马立克氏病病毒超强毒感染鸡羽髓蛋白质组分析

【目的】羽毛是细胞游离马立克氏病病毒(Marek’s disease virus,MDV)释放的部位,为了解感染MDV后鸡羽中宿主基因表达的变化及对病毒感染的应答,进行了MDV感染鸡的羽髓蛋白质组学分析。【方法】1日龄无特定病原体(specific pathogen free,SPF)鸡人工感染MDV超强毒RB1B株(1000PFU),感染后21d采集鸡羽毛,提取羽髓蛋白,以17cm,pH5-8的IPG胶条进行二维电泳,以未感染病毒的SPF鸡羽髓蛋白为对照,使用PDQuest软件对二维电泳图谱进行差异蛋白分析,并选取部分差异斑点进行质谱鉴定。【结果】PDQuest软件分析发现攻毒组和对照组表达差异大于两倍的蛋白点有41个,其中攻毒组表达上调的蛋白点25个,下调的蛋白点7个,新出现的蛋白点有9个。质谱分析共成功鉴定了21个斑点,对应于20个蛋白。如载脂蛋白AI(apolipoprotein AI)、14-3-3 sigma(两个斑点均为该蛋白)、癌蛋白18(stathmin)等。【结论】功能预测表明这些蛋白涉及到宿主的抗病毒应答、物质代谢、细胞骨架成分、细胞增殖相关等方面。     

球囊菌胁迫中华蜜蜂幼虫肠道过程中病原的转录组学研究

【目的】本研究利用RNA-seq技术对球囊菌胁迫的中华蜜蜂(中蜂)幼虫肠道进行深度测序,经趋势分析得到差异表达基因(DEGs)的显著表达模式,进而对胁迫过程中的球囊菌进行转录组学分析。【方法】利用Illumina HiSeq 2500平台对球囊菌胁迫的中蜂幼虫肠道进行深度测序,并利用相关软件进行了深入分析。最后,通过RT-qPCR对RNA-seq数据进行了验证。【结果】本研究共得到球囊菌的41133932条高质量clean reads。22865个DEGs共聚类为8个基因表达模式,其中,16769个DEGs聚类为2个显著上调趋势与2个显著下调趋势。GO富集分析结果显示,显著上调与显著下调趋势中的DEGs分别富集于40与37个GO term,基因富集数最多的为细胞进程(2486 unigenes)。KEGG代谢通路(pathway)富集分析结果显示,显著上调与显著下调趋势中的DEGs分别富集于119和112个pathway,基因富集数最多的分别是氨基酸生物合成(127 unigenes)与核糖体(98 unigenes)。进一步分析表明球囊菌在胁迫中蜂幼虫肠道的过程中通过提高物质合成促进其增殖,而宿主通过抑制球囊菌的蛋白合成抵御病原入侵。富集在MAPK信号通路的11个DEGs的表达水平随着胁迫时间的延长而逐渐下降,推测中蜂幼虫通过抑制该通路而阻遏球囊菌增殖。【结论】本研究不仅为揭示白垩病过程中的球囊菌-中蜂幼虫互作提供了重要信息,也为阐明不同抗性蜂种的球囊菌抗性差异奠定了基础。     

植物表达载体pKC—3的构建及大分子DNA连接策略

以根癌农杆菌双元载体ｐＣＡＭＢＬＡ１３００为基础,先后连接含有马铃薯蛋白酶抑制剂－Ⅱ基因（ＰⅡ）、苏云金杆菌毒蛋白基因（Ｂ．ｔ ｃｒｙＩ（Ａ））及雪花莲外源凝集素基因（ＧＮＡ）的完整表达片段,构建了抗多种稻田害虫的表达载体ｐＫＣ－３,将其导入根癌农杆菌ＬＢＡ４４０４,可进一步用于水稻抗虫基因转化的研究。载体构建进程采用多次的大分子ＤＮＡ片段连接,总结出了一套适合大片段连接转化的可行策略。     

Retrolinkin cooperates with endophilin A1 to mediate BDNF-TrkB early endocytic trafficking and signaling from early endosomes

Brain-derived neurotrophic factor (BDNF) binds to its cell surface receptor TrkB to regulate differentiation, development, synaptic plasticity, and functional maintenance of neuronal cells. Binding of BDNF triggers TrkB dimerization and autophosphorylation, which provides docking sites for adaptor proteins to recruit and activate downstream signaling molecules. The molecular mechanisms underlying BDNF-TrkB endocytic trafficking crucial for spatiotemporal control of signaling pathways remain to be elucidated. Here we show that retrolinkin, a transmembrane protein, interacts with endophilin A1 and mediates BDNF-activated TrkB (pTrk) trafficking and signaling in CNS neurons. We find that activated TrkB colocalizes and interacts with the early endosome marker APPL1. Both retrolinkin and endophilin A1 are required for BDNF-induced dendrite development and acute extracellular signal-regulated kinase activation from early endosomes. Suppression of retrolinkin expression not only blocks BDNF-triggered TrkB internalization, but also prevents recruitment of endophilin A1 to pTrk vesicles trafficking through APPL1-positive endosomes. These findings reveal a novel mechanism for BDNF-TrkB to regulate signaling both in time and space through a specific membrane trafficking pathway.     

家蚕二分浓核病毒VD1-ORF4基因截短多肽的表达和抗体制备

通过PCR扩增家蚕二分浓核病毒(Bombyx mori bidensovirus,BmBDV)VD1-ORF4基因序列中的两个DNA片段,将测序正确的两个目的片段分别亚克隆到原核表达载体pET-30a上,通过不同浓度的IPTG对含有重组质粒的大肠杆菌BL21(DE3)进行诱导,对诱导产物进行SDS-PAGE和Westen blot分析.结果表明,这两个截短多肽都获得了表达,其N-端融合有6个组氨酸.将割胶纯化的蛋白多肽与佐剂充分研磨,以研磨后的匀浆液对昆明小鼠进行皮下多点注射.获得的抗血清分别对原核诱导表达产物进行Western blot分析,结果表明,在特定的位置都能杂交到一条特异的蛋白带,表明制备的两个多抗能为深入研究VD1-ORF4基因的功能提供基础.     

江西省中华秋沙鸭越冬种群现状调查与胁迫因素分析

于2009年2月对江西省五大水系:赣江、抚河、信江、饶河和修水的越冬中华秋沙鸭种群展开专项调查,分析了江西省越冬中华秋沙鸭种群的数量、性比、空间分布格局、生境特征及其胁迫因素.主要采用样线法在五大水系的主要河道展开同步调查,调查样线总长度400 km.结果发现,江西省越冬中华秋沙鸭有26个生活群255只个体,雌雄性比为119:117,其生活群雌雄性比也多接近1:1.江西省五大水系中,除赣江外,在其他4个水系均发现有中华秋沙鸭越冬群体.除了在曾有中华秋沙鸭越冬记录的婺源、贵溪、弋阳、浮梁、修水发现其越冬群体外,还首次在江西省境内的宜黄、武宁和龙虎山地区发现有较大的中华秋沙鸭越冬群体,而且龙虎山泸溪河段分布有本次调查中个体数量最大的有53只个体的群体.人类采砂活动、水质污染、非法捕鱼和家禽饲养是目前影响该种群生存的主要胁迫因素.     

Molecular characterization of avian-like H1N1 swine influenza a viruses isolated in Eastern China, 2011

Currently, three predominant subtypes of influenza virus are prevalent in pig populations worldwide: H1N1, H3N2, and H1N2. European avian-like H1N1 viruses, which were initially detected in European pig populations in 1979, have been circulating in pigs in eastern China since 2007. In this study, six influenza A viruses were isolated from 60 swine lung samples collected from January to April 2011 in eastern China. Based on whole genome sequencing, molecular characteristics of two isolates were determined. Phylogenetic analysis showed the eight genes of the two isolates were closely related to those of the avian-like H1N1 viruses circulating in pig populations, especially similar to those found in China. Four potential glycosylation sites were observed at positions 13, 26, 198, 277 in the HA1 proteins of the two isolates. Due to the presence of a stop codon at codon 12, the isolates contained truncated PB1-F2 proteins. In this study, the isolates contained 591Q, 627E and 701N in the polymerase subunit PB2, which had been shown to be determinants of virulence and host adaptation. The isolates also had a D rather than E at position 92 of the NS1, a marker of mammalian adaptation. Both isolates contained the GPKV motif at the PDZ ligand domain of the 3′ end of the NS1, a characteristic marker of the European avian-like swine viruses since about 1999, which is distinct from those of avian, human and classical swine viruses. The M2 proteins of the isolates have the mutation (S31N), a characteristic marker of the European avian-like swine viruses since about 1987, which may confer resistance to amantadine and rimantadine antivirals. Our findings further emphasize the importance of surveillance on the genetic diversity of influenza A viruses in pigs, and raise more concerns about the occurrence of cross-species transmission events.     

紫茎泽兰根区土壤酚酸类物质组成及其对土传病菌的影响

紫茎泽兰根系释放的化感物质是导致其土壤环境发生变化的重要因素,可为其成功入侵提供有利条件.以空白土样为对照,提取健康和感染棉蚜的紫茎泽兰根区土壤酚酸类化感物质,采用GC-MS分析其组分和相对含量.结果表明：不同处理根区土壤酚酸类化感物质的种类没有变化,但相对含量发生了改变.其中,虫害处理土样中苯甲酸和对羟基苯甲酸的相对含量均显著高于对照,但对羟基肉桂酸的相对含量显著低于对照,且苯甲酸、对羟基苯甲酸和对羟基肉桂酸的相对含量在3种处理土样中的比例存在差异.用上述3种物质单体及它们在3个处理土样的比例复配进行抑菌试验发现：在较低浓度（50～150 mg·L^-1）下,3种化感物质对5种土传性病菌有显著的抑制作用,但复配后对5种病菌的抑制作用各不相同,虫害处理的复配对其中3种病原菌的抑制作用均低于对照和健康植株.     

The Role of Monoacylglycerol Lipase (MAGL) in the Cancer Progress

In recent years, metabolism is talked highly important in cancer research, especially the lipid metabolism. Researchers believe that the de novo fatty acid synthesis plays an important role in tumor development, while many studies illustrated that Endocannabinoids have anti-tumorigenic actions, including anti-proliferation, apoptosis induction, and anti-metastatic effects, MAGL as an important decomposing enzyme of both lipid metabolism and endocannabinoids system, additionally as a part of a gene expression signature contributes to different aspects of tumourigenesis.     

喀斯特关键带植被时空变化及其驱动因素

中国南方喀斯特地区广泛面临着生态问题,植被的保护与恢复倍受关注,对这一区域植被覆盖的进行监测和预测是非常必要的。以MODIS-NDVI为数据源,分析2000—2016年间,研究区不同地质背景,多种土地覆被类型的NDVI时空变化特征及驱动因素。结果表明:(1)从2000—2016年间,研究区植被覆盖整体呈增长趋势;其中喀斯特区域增长情况略优于非喀斯特区域。植被覆盖在空间上呈现东高西低;其中林地的NDVI值最高,耕地次之,依次草地,居民用地,水域,未利用地最低;在林地和耕地中,非喀斯特区域的NDVI值比喀斯特高,其余的土地覆被类型中都比喀斯特区域低。(2)研究区植被覆盖改善的地区占60.19%,退化地区占17.06%;草地,耕地区改善明显,退化主要在水域和建设用地; Hurst指数显示在研究区持续性改善的NDVI大于持续性退化;相比非喀斯特区域,喀斯特区域改善及持续性改善情况更佳。(3)整体而言,海拔对NDVI的空间分布影响力最大,温度次之,依次为降雨,夜间灯光指数;相比而言,非喀斯特区域NDVI空间分布更易受地形因子影响;喀斯特区域NDVI空间分布更易受气候差异及人类活动影响。(4)研究区分别有49%,45%,61%的NDVI与气温,降雨,日照的相关系数通过a=0.05的显著性检验;相比非喀斯特而言,喀斯特区域植被生长更易受气候变化的影响。