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The present study aimed to recognize the microscopic characteristics of black aspergilli species which exhibit similar characteristics on culture media. Forty eight black aspergilli isolates were obtained from clinical wastes and purified using single spore technique on six different culture media. The ultrastructure of fungal conidiophore and spores was detected by using Scanning Electron Microscope (SEM). The fungal isolates were identified within five species included A. niger, A. tubingensis, Aspergillus section Nigri, A. violaceofuscus, A. neoniger. Besides, two isolates identified as Aspergillus sp. strain no. 39, Aspergillus sp. strain no. 53 appear as new strains based on the structure of conidiophore and spores. The fungi species have similar culture characteristics. However, SEM observation demonstrated that they have quite different conidiophore and spores morphology. The study revealed that the microstructure of the fungal spores and conidiophores plays an important role in the identification of fungi species based on the phenotypic method.

Noriyuki Kanzaki,1 Yuta Otsuka,1 Takayuki Izumo,1 Hiroshi Shibata,1 Hideyuki Nagao,2 Keita Ogawara,3 Hiroshi Yamada,3 Seiji Miyazaki,3 Yutaka Nakamura3 1Institute for Health Care Science, Suntory Wellness Ltd, Seika-cho, Soraku-gun, Kyoto, Japan; 2Research Institute of Sports Medical Science, Tokai University, Hiratsuka, Kanagawa, Japan; 3School of Physical Education, Tokai University, Hiratsuka, Kanagawa, Japan Background: Previously, we demonstrated that glucosamine-containing supplementation was effective for improving locomotor functions, especially walking speed. However, the biomechanical mechanism of efficacy has not been elucidated. This study aimed to address this challenge in subjects with knee pain, using a motion capture system. Methods: An open label study was conducted in 30 Japanese subjects with knee pain. The subjects were administered a daily supplement containing 1,200 mg of glucosamine hydrochloride, 60 mg of chondroitin sulfate, 45 mg of type II collagen peptides, 90 mg of quercetin glycosides, 10 mg of imidazole peptides, 1 mg of proteoglycan, and 5 µg of vitamin D (GCQID). The intervention continued for 16 weeks. Efficacy for locomotor functions involving the knee joint was evaluated mainly using the Japanese Knee Osteoarthritis Measure (JKOM) and the 5-question Geriatric Locomotive Function Scale (GLFS-5). To examine the biomechanical mechanism of efficacy for locomotor functions, motions of subjects in a normal walking state were captured. Gait analysis was conducted and efficacy for gait parameters such as normal walking speed, stride length, cadence, and angle of soles was evaluated. Results: GCQID significantly improved total scores on the JKOM and GLFS-5. In gait analysis, normal walking speed, stride length, and angle of soles at the end of the stance phase were all significantly increased, but cadence did not change significantly during the intervention period. There were significant intercorrelations of changes in normal walking speed, stride length, and angle of soles at the end of the stance phase, and between changes in stride length and total JKOM score. Conclusion: A GCQID supplement may increase walking speed through increased stride length and angle of kicking from the ground during steps, which might be mainly associated with alleviated knee pain. Keywords: locomotive syndrome, GLFS-5, normal walking speed, stride length, toe angle

Seven near isogenic lines (NILs) for six partial blast-resistance genes—Pi35, pi21, Pi37(t)(Pi37-KRIL17), Pb1, and Pi34 (two: Pi34-Chubu32 and Pi34-Chugoku40, originating from different Japonica Group rice lines, ‘Chubu 32’ and ‘Chugoku 40’, respectively)—and one quantitative trait locus (QTL), PiPHL9, derived from Japonica Group line ‘Hokkai PL9’, were developed in rice (Oryza sativa L.). These NILs have the genetic background of an Indica Group susceptible line ‘US-2’, which does not harbor complete resistance genes and can clarify the effects of partial resistance genes and the QTL. These NILs and US-2 were used to determine the effects of the partial resistance genes and the QTL based on the results of an inoculation test for leaf blast using international standard differential blast isolates in the young seedling stage and a natural infection test in the mature stage after heading at a blast field. The NILs produced mainly moderate reactions in the inoculation test and rarely exhibited “resistant” and “susceptible” reactions. Each of the partial resistance genes and the QTL had different effects in panicle blast. Two genes, Pi35 and Pb1, were the most effective with respect to resistance, followed by Pi37-KRIL17. Pi34-Chugoku40 was the weakest. The effects of pi21, Pi34-Chubu32, and PiPHL9 varied among cultivation seasons and could be easily affected by environmental conditions. This is the first report clarifying the effects of partial resistance genes without the major complete genes, which will be helpful for advanced genetic and pathological studies and as a source of genes for rice breeding. [Figure: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

The aim of the current study was to describe the ultrastructure of Aspergillus spp. spores and conidiophores using Scanning Electron Microscope (SEM) and their role in contaminating of clinical solid wastes. Aspergillus spp. were described on, Czapek Yeast Extract Agar (CYA), Malt Extract Agar (MEA), Sabouraud Dextrose Agar (SDA), Czapek-Dox Agar (CZ) and Potato Dextrose Agar (PDA), while the ultrastructure of spores and conidiophores were observed using light Microscope and SEM. Fungal isolates were identified as A. caesiellus, A. terreus var. terreus; A. flavus, A. parasiticus, A. sydowii and A. Fumigatus with two new strains identified as Aspergillus sp. strain no. 145, Aspergillus sp. strain no. 311. SEM analysis for Aspergillus spp. conidiophores and spores were more useful in the taxonomy of fungi. Microstructures of the spores play important role in the contamination of clinical wastes, where the spiny fungal spores surface have high adherence for the solid wastes.

Rice blast, caused by Magnaporthe oryzae, poses a significant threat to rice production. Rice blast susceptibility has been observed in Japanese rice varieties with excellent eating quality. Enhancing blast resistance is essential to ensure minimal use of agricultural chemicals. Two types of blast resistance are observed: True resistance, which is a type of qualitative resistance expressed by a major gene, and field resistance, which is a type of quantitative resistance expressed by multiple micro-acting genes. ‘Resistance collapse’, in which a variety with a true resistance gene becomes diseased by blast fungus races compatible with the resistance gene, has been observed. Varieties carrying blast-resistance genes, such as Pb1 (panicle blast resistance 1), have been developed through DNA marker-assisted selection. In this review, we focus on the Pb1, which expresses strong quantitative resistance to panicle blast and has been widely used in Japan without showing ‘resistance collapse’ for 40 years. Pb1 is an ‘adult plant resistance gene’ that does not exert strong selection pressure on the blast population during the leaf blast stage, thus preventing the selective multiplication of Pb1-compatible blast strains. This epidemiological mechanism prevents ‘resistance collapse’. Interdisciplinary research and breeding are required to sustainably use genes that induce high field resistance.

Abstract Background Rice blast is the most serious disease afflicting rice and there is an urgent need for the use of disease resistance (R) genes in blast tolerance breeding programs. Pb1 is classified as a quantitative resistance gene and it does not have fungal specificity. Pb1-mediated resistance develops in the latter stages of growth. However, some cultivars, such as Kanto209 (K209), cultivar name Satojiman, despite possessing Pb1, do not exert resistance to rice blast during the reproductive stage. Results We found that the expression of WRKY45 gene downstream of Pb1 was weakly induced by rice blast inoculation at the full heading stage in K209. Genetic analysis using the SNP-based Golden Gate assay of K209 crossing with Koshihikari Aichi SBL (KASBL) found at least four regions related to the resistance in the rice genome (Chr8, Chr9, Chr7, Chr11). Mapping of QTL related to Chr7 confirmed the existence of factors that were required for the resistance of Pb1 in the 22 to 23 Mbp region of the rice genome. Conclusion We clarified how the K209 cultivar is vulnerable to the blast disease despite possessing Pb1 and found the DNA marker responsible for the quantitative resistance of Pb1. We identified the QTL loci required for Pb1-mediated resistance to rice panicle blast. Pb1 was negatively dependent on at least three QTLs, 7, 9 and 11, and positively dependent on one, QTL 8, in the K209 genome. This finding paves the way for creating a line to select optimal QTLs in order to make use of Pb1-mediated resistance more effectively.

Plant hormones play pivotal signaling roles in plant–pathogen interactions. Here, we report characterization of an antagonistic interaction of abscisic acid (ABA) with salicylic acid (SA) signaling pathways in the rice–Magnaporthe grisea interaction. Exogenous application of ABA drastically compromised the rice resistance to both compatible and incompatible M. grisea strains, indicating that ABA negatively regulates both basal and resistance gene–mediated blast resistance. ABA markedly suppressed the transcriptional upregulation of WRKY45 and OsNPR1, the two key components of the SA signaling pathway in rice, induced by SA or benzothiadiazole or by blast infection. Overexpression of OsNPR1 or WRKY45 largely negated the enhancement of blast susceptibility by ABA, suggesting that ABA acts upstream of WRKY45 and OsNPR1 in the rice SA pathway. ABA-responsive genes were induced during blast infection in a pattern reciprocal to those of WRKY45 and OsPR1b in the compatible rice–blast interaction but only marginally in the incompatible one. These results suggest that the balance of SA and ABA signaling is an important determinant for the outcome of the rice–M. grisea interaction. ABA was detected in hyphae and conidia of M. grisea as well as in culture media, implying that blast-fungus-derived ABA could play a role in triggering ABA signaling at host infection sites.

We obtained three Magnaporthe grisea morphological mutants that had the LINE transposon MGL inserted into the ACR1 locus. Sequence analysis revealed that ACR1 is homologous to medA, a developmental regulator of Aspergillus nidulans conidiation. These results demonstrated that MGL elements could transpose and cause insertional mutagenesis in M. grisea.

Abstract Background The Hedgehog signaling pathway functions as an organizer in embryonic development. Recent studies have demonstrated constitutive activation of Hedgehog pathway in various types of malignancies. However, it remains unclear how Hedgehog pathway is involved in the pathogenesis of osteosarcoma. To explore the involvement of aberrant Hedgehog pathway in the pathogenesis of osteosarcoma, we investigated the expression and activation of Hedgehog pathway in osteosarcoma and examined the effect of SMOOTHENED (SMO) inhibition. Results To evaluate the expression of genes of Hedgehog pathway, we performed real-time PCR and immunohistochemistry using osteosarcoma cell lines and osteosarcoma biopsy specimens. To evaluate the effect of SMO inhibition, we did cell viability, colony formation, cell cycle in vitro and xenograft model in vivo. Real-time PCR revealed that osteosarcoma cell lines over-expressed Sonic hedgehog, Indian hedgehog, PTCH1, SMO, and GLI. Real-time PCR revealed over-expression of SMO, PTCH1, and GLI2 in osteosarcoma biopsy specimens. These findings showed that Hedgehog pathway is activated in osteosarcomas. Inhibition of SMO by cyclopamine, a specific inhibitor of SMO, slowed the growth of osteosarcoma in vitro. Cell cycle analysis revealed that cyclopamine promoted G1 arrest. Cyclopamine reduced the expression of accelerators of the cell cycle including cyclin D1, cyclin E1, SKP2, and pRb. On the other hand, p21cip1 wprotein was up-regulated by cyclopamine treatment. In addition, knockdown of SMO by SMO shRNA prevents osteosarcoma growth in vitro and in vivo. Conclusions These findings suggest that inactivation of SMO may be a useful approach to the treatment of patients with osteosarcoma.

Aims: The effect of different types of fungal inocula to the cellulase activity measured on palm kernel cake (PKC) was studied. Methodology and Results: Isolate Pro-A1 which was identified as Trichoderma sp. was selected as a potential producer of cellulase via solid state fermentation technique (SSF). Two types of PKCs were used; raw PKC (containing residual oil) and defatted PKC. The PKCs were inoculated with different concentrations of conidia and varying amounts (g) of solid mycelia plugs (SMP) for SSF. The effect of ultrafiltered crude fungal filtrate (CFF) as inocula was also being tested. The highest cellulase activity of 2.454 FPU/mL was detected with 60% (wt/wt) SMP applied to the raw PKC. Conversely, 2.059 FPU/mL of cellulase activity was measured when 80% (wt/wt) of SMP was applied to the defatted PKC which is 62.3% higher than the untreated defatted PKC; and more than 100% increase in enzymatic activity compared to raw PKC. The cellulase activity in the SSF inoculated with 8 x 106 conidia /mL and 12 x 106 conidia /mL were 1.704 FPU/mL for raw PKC and 1.856 FPU/mL for defatted PKC, an enhancement of about 46% from uninoculated batch. Inoculation with CFF bears corresponding maximum improvement of the cellulase activity on both PKCs of 13.58% (raw) and 2.86% (defatted). Conclusion, significance and impact of study: The current study proves that Trichoderma sp. in the form of SMP can enhance the cellulase activity on PKCs effectively with more than 100% increment. Fungal conidia are also a better choice in enhancing cellulase activity of Trichoderma sp. permitted that the PKC used is devoid of oil. From this study, Trichoderma sp. holds the potential of converting lignocellulosic materials into products of commercial and industrial values such as glucose and other biofuels.