Construction of a Homologous Selectable Marker Gene for Lentinula edodes Transformation

We cloned a gene for the iron sulfur protein (Ip) subunit from an edible mushroom, Lentinula edodes, and introduced a point mutation that confers carboxin resistance into it. The mutant gene successfully transformed L. edodes with high efficiency (9 transformants/2.5 μg vector DNA). Restriction enzyme-mediated integration (REMI) increased the transformation efficiency by about two-fold.     

Scale‐down simulators for mammalian cell culture as tools to access the impact of inhomogeneities occurring in large‐scale bioreactors

During the scale‐up of a bioprocess, not all characteristics of the process can be kept constant throughout the different scales. This typically results in increased mixing times with increasing reactor volumes. The poor mixing leads in turn to the formation of concentration gradients throughout the reactor and exposes cells to varying external conditions based on their location in the bioreactor. This can affect process performance and complicate process scale‐up. Scale‐down simulators, which aim at replicating the large‐scale environment, expose the cells to changing environmental conditions. This has the potential to reveal adaptation mechanisms, which cells are using to adjust to rapidly fluctuating environmental conditions and can identify possible root causes for difficulties maintaining similar process performance at different scales. This understanding is of utmost importance in process validation. Additionally, these simulators also have the potential to be used for selecting cells, which are most robust when encountering changing extracellular conditions. The aim of this review is to summarize recent work in this interesting and promising area with the focus on mammalian bioprocesses, since microbial processes have been extensively reviewed.     

Development,characterization, and application of a 2‐Compartment system to investigate the impact of pH inhomogeneities in large‐scale CHO‐based processes

Large‐scale bioreactors for the production of monoclonal antibodies reach volumes of up to 25 000 L. With increasing bioreactor size, mixing is however affected negatively, resulting in the formation of gradients throughout the reactor. These gradients can adversely affect process performance at large scale. Since mammalian cells are sensitive to changes in pH, this study investigated the effects of pH gradients on process performance. A 2‐Compartment System was established for this purpose to expose only a fraction of the cell population to pH excursions and thereby mimicking a large‐scale bioreactor. Cells were exposed to repeated pH amplitudes of 0.4 units (pH 7.3), which resulted in decreased viable cell counts, as well as the inhibition of the lactate metabolic shift. These effects were furthermore accompanied by increased absolute lactate levels. Continuous assessment of molecular attributes of the expressed target protein revealed that subunit assembly or N‐glycosylation patterns were only slightly influenced by the pH excursions. The exposure of more cells to the same pH amplitudes further impaired process performance, indicating this is an important factor, which influences the impact of pH inhomogeneity. This knowledge can aid in the design of pH control strategies to minimize the effects of pH inhomogeneity in large‐scale bioreactors.     

Membrane‐Free Zn/MnO2 Flow Battery for Large‐Scale Energy Storage

The traditional Zn/MnO₂ battery has attracted great interest due to its low cost, high safety, high output voltage, and environmental friendliness. However, it remains a big challenge to achieve long‐term stability, mainly owing to the poor reversibility of the cathode reaction. Different from previous studies where the cathode redox reaction of MnO₂/MnOOH is in solid state with limited reversibility, here a new aqueous rechargeable Zn/MnO₂ flow battery is constructed with dissolution–precipitation reactions in both cathodes (Mn²⁺/MnO₂) and anodes (Zn²⁺/Zn), which allow mixing of anolyte and catholyte into only one electrolyte and remove the requirement for an ion selective membrane for cost reduction. Impressively, this new battery exhibits a high discharge voltage of ≈1.78 V, good rate capability (10C discharge), and excellent cycling stability (1000 cycles without decay) at the areal capacity ranging from 0.5 to 2 mAh cm^‐2. More importantly, this battery can be readily enlarged to a bench scale flow cell of 1.2 Ah with good capacity retention of 89.7% at the 500th cycle, displaying great potential for large‐scale energy storage.     

2D Materials for Large‐Area Flexible Thermoelectric Devices

The rapid development of the concept of the “Internet of Things (IoT)” requires wearable devices with maintenance‐free batteries, and thermoelectric energy conversion based on large‐area flexible materials has attracted much attention. Among large‐area flexible materials, 2D materials, such as graphene and related materials, are promising for thermoelectric applications due to their excellent transport properties and large power factors. In this Review, both single‐crystalline and polycrystalline 2D materials are surveyed using the experimental reports on thermoelectric devices of graphene, black phosphorus, transition metal dichalcogenides, and other 2D materials. In particular, their carrier‐density dependent thermoelectric properties and power factors maximized by Fermi level tuning techniques are focused. The comparison of the relevant performances between 2D materials and commonly used thermoelectric materials reveals the significantly enhanced power factors in 2D materials. Moreover, the current progress in thermoelectric module applications using large‐area 2D material thin films is summarized, which consequently offers great potential for the use of 2D materials in large‐area flexible thermoelectric device applications. Finally, important remaining issues and future perspectives, such as preparation methods, thermal transports, device designs, and promising effects in 2D materials, are discussed.     

Effect of litter size,milk replacer and housing on production results of hyper-prolific sows

The modern hyper-prolific sow gives birth to more piglets than she has functional teats (in the following called supernumerary piglets). The aim of the present study was (1) to investigate the production consequences of hyper-prolific sows rearing supernumerary piglets equal to the mean live-born litter size, and (2) investigate whether potential negative effects on survival and growth could be alleviated by providing access to milk replacer and/or providing easier access to the udder (by loose housing). At day 1 (D1) postpartum (pp), 93 litters were standardised to 14 or 17 piglets (LS14/LS17) after which no piglets were moved between sows leading to decreased litter size if piglets died. Litters were provided with or without milk replacer in milk cups (+MILK/−MILK), and sows were either crated or loose housed (CRATE/LOOSE) in a 2 × 2 × 2 factorial design. Piglet mortality was higher in LS17 compared to LS14 (P < 0.01; OR = 2.0), higher in −MILK compared to +MILK (P = 0.01; OR = 1.2) and higher in LOOSE compared to CRATE (P = 0.02; OR = 1.8). This study showed that sow rearing of supernumerary piglets while supplying with milk replacer can increase piglet survival. It also showed that early mortality before piglets learned to drink milk replacer posed a challenge using this automatic milk replacer system. An interaction between access to milk replacer and the standardised litter size D1 affected litter weight (P < 0.01) and piglet weight day 28 (D28) (P = 0.03). The highest litter weight D28 was found in LS17 +MILK (P < 0.01) but with a lower individual piglet weight than in LS14 −MILK. Piglet weight D28 was higher in LS14 −MILK compared to LS17 regardless of access to milk replacer. Heterogeneity in piglet weight within litters D28 was larger in LS17 (P = 0.03) but could be reduced with +MILK in CRATE (P < 0.01). No effects were found on sow weight loss and feed intake (P > 0.05). In conclusion, the results showed that sows cannot rear the supernumerary piglets without further management interventions to reduce mortality. Supplying supernumerary piglets equal to the mean live-born litter size of hyper-prolific sows with milk replacer can from results of this study be an alternative strategy to the use of nurse sows.     

Leaf size of woody dicots predicts ecosystem primary productivity

A key challenge in ecology is to understand the relationships between organismal traits and ecosystem processes. Here, with a novel dataset of leaf length and width for 10 480 woody dicots in China and 2374 in North America, we show that the variation in community mean leaf size is highly correlated with the variation in climate and ecosystem primary productivity, independent of plant life form. These relationships likely reflect how natural selection modifies leaf size across varying climates in conjunction with how climate influences canopy total leaf area. We find that the leaf size?primary productivity functions based on the Chinese dataset can predict productivity in North America and vice‐versa. In addition to advancing understanding of the relationship between a climate‐driven trait and ecosystem functioning, our findings suggest that leaf size can also be a promising tool in palaeoecology for scaling from fossil leaves to palaeo‐primary productivity of woody ecosystems.     

Increased Rubisco content in maize mitigates chilling stress and speeds recovery

Many C₄ plants, including maize, perform poorly under chilling conditions. This phenomenon has been linked in part to decreased Rubisco abundance at lower temperatures. An exception to this is chilling‐tolerant Miscanthus, which is able to maintain Rubisco protein content under such conditions. The goal of this study was to investigate whether increasing Rubisco content in maize could improve performance during or following chilling stress. Here, we demonstrate that transgenic lines overexpressing Rubisco large and small subunits and the Rubisco assembly factor RAF1 (RAF1‐LSSS), which have increased Rubisco content and growth under control conditions, maintain increased Rubisco content and growth during chilling stress. RAF1‐LSSS plants exhibited 12% higher CO₂ assimilation relative to nontransgenic controls under control growth conditions, and a 17% differential after 2 weeks of chilling stress, although assimilation rates of all genotypes were ~50% lower in chilling conditions. Chlorophyll fluorescence measurements showed RAF1‐LSSS and WT plants had similar rates of photochemical quenching during chilling, suggesting Rubisco may not be the primary limiting factor that leads to poor performance in maize under chilling conditions. In contrast, RAF1‐LSSS had improved photochemical quenching before and after chilling stress, suggesting that increased Rubisco may help plants recover faster from chilling conditions. Relatively increased leaf area, dry weight and plant height observed before chilling in RAF1‐LSSS were also maintained during chilling. Together, these results demonstrate that an increase in Rubisco content allows maize plants to better cope with chilling stress and also improves their subsequent recovery, yet additional modifications are required to engineer chilling tolerance in maize.     

Photorespiration in the context of Rubisco biochemistry,CO2 diffusion and metabolism

Photorespiratory metabolism is essential for plants to maintain functional photosynthesis in an oxygen‐containing environment. Because the oxygenation reaction of Rubisco is followed by the loss of previously fixed carbon, photorespiration is often considered a wasteful process and considerable efforts are aimed at minimizing the negative impact of photorespiration on the plant’s carbon uptake. However, the photorespiratory pathway has also many positive aspects, as it is well integrated within other metabolic processes, such as nitrogen assimilation and C₁ metabolism, and it is important for maintaining the redox balance of the plant. The overall effect of photorespiratory carbon loss on the net CO₂ fixation of the plant is also strongly influenced by the physiology of the leaf related to CO₂ diffusion. This review outlines the distinction between Rubisco oxygenation and photorespiratory CO₂ release as a basis to evaluate the costs and benefits of photorespiration.