Expression of Anti-Lipopolysaccharide Factor Isoform 3 in Chlamydomonas reinhardtii Showing High Antimicrobial Activity

Antimicrobial peptides are a class of proteins with antibacterial functions. In this study, the anti-lipopolysaccharide factor isoform 3 gene (ALFPm3), encoding an antimicrobial peptide from Penaeus monodon with a super activity was expressed in Chlamydomonas reinhardtii, which would develop a microalga strain that can be used for the antimicrobial peptide production. To construct the expression cluster, namely pH2A-Pm3, the codon optimized ALFPm3 gene was fused with the ble reporter by 2A peptide and inserted into pH124 vector. The glass-bead method was performed to transform pH2A-Pm3 into C. reinhardtii CC-849. In addition to 8 μg/mL zeocin resistance selection, the C. reinhardtii transformants were further confirmed by genomic PCR and RT-PCR. Western blot analysis showed that the C. reinhardtii-derived ALFPm3 (cALFPm3) was successfully expressed in C. reinhardtii transformants and accounted for 0.35% of the total soluble protein (TSP). Furthermore, the results of antibacterial assay revealed that the cALFPm3 could significantly inhibit the growth of a variety of bacteria, including both Gram-negative bacteria and Gram-positive bacteria at a concentration of 0.77 μM. Especially, the inhibition could last longer than 24 h, which performed better than ampicillin. Hence, this study successfully developed a transgenic C. reinhardtii strain, which can produce the active ALFPm3 driven from P. monodon, providing a potential strategy to use C. reinhardtii as the cell factory to produce antimicrobial peptides.     

基于应用型人才培养教学中思政教育的融入策略实施

思政教育是高职院校教学的重点内容,对学生的思想、行为会产生直接影响,同时与工匠精神存在紧密关系,需要对其予以融合。为了提升高职机械类专业教学的质量,引入思政教育极为重要,其在推动学生全面发展方面也发挥出了重要作用。基于此,主要以机械基础类专业教学为例,分析了高职机械类专业课程与思政教育的联系为切入点。同时,通过调整教学思路与路径,结合机械发展史展开思政教育,引入大国重器实际研发案例,落实全面评价这些思政教育的融入策略。     

Exogenous application of allelopathic water extracts helps improving tolerance against terminal heat and drought stresses in bread wheat (Triticum aestivum L. Em. Thell.)

The allelopathic water extracts (AWEs) may help improve the tolerance of crop plants against abiotic stresses owing to the presence of the secondary metabolites (i.e., allelochemicals). We conducted four independent experiments to evaluate the influence of exogenous application of AWEs (applied through seed priming or foliage spray) in improving the terminal heat and drought tolerance in bread wheat. In all the experiments, two wheat cultivars, viz. Mairaj‐2008 (drought and heat tolerant) and Faisalabad‐2008 (drought and heat sensitive), were raised in pots. Both wheat cultivars were raised under ambient conditions in the wire house till leaf boot stage (booting) by maintaining the pots at 75% water‐holding capacity (WHC). Then, managed drought and heat stresses were imposed by maintaining the pots at 35% WHC, or shifting the pots inside the glass canopies (at 75% WHC), at booting, anthesis and the grain filling stages. Drought stress reduced the grain yield of wheat by 39%–49%. Foliar application of AWEs improved the grain yield of wheat by 26%–31%, while seed priming with AWEs improved the grain yield by 18%–26%, respectively, than drought stress. Terminal heat stress reduced the grain yield of wheat by 38%. Seed priming with AWEs improved the grain yield by 21%–27%; while foliar application of AWEs improved the grain yield by 25%–29% than the heat stress treatment. In conclusion, the exogenous application of AWEs improved the stay green, accumulation of proline, soluble phenolics and glycine betaine, which helped to stabilize the biological membranes and improved the tolerance against terminal drought and heat stresses.     

Calibration and validation of optical chlorophyll‐measuring devices for use in predicting crude protein concentration in tropical grass herbage

The objective of this experiment was to evaluate the Fieldscout CM 1000 NDVI and Yara N‐Tester as easy‐to‐use and cost‐effective tools for predicting foliar chlorophylls (a, b and total) and crude protein (CP) concentrations in herbage from three tropical grass species. Optical chlorophyll measurements were taken at three stages (4, 8 and 12 weeks) of regrowth maturity in Guinea grass (Panicum maximum) and Mulato II (Brachiaria hybrid) and at 6 and 12 weeks maturity in Paspalum spp (Paspalum atratum). Grass samples were harvested subsequent to optical measurements for laboratory analysis to determine CP and solvent‐extractable chlorophylls (a, b and total) concentrations. Optical chlorophyll measurements and CP concentrations were highly correlated (Yara N‐Tester: r² = 0·77–0·89; Fieldscout CM 1000 NDVI: r² = 0·52–0·84). Crude protein prediction models from the Yara N‐Tester and Fieldscout CM 1000 NDVI accounted for 70–89% and 44–73% CP variability, respectively, in Mulato II and Guinea grass. The Yara N‐tester produced more accurate and reliable CP estimates based on very high concordance correlation coefficient [CCC (0·73–0·91)] and low rMSPE, mean and regression bias. It is concluded that the Yara N‐Tester produces more accurate and reliable CP estimates of tropical pastures.     

Impact of grassland contract policy on soil organic carbon losses from alpine grassland on the Qinghai–Tibetan Plateau

Carbon storage in the soils on the Qinghai–Tibetan Plateau plays a very important role in the global carbon budget. In the 1990s, a policy of contracting collective grasslands to smaller units was implemented, resulting in a change from the traditional collective grassland management to two new management patterns: a multi‐household management pattern (MMP: grassland shared by several households without enclosures) and a single‐household management pattern (SMP: grassland enclosed and used by only one household). In 2016, 50 MMP and 54 SMP winter pastures on the Qinghai–Tibetan Plateau were sampled to assess the differences in soil organic carbon (SOC) between the two management patterns. Results showed that average SOC was significantly greater under MMP than under SMP, with an estimated 0.41 Mg C/ha/yr lost due to SMP following the new grassland contract. Based on the government's grassland policy, four grassland utilization scenarios were developed for both summer and winter pastures. We found that if the grassland were managed under SMP, likely C losses ranged between 0.31 × 10⁷ and 6.15 × 10⁷ Mg C/yr across the Qinghai–Tibetan Plateau relative to MMP, which more closely resembles pre‐1990s grassland management. Previous estimates of C losses have only considered land use change (with cover change) and ignored the impacts driven by land management pattern changes (without cover change). The new data suggest that C losses from the Qinghai–Tibetan Plateau are greater than previously estimated, and therefore that the grassland contract policy should be reviewed and SMP households should be encouraged to reunite into the MMP. These findings have potential implications for land management strategies not only on the Qinghai–Tibetan Plateau but also other grazing regions globally where such practices may exist.     

Utilizing a DUALEM‐421 and inversion modelling to map baseline soil salinity along toposequences in the Hunter Valley Wine district

In the oldest commercial wine district of Australia, the Hunter Valley, there is the threat of soil salinization because marine sediments underlie the area. To understand the risk requires information about the spatial distribution of soil properties. Electromagnetic (EM) induction instruments have been used to identify and map the spatial variation of average soil salinity to a certain depth. However, soils vary with depth dependent on soil forming factors. We collected data from a single‐frequency and multiple‐coil DUALEM‐421 along a toposequence. We inverted this data using EM4Soil software and evaluated the resultant 2‐dimensional model of true electrical conductivity (σ – mS/m) with depth against electrical conductivity of saturated soil pastes (EC_p – dS/m). Using a fitted linear regression (LR) model calibration approach and by varying the forward model (cumulative function‐CF and full solution‐FS), inversion algorithm (S1 and S2), damping factor (λ) and number of arrays, we determined a suitable electromagnetic conductivity image (EMCI), which was optimal (R² = 0.82) when using the full solution, S2, λ = 3.6 and all six coil arrays. We conducted an uncertainty analysis of the LR model used to estimate the electrical conductivity of the saturated soil‐paste extract (EC_e – dS/m). Our interpretation based on estimates of EC_e suggests the approach can identify differences in salinity, how these vary with parent material and how topography influences salt distribution. The results provide information leading to insights into how soil forming factors and agricultural practices influence salinity down a toposequence and how this can guide soil management practices.     

Comparative effect of advanced soy products or corn protein concentrate with porcine meal on growth,body composition,and distal intestine histology of Florida pompano,Trachinotus carolinus

The present study was designed to investigate the effects of diets containing advanced soy products (enzyme‐treated soy and fermented soy) or corn protein concentrate (CPC) in combination with porcine meal (PM) to completely replace poultry byproduct meal (PBM) on growth performance, body composition, and distal intestine histology of Florida pompano, Trachinotus carolinus. Four experimental diets were formulated to be isonitrogenous and isolipidic, to contain 400 g/kg crude protein and 80 g/kg lipid. A reference diet (PBM diet [PBMD]) contained 150 g/kg PBM and 495 g/kg soybean meal (SBM), and three test diets were formulated replacing PBM with 15 g/kg of CPC (CPC diet [CPCD]) or replacing all SBM and PBM with 535 g/kg fermented soy (fermented soybean meal diet [FSBMD]) or 451.3 g/kg enzyme‐treated soy (enzyme‐treated soybean meal diet [ESBMD]). All three test diets were supplemented with 38 g/kg of PM. Diets were fed based on a percentage of bodyweight adjusted after sampling the fish every 2 weeks to triplicate groups of Florida pompano juveniles (mean weight 8.06 ± 0.22 g). After 8 weeks of feeding, fish fed CPCD and ESBMD performed equally well in terms of final body weight, thermal growth coefficient, and percentage weight gain in comparison to fish fed PBMD. In all cases, feeding FSBMD resulted in poor feed conversion and lower feed intake compared to other treatments. Protein retention efficiency, whole‐body proximate composition, phosphorus, sulfur, potassium, magnesium, calcium, sodium, and zinc contents were not significantly influenced by the dietary treatments. The results obtained in the present histological study showed no significant differences in the thickness of serous layer, muscular layer, and submucosal layer of the intestine among treatments. Fish fed CPCD showed a significant widening of the lamina propria with an increase of cellular infiltration and higher presence of goblet cells compared to other dietary treatment. Based on these results, 451 g/kg ESBM or combination of 150 g/kg of CPC and 495 g/kg SBM supplemented with 38 g/kg PM can be utilized to develop a practical diet for juvenile Florida pompano without impacting growth, nutritive parameters, and several distal intestine health parameters.     

Beaver dams,streamflow complexity,and the distribution of a rare minnow,Lepidomeda copei

Freshwater fishes are threatened globally, and often too little is known about threatened species to effectively guide their conservation. Habitat complexity is linked to fish species diversity and persistence, and degraded streams often lack habitat complexity. Beaver Castor spp., in turn, have been used to restore streams and increase habitat complexity. The northern leatherside chub Lepidomeda copei is a rare, small‐bodied, drift‐feeding minnow that has anecdotally been observed to use complex habitats associated with beaver dams in the western United States. To investigate this anecdote, we conducted fish and habitat surveys, the latter focusing on quantifying habitat complexity, in a sub‐basin of the Upper Snake River Basin in the USA. Complementary generalised linear model and path analyses revealed that northern leatherside chub occurred more often at sites with complex streamflows, and streamflows were more complex when beaver dams were present and pools were deeper. Northern leatherside chubs were also more likely to occur when temperatures were warmer, aquatic macrophytes were abundant and stream channels were narrow and deep. The linkage between chubs, complex streamflows and beaver dams needs to be evaluated more broadly to completely understand its role in the rangewide status of the species. However, it does suggests that increased use of beaver reintroductions and dam analogues for stream restoration could be a boon for the northern leatherside chub, but such efforts should be monitored to determine their effectiveness to help adapt beaver‐based restoration approaches to best benefit the species.     

Carbohydrate and crude protein fractions in perennial ryegrass as affected by defoliation frequency and nitrogen application rate

The objective of this study was to evaluate the effects of defoliation frequency (either at two‐ or three‐leaf stage) and nitrogen (N) application rate (0, 75, 150, 300, 450 kg N ha^?1 year^?1) on herbage carbohydrate and crude protein (CP) fractions, and the water‐soluble carbohydrate‐to‐protein ratio (WSC:CP) in perennial ryegrass swards. Crude protein fractions were analysed according to the Cornell carbohydrate and protein system. Carbohydrate fractions were analysed by ultra‐high‐performance liquid chromatography. Sward defoliation at two‐leaf stage increased the total CP, reduced the buffer‐soluble CP fractions and decreased carbohydrate fractions of herbage (P < 0·001). The effect of defoliation frequency was less marked during early spring and autumn (P < 0·001) than for the rest of the seasons. An increase in N application rate was negatively associated with WSC, fructans and neutral detergent fibre (P < 0·001), and positively associated with CP and nitrate (N‐NO₃) contents of herbage. Nitrogen application rate did not affect CP fractions of herbage (P > 0·05). The fluctuations in CP and WSC contents of herbage resulted in lower WSC:CP ratios during early spring and autumn (0·45:1 and 0·75:1 respectively) than in late spring (1·11:1). The herbage WSC:CP ratio was greater (P < 0·001) at the three‐leaf than the two‐leaf defoliation stage and declined as the N application increased in all seasons (P < 0·001). The results of this study indicate that CP and carbohydrate fractions of herbage can be manipulated by sward defoliation frequency and N application rate. The magnitude of these effects, however, may vary with the season.     

Neuroprotective Metabolites from Vietnamese Marine Derived Fungi of Aspergillus and Penicillium Genera

Low molecular weight secondary metabolites of marine fungi Aspergillus flocculosus, Aspergillus terreus and Penicillium sp. from Van Phong and Nha Trang Bays (Vietnam) were studied and a number of polyketides, bis-indole quinones and terpenoids were isolated. The structures of the isolated compounds were determined by 1D and 2D NMR and HR-ESI-MS techniques. Stereochemistry of some compounds was established based on ECD data. A chemical structure of asterriquinone F (6) was thoroughly described for the first time. Anthraquinone (13) was firstly obtained from a natural source. Neuroprotective influences of the isolated compounds against 6-OHDA, paraquat and rotenone toxicity were investigated. 4-Hydroxyscytalone (1), 4-hydroxy-6-dehydroxyscytalone (2) and demethylcitreoviranol (3) have shown significant increasing of paraquat- and rotenone-treated Neuro-2a cell viability and anti-ROS activity.