Reconstituted low density lipoprotein: A vehicle for the delivery of hydrophobic fluorescent probes to cells

Previous studies have shown that the cholesteryl ester core of plasma low density lipoprotein (LDL) can be extracted with heptane and replaced with a variety of hydrophobic molecules. In the present report we use this reconstitution technique to incorporate two fluorescent probes, 3-pyrenemethyl-23, 24-dinor-5-cholen-22-oate-3β-yl oleate (PMCA oleate) and dioleyl fluorescein, into heptane-extracted LDL. Both fluorescent lipoprotein preparations were shown to be useful probes for visualizing the receptor-mediated endocytosis of LDL in cultured human fibroblasts. When normal fibroblasts were incubated at 37°C with either of the fluorescent LDL preparations, fluorescent granules accumulated in the perinuclear region of the cell. In contrast, fibroblasts from patients with the homozygous form of familial hypercholesterolemia (FH) that lack functional LDL receptors did not accumulate visible fluorescent granules when incubated with the fluorescent reconstituted LDL. A fluorescence-activated cell sorter was used to quantify the fluorescence intensity of individual cells that had been incubated with LDL reconstituted with dioleyl fluorescein. With this technique a population of normal fibroblasts could be distinguished from a population of FH fibroblasts. The current studies demonstrate the feasibility of using fluorescent reconstituted LDL in conjunction with the cell sorter to isolate mutant cells lacking functional LDL receptors.     

Chromosome-level genome assembly and characterization of Sophora Japonica

Sophora japonica is a medium-size deciduous tree belonging to Leguminosae family and famous for its high ecological, economic and medicinal value. Here, we reveal a draft genome of S. japonica, which was ∼511.49 Mb long (contig N50 size of 17.34 Mb) based on Illumina, Nanopore and Hi-C data. We reliably assembled 110 contigs into 14 chromosomes, representing 91.62% of the total genome, with an improved N50 size of 31.32 Mb based on Hi-C data. Further investigation identified 271.76 Mb (53.13%) of repetitive sequences and 31,000 protein-coding genes, of which 30,721 (99.1%) were functionally annotated. Phylogenetic analysis indicates that S. japonica separated from Arabidopsis thaliana and Glycine max ∼107.53 and 61.24 million years ago, respectively. We detected evidence of species-specific and common-legume whole-genome duplication events in S. japonica. We further found that multiple TF families (e.g. BBX and PAL) have expanded in S. japonica, which might have led to its enhanced tolerance to abiotic stress. In addition, S. japonica harbours more genes involved in the lignin and cellulose biosynthesis pathways than the other two species. Finally, population genomic analyses revealed no obvious differentiation among geographical groups and the effective population size continuously declined since 2 Ma. Our genomic data provide a powerful comparative framework to study the adaptation, evolution and active ingredients biosynthesis in S. japonica. More importantly, our high-quality S. japonica genome is important for elucidating the biosynthesis of its main bioactive components, and improving its production and/or processing.     

基于RS/GIS公路路域水土流失动态变化的研究——以榆靖高速公路为例

基于RS数据源,利用ERDAS和ArcGIS软件技术,对榆靖高速公路两侧300m范围水土流失动态变化开展研究。通过遥感图像数字处理方法,提取对水土流失起主导作用的植被覆盖度、土地利用类型、沟谷密度等因子;在矢量化等高线数据的基础上生成数字高程模型(DEM),提取地形坡度因子,将这4个因子在GIS中进行空间叠加分析。参照《土壤侵蚀分级标准》,生成研究区不同时段水土流失强度等级图,利用GIS属性统计功能对公路沿线各等级水土流失区面积进行统计,参照研究区不同侵蚀类型的土壤侵蚀模数,得出1996年和2006年公路沿线各流失区的水土流失量,并对公路沿线水土流失动态变化进行分析。结果表明：榆靖高速公路修建后沿线600m范围水土流失总量减少19.26万t/a,水土流失呈减弱趋势。公路施工期导致水土流失增强的各种因素随着营运期固化路面的形成而逐渐消失,防沙固沙和生态恢复等措施有效改善了公路路域生态环境,对公路沿线水土流失防治起到了积极作用。     

Rapid improvement of rice eating and cooking quality through gene editing toward glutelin as target

Rice eating and cooking quality(ECQ) is a major concern of breeders and consumers, determining market competitiveness worldwide. Rice grain protein content(GPC) is negatively related to ECQ,making it possible to improve ECQ by manipulating GPC. However, GPC is genetically complex and sensitive to environmental conditions; therefore, little progress has been made in traditional breeding for ECQ. Here, we report that CRISPR/Cas9-mediated knockout of genes encoding the grain storage protein gluteli...     

三峡库区9种植物种子萌发特性及其在植被恢复中的意义

三峡大坝蓄水后形成的库区消涨带面临植被消亡、生态退化等问题。为了筛选出适用于库区消涨带植被恢复的植物, 将9种1年生植物种子放置在库区消涨带不同海拔进行水淹(W 165-8 m, 121 d;W 155-18 m, 230 d;W 147-26 m, 271 d), 然后在实验室条件下进行萌发, 研究在消涨带淹水胁迫下这些种子的萌发特性。结果表明: (1) 除马唐(Digitaria sanguinalis)、小蓬草(Conyza canadensis)、金色狗尾草(Setaria glauca)种子在各条件下萌发率都较低外, 不同水淹条件对萌发率影响不同: 与新鲜种子相比, W 165水淹后, 旱稗(Echinochloa hispidula)和婆婆针(Bidens bipinnata)种子萌发率显著上升, 其余种子萌发率均显著下降; W 155水淹后, 所有种子的萌发率都显著下降且只有鱧肠(Eclipta prostrate)、黄花蒿(Artemisia annua)、合萌(Aeschynomene indica)3个物种有萌发, 萌发率分别为11.0%、7.3%和2.7%; W 147水淹后, 旱稗和婆婆针种子萌发率显著上升, 鱧肠种子无显著差异, 其余种子萌发率显著下降。(2) 鱧肠、黄花蒿、婆婆针和旱稗种子比其它物种更耐水淹。W 165水淹后, 鱧肠、黄花蒿、婆婆针、旱稗种子萌发率分别为44.7%、42%、20.7%和4.3%, W 147水淹后分别为76.3%、23%、15%和26.3%, 高于其他物种。(3) 水淹后种子萌发时间格局也受到影响, 大部分种子起始萌发时间推迟、萌发速度变慢。鱧肠、黄花蒿、婆婆针和旱稗的种子对三峡库区消涨带的水淹胁迫具有一定的适应能力, 可根据它们对水淹条件适应能力的差异在消涨带不同海拔高度进行植被恢复。     

Circular RNA circ‐RCCD promotes cardiomyocyte differentiation in mouse embryo development via recruiting YY1 to the promoter of MyD88

Congenital heart disease (CHD) is the most common birth defect, affecting approximately 1% of live births. Genetic and environmental factors are leading factors to CHD, but the mechanism of CHD pathogenesis remains unclear. Circular RNAs (circRNAs) are kinds of endogenous non‐coding RNAs (ncRNAs) involved in a variety of physiological and pathological processes, especially in heart diseases. In this study, three significant differently expressed circRNA between maternal embryonic day (E) E13 and E17 was found by microarray assay. Among them, the content of circ‐RCCD increases with the development of heart and was enriched in primary cardiomyocytes of different species, which arouses our attention. Functional experiments revealed that inhibition of circ‐RCCD dramatically suppressed the formation of beating cell clusters, the fluorescence intensity of cardiac differentiation marker MF20, and the expression of the myocardial‐specific markers CTnT, Mef2c, and GATA4. Next, we found that circ‐RCCD was involved in cardiomyocyte differentiation through negative regulation of MyD88 expression. Further experiments proved that circ‐RCCD inhibited MyD88 levels by recruiting YY1 to the promoter of MyD88; circ‐RCCD inhibited nuclear translocation of YY1. These results reported that circ‐RCCD promoted cardiomyocyte differentiation by recruiting YY1 to the promoter of MyD88. And, this study provided a potential role and molecular mechanism of circ‐RCCD as a target for the treatment of CHD.     

A secreted ribonuclease effector from Verticillium dahliae localizes in the plant nucleus to modulate host immunity

The arms race between fungal pathogens and plant hosts involves recognition of fungal effectors to induce host immunity. Although various fungal effectors have been identified, the effector functions of ribonucleases are largely unknown. Herein, we identified a ribonuclease secreted by Verticillium dahliae (VdRTX1) that translocates into the plant nucleus to modulate immunity. The activity of VdRTX1 causes hypersensitive response (HR)‐related cell death in Nicotiana benthamiana and cotton. VdRTX1 possesses a signal peptide but is unlikely to be an apoplastic effector because its nuclear localization in the plant is necessary for cell death induction. Knockout of VdRTX1 significantly enhanced V. dahliae virulence on tobacco while V. dahliae employs the known suppressor VdCBM1 to escape the immunity induced by VdRTX1. VdRTX1 homologs are widely distributed in fungi but transient expression of 24 homologs from other fungi did not yield cell death induction, suggesting that this function is specific to the VdRTX1 in V. dahliae. Expression of site‐directed mutants of VdRTX1 in N. benthamiana leaves revealed conserved ligand‐binding sites that are important for VdRTX1 function in inducing cell death. Thus, VdRTX1 functions as a unique HR‐inducing effector in V. dahliae that contributes to the activation of plant immunity.     

TVDI模型的农业旱情时空变化遥感应用

温度植被干旱指数(TVDI)是一种通过反演土壤湿度来反映农业干旱状况的重要方法,其中能量平衡和植被指数的变化是影响TVDI模型精度的主要因子。在研究比值植被指数(RVI)、归一化植被指数(NDVI)、增强型植被指数(EVI)和修正土壤调节植被指数(MSAVI)和下垫面温度(Ts)的基础上,引入DEM数据并对Ts做地形校正,减少了地形起伏对能量平衡的影响,建立不同植被指数的Ts-Ⅵ特征空间,选择与土壤湿度相关性最好的温度植被干旱指数(TVDI),获取研究区2005年作物生长季(5—9月)的干旱状况,并用同步的气象信息对干旱状况进行验证。结果表明:根据不同时期采用不同植被指数的TVDI模型,经过地形校正后能够更好地反映研究区的农业干旱状况。     

干旱区绿洲城镇景观生态风险时空变化分析——以张掖绿洲乡镇为例

科学评估与分析干旱区绿洲乡镇景观生态风险,对优化绿洲土地资源管控和维护干旱区绿洲生态系统稳定性具有重要现实意义。以张掖市绿洲乡镇为对象,基于土地利用数据和景观格局指数,结合ArcGIS空间分析功能,研究其景观生态风险时空特征及影响因素。结果表明：1987—2018年,耕地和建设用地变化明显,分别增加429.03 km~2和236.26 km~2;整体景观生态风险呈下降趋势,并呈“南北高、中间低”格局特征;各乡镇风险水平在空间上正向聚集,位于绿洲边缘地区的乡镇风险变化明显;人为活动干扰是近年风险水平变化的主要因素。研究成果可为西北干旱区绿洲生态保护与治理提供决策参考。