Morphological and molecular analyses reveal two new species of Microcera (Nectriaceae, Hypocreales) associated with scale insects on walnut in China.

The fungal genus Microcera consists of species mostly occurring as parasites of scale insects, but are also commonly isolated from soil or lichens. In the present study, we surveyed the diversity and assess the taxonomy of entomopathogenic fungi in Sichuan Province, China. Two new species of Microcera, viz. M.chrysomphaludis and M.pseudaulacaspidis, were isolated from scale insects colonising walnut (Juglansregia). Maximum Likelihood and Bayesian Inference analyses of ITS, LSU, tef1-α, rpb1, rpb2, acl1, act, tub2, cmdA and his3 sequence data provide evidence for the validity of the two species and their placement in Nectriaceae (Hypocreales). Microcerapseudaulacaspidis primarily differs from similar species by having more septate and smaller cylindrical macroconidia, as well as DNA sequence data. Meanwhile, Microcerachrysomphaludis has elliptical, one-septate ascospores with acute ends and cylindrical, slightly curved with 4-6 septate macroconidia up to 78 µm long. Morphological descriptions with illustrations of the novel species and DNA-based phylogeny generated from analyses of multigene dataset are also provided to better understand species relationships.

Morphology and phylogeny of ascomycetes associated with walnut trees (Juglans regia) in Sichuan province, China.

In Sichuan province, walnuts, consisting of Juglans regia, Juglans sigillata, and the hybrid J. regia × J. sigillata, are commercially important edible nuts, and J. regia is the most widespread plant. To date, the diversity and distribution of fungi inhabiting on Juglans have not received enough attention, although there have been studies focusing on pathogens from fruit and stem. In order to update the checklist of fungi associated with Sichuan walnuts, a survey on fungi associated with the three Juglans species from 15 representative regions in Sichuan was conducted. In this article, ten fungi distributed in two classes of Ascomycota (Dothideomycetes and Sordariomycetes) were described based on morpho-molecular analyses, and two novel species, Neofusicoccum sichuanense and Sphaerulina juglandina, a known species of Ophiognomonia leptostyla, and seven new hosts or geographical records of Cladosporium tenuissimum, Diatrypella vulgaris, Helminthosporium juglandinum, Helminthosporium velutinum, Loculosulcatispora hongheensis, Periconia byssoides, and Rhytidhysteron subrufulum were included. Morphological descriptions and illustrations of these fungi are provided.

Morpho-Molecular Characterization of Microfungi Associated with Phyllostachys (Poaceae) in Sichuan, China.

In the present study, we surveyed the ascomycetes from bamboo of Phyllostachys across Sichuan Province, China. A biphasic approach based on morphological characteristics and multigene phylogeny confirmed seven species, including one new genus, two new species, and five new host record species. A novel genus Paralloneottiosporina is introduced to accommodate Pa. sichuanensis that was collected from leaves of Phyllostachys violascens. Moreover, the newly introduced species Bifusisporella sichuanensis was isolated from leaves of P. edulis, and five species were newly recorded on bamboos, four species belonging to Apiospora, viz. Ap. yunnana, Ap. neosubglobosa, Ap. jiangxiensis, and Ap. hydei, and the last species, Seriascoma yunnanense, isolated from dead culms of P. heterocycla. Morphologically similar and phylogenetically related taxa were compared. Comprehensive descriptions, color photo plates of micromorphology are provided.

Insight into the Systematics of Novel Entomopathogenic Fungi Associated with Armored Scale Insect, Kuwanaspis howardi (Hemiptera: Diaspididae) in China.

This study led to the discovery of three entomopathogenic fungi associated with Kuwanaspis howardi, a scale insect on Phyllostachys heteroclada (fishscale bamboo) and Pleioblastus amarus (bitter bamboo) in China. Two of these species belong to Podonectria: P. kuwanaspidis X.L. Xu & C.L. Yang sp. nov. and P. novae-zelandiae Dingley. The new species P. kuwanaspidis has wider and thicker setae, longer and wider asci, longer ascospores, and more septa as compared with similar Podonectria species. The morphs of extant species P. novae-zelandiae is confirmed based on sexual and asexual morphologies. Maximum likelihood and Bayesian inference analyses of ITS, LSU, SSU, tef1-α, and rpb2 sequence data provide further evidence for the validity of the two species and their placement in Podonectriaceae (Pleosporales). The second new species, Microcera kuwanaspidis X.L. Xu & C.L. Yang sp. nov., is established based on DNA sequence data from ITS, LSU, SSU, tef1-α, rpb1, rpb2, acl1, act, cmdA, and his3 gene regions, and it is characterized by morphological differences in septum numbers and single conidial mass.

Morpho-molecular diversity of Linocarpaceae (Chaetosphaeriales): Claviformispora gen. nov. from decaying branches of Phyllostachys heteroclada.

In this paper, Claviformispora gen. nov. in Linocarpaceae is introduced from Phyllostachys heteroclada in Sichuan Province, China. The new genus is characterised by its distinct morphological characters, such as ostiole with periphyses, asci with a thick doughnut-shaped, J- apical ring and clavate ascospore without septum-like band and appendage. Maximum Likelihood and Bayesian Inference phylogenetic analyses, based on DNA sequence data from ITS, LSU, SSU and TEF-1α regions, provide further evidence that the fungus is a distinct genus within this family. The new genus is compared with similar genera, such as Linocarpon and Neolinocarpon. Descriptions, illustrations and notes are provided for the new taxon.

The complete mitochondrial genome of the fig weevil, Aclees cribratus (Coleoptera: Curculionidae).

Aclees cribratus Gyllenhyl (Coleoptera: Curculionidae) is an important pest of fig. In this study, the complete mitogenome of A. cribratus was determined, which was 17,329 bp in length and contained 37 genes, including 13 protein-coding genes (PCGs), 2 rRNA, 22 tRNA genes, and 2 control regions. The phylogenetic analysis based on mitogenomes showed that A. cribratus is the sister group of Molytinae.

Neostagonosporellasichuanensis gen. et sp. nov. (Phaeosphaeriaceae, Pleosporales) on Phyllostachysheteroclada (Poaceae) from Sichuan Province, China.

Neostagonosporellasichuanensis sp. nov. was found on Phyllostachysheteroclada collected from Sichuan Province in China and is introduced in a new genus Neostagonosporella gen. nov. in this paper. Evidence for the placement of the new taxon in the family Phaeosphaeriaceae is supported by morphology and phylogenetic analysis of a combined LSU, SSU, ITS and TEF 1-α DNA sequence dataset. Maximum-likelihood, maximum-parsimony and Bayesian inference phylogenetic analyses support Neostagonosporella as a distinct genus within this family. The new genus is compared with related genera of Phaeosphaeriaceae and full descriptions and illustrations are provided. Neostagonosporella is characterised by its unique suite of characters, such as multiloculate ascostromata and cylindrical to fusiform, transversely multiseptate, straight or curved ascospores, which are widest at the central cells. Conidiostromata are multiloculate, fusiform to long fusiform or rhomboid, with two types conidia; macroconidia vermiform or subcylindrical to cylindrical, transversely multiseptate, sometimes curved, almost equidistant between septa and microconidia oval, ellipsoidal or long ellipsoidal, aseptate, rounded at both ends. An updated phylogeny of the Phaeosphaeriaceae based on multigene analysis is provided.

Massively Parallel Sequencing of Forensic STRs Using the Ion Chef™ and the Ion S5™ XL Systems.

Next-generation sequencing (NGS) has been used to genotype forensic short tandem repeat (STR) markers for individual identification and kinship analysis. STR data from several NGS platforms have been published, but forensic application trials using the Ion S5™ XL system have not been reported. In this work, we report sensitivity, reproducibility, mixture, simulated degradation, and casework sample data on the Ion Chef™ and S5™ XL systems using an early access 25-plex panel. Sensitivity experiments showed that over 97% of the alleles were detectable with down to 62 pg input of genomic DNA. In mixture studies, alleles from minor contributors were correctly assigned at 1:9 and 9:1 ratios. NGS successfully gave 12 full genotype results from 13 challenging casework samples, compared with five full results using the CE platform. In conclusion, the Ion Chef™ and the Ion S5™ XL systems provided an alternative and promising approach for forensic STR genotyping.

DNA profiling of spermatozoa by laser capture microdissection and low volume-PCR.

Genetic profiling of sperm from complex biological mixtures such as sexual assault casework samples requires isolation of a pure sperm population and the ability to analyze low abundant samples. Current standard procedure for sperm isolation includes preferential lysis of epithelial contaminants followed by collection of intact sperm by centrifugation. While effective for samples where sperm are abundant, this method is less effective when samples contain few spermatozoa. Laser capture microdissection (LCM) is a proven method for the isolation of cells biological mixtures, even when found in low abundance. Here, we demonstrate the efficacy of LCM coupled with on-chip low volume PCR (LV-PCR) for the isolation and genotyping of low abundance sperm samples. Our results indicate that this method can obtain complete profiles (13-16 loci) from as few as 15 sperm cells with 80% reproducibility, whereas at least 40 sperm cells are required to profile 13-16 loci by standard 'in-tube' PCR. Further, LCM and LV-PCR of a sexual assault casework sample generated a DNA genotype that was consistent with that of the suspect. This method was unable, however, to analyze a casework sample from a gang rape case in which two or more sperm contributors were in a mixed population. The results indicate that LCM and LV-PCR is sensitive and effective for genotyping sperm from sperm/epithelial cell mixtures when epithelial lysis may be insufficient due to low abundance of sperm; LCM and LV-PCR, however, failed in a casework sample when spermatozoa from multiple donors was present, indicating that further study is necessitated.

A new method of extrapolating the sweep pattern visual evoked potential acuity.

PURPOSE: In order to compare the amplitude-spatial frequency (A-SP) regression method with amplitude-logVA (A-logVA) regression methods in extrapolating the sweep pattern visual evoked potential (SPVEP) acuity. METHODS: We measured SPVEPs in 21 children and three adults using sinusoidally-modulated horizontal gratings as stimuli. The responses were averaged and displayed through discrete Fourier transformations. SPVER acuity was then estimated by using both the SPVEP amplitude- spatial frequency function (A-SP function regression method) and the SPVEP amplitude-log visual-angle function (A-logVA function regression method). Furthermore, the Bailey Lovie logMAR chart was employed to define visual acuity. Curve estimates were calculated to derive a correlation index (R) for each method. RESULTS: There are significant differences (t = 2.71, P < 0.05) between the correlation indices of curves obtained using the A-logVA function (logarithmic model, 0.95 +/- 0.01) and that obtained by the A-SP function (inverse model, 0.92 +/- 0.02). The overall correlation coefficient (r) between logMAR acuity and acuity calculated by the A-logVA regression method was 0.32 (P < 0.05). The overall correlation coefficient (r) between logMAR acuity and acuity calculated by the A-SP regression method was 0.41 (P < 0.05). Paired t-tests show that SPVEP acuity from the A-logVA function was not significantly different from acuities of the logMAR function (t = 1.77, P = 0.09). The difference in their mean values is 0.14 +/- 0.08. However, SPVEP acuity calculated using the A-SP function regression method is significantly different from the acuity calculated from the logMAR function (t = 10.09, P < 0.01). The difference in their mean values is 0.41 +/- 0.04. CONCLUSIONS: The amplitude-logVA function regression method is more accurate in estimating SPVEP acuity in normal subjects with good visual acuity.

[A comparative study of various diagnostic procedures in macular edema].

OBJECTIVE: To compare the results for diagnosis of macular edema between Heidelberg retina tomography (HRT-II macular edema module) and optical coherence tomography (OCT). METHODS: Two hundred and twelve eyes of 106 macular edema patients were diagnosed with fundus fluorescein angiography (FFA). HRT-II was used to obtain Edema Index and OCT was used to measure the retina thickness in the fovea. The results were compared with FFA. RESULTS: As FFA for gold standard, we compared with other diagnosis method. With regard to the diagnosis of macular edema by OCT, the Kappa value was 0.553, the odds ratio (OR) was 8.519. The Kappa value of HRT-II was 0.403 and OR was 3.210. In two groups of patients with and without macular edema, which were diagnosed by FFA, the difference of Macular edema Index P assayed by HRT-II between these two groups was significant (P = 0.00); while the difference of fovea retina thickness P assayed by OCT between these two groups was significant, too (P = 0.00). The best critical value of E was 2.00 and the ROC curve's area was 0.673 in the detection of macular edema by HRT-II. The best critical value of retina thickness for the diagnosis of macular edema was 170 microm and the ROC curve's area was 0.774 in the detection of macular edema by OCT. CONCLUSIONS: In regard to the diagnosis of macular edema, the sensitivity of HRT-II is better than that from OCT, with a lower false-negative result. However, HRT-II has a relatively low specialty with a higher false- positive result. As for the localization of macular edema, HRT-II has its advantage. Compared with FFA, the consistence of OCT for the diagnosis of macular edema is better than that of HRT-II. We can conclude from the OR value that OCT has greater influence than HRT-II in the diagnosis of macular edema. In addition, as for the diagnosis and different diagnosis, the clinical value of OCT is higher than that of HRT-II. Combination of these two different diagnostic methods might provide an accurate quantitative analysis of macular edema.