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CRISPR/Cas9 has potential for efficient molecular breeding. Recently, a foreign-DNA-free gene-targeting technology was established by introducing a preassembled Cas9 ribonucleoprotein (RNP) complex into the oyster mushroom Pleurotus ostreatus. However, the target gene was restricted to such a gene like pyrG, since screening of a genome-edited strain was indispensable and could be performed via examination of 5-fluoroorotic acid (5-FOA) resistance caused by the disruption of the target gene. In this study, we simultaneously introduced the Cas9 RNP complex targeting fcy1, a mutation that conferred P. ostreatus resistance to 5-fluorocytosine (5-FC), together with that targeting pyrG. A total of 76 5-FOA resistant strains were isolated during the first screening. Subsequently, a 5-FC resistance examination was conducted, and three strains exhibited resistance. Genomic PCR experiments followed by DNA sequencing revealed that mutations were successfully introduced into fcy1 and pyrG in the three strains. The results indicated that double gene-edited mutants could be obtained in one experiment employing 5-FOA resistance screening for strains with Cas9 RNP incorporation. This work may pave the way for safe CRISPR/Cas9 technology to isolate mutant strains in any gene of interest without an ectopic marker gene.

Clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9)-assisted gene targeting is a promising method used in molecular breeding. We recently reported the successful introduction of this method in the monokaryotic Pleurotus ostreatus (oyster mushroom), PC9. However, considering their application in mushroom breeding, dikaryotic strains (with targeted gene mutations in both nuclei) need to be generated. This is laborious and time-consuming because a classical crossing technique is used. Herein, we report a technique that targets both nuclei of dikaryotic P. ostreatus, PC9×#64 in a transformation experiment using plasmid-based CRISPR/Cas9, with the aim of developing a method for efficient and rapid molecular breeding. As an example, we targeted strains with low basidiospore production ability through the meiosis-related genes mer3 or msh4. Four different plasmids containing expression cassettes for Cas9 and two different gRNAs targeting mer3 or msh4 were constructed and separately introduced into PC9×#64. Eight of the 38 dikaryotic transformants analyzed produced no basidiospores. Genomic PCR suggested that msh4 or mer3 mutations were introduced into both nuclei of seven out of eight strains. Thus, in this study, we demonstrated simultaneous gene targeting using our CRISPR/Cas9 system, which may be useful for the molecular breeding of cultivated agaricomycetes.

Pleurotus species are commercially essential mushrooms and widely cultivated throughout the world. The production of Pleurotus mushrooms alone accounts for around 25% of that total cultivated mushrooms globally. In America and Europe, Pleurotus species are considered specialty mushrooms, whereas, in Korea, their cultivation is economically profitable, and it is one of the highly consumed species. Pleurotus species are predominantly found in tropical forests and often grow on fallen branches, dead and decaying tree stumps, and wet logs. Biographical studies have shown that the Pleurotus genus is among the more conspicuous fungi that induce wood decay in terrestrial ecosystems worldwide due to its formidable lignin-modifying enzymes, including laccase and versatile peroxidases. Pleurotus species can be grown easily due to their fast colonization nature on diversified agro-substrates and their biological efficiency 100%. Pleurotus mushrooms are rich in proteins, dietary fiber, essential amino acids, carbohydrates, water-soluble vitamins, and minerals. These mushrooms are abundant in functional bioactive molecules, though to influence health. Pleurotus mushrooms are finding unique applications as flavoring, aroma, and excellent preservation quality. Apart from its unique applications, Pleurotus mushrooms have a unique status delicacy with high nutritional and medicinal values. The present review provides an insight into the cultivation of Pleurotus spp. using different agro-waste as growth substances paying attention to their effects on the growth and chemical composition.

Obesity is associated with chronic low back pain (CLBP), but the association between fat distribution and CLBP is unclear. This cross-sectional study evaluated the relationship using the Korean National Health and Nutrition Examination Survey data. A total of 10,606 adults (average age: 45.4, female: 57.1%) were included. We estimated the regional fat distribution, waist circumference, and body fat proportion, compared the values in people with and without CLBP, and stratified the estimates by sex and obesity status using a multivariable linear model. There were no statistically significant differences in the average waist circumference between the people with and without CLBP (p = 0.731) and the average fat proportion between those with and without CLBP (p = 0.731). The average regional fat distribution was significantly higher in the people with CLBP than in those without CLBP, in the upper limbs (11.4%, 95% confidence interval [CI]: [11.3, 11.5] vs. 11.2%, 95% CI: [11.1, 11.3], p < 0.05) and in the lower limbs (31.9%, 95% CI: [31.6, 32.2] vs. 31.4%, 95% CI: [31.2, 31.6], p < 0.01). More obvious among men, fat distribution in the lower limbs is higher than in people without obesity (p < 0.001). People with CLBP tend to have a higher fat distribution in the limbs than those without it and obese people with CLBP would need to reduce the fat in the lower limbs.

Interspecific hybridization between Ganoderma lingzhi and G. applanatum was attempted through polyethylene glycol (PEG) induced fusion technique. The protoplast isolation procedure was simplified, and we obtained a significant number of protoplasts from both Ganoderma species. The number of protoplasts obtained was 5.27 ± 0.31 × 107/mL in G. lingzhi and 5.57 ± 0.49 × 106/mL in G. applanatum. Osmotic stabilizer NaCl (0.4 M) at pH 5.8 and enzymolysis time 3.5 h have supported high frequency of protoplast regeneration. G. lingzhi and G. applanatum regeneration frequency was 1.73 ± 0.04% and 0.23 ± 0.02%, respectively. 40% of PEG induced high number of protoplast fusion the regeneration frequency was 0.09% on a minimal medium. Two hundred fifty-two fusant colonies were isolated from the following four individual experiments. Among them, ten fusants showed the mycelial morphological difference compared to their parents and other fusant isolates. The fruiting body could be generated on oak sawdust and wheat bran substrate, and a few of them showed recombined morphology of the parental strains. The highest yield and biological efficacy (BE) were recorded in GF248, while least in GF244. The hybridity of the fusant was established based on mycelia, fruiting morphology, and PCR fingerprinting. ISSR and RAPD profile analysis of ten fusants and parents depicted that fusants contained polymorphic bands, which specified the rearrangement and deletion of DNA in the fusants. A Dendrogram was constructed based on the RAPD profile, and the clustering data exhibited two major clusters: cluster I included the G. lingzhi and Cluster II, including the G. applanatum and fusant lines. Total polysaccharide (α, β and total glucan) content was compared with fusants and parental strains. The present study highlighted the efficient methods for protoplast isolation from Ganoderma species. PEG-induced fusants showed high polymorphic frequency index, while the phenotypic characters showed high similarity to G. applanatum. A significant difference was observed in the mushroom yield and its total polysaccharide between the fusants and parental strains.

The production scale of mushrooms in Korea is approximately 600 billion won, which is 1.6% of Korea’s gross agricultural output. In Korea, ca. 190,000 tons of mushrooms are harvested annually. Although the numbers of mushroom farms and cultivators are constantly decreasing, total mushroom yields are increasing owing to large- scale cultivation facilities and automation. The recent expansion of the well-being trend has caused an increase in mushroom consumption in Korea: the annual per capita mushroom was 3.9 kg (’13), whichis a little higher than that in Europe. Thus, mushroom export, mainly Flammulina velutipes and Pleurotus ostreatus, has increased since the mid-2000s. Recently, however, it is slightly reduced. Nevertheless, Vietnam, Hong Kong, the United States, and the Netherlands continue to export mushrooms, and Korea has increased its export to Australia, Canada, Southeast Asia, etc. Canned Agaricus bisporus, the first export of the Korean mushroom industry, reached it speak sales in 1977-1978. When Korea initiated trade with China in 1980, the international prices of mushrooms fell sharply, leading to shrinkage of the domestic markets. Spurred by the high demand to develop substitute goods for A. bisporus, the oyster mushroom (P. ostreatus) gained attention since it seemed to suit the taste of Korean consumers. Although the log cultivation technique for oyster mushroom was developed in the early 1970s, it required a great deal of labor. Thus, we developed the shelf cultivation technique, which is easier to manage and allows for mass production. In this technique, the growing shelf is made mafrom fermented rice straw, whichis the only P. ostreatus medium in the world and isused only in South Korea. After then, the use of cotton wastes as an additional material of medium, the productivity. Currently, we are developing a standard cultivation technique and environmental control system that can stably produce mushrooms throughout the year. The increase of oyster mushroom production may boostthe domestic market and contribute to industrial development. In addition, oyster mushroom production technology played a role in forming the basis for the development of bottle cultivation, which made mass production . In particular, bottle cultivation using liquid spawn could allow for the export of F. velutipes and Pleurotus eryngii. In addition, the white varieties of F. velutipes were second developed in the world after Japan. We also developed the new A. bisporus cultivar ‘Saeah’, which is easy to grow in Korea. In hopes to advance the mushroom industry, we will continue to develop cultivars with international competitive power and to improve cultivation techniques.

Dynamic adsorption studies of vaporous benzene (3000 ppm) on a hierarchically mesoporous/microporous beta zeolite-templated carbon (HBTC) was compared to those of commercial activated carbon (AC) and solely microporous beta zeolite-templated carbon (BTC). The BTC had periodic micropores with a narrow distribution of 1 nm in size and a high micropore surface area of ~2870 m2 g−1, thereby showing a much higher dynamic adsorption capacity (653 mg g−1) for benzene than that exhibited by the AC (264 mg g−1). Nevertheless, the BTC exhibited a lower breakthrough time (3.26 mg) for benzene than that shown by the AC (4.9 mg) owing to severe diffusion constraints. The diffusion problem can be solved by introducing mesoporosity into microporous BTC to form a hierarchically mesoporous/microporous structure. Isosteric heat and benzene diffusivity results indicated the strong affinity and facile diffusion exhibited by the hierarchical BTC, which were attributed to the graphitic nature of the carbon framework, uniform micropores, and hierarchically mesoporous/microporous structures. Therefore, the HBTC exhibited a benzene breakthrough time of 11.4 mg and a dynamic adsorption capacity of 472 mg g−1 (100% breakthrough) that was larger than those of commercial AC. Furthermore, the adsorption capacity and partition coefficient (PC) of the tested adsorbents were evaluated at 10% breakthrough. Among all tested carbons, HBTC exhibited the best performance for benzene capture with adsorption capacity of 332 mg g−1 and PC value of 0.140 mol Kg−1 Pa−1. The effects of adsorption temperature and relative humidity on the benzene adsorption capacities of HBTC and BTC were also explored. Furthermore, desorption and subsequent regeneration studies demonstrated that the adsorption capacity of HBTC can be completely restored by heat treatment. These findings indicate that HBTC is a promising candidate for VOC adsorption that can replace the currently employed commercial AC.

The mapping of galaxy clustering from real space to redshift space introduces the anisotropic property to the measured galaxy density power spectrum in redshift space, known as the redshift space distortion (RSD) effect. The mapping formula is intrinsically non-linear, which is complicated by the higher order polynomials due to indefinite orders of cross correlations between density and velocity fields, and the Finger--of--God (FoG) effect due to the randomness of the galaxy peculiar velocity field. In previous works, we have verified the robustness of advanced TNS mapping formula in our hybrid RSD model in dark matter case, where the halo bias models are not taken into account for the halo mapping formula in redshift space. Using 100 realizations of halo catalogs in N-body simulations, we find that our halo RSD model with the known halo bias model and the effective FoG function accurately predicts the halo power spectrum measurements, within 1$\sim$2% accuracy up to $k\sim 0.2h$/Mpc, depending on different halo masses and redshifts., 33 pages, 12 figures. Results of one more halo mass bin are added. Influence of non-vanishing halo velocity bias is tested and discussed. Main conclusions unchanged. Accepted by JCAP

The observed power spectrum in redshift space appears distorted due to the peculiar motion of galaxies, known as redshift-space distortions (RSD). While all the effects in RSD are accounted for by the simple mapping formula from real to redshift spaces, accurately modeling redshift-space power spectrum is rather difficult due to the non-perturbative properties of the mapping. Still, however, a perturbative treatment may be applied to the power spectrum at large-scales, and on top of a careful modeling of the Finger-of-God effect caused by the small-scale random motion, the redshift-space power spectrum can be expressed as a series of expansion which contains the higher-order correlations of density and velocity fields. In our previous work [JCAP 8 (Aug., 2016) 050], we provide a perturbation-theory inspired model for power spectrum in which the higher-order correlations are evaluated directly from the cosmological $N$-body simulations. Adopting a simple Gaussian ansatz for Finger-of-God effect, the model is shown to quantitatively describe the simulation results. Here, we further push this approach, and present an accurate power spectrum template which can be used to estimate the growth of structure as a key to probe gravity on cosmological scales. Based on the simulations, we first calibrate the uncertainties and systematics in the pertrubation theory calculation in a fiducial cosmological model. Then, using the scaling relations, the calibrated power spectrum template is applied to a different cosmological model. We demonstrate that with our new template, the best-fitted growth functions are shown to reproduce the fiducial values in a good accuracy of 1 \% at $k, 24 pages

Lovastatin is a 3-hydroxy-3-methylglutaryl-CoA reductase inhibitor that is clinically used for the prevention of cardiovascular diseases. Although it has been reported that lovastatin has anti-inflammatory properties in several studies, how lovastatin regulates the inflammation is still unclear. To evaluate the effect of lovastatin on nitric oxide production (NO) in RAW264.7 macrophages, NO production assay was performed. Also, cell viability was measured to confirm cytotoxicity. Level of tumor necrosis factor-α (TNF-α) transcription was measured by reverse transcription polymerase chain reaction (RT-PCR) from total RNA in RAW264.7 cells. Western blot analysis and immunofluorescence staining were used to investigate the regulation of lovastatin on the expression, phosphorylation, and nuclear translocation of cellular proteins. The results of the present study revealed that lovastatin reduced nitric oxide production via the reduction of inducible nitric oxide synthase (iNOS) expression in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. The mRNA level of TNF-α was reduced in presence of lovastatin. In addition, lovastatin downregulated histone deacetylase 1 (HDAC1), resulting in the accumulation of acetylated histone H3 and heat shock protein 70. Furthermore, the expression of phosphoinositide 3-kinase catalytic subunits α and β was reduced under lovastatin treatment, and the phosphorylation of Akt and mammalian target of rapamycin was consequently inhibited. Lovastatin also inhibited the phosphorylation of inhibitor of nuclear factor (NF)-κBα and the translocation of NF-κB into the nucleus. Therefore, the present study demonstrates that lovastatin inhibits the expression of pro-inflammatory mediators, including iNOS and TNF-α, through the suppression of HDAC1 expression, PI3K/Akt phosphorylation and NF-κB translocation in LPS-stimulated RAW264.7 macrophage cells.

Both multi-streaming (random motion) and bulk motion cause the Finger-of-God (FoG) effect in redshift space distortion (RSD). We apply a direct measurement of the multi-streaming effect in RSD from simulations, proving that it induces an additional, non-negligible FoG damping to the redshift space density power spectrum. We show that, including the multi-streaming effect, the RSD modelling is significantly improved. We also provide a theoretical explanation based on halo model for the measured effect, including a fitting formula with one to two free parameters. The improved understanding of FoG helps break the $f\sigma_8-\sigma_v$ degeneracy in RSD cosmology, and has the potential of significantly improving cosmological constraints., Comment: 19 pages, 4 figures. Match the published version. Typos corrected

The anisotropic galaxy clustering of large scale structure observed by the Baryon Oscillation Spectroscopic Survey Data Release 11 is analyzed to probe the sum of neutrino mass in the small $m_\nu < 1eV$ limit in which the early broadband shape determined before the last scattering surface is immune from the variation of $m_\nu$. The signature of $m_\nu$ is imprinted on the altered shape of the power spectrum at later epoch, which provides an opportunity to access the non--trivial $m_\nu$ through the measured anisotropic correlation function in redshift space (hereafter RSD instead of Redshift Space Distortion). The non--linear RSD corrections with massive neutrinos in the quasi linear regime are approximately estimated using one-loop order terms computed by tomographic linear solutions. We suggest a new approach to probe $m_\nu$ simultaneously with all other distance measures and coherent growth functions, exploiting this deformation of the early broadband shape of the spectrum at later epoch. If the origin of cosmic acceleration is unknown, $m_\nu$ is poorly determined after marginalising over all other observables. However, we find that the measured distances and coherent growth functions are minimally affected by the presence of mild neutrino mass. Although the standard model of cosmic acceleration is assumed to be the cosmological constant, the constraint on $m_\nu$ is little improved. Interestingly, the measured CMB distance to the last scattering surface sharply slices the degeneracy between the matter content and $m_\nu$, and the hidden $m_\nu$ is excavated to be $m_\nu=0.19^{+0.28}_{-0.17} eV$ which is different from massless neutrino more than 68% confidence., Comment: 11 pages, 4 figures, 2 tables

We analyze the clustering of large scale structure in the Universe in a model independent method, accounting for anisotropic effects along and transverse to the line of sight. The Baryon Oscillation Spectroscopy Survey Data Release 11 provides a large sample of 690,000 galaxies, allowing determination of the Hubble expansion H, angular distance D_A, and growth rate G_T at an effective redshift of z=0.57. After careful bias and convergence studies of the effects from small scale clustering, we find that cutting transverse separations below 40 Mpc/h delivers robust results while smaller scale data leads to a bias due to unmodelled nonlinear and velocity effects. The converged results are in agreement with concordance LCDM cosmology, general relativity, and minimal neutrino mass, all within the 68% confidence level. We also present results separately for the northern and southern hemisphere sky, finding a slight tension in the growth rate -- potentially a signature of anisotropic stress, or just covariance with small scale velocities -- but within 68% CL., 9 pages, 6 figures

Our observations of the Universe are fundamentally anisotropic, with data from galaxies separated transverse to the line of sight coming from the same epoch while that from galaxies separated parallel to the line of sight coming from different times. Moreover, galaxy velocities along the line of sight change their redshift, giving redshift space distortions. We perform a full two-dimensional anisotropy analysis of galaxy clustering data, fitting in a substantially model independent manner the angular diameter distance D_A, Hubble parameter H, and growth rate ddelta/dln a without assuming a dark energy model. The results demonstrate consistency with LCDM expansion and growth, hence also testing general relativity. We also point out the interpretation dependence of the effective redshift z_eff, and its cosmological impact for next generation surveys., Comment: 12 pages, 14 figures