基于Golden Gate技术的齐整小核菌转录单元高效组装系统

【目的】为齐整小核菌代谢工程研究建立高效的转录单元组装系统。【方法】通过应用Golden Gate技术,以mobius assembly为基础,分别设计并构建DNA元件标准化接口改造、单转录单元组装、应用质粒(多转录单元)组装等功能的载体,从而形成一套完整的多转录单元组装系统。【结果】构建了2个用于DNA元件标准化接口改造的Level 0载体,4个用于单转录单元组装的Level 1载体,4个用于应用质粒组装的Level 2载体和13个应用质粒组装的辅助质粒。然后应用此系统为齐整小核菌组装了若干经过标准化接口改造的DNA元件质粒、单转录单元质粒和硬葡聚糖相关基因的功能分析质粒。所构建的最终应用质粒可以同时适用于齐整小核菌的根癌农杆菌介导转化法、电穿孔转化法和原生质体转化法。【结论】此质粒系统具有强大的DNA设计、组装和容纳能力,为未来齐整小核菌代谢工程和功能基因组学研究提供了高效的质粒构建技术平台。     

The northern corn leaf blight resistance gene Ht1 encodes an nucleotide-binding,leucine-rich repeat immune receptor

Northern corn leaf blight, caused by the fungal pathogen Exserohilum turcicum, is a major disease of maize. The first major locus conferring resistance to E. turcicum race 0, Ht1, was identified over 50 years ago, but the underlying gene has remained unknown. We employed a map-based cloning strategy to identify the Ht1 causal gene, which was found to be a coiled-coil nucleotide-binding, leucine-rich repeat (NLR) gene, which we named PH4GP-Ht1. Transgenic testing confirmed that introducing the native PH4GP-Ht1 sequence to a susceptible maize variety resulted in resistance to E. turcicum race 0. A survey of the maize nested association mapping genomes revealed that susceptible Ht1 alleles had very low to no expression of the gene. Overexpression of the susceptible B73 allele, however, did not result in resistant plants, indicating that sequence variations may underlie the difference between resistant and susceptible phenotypes. Modelling of the PH4GP-Ht1 protein indicated that it has structural homology to the Arabidopsis NLR resistance gene ZAR1, and probably forms a similar homopentamer structure following activation. RNA sequencing data from an infection time course revealed that 1 week after inoculation there was a threefold reduction in fungal biomass in the PH4GP-Ht1 transgenic plants compared to wild-type plants. Furthermore, PH4GP-Ht1 transgenics had significantly more inoculation-responsive differentially expressed genes than wild-type plants, with enrichment seen in genes associated with both defence and photosynthesis. These results demonstrate that the NLR PH4GP-Ht1 is the causal gene underlying Ht1, which represents a different mode of action compared to the previously reported wall-associated kinase northern corn leaf blight resistance gene Htn1/Ht2/Ht3.     

Genomic insights into selection for heterozygous alleles and woody traits in Populus tomentosa

Heterozygous alleles are widespread in outcrossing and clonally propagated woody plants. The variation in heterozygosity that underlies population adaptive evolution and phenotypic variation, however, remains largely unknown. Here, we describe a de novo chromosome-level genome assembly of Populus tomentosa, an economic and ecologically important native tree in northern China. By resequencing 302 natural accessions, we determined that the South subpopulation (Pop_S) encompasses the ancestral strains of P. tomentosa, while the Northwest subpopulation (Pop_NW) and Northeast subpopulation (Pop_NE) experienced different selection pressures during population evolution, resulting in significant population differentiation and a decrease in the extent of heterozygosity. Analysis of heterozygous selective sweep regions (HSSR) suggested that selection for lower heterozygosity contributed to the local adaptation of P. tomentosa by dwindling gene expression and genetic load in the Pop_NW and Pop_NE subpopulations. Genome-wide association studies (GWAS) revealed that 88 single nucleotide polymorphisms (SNPs) within 63 genes are associated with nine wood composition traits. Among them, the selection for the homozygous AA allele in PtoARF8 is associated with reductions in cellulose and hemicellulose contents by attenuating PtoARF8 expression, and the increase in lignin content is attributable to the selection for decreases in exon heterozygosity in PtoLOX3 during adaptive evolution of natural populations. This study provides novel insights into allelic variations in heterozygosity associated with adaptive evolution of P. tomentosa in response to the local environment and identifies a series of key genes for wood component traits, thereby facilitating genomic-based breeding of important traits in perennial woody plants.     

Avicularin alleviates acute liver failure by regulation of the TLR4/MyD88/NF-κB and Nrf2/HO-1/GPX4 pathways to reduce inflammation and ferroptosis

Acute liver failure (ALF) is an inflammation-mediated hepatocyte death process associated with ferroptosis. Avicularin (AL), a Chinese herbal medicine, exerts anti-inflammatory and antioxidative effects. However, the protective effect of AL and the mechanism on ALF have not been reported. Our in vivo results suggest that AL significantly alleviated lipopolysaccharide (LPS)/D-galactosamine (D-GalN)-induced hepatic pathological injury, liver enzymes, inflammatory cytokines, reactive oxygen species and iron levels and increased the antioxidant enzyme activities (malondialdehyde and glutathione). Our further in vitro experiments demonstrated that AL suppressed inflammatory response in LPS-stimulated RAW 264.7 cells via blocking the toll-like receptor 4 (TLR4)/myeloid differentiation protein-88 (MyD88)/nuclear factor kappa B (NF-κB) pathway. Moreover, AL attenuated ferroptosis in D-GalN-induced HepG2 cells by activating the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1)/glutathione peroxidase 4 (GPX4) pathway. Therefore, AL can alleviate inflammatory response and ferroptosis in LPS/D-GalN-induced ALF, and its protective effects are associated with blocking TLR4/MyD88/NF-κB pathway and activating Nrf2/HO-1/GPX4 pathway. Moreover, AL is a promising therapeutic option for ALF and should be clinically explored.     

北部湾鱼类碳、氮、磷生态化学计量特征

为探究北部湾鱼类生态化学计量特征,于2021年3月—4月采集分析了北部湾沿岸9个渔港的79种鱼类(382尾)碳(C)、氮(N)、磷(P)含量特征。结果表明北部湾鱼类C、N、P元素含量变化范围分别为33.87%—58.34%、6.31%—14.92%和0.77%—4.67%;C∶N、C∶P、N∶P的变化范围分别为3.43—9.72、19.15—173.06和5.04—33.68,其中P元素含量变化最大,导致C∶P和N∶P的变化。不同物种在科间的差异较大,科内的差异较小。不同体型和食性鱼类之间的C、N、P含量及比值具有显著差异(P<0.05)。鱼类的C∶P和N∶P与P含量呈显著负相关(R²=0.966,P<0.01;R²=0.877,P<0.01),P含量和Ca含量呈显著正相关(R²=0.919,P<0.01),P含量的变化可能与鱼类骨骼和鳞片的形成有关。总之,北部湾鱼类未保持严格的生态化学计量平衡,鱼类生态化学计量特征存在显著的种间差异,主要因鱼类不同的体型和食性差异所致。     

基于土地利用变化的东海区大陆海岸带生境质量时空演变研究

人类活动对生态环境的影响主要表现为土地利用方式的变化。土地利用变化影响下的生境质量时空格局演变,对优化区域国土生态布局具有重要意义。利用DEM数据、气象数据、社会经济数据与1980、1990、2000、2010、2018年5期土地利用数据,通过InVEST模型、景观格局指数、地理探测器评估、描述、分析东海区大陆海岸带生境质量时空演变特征。结果表明：(1)1980—2018年东海区大陆海岸带陆地面积增加,土地利用中林地所占比重最大,耕地、建设用地面积变化最明显,2000—2010年土地利用转化最显著。土地变化北部大于南部、东侧大于西侧。(2)1980—2018年生境质量均值为0.75,总体生境质量较好,但呈下降趋势,2000—2010年生境质量下降最明显,2010年后下降速度减缓。生境质量斑块趋于破碎化、多样化、均匀化。(3)生境质量总体上从沿海向内陆下降,受岸线性质与围垦影响,北部生境质量由陆向海退化,南部生境退化沿海向陆推进。低等生境质量区呈现点—面—带的扩张趋势。(4)建设用地指数、高程、夜间灯光亮度、土壤类型、坡度为影响生境质量分布的主要因素,38年间主导因素由高程变为建设用地指...     

毛乌素沙地油蒿叶性状对光能利用效率动态的影响

目前对于荒漠灌木光能利用效率（LUE）的季节变异及其调控因素,尤其是其生物调控因素的认识非常有限,导致了荒漠生态系统生产力模型的不确定性。拟验证假设：长期干旱环境下,典型荒漠灌木油蒿光能利用效率日均值（LUE_day）的动态变化与叶片性状的季节性调整有关。试验采用Li-6400便携式光合仪定期测量了油蒿生长季叶片LUE_day的季节动态及相关叶性状指标,探究叶性状对LUE_day的影响。结果表明：LUE_day的季节波动范围为0.003-0.017 mol/mol,整体变异系数（CV）为38.75%。完全展叶期LUE_day均值相比生长季平均值降低17.37%,相比展叶期和落叶期时降低30%;8个叶性状的季节变异幅度差异较大,其中总叶绿素含量（Chl）、类胡萝卜素含量（Car）和叶氮含量（LNC）均表现出较大的季节变异性（CV ≥ 20%）,叶碳含量（LCC）和叶片相对含水量（LRWC）的变异程度最低（CV<7%）。LRWC与所有叶片化学性状（Chl、Chl a/b、Car、LNC和LCC）均存在显著相关,表明其变化与叶片的养分吸收、光合色素合成以及碳同化的运输过程密切相关;油蒿LUE_day的相对变化与LRWC、Chl a/b和LNC显著正相关,而LRWC和LNC的季节动态受空气温度（T_a）和土壤含水量（VWC）的共同调节,Chl a/b的季节波动主要由浅层土壤含水量（10 cm VWC）控制。以上研究结果强调,在未来预计极端的气候事件（如极端干旱和持续热浪事件）发生更频繁的旱地场景中,时间尺度植物叶性状对于土壤干旱和高温的适应性调整应当被充分考虑到旱地生态系统的通量建模方案中。该结果将为构建叶片尺度的光合生理模型与厘清LUE的生物调控机制提供理论依据。     

中国遥感干旱指数时空特征及其对气候和地表覆盖变化的响应

干旱严重影响植被生长,威胁粮食安全,基于遥感计算的植被状态指数（Vegetation Condition Index,VCI）、温度状态指数（Temperature Condition Index,TCI）和植被健康指数（Vegetation Health Index,VHI）是常用的干旱指数,被广泛应用于干旱监测。为了探究近年来我国干旱特征及其对气候和地表覆盖变化的响应,分析了2003-2016年期间VCI、TCI和VHI的时空变化特征;采用最小二乘（OLS）和偏相关分析方法分析了这些指数对气候和地表覆盖变化的响应。基于上述干旱指数计算的干旱频率表明,中温带中部和南温带等地区干旱发生频率高,干旱指数变化趋势表明在2003-2016年期间中国大部分地区干旱缓解,但在中温带、南温带和高原气候区等局部地区干旱加剧;总体而言,干旱指数随着年平均温度的上升和年降水量的降低而减小,VHI与温度和降水量的相关性在不同气候区的一致性优于VCI和TCI;裸土的减少和植被的增加导致干旱指数增大,树木转变为低矮植被干旱指数降低。