Combined use of dbcAMP and IBMX minimizes the damage induced by a long‐term artificial meiotic arrest in mouse germinal vesicle oocytes

Phosphodiesterase (PDE)‐mediated reduction of cyclic adenosine monophosphate (cAMP) activity can initiate germinal vesicle (GV) breakdown in mammalian oocytes. It is crucial to maintain oocytes at the GV stage for a long period to analyze meiotic resumption in vitro. Meiotic resumption can be reversibly inhibited in isolated oocytes by cAMP modulator forskolin, cAMP analog dibutyryl cAMP (dbcAMP), or PDE inhibitors, milrinone (Mil), Cilostazol (CLZ), and 3‐isobutyl‐1‐methylxanthine (IBMX). However, these chemicals negatively affect oocyte development and maturation when used independently. Here, we used ICR mice to develop a model that could maintain GV‐stage arrest with minimal toxic effects on subsequent oocyte and embryonic development. We identified optimal concentrations of forskolin, dbcAMP, Mil, CLZ, IBMX, and their combinations for inhibiting oocyte meiotic resumption. Adverse effects were assessed according to subsequent development potential, including meiotic resumption after washout, first polar body extrusion, early apoptosis, double‐strand DNA breaks, mitochondrial distribution, adenosine triphosphate levels, and embryonic development. Incubation with a combination of 50.0 μM dbcAMP and 10.0 μM IBMX efficiently inhibited meiotic resumption in GV‐stage oocytes, with low toxicity on subsequent oocyte maturation and embryonic development. This work proposes a novel method with reduced toxicity to effectively arrest and maintain mouse oocytes at the GV stage.     

Changes in soil properties and resistance to concentrated flow across a 25‐year passive restoration chronosequence of grasslands on the Chinese Loess Plateau

Revegetation represents an effective measure for preventing soil erosion on the Loess Plateau. However, the effects of revegetation‐induced changes in soil and root properties on soil resistance to concentrated flow erosion (SRC) remain unclear. This study sampled soils and roots across a 25‐year chronosequence from farmland to grasslands of different ages (3, 7, 10, 18, and 25 years) to quantify variations in soil and root properties (soil bulk density, SBD; soil disintegration rate, SDR; saturated hydraulic conductivity, SHC; organic matter content, OMC; water‐stable aggregate, WSA; mean weight diameter, MWD; root mass density, RMD; root length density, RLD; and root surface area density, RSAD) and their effects on SRC. Farmland and grassland SRCs were obtained using a hydraulic flume. Soil properties and root density gradually improved with restoration time. In terms of the comprehensive soil property index calculated via principal component analysis, grassland values were 0.66 to 1.94 times greater than farmland values. Grassland SRCs increased and gradually stabilized (>18 years) over time and were 1.60 to 8.26 times greater than farmland SRC. SRC improvement was significantly related to increases in OMC, SHC, WSA, and MWD and decreases in SBD and SDR over time. SRC was effectively simulated by the Hill curve of RMD, RLD, and RSAD. SDR, SHC, and RMD (0.5–1.0 mm) affected SRC the most. This study scientifically describes how revegetation improves soil quality and soil resistance to flow erosion, and suggests that vegetations rich in 0.5–1.0 mm roots should be preferred during revegetation.     

Platelet‐rich plasma inhibits osteoblast apoptosis and actin cytoskeleton disruption induced by gingipains through upregulating integrin β1

The aim of this study was to explore the effects of platelet‐rich plasma on gingipain‐caused changes in cell morphology and apoptosis of osteoblasts. Mouse osteoblasts MC3T3‐E1 cells were treated with gingipain extracts from Porphyromonas gingivalis in the presence or absence of platelet‐rich plasma. Apoptosis was detected with terminal deoxynucleotidyl transferase‐mediated dUTP nick‐end labeling staining. F‐actin was determined by phalloidin‐fluorescent staining and observed under confocal microscopy. Western blot analysis was used to detect integrin β1, F‐actin, and G‐actin protein expressions. A knocking down approach was used to determine the role of integrin β1. The platelet‐rich plasma protected osteoblasts from gingipain‐induced apoptosis in a dose‐dependent manner, accompanied by upregulation of integrin β1. Platelet‐rich plasma reversed the loss of F‐actin integrity and decrease of F‐actin/G‐actin ratio in osteoblasts in the presence of gingipains. By contrast, the effects of platelet‐rich plasma were abrogated by knockdown of integrin β1. The platelet‐rich plasma failed to reduce cell apoptosis and reorganize the cytoskeleton after knockdown of integrin β1. In conclusion, platelet‐rich plasma inhibits gingipain‐induced osteoblast apoptosis and actin cytoskeleton disruption by upregulating integrin β1 expression.     

Warming counteracts defoliation‐induced mismatch by increasing herbivore‐plant phenological synchrony

Climate change is altering phenology; however, the magnitude of this change varies among taxa. Compared with phenological mismatch between plants and herbivores, synchronization due to climate has been less explored, despite its potential implications for trophic interactions. The earlier budburst induced by defoliation is a phenological strategy for plants against herbivores. Here, we tested whether warming can counteract defoliation‐induced mismatch by increasing herbivore‐plant phenological synchrony. We compared the larval phenology of spruce budworm and budburst in balsam fir, black spruce, and white spruce saplings subjected to defoliation in a controlled environment at temperatures of 12, 17, and 22°C. Budburst in defoliated saplings occurred 6–24 days earlier than in the controls, thus mismatching needle development from larval feeding. This mismatch decreased to only 3–7 days, however, when temperatures warmed by 5 and 10°C, leading to a resynchronization of the host with spruce budworm larvae. The increasing synchrony under warming counteracts the defoliation‐induced mismatch, disrupting trophic interactions and energy flow between forest ecosystem and insect populations. Our results suggest that the predicted warming may improve food quality and provide better growth conditions for larval development, thus promoting longer or more intense insect outbreaks in the future.     

Overexpression of the persimmon abscisic acid β‐glucosidase gene (DkBG1) alters fruit ripening in transgenic tomato

β‐Glucosidases (BG) are present in many plant tissues. Among these, abscisic acid (ABA) β‐glucosidases are thought to take part in the adjustment of cellular ABA levels, however the role of ABA‐BG in fruits is still unclear. In this study, through RNA‐seq analysis of persimmon fruit, 10 full‐length DkBG genes were isolated and were all found to be expressed. In particular, DkBG1 was highly expressed in persimmon fruits with a maximum expression 95 days after full bloom (DAFD). We verified that, in vitro, DkBG1 protein can hydrolyze ABA‐glucose ester (ABA‐GE) to release free ABA. Compared with wild‐type, tomato plants that overexpressed DkBG1 significantly upregulated the expression of ABA receptor PYL3/7 genes and showed typical symptoms of ABA hypersensitivity in fruits. DkBG1 overexpression (DkBG1‐OE) accelerated fruit ripening onset by 3–4 days by increasing ABA levels at the pre‐breaker stage and induced early ethylene release compared with wild‐type fruits. DkBG1‐OE altered the expression of ripening regulator NON‐RIPENING (NOR) and its target genes; this in turn altered fruit quality traits such as coloration. Our results demonstrated that DkBG1 plays an important role in fruit ripening and quality by adjusting ABA levels via hydrolysis of ABA‐GE.     

基于无人机低空遥感的荒漠植被覆盖度与归一化植被指数验证及其对水热梯度的响应

为了验证在荒漠地区MODIS-NDVI产品的精度以及为在气候变化背景下荒漠草地的科学管理提供依据,本文利用无人机低空遥感研究了干旱荒漠地区植被覆盖度(FVC)和归一化植被指数(NDVI)对水、热梯度的响应规律。在内蒙古阿拉善荒漠地区的100个样点采用GreenSeeker手持光谱仪获得NDVI值(NDVI_R),通过MODIS-NDVI数据产品提取每个样点的NDVI(NDVI_M),借助NDVI_R验证NDVI_M的精确度;通过无人机遥感手段获得每个采样点的FVC(FVC_U),利用像元二分模型反演每个样点的FVC(FVC_M),借助FVC_U验证FVC_M的精确度;并结合气象数据探讨基于无人机低空遥感的荒漠地区FVC和NDVI对水热梯度的响应。结果表明: MODIS-NDVI数据产品能够反映阿拉善地区的NDVI,精确度为84.2%,但比真实值高15.7%;FVC_M能够反映阿拉善地区的FVC状况,精确度为83.1%,但比真实值低14.8%;不同采集方式获得的NDVI受气象因子的影响程度不同,NDVI不仅受气温和降雨的影响,也受地温、蒸发量以及两者相互作用的影响,由于受大气影响程度不同, NDVI_M受地温、蒸发量、降水量的影响比NDVI_R大,NDVI_R受气温的影响比NDVI_M大。在阿拉善地区研究FVC随水热梯度的变化不仅要考虑降水量和气温,还应考虑蒸发量、地温以及气象因子之间相互作用的影响,其中,气温与降雨、蒸发量与地温以及气温与蒸发量之间相互作用对FVC_U的影响较大。     

Efficient deletion of LoxP-flanked selectable marker genes from the genome of transgenic pigs by an engineered Cre recombinase

Genetically modified (GM) pigs hold great promises for pig genetic improvement, human health and life science. When GM pigs are produced, selectable marker genes (SMGs) are usually introduced into their genomes for host cell or animal recognition. However, the SMGs that remain in GM pigs might have multiple side effects. To avoid the possible side effects caused by the SMGs, they should be removed from the genome of GM pigs before their commercialization. The Cre recombinase is commonly used to delete the LoxP sites-flanked SMGs from the genome of GM animals. Although SMG-free GM pigs have been generated by Cre-mediated recombination, more efficient and cost-effective approaches are essential for the commercialization of SMG-free GM pigs. In this article we describe the production of a recombinant Cre protein containing a cell-penetrating and a nuclear localization signal peptide in one construct. This engineered Cre enzyme can efficiently excise the LoxP-flanked SMGs in cultured fibroblasts isolated from a transgenic pig, which then can be used as nuclear donor cells to generate live SMG-free GM pigs harboring a desired transgene by somatic cell nuclear transfer. This study describes an efficient and far-less costly method for production of SMG-free GM pigs.