of its host. Inferences were drawn from multilocus microsatellite data obtained from samples collected on (i) the Central Asian Malus sieversii, the main progenitor of apple, (ii) the European crabapple, Malus sylvestris, a secondary progenitor of apple, and (iii) the cultivated apple, Malus x domestica, in orchards from Europe and Central Asia. Using clustering methods, we identified three distinct populations: (i) a large European population on domesticated and wild apples, (ii) a large Central Asian population on domesticated and wild apples in urban and agricultural areas, and (iii) a more geographically restricted population in M. sieversii forests growing in the eastern mountains of Kazakhstan. Unique allele richness and divergence time estimates supported a host-tracking co-evolutionary scenario in which this latter population represents a relict of the ancestral populations from which current populations found in human-managed habitats were derived. Our analyses indicated that the domestication of apple induced a significant change in the genetic differentiation of populations of V. inaequalis in its centre of origin, but had little impact on its population dynamics and mating system. We discuss how the structure of the apple-based agrosystem may have restricted changes in the population structure of the fungus in response to the domestication of its host.

New species and records of Shrungabeeja from southern China.

Three species of the anamorphic fungus Shrungabeeja were collected from tropical forests in Yunnan Province, southern China. S. melicopeae sp. nov. and S. begoniae sp. nov. are described and illustrated from specimens collected respectively on dead branches of Melicope triphylla and Begonia semperflorens. S. vadirajensis is recorded for the first time from China. A key to the three known species of Shrungabeeja is provided.

Three new species of Piricaudiopsis from southern China.

Piricaudiopsis punicae sp. nov., P. rosae sp. nov. and P. rhaphidophorae sp. nov., occurring respectively on dead branches of Punica granatum, Rosa chinensis and Rhaphidophora decursiva were collected from tropical forest in southern China. They differ from the three previously described Piricaudiopsis species in conidial morphology and proliferation of the conidiogenous cell. The presence or absence of percurrent proliferation of the conidiogenous cells and conidial appendages and the height of conidia are recognized as important characters in the delimitation of species of Piricaudiopsis. A key to Piricaudiopsis species is provided.

Functional analysis of Pcipg2 from the straminopilous plant pathogen Phytophthora capsici.

Phytophthora capsici causes serious diseases in numerous crop plants. Polygalacturonases (PGs) are cell wall-degrading enzymes that play an important role in pathogenesis in straminopilous pathogens. To understand PGs as they relate to the virulence of P. capsici, Pcipg2 was identified from a genomic library of a highly virulent P. capsici strain. Pcipg2 was strongly expressed during symptom development after the inoculation of pepper leaves with P. capsici. The wild protein (PCIPGII) was obtained from the expression of pcipg2 and found that increasing activity of PGs in PCIPGII-treated pepper leaves was consistent with increasing symptom development. Asp residues in active sites within pcipg2 affected PCIPGII activity or its virulence on pepper leaves. Results show that pcipg2 is an important gene among pcipg genes, and illustrate the benefit of analyzing mechanisms of pathogenicity during the period of host/parasite interaction.

Two new species of Ulocladium from Southwest China.

Ulocladium subcucurbitae and U. brassicae are described and illustrated. These species were isolated from diseased leaves of Chenopodium glaucum and Brassica pekinensis from Guizhou and Yunnan provinces of China respectively.