26S蛋白酶体的结构生物学研究进展

26S蛋白酶体是真核细胞内负责蛋白质降解的主要分子机器,通过特异性降解目的蛋白质,几乎参与了生物体的绝大多数生命活动.26S蛋白酶体在结构上可分为19S调节颗粒和20S核心颗粒两部分.19S调节颗粒负责识别带有泛素链标记的蛋白质底物及对其进行去折叠,并最终将去折叠的蛋白质底物传送至20S核心颗粒中进行降解.由于26S蛋白酶体的结构组成复杂,分子量十分巨大,现有的X-ray技术和NMR技术对其完整结构的解析都无能为力,仅能解析出部分单个蛋白成员或分子量较低的亚复合物晶体结构.而冷冻电镜技术在相当一段时间内处于发展的初级阶段,导致其三维结构的研究进展曾经十分缓慢,严重阻碍了人们对其结构和功能的了解.近年来,随着在X-ray技术领域对大分子复合物结构解析的经验积累和冷冻电镜技术领域的技术革命,完整的26S蛋白酶体三维结构解析取得了飞速的发展.本文回顾了近几年在26S蛋白酶体结构生物学领域的重要进展,并展望了该领域未来的发展及面临的挑战.     

基于RNA-Seq高通量测序技术的牦牛卵巢转录组研究:进一步完善牦牛基因结构及挖掘与繁殖相关新基因

RNA-Seq作为近年来新发展起来的高通量转录组测序技术,为大规模转录组学研究提供了一种全新的且更为有效的方法.目前该技术已广泛应用于转录组学中多方面的研究,尤其近年来,该技术在进一步完善基因结构信息及挖掘新转录本及新基因方面的功能也逐渐受到关注.本研究以牦牛卵巢组织作为研究对象,应用RNA-Seq技术对其进行高通量转录组测序分析,经测序后得到了一个包含26826516条过滤后测序读数,4828772880 bp的卵巢测序文库,比对分析显示,有16992条牦牛基因发生表达,其中3734条基因存在不同类型的可变剪接.功能分析表明,这些表达基因涉及多种GO分类及KEGG通路.进一步分析转录组数据发现,共有7340个基因的5′或3′端在原有基因组的位置基础上发生了延伸,同时还发现了6321个新转录本,定位回基因组预测显示,外显子数量为1~84个,其中2267个新转录本预测具有编码蛋白的能力.比对分析显示,共有1200~4993条新转录本分别与Nt数据库、Nr数据库及SwissProt数据库中的基因比对上,其中与牛相似性基因最多(41.4%),其次为野牦牛(33.0%)、绵羊(6.3%)、人类(2.8%)及小鼠(1.6%)等其他物种.进一步对新转录本进行GO分类注释,结果显示,与繁殖发育相关的GO分类占有较大比例,其中繁殖类别(reproduction)所涉及的新转录本最多.本研究结果为描绘牦牛卵巢正常转录组图谱及进一步探析牦牛繁殖性能提供了基础,同时证实RNA-Seq高通量转录组技术在完善基因结构及挖掘新转录本及新基因方面的具有强大的优势,为进一步完善牦牛基因组结构信息及挖掘潜在的新基因提供了丰富数据.     

“青县大棚”结构设计与建造技术

为提升青县大棚设施生产性能和抗灾能力,对青县大棚设施结构进行调研,并提出适宜青县乃至周边地区发展设施蔬菜的标准化"青县大棚"结构设计及建造方法。     

植物防御素结构与功能及其抗真菌机制研究进展

植物防御素是一类分子量约为5 kD的阳离子肽,是植物内免疫系统的主要成分之一,参与了多种植物生理生化活动。植物防御素含8个保守半胱氨酸,形成4对链内二硫键来稳定其3条反向平行的β折叠片和1个α螺旋,构成所谓的Csαβ模体结构。植物防御素具有抑制真菌和细菌生长、抑制酶的活性、抑制癌细胞增殖和作为离子通道阻断剂等功能。植物防御素抗真菌机制比较复杂,但它可能首先必须和真菌细胞质膜外侧的鞘脂等复杂脂类结合,引起真菌细胞发生一系列的反应,包括细胞膜通透性增强、胞内活性氧水平增高和诱导真菌细胞凋亡等。但是植物防御素详细的抗真菌机制及其在真菌细胞内作用靶点等有待于深入研究。综述了近年来植物防御素结构、功能及其抗真菌机制等方面的最新研究进展。     

Bacterial microbiome of Coptotermes curvignathus （Isoptera： Rhinotermitidae） reflects the coevolution of species and dietary pattern

Coptotermes curvignathus Holmgren is capable of feeding on living trees. This ability is attributed to their effective digestive system that is furnished by the termite＇s own cellulolytic enzymes and cooperative enzymes produced by their gut microbes. In this study, the identity of an array of diverse microbes residing in the gut of C. curvignathus was revealed by sequencing the near-full-length 16S rRNA genes. A total of 154 bacterial phylotypes were found. The Bacteroidetes was the most abundant phylum and accounted for about 65% of the gut microbial profile. This is followed by Firmicutes, Actinobacteria, Spirochetes, Proteobacteria, TM7, Deferribacteres, Planctomycetes, Ferrucomicrobia, and Termite Group 1. Based on the phylogenetic study, this symbiosis can be a result of long coevolution of gut enterotypes with the phylogenic distribution, strong selection pressure in the gut, and other speculative pressures that determine bacterial biome to follow. The phylogenetic distribution of cloned rRNA genes in the bacterial domain that was considerably different from other termite reflects the strong selection pressures in the gut where a proportional composition of gut microbiome of C. curvignathus has established. The selection pressures could be linked to the unique diet preference of C. curvignathus that profoundly feeds on living trees. The delicate gut microbiome composition may provide available nutrients to the host as well as potential protection against opportunistic pathogen.     

Morphological traits and physiological characteristics in drought tolerance in 20 shrub species on the Qinghai-Xizang Plateau

Aims Providing indispensably theoretical evidence for establishing indices of evaluation on drought tolerance in shrubs, and screening for tree species that are drought tolerant for afforestation in arid regions or for matching their characteristics with suitable habitat conditions are the key to vegetation restoration in the Qinghai-Xizang Plateau. However, these issues are not adequately addressed in recent research due to lack of systematic methods. Therefore, our objective was to make a comprehensive evaluation on drought tolerance in 20 shrub species collected from different areas in Qinghai-Xizang Plateau, and to study their underlying mechanisms in drought tolerance.
Methods We made measurements on variables depicting root characteristics, including the root length (TRL), surface area (TRSA), volume (TRV), and tips number (TRTN) of all roots, the root length (FRL), surface area (FRSA), volume (FRV), and tips number (FRTN) of fine roots (d≤2 mm), and derived plant characteristic indices including thickness of cuticle (CT), thickness of palisade tissue (TPT), thickness of spongy tissue (TST), TPT/TST, thickness of leaf (LT), palisable tissue cell density, and tissue structural tense ratio (CTR = TPT/LT × 100%) and spongy tissue loosened ratio (SR = TST/LT × 100%) of leaf anatomical structure, root to shoot ratio (RSR), leaf transpiration rate (T_r), instantaneous water use efficiency (WUE_i), and carbon isotopic composition (δ¹³C) of the 20 shrub species through field experiments. Correlation analysis and principal component analysis were performed on the 19 variables and indices.
Important findings Different shrubs had different mechanisms of drought tolerance. In this study, the character- istics of drought tolerance were mainly categorized into 6 types, involving modifications of (1) root systems, (2) leaf anatomical structure, (3) leaf pattern, and (4) biomass allocation, or via (5) low water-consumption and (6) high WUE_i. Different genera or different tree species within the same genus clearly differed in drought tolerance. The species of the genus Hippophae were relatively poorly tolerant to drought, whereas several shrubs including Potentilla fruticosa, Berberis julianae, Caragana arborescens, Spiraea salicifolia and Hippophae rhamnoides ssp. mongolica occurred to be more drought tolerant than other shrub species investigated in this study. On the other hand, there were highly significant correlations among the characteristics of root systems and among characteristics of leaf anatomical structure. The results of principal component analysis on 19 variables and indices showed that TRL, TRSA, TRV, TRTN, FRL, FRSA, FRV, FRTN, CT, TPT, TST and WUE_i could be effective indicators of drought tolerance of shrubs in the Qinghai-Xizang Plateau. In addition, the drought tolerance of shrubs had a close connection with their origin of collections; the shrubs collected from Xining prefecture in Qinghai Province were more drought tolerant than those from Tianshui Prefecture in Gansu and Lasa Prefecture in Xizang.     

南岭大东山长柄双花木群落物种组成与种群结构特征分析

采用样方法以及用基径结构代替年龄结构的方法,对位于南岭大东山不同海拔的3个长柄双花木(Disanthus cercidifolius var.longipes Chang)样带(包括小洞、大洞和鸡子场样带,海拔分别为1 010、1 170和1 260 m)进行了调查,并编制了种群静态生命表和生存曲线,对其群落物种组成、种群年龄结构和生殖力现状进行了分析。结果显示:3个样带群落乔木层优势种数量及种类组成差异明显,共包含26个种类,但共有种仅3种,除长柄双花木外还有多花杜鹃(Rhododendron cavaleriei Lévl.)和两广杨桐(Adinandra glischroloma Hand.-Mazz.)2种;小洞样带长柄双花木种群的重要值(46.116%)明显高于另2个样带。根据基径可将长柄双花木种群分为68个龄级,3个样带的种群年龄结构基本一致,但小洞样带个体数量更多且基径31.0 mm以下的幼体所占比例最高;总体上长柄双花木种群中5 a以下的幼苗数量偏少,其他龄级的个体数量呈现随株龄增大波动但整体下降的趋势。种群的个体死亡率在5~10 a和10~15 a龄级较高(分别为20.30%和20.50%),并随年龄增长总体呈逐渐下降的趋势;且不同龄级个体生命期望值为4.49~0.50,总体偏低。种群生存曲线为Ⅰ型,种群净增殖率、内禀增长率、周限增长率和世代平均周期分别为0.504、-0.018、0.982和38.351 a,显示该种群在当前状态下自我更新不良,属缓慢的负增长型种群。根据研究结果,对该区域长柄双花木种群提出了相应的保护对策和建议。     

蝎β型昆虫毒素的结构及其与钠通道作用机制

蝎毒素是蝎为防卫的需要而产生的一系列活性短肽.其中蝎昆虫特异性毒素可特异性结合并调控昆虫可兴奋细胞膜上的钠离子通道,是研究离子通道结构与功能的首选探针,并在转基因抗虫植物及生物杀虫剂研究方面具有潜在的应用价值.本文对蝎β型昆虫毒素的结构与功能及其对钠离子通道的作用方式和β毒素的电压传感器捕获(voltage sensor-trapping)模型做一综述,为进一步揭示蝎β毒素的结构与功能的关系和在农作物抗虫领域的应用提供依据.     

濒危植物南方红豆杉不同种群的结构和动态变化

为了解南方红豆杉(Taxus chinensis var.mairei)不同种群的结构变化,于2007年、2008年、2012年分别对皖南仙寓山双坑村、浙江天目山桐坑村及福建梅花山的南方红豆杉天然种群进行了研究。结果表明,仙寓山的1214株南方红豆杉中,幼苗占80%;天目山有510株,幼苗占39.61%,其次是9 m以上的古树,占22.16%;梅花山有174株,幼苗占70.11%,古树占20.11%。3个种群的天然更新都呈缓慢增长状态,存活曲线为Deevey-Ⅲ型,空间分布均为聚集分布,但从幼苗到幼树的生长阶段死亡率高。因此,对濒危植物南方红豆杉的保护重点应放在幼苗和幼树阶段。     

A型流感病毒NS1与突触后密度蛋白-95的相互作用

目的 A型流感病毒NS1蛋白是一种多功能的致病因子,能够与被感染细胞中的多种蛋白相互结合,影响并干扰宿主细胞内的信号转导、蛋白质合成及抗病毒反应。突触后密度蛋白(Postsynaptic density protein95,PSD-95)主要存在于神经元及SH-SY-5Y等神经来源的细胞株中。假设NS1能够与PSD-95结合,则更有利于了解A型流感病毒对神经元及相关细胞的作用机制。方法通过酵母双杂交,GST-pull down及免疫荧光技术分别从体外和体内两方面检测NS1与PSD-95的相互作用。结果酵母双杂交表明,仅转染PGAD-NS51/PGBK-PSD-95的QDO有菌落生长,且α-半乳糖苷酶活性显著高于阳性对照;而转染PGAD-NS32/PGBK-PSD-95的QDO无菌落生长;GST-pull down表明仅NS51与PSD-95孵育后,能够被Western-blot检测到;免疫荧光表明NS51与PSD-95可能存在共定位,而NS32与PSD-95则不存在共定位。结论 H5N1(A/chicken/Guangdong/1/2005)的NS1能够与PSD-95结合;反之,H3N2(A/Shantou/602/06)的NS1则不能。