Aspergillus fumigatus来源植酸酶的Q23L和Q23LG272E突变研究

Aspergillus fumigatus来源的植酸酶具有热稳定性好、pH作用范围广的优点, 但其比活性很低。设计的植酸酶Q23L突变能在pH4.5～7.0范围内大幅提高比活性,但pH稳定性却显著下降, 为了进一步改良Q23L的pH稳定性, 在Q23L分子上加入了G272E突变。将原酶、突变酶Q23L和突变酶Q23LG272E分别在毕赤酵母GS115中表达,表达酶经纯化后进行酶学性质比较分析,结果表明：突变酶Q23L的比活性比原酶显著提高, 在pH5.5比活性由51u/mg提高到109u/mg, 但其pH稳定性, 尤其是在pH3.0～4.0酸性条件下的稳定性却显著降低,低于80%。突变酶Q23LG272E在pH3.0～4.5和pH6.5～7.0时的稳定性比Q23L有所提高, 恢复到原酶的水平,而比活性基本维持在Q23L的水平。通过一级序列和三维结构比较,分析了可能影响Q23LG272E酶学性质的因素,为进一步研究植酸酶的结构与功能提供了材料。     

泰山4种优势造林树种细根分解对细菌群落结构的影响

为了理解细菌群落结构和多样性对森林生态系统细根凋落物分解的影响, 该研究以泰山4种主要优势造林树种刺槐(Robinia pseudoacacia)、麻栎(Quercus acutissima)、油松(Pinus tabulaeformis)和赤松(Pinus densiflora)为研究对象, 采用凋落物分解袋法及Illumina Miseq测序平台对细菌16S rDNA V4-V5区扩增产物进行双端测序, 分析了4种树种细根分解对细菌群落结构及多样性的影响。结果表明: (1) 4种植物细根分解速率差异显著, 阔叶树种分解速率显著高于针叶树种, 表现为刺槐>麻栎>油松>赤松。(2) 4个树种细菌序列操作分类单元(OTU)、观测到的物种数、Ace指数和系统发育多样性之间差异显著, 且阔叶树种刺槐和麻栎显著低于针叶树种赤松和油松。4种细根分解的细菌群落结构存在极显著差异。细根初始碳(C)含量、木质素:氮(N)和C:N对细菌群落结构的影响较大。(3)细菌群落相对丰度在5%以上的优势类群是变形菌门、放线菌门、拟杆菌门、酸杆菌门, 且变形菌门、酸杆菌门在4个树种之间差异显著, 特别是阔叶树种变形菌门显著高于针叶树种。在纲水平上, α-变形菌纲、β-变形菌纲、γ-变形菌纲、不明放线菌纲、鞘脂杆菌纲为主要的优势纲, 其中α-变形菌纲、不明放线菌纲在4个树种之间差异显著。(4) Pearson相关性分析表明, 细菌优势门和纲相对丰度受到凋落物初始化学性质的影响, 特别是变形菌门和α-变形菌纲; 变形菌门和α-变形菌纲相对丰度与细根分解速率显著正相关。冗余分析结果也显示, 细根初始N、磷(P)含量和木质素含量对细菌群落结构的影响较大。研究结果有助于理解细菌群落结构和多样性对森林生态系统细根凋落物分解的影响。     

Acyl-ACPs的规模化合成

脂酰-酰基载体蛋白(fatty acyl-acyl carrier protein, acyl-ACP)是多种生物合成途径中的酰基供体。因供给限制,体外研究常用类似物acyl-CoA替代,而CoA部分和ACP有较大差异,限制了相关酶对底物识别的认识。因此稳定获得大量acyl-ACP是体外研究相关酶的催化机制及其代谢途径的关键。研究以holo-ACP和C4~C18链长脂肪酸为底物,在哈氏弧菌acyl-ACP合成酶(Vibrio harveyi acyl-ACP synthetase, VhAasS)催化下合成不同碳链长度的acyl-ACP;通过高效液相色谱(HPLC)方法,确定不同碳链长度acyl-ACP的合成产率。结果表明:碳链为C4~C14的acyl-ACP产率均高于90.0%,16:0-ACP产率为85.9%,18:1-ACP产率仅为25.7%。通过加入Li ⁺优化反应体系,16:0-ACP、18:1-ACP的产率达90.0%。进一步优化扩大反应体系可稳定获得20mg以上acyl-ACP;最后,把合成的acyl-ACP应用到甘油-3-磷酸酰基转移酶催化的反应体系中。不同链长acyl-ACP的规模化合成研究,为体外研究相关酶的催化机制提供重要基础。     

Tecia solanivora infestation increases tuber starch accumulation in Pastusa Suprema potatoes

In response to infestation with larvae of the Guatemalan tuber moth(Tecia solanivora), some Solanum tuberosum(potato) varieties exhibit an overcompensation response, whereby the total dry mass of uninfested tubers is increased. Here, we describe early responses,within the first few days, of T. solanivora feeding, in the Colombian potato variety Pastusa Suprema. Nontargeted metabolite profiling showed significant secondary metabolism changes in T. solanivora-infested tubers,but not in uninfested systemic tubers. In contrast,changes in primary metabolism were greater in uninfested systemic tubers than in the infested tubers, with a notable 80% decline in systemic tuber sucrose levels within 1 d of T. solanivora infestation. This suggested either decreased sucrose transport from the leaves orincreased sink strength, i.e., more rapid sucrose to starch conversion in the tubers. Increased sucrose synthesis was indicated by higher rubisco activase and lower starch synthase gene expression in the leaves of infested plants.Elevated sink strength was demonstrated by 45% more total starch deposition in systemic tubers of T. solanivorainfested plants compared to uninfested control plants.Thus, rather than investing in increased defense of uninfested tubers, Pastusa Suprema promotes deposition of photoassimilates in the form of starch as a response to T. solanivora infestation.     

The evolution of plant microRNAs: insights from a basal eudicot sacred lotus

microRNAs (miRNAs) are important noncoding small RNAs that regulate mRNAs in eukaryotes. However, under which circumstances different miRNAs/miRNA families exhibit different evolutionary trajectories in plants remains unclear. In this study, we sequenced the small RNAs and degradome from a basal eudicot, sacred lotus (Nelumbo nucifera or lotus), to identify miRNAs and their targets. Combining with public miRNAs, we predicted 57 pre‐eudicot miRNA families from different evolutionary stages. We found that miRNA families featuring older age, higher copy and target number tend to show lower propensity for miRNA family loss (PGL) and stronger signature of purifying selection during divergence of temperate and tropical lotus. Further analyses of lotus genome revealed that there is an association between loss of miRNA families in descendent plants and in duplicated genomes. Gene dosage balance is crucial in maintaining those preferentially retained MIRNA duplicates by imposing stronger purifying selection. However, these factors and selection influencing miRNA family evolution are not applicable to the putative MIRNA‐likes. Additionally, the MIRNAs participating in lotus pollen–pistil interaction, a conserved process in angiosperms, also have a strong signature of purifying selection. Functionally, sequence divergence in MIRNAs escalates expression divergence of their target genes between temperate and tropical lotus during rhizome and leaf growth. Overall, our study unravels several important factors and selection that determine the miRNA family distribution in plants and duplicated genomes, and provides evidence for functional impact of MIRNA sequence evolution.     

Brefeldin A enhances docetaxel-induced growth inhibition and apoptosis in prostate cancer cells in monolayer and 3D cultures

Docetaxel is a commonly used chemotherapeutic drug for patients with late stage prostate cancer. However, serious side effect and drug resistance limit its clinical success. Brefeldin A is a 16-membered macrolide antibiotic from mangrove-derived Fungus Aspergillus sp. (9Hu), which exhibited potent cytotoxicity against human cancer cells. In the present study, we determined the effect of brefeldin A on docetaxel-induced growth inhibition and apoptosis in human prostate cancer PC-3 cells. Brefeldin A in combination with docetaxel inhibited the growth of PC-3 cells in monolayer and in three dimensional cultures. The combination also potently stimulated apoptosis in PC-3 cells as determined by propidium iodide staining and morphological assessment. Mechanistic studies showed that growth inhibition and apoptosis in PC-3 cells treated with brefeldin A and docetaxel were associated with decrease in the level of Bcl-2. The present study indicates that combined brefeldin A with docetaxel may represent a novel approach for improving the efficacy of docetaxel, and Bcl-2 may serve as a target for brefeldin A to enhance the effects of docetaxel chemotherapy.     

甘南尕海湿地退化过程中植被生物量变化及其季节动态

为探讨尕海湿地退化过程中植被生物量变化规律,以尕海泥炭沼泽和沼泽化草甸为例,采用定位样地调查方法,研究了不同退化程度湿地植被生物量的时空分布格局。结果表明,1)随着湿地退化演替,两类湿地植被地上生物量逐渐减小,泥炭沼泽未退化(PⅠ)、退化阶段(PⅡ)地上生物量依次为334.19,290.72 g/m~2,沼泽化草甸未退化(SⅠ)、轻度退化(SⅡ)、中度退化(SⅢ)地上生物量依次为378.40,308.07,261.21 g/m~2;地上生物量季节动态规律均为单峰型,8月中下旬达到峰值;同一湿地类型各退化阶段地上生物量绝对增长率(AGR)和相对增长率(RGR)在同一年份变化趋势基本相同,但不同年份间存在差异,而同一湿地类型不同阶段AGR和RGR的大小存在差异。2)地下生物量也随退化程度加剧显著减小(P0.05),PⅠ,PⅡ地上生物量依次为23081.46,12607.72 g/m~2,SⅠ,SⅡ,SⅢ地下生物量依次为4583.16,3008.63,1290.73 g/m~2;地下生物量季节变化均表现出愈接近生长季始末值愈大;地下生物量由土壤表层向深层显著下降(P0.05),总体呈"T"形分布,0—10cm土层,泥炭沼泽、沼泽化草甸地下生物量都最大,分别占各自总地下生物量50%和70%以上。3)尕海2类高寒湿地5—9月平均根冠比均表现未退化高于退化,根冠比季节动态为越接近生长季始末值越大,生长旺盛季值越小。     

高寒生态脆弱区农户的气候变化适应策略评价——以甘南高原为例

气候变化加剧了高寒生态脆弱区农户的生计脆弱性,为应对气候变化,农户已采取了各种响应措施,当前急需评估农户所采取适应策略的效果,以便选择更有效的适应策略。以甘南高原为研究区,基于入户调查数据分析了农户所采取适应策略的特征,采用模糊综合评价法分析现行适应策略的效果,并利用多准则决策模型确定了最优的气候变化适应策略。结果显示:(1)甘南高原农户多采取组合型策略应对气候变化,尤以调整+扩张型策略为主;(2)甘南高原农户所采取气候变化适应策略的效果较好,效益指数为3.43。其中,农区农户的适应策略效果最好、半农半牧区次之、纯牧区最差;(3)甘南高原不同区域农户筛选的最优策略存在差异。其中,纯牧区和半农半牧区农户筛选的最优策略为调整农牧业结构,而农区农户为完善农牧业基础设施。最后,提出了提高农户适应策略效果的对策建议及未来研究中需进一步关注的问题。     

北京地区典型落叶阔叶乔木叶片含氮量和δ15N值对大气氮沉降的响应

为探究植物对大气氮沉降的响应和对这部分氮素的来源指示作用,本研究通过对北京地区198个采样点,典型落叶阔叶乔木杨属(Populus)和柳属(Salix)植物叶片进行采样,测定其叶片样品含氮量和δ~(15)N值。结果表明:北京地区杨属植物叶片含氮量为16.5—38.6g/kg,平均(24.0±4.0)g/kg;柳属植物叶片含氮量为17.2—36.2g/kg,平均(25.9±4.1)g/kg。研究区域范围内杨属、柳属植物叶片的含氮量均呈现出西北低、东南高的对角线型分布,与该区域大气氮沉降的空间变异相吻合。由于研究区域范围内气候因子无明显的变异,植物叶片的含氮量变化反应了大气氮沉降对植物元素化学计量特征的影响和植物对大气氮沉降的响应。北京地区杨属植物叶片δ~(15)N值为-3.95‰—8.10‰,平均(1.15±2.48)‰;柳属植物叶片δ~(15)N值为-3.04‰—9.73‰,平均(2.31±2.60)‰。杨属和柳属植物叶片的δ~(15)N值均呈现出西北高、中部高、东南低的空间分布,与叶片含氮量空间分布趋势相反。中部城区较高的δ~(15)N值反应了交通污染对大气含氮化合物增加的影响;西北部较高的δ~(15)N值反应了该区域受人为活动排放源的影响较少,自然的氮循环是其较高δ~(15)N值的主要原因;东南部较低的δ~(15)N值则有可能是由农业活动和交通共同作用的结果。