Additive screening and formula optimization of microbial inhibitor having disease prevention and growth promotion effects on Avena sativa.

In order to develop environment friendly microbial inhibitor that can also control disease and promote oat (Avena sativa) growth, the growth rate method and response surface methodology were used to screen wetting agents, preservatives and protective agents at optimal concentrations in this study. Antagonistic activity of the tested bacterium and cell-free fermentation liquid against pathogenic fungi was evaluated on potato dextrose agar (PDA) substratum plates by dual culture technique. Oxford cup method was used to measure antagonistic reaction between screened bacteria. According to each screened bacteria with 50 mL were mixed and cultured in Luria-bertani (LB) substratum. Additives of Wetting agents, UV-protectors, and preservatives were screened by single factor test on the growth concentration of screened mixed bacteria. Afterwards, the optimal additives and concentrations were screened by Box-Behnken method. The microbial inhibitor was detected according to national standards GB20287-2006 and tested on oat in a pot experiment. The results showed that: (1) Functional bacteria which including Bacillus velezensis and Brevundimonas faecalis had control effects of 50.00% to 83.29% on three pathogenic fungi, and their cell free-fermentation liquid could inhibit the growth of pathogenic fungi from 23.51% to 39.90%; (2) Tween-80 was most suitable as wetting agents for Mix biocontrol bacteria (MBB) with 1.00% mass fraction; Sorbitol was selected as UV protective agents for MBB with 0.50% mass fraction. And methyl paraben was used as a preservative for MBB, with 0.50% mass fraction; (3) The most effective adjuvant contained 14.96 mL/L Tween-80, 5.12 g/L methylparaben and 5.6 g/L sorbitol; and (4) The microbial inhibitor controlled 45.57% of oat root rot and increased plant height, root length and seedling biomass. This study provides a suitable environment for the protection of mixed biocontrol bacteria, and lays a foundation for the prevention and control of oat diseases, the promotion of growth and the improvement of quality.

Grazing exclusion alters soil methane flux and methanotrophic and methanogenic communities in alpine meadows on the Qinghai-Tibet Plateau.

Grazing exclusion (GE) is an effective measure for restoring degraded grassland ecosystems. However, the effect of GE on methane (CH4) uptake and production remains unclear in dominant bacterial taxa, main metabolic pathways, and drivers of these pathways. This study aimed to determine CH4 flux in alpine meadow soil using the chamber method. The in situ composition of soil aerobic CH4-oxidizing bacteria (MOB) and CH4-producing archaea (MPA) as well as the relative abundance of their functional genes were analyzed in grazed and nongrazed (6 years) alpine meadows using metagenomic methods. The results revealed that CH4 fluxes in grazed and nongrazed plots were -34.10 and -22.82 µg‧m-2‧h-1, respectively. Overall, 23 and 10 species of Types I and II MOB were identified, respectively. Type II MOB comprised the dominant bacteria involved in CH4 uptake, with Methylocystis constituting the dominant taxa. With regard to MPA, 12 species were identified in grazed meadows and 3 in nongrazed meadows, with Methanobrevibacter constituting the dominant taxa. GE decreased the diversity of MPA but increased the relative abundance of dominated species Methanobrevibacter millerae from 1.47 to 4.69%. The proportions of type I MOB, type II MOB, and MPA that were considerably affected by vegetation and soil factors were 68.42, 21.05, and 10.53%, respectively. Furthermore, the structural equation models revealed that soil factors (available phosphorus, bulk density, and moisture) significantly affected CH4 flux more than vegetation factors (grass species number, grass aboveground biomass, grass root biomass, and litter biomass). CH4 flux was mainly regulated by serine and acetate pathways. The serine pathway was driven by soil factors (0.84, p < 0.001), whereas the acetate pathway was mainly driven by vegetation (-0.39, p < 0.05) and soil factors (0.25, p < 0.05). In conclusion, our findings revealed that alpine meadow soil is a CH4 sink. However, GE reduces the CH4 sink potential by altering vegetation structure and soil properties, especially soil physical properties.

Cross-Breeding Improvement and Performance Analysis of Dominant Production Traits in Grazing-Type Alfalfa (Medicago sativa L.).

Rhizome-rooted Medicago sativa L. "Qingshui" is an excellent germplasm for establishing grazing and ecological grasslands but inferior in yield, in which both high production and ecological values can be achieved by cross-breeding. We have obtained valuable rhizome-rooted hybrid strains (RSA-01, RSA-02, and RSA-03) by crossing of Qingshui and the high-yielding Medicago sativa L. "WL168." In this study, the Qingshui plants with low production performance were crossed for improvement, and progenies with better production and higher quality than those of Qingshui were selected. The results reveal that the branch number, crude protein (CP) content, and relative feed value (RFV) of RSA-01; the stem thickness, CP content, and ether extract (EE) content of RSA-02; and the plant height, stem thickness, branch number, and dry hay yield of RSA-03 were higher than those of Qingshui. Except for the leaf/stem ratio and plant height of RSA-01, leaf/stem ratio of RSA-02, and plant height of RSA-03, the coefficient of variation (CV) of yield traits of the hybrid strains was lower than those of Qingshui, ranging from 0.1% to 4.28%. In addition to the lignin and acid detergent fiber content of RSA-01 as well as EE content of RSA-02 and RSA-03, the CV of the nutritional traits of the hybrid strains was low, ranging from 0.60% to 3.43%. The tested samples were ranked as follows based on yield performance and nutritional values: WL168 > RSA - 03 > RSA - 01 > RSA - 02 > Qingshui and RSA - 01 > WL168 > RSA - 03 > Qingshui > RSA - 02, respectively. Compared with parental Qingshui, RSA-01, RSA-02, and RSA-03 show better yield performance; meanwhile, RSA-01 and RSA-03 had higher nutritional traits. RSA-01 shows heterosis in branch number, CP content, and RFV; RSA-02 shows heterosis in stem thickness and RSA-03 in plant height, stem thickness, branch number, fresh yield, dry hay yield, and CP content. Notably, the low production performance of Qingshui was improved after crossing it with WL168, substantially resulting in an abundant rhizome-rooted germplasm resource for the establishment of grazing grasslands.

Estimating intervention effects on infectious disease control: The effect of community mobility reduction on Coronavirus spread.

Understanding the effects of interventions, such as restrictions on community and large group gatherings, is critical to controlling the spread of COVID-19. Susceptible-Infectious-Recovered (SIR) models are traditionally used to forecast the infection rates but do not provide insights into the causal effects of interventions. We propose a spatiotemporal model that estimates the causal effect of changes in community mobility (intervention) on infection rates. Using an approximation to the SIR model and incorporating spatiotemporal dependence, the proposed model estimates a direct and indirect (spillover) effect of intervention. Under an interference and treatment ignorability assumption, this model is able to estimate causal intervention effects, and additionally allows for spatial interference between locations. Reductions in community mobility were measured by cell phone movement data. The results suggest that the reductions in mobility decrease Coronavirus cases 4 to 7 weeks after the intervention.

Dry matter recovery, ensiling characteristics and aerobic stability of oat silage treated with microbial inoculants at different temperatures.

To evaluate the effects of temperature and lactic acid bacteria (LAB) inoculants on oat silage in Loess Plateau of China, oat was harvested at dough stage, inoculated without (Control) or with LAB inoculants Synlac I (SLI, Lactobacillus plantarum and Pedioccocus acidilactici) and a selected strain HT1 (L. rhamnosus) and ensiled at 25°C (T25), 35°C (T35) and 45°C (T45). The fermentation quality was measured after 60 d of ensiling and the aerobic exposure was conducted at 30°C for 9 d. The results showed that control silage (stored at 25°C) had better fermentation quality than that ensiled at 35°C or 45°C. High temperature of 45°C resulted in sharp decreases in LAB counts and lactic acid concentration and increases in pH and NH3-N concentration in the control group. Inoculation improved the fermentation quality, and HT1 was more effective than SLI at 35°C and 45°C, while SLI showed better performance at 25°C. All silages displayed mild fluctuation for all treatments at the first 3 d of aerobic exposure, and significant differences were observed among treatments after that. Both control and inoculated silages stored at 25°C showed a sharp pH increase, while HT1 treated silages stored at 35°C and 45°C maintained stable pH and better fermentation quality during the aerobic exposure. In conclusion, SLI was suitable for oat silage fermentation at normal atmospheric temperature (25°C), while HT1 was more effective in improving DM recovery, fermentation quality and aerobic stability of oat silage at high temperature during summer in the Loess Plateau of China.

Unveiling the Complexity of Red Clover (Trifolium pratense L.) Transcriptome and Transcriptional Regulation of Isoflavonoid Biosynthesis Using Integrated Long- and Short-Read RNAseq.

Red clover (Trifolium pratense L.) is used as forage and contains a high level of isoflavonoids. Although isoflavonoids in red clover were discovered a long time ago, the transcriptional regulation of isoflavonoid biosynthesis is virtually unknown because of the lack of accurate and comprehensive characterization of the transcriptome. Here, we used a combination of long-read (PacBio Iso-Seq) and short-read (Illumina) RNAseq sequencing to develop a more comprehensive full-length transcriptome in four tissues (root, stem, leaf, and flower) and to identify transcription factors possibly involved in isoflavonoid biosynthesis in red clover. Overall, we obtained 50,922 isoforms, including 19,860 known genes and 2817 novel isoforms based on the annotation of RefGen Tp_v2.0. We also found 1843 long non-coding RNAs, 1625 fusion genes, and 34,612 alternatively spliced events, with some transcript isoforms validated experimentally. A total of 16,734 differentially expressed genes were identified in the four tissues, including 43 isoflavonoid-biosynthesis-related genes, such as stem-specific expressed TpPAL, TpC4H, and Tp4CL and root-specific expressed TpCHS, TpCHI1, and TpIFS. Further, weighted gene co-expression network analysis and a targeted compound assay were combined to investigate the association between the isoflavonoid content and the transcription factors expression in the four tissues. Twelve transcription factors were identified as key genes for isoflavonoid biosynthesis. Among these transcription factors, the overexpression of TpMYB30 or TpRSM1-2 significantly increased the isoflavonoid content in tobacco. In particular, the glycitin was increased by 50-100 times in the plants overexpressing TpRSM1-2, in comparison to that in the WT plants. Our study provides a comprehensive and accurate annotation of the red clover transcriptome and candidate genes to improve isoflavonoid biosynthesis and accelerate research into molecular breeding in red clover or other crops.

Multivariate Spatial Prediction of Air Pollutant Concentrations with INLA.

Estimates of daily air pollution concentrations with complete spatial and temporal coverage are important for supporting epidemiologic studies and health impact assessments. While numerous approaches have been developed for modeling air pollution, they typically only consider each pollutant separately. We describe a spatial multipollutant data fusion model that combines monitoring measurements and chemical transport model simulations that leverages dependence between pollutants to improve spatial prediction. For the contiguous United States, we created a data product of daily concentration for 12 pollutants (CO, NOx, NO2, SO2, O3, PM10, and PM2.5 species EC, OC, NO3, NH4, SO4) during the period 2005 to 2014. Out-of-sample prediction showed good performance, particularly for daily PM2.5 species EC (R2 = 0.64), OC (R2 = 0.75), NH4 (R2 = 0.84), NO3 (R2 = 0.73), and SO4 (R2 = 0.80). By employing the integrated nested Laplace approximation (INLA) for Bayesian inference, our approach also provides model-based prediction error estimates. The daily data product at 12km spatial resolution will be publicly available immediately upon publication. To our knowledge this is the first publicly available data product for major PM2.5 species and several gases at this spatial and temporal resolution.

Coniferyl ferulate exerts antidepressant effect via inhibiting the activation of NMDAR-CaMKII-MAPKs and mitochondrial apoptotic pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Xiaoyaosan (XYS), composed of Radix Bupleuri, Radix Angelicae Sinensis, Radix Paeoniae Alba, Rhizoma Atractylodis Macrocephalae, Poria, Herba Menthae, Rhizoma Zingiberis Recens and Radix Glycyrrhizae, is a valuable traditional Chinese medicine (TCM) which is used for the treatment of depression in China. In our previous experiments, we found that coniferyl ferulate (CF) was the main active constituent of Xiaoyaosan based on UPLC-PDA guided isolation technique. However, the antidepressant effect and mechanisms of CF is still unknown. AIM OF THE STUDY: In the current study, we aim to explore the possible mechanisms involved in the neuroprotective effect of CF in glutamate-injured PC12 cells, and further to confirm the anti-depressant effect of CF on the model of behavioral despair in vivo. MATERIAL AND METHODS: The model of glutamate-injured PC12 cells was employed to investigate the possible mechanisms involved in the neuroprotective effect of CF. The model of behavioral despair was carried out to examine the in vivo anti-depressant effect of CF. RESULTS: The results showed that CF significantly attenuated the decrease of cell viability, the release of lactate dehydrogenase (LDH), and the increase of apoptosis rates induced by glutamate. CF could also suppress the influx of Ca2+ and the elevation of p-NR2B, p-CaMK II, p-JNK, and p-p38 level induced by glutamate. Besides, CF could also inhibit the generation of reactive oxygen species (ROS), the decrease of SOD activity, the elevation of malondialdehyde (MDA) level, and suppress the loss of mitochondrial membrane potential (MMPs) and the activation Bcl-2/Bax mediated apoptotic pathways induced by glutamate. Furthermore, CF obviously decreased the immobility time in tail suspension test (TST) and forced swimming test (FST). CONCLUSION: In conclusion, CF exert the indeed anti-depressant effect. The inhibition of NMDAR-CaMKII-MAPKs signaling pathway, oxidative stress, and mitochondrial apoptotic pathways were involved in the anti-depressant effect of CF.

Structural and Binding Properties on Aß Mature Fibrils Due to the Histidine Tautomeric Effect.

Many studies have focused on histidine behaviors in misfolding diseases. However, histidine behaviors on mature fibrils are still unknown. In the current study, we investigated mature fibrils with various histidine states to understand the structural properties of the histidine tautomeric effect on mature fibrils. Our results show that substituting chain 1 with different histidine states affects Aß structural properties in A2, D7-G9, H14-Q15, S26-N27, and G33-G37 regions. The binding free energies with substituted fibrils were influenced not only along the axial direction, but also between duplex fibrils. Our results suggest that substituted (εδδ) preferentially disturbed the stability among the current mature fibrils. Further, H-bonded network differences indicate that twisted morphologies in mature fibrils are derived from the position and orientation of the imidazole ring in histidines. Our current study helps to elucidate histidine behaviors on mature fibrils, which will present opportunities to understand the misfolding mechanisms.

Genome-wide identification and comparative analysis of alternative splicing across four legume species.

MAIN CONCLUSION: Alternative splicing EVENTS were genome-wide identified for four legume species, and nitrogen fixation-related gene families and evolutionary analysis was also performed. Alternative splicing (AS) is a key regulatory mechanism that contributes to transcriptome and proteome diversity. Investigation of the genome-wide conserved AS events across different species will help with the understanding of the evolution of the functional diversity in legumes, allowing for genetic improvement. Genome-wide identification and characterization of AS were performed using the publically available mRNA, EST, and RNA-Seq data for four important legume species. A total of 15,165 AS genes in Glycine max, 6077 in Cicer arietinum, 7240 in Medicago truncatula, and 7358 in Lotus japonicus were identified. Intron retention (IntronR) was the dominant AS type among the identified events, with IntronR occurring from 53.76% in M. truncatula to 43.91% in C. arietinum. We identified 1159 AS genes that were conserved among four species. Furthermore, nine nitrogen fixation-related gene families with 237 genes were identified, and 80 of them were AS, accounting for the 43.48% in G. max and 27.78% in C. arietinum. An evolutionary analysis showed that these AS genes tended to be located adjacent to each other in the evolutionary tree and are unbalanced in the distribution in the sub-family. This study provides a foundation for future studies on transcription complexity, evolution, and the role of AS on plant functional regulation.

Development and application of EST-SSRs markers for analysis of genetic diversity in erect milkvetch (Astragalus adsurgens Pall.).

Erect milkvetch (Astragalus adsurgens Pall.) is a major legume forage plant widely grown in Northern China. However, the lack of molecular markers has limited its research into its genetic diversity and work on germplasm improvement. In this study, a total of 39,163 EST-SSR loci were identified from 30,262 unigene sequences in the erect milkvetch transcriptome using Illumina sequencing. Moreover, 22,367 EST-SSR primer pairs (PPs) were successfully designed. In addition, 100 PPs were synthesized and preliminarily screened in two accessions; of these, 90 were determined to be clear and stable EST-SSR markers. Fifty-one PPs were randomly selected in order to assess the genetic diversity of 27 erect milkvetch accessions. The average polymorphism information content of the 51 PPs was 0.682. Greater genetic diversity was detected in accessions from Inner Mongolia and in the group of landrace and wild erect milkvetch accessions. This study provides an important resource for germplasm improvement and genetic diversity analysis in erect milkvetch.