草麻黄高通量转录组分析及黄酮类代谢途径相关基因的鉴定

草麻黄（Ephedra sinica Stapf）植物富含黄酮类和生物碱类化合物,具有重要药用价值,并且适应性强,在防风固沙、改善沙漠环境等方面具有重要作用。通过对已测序草麻黄转录组的原始数据进行重新组装、拼接等生物信息学分析,共得到7 947 954个clean reads,13 389条unigene序列。经过Nr,KOGs,GO,KEGG和Swiss Prot等数据库分析后,获得10 481个Nr注释,4 533个KOG功能注释,7 121个GO功能注释,5 833个KEGG注释,8 039个 Swiss Prot注释,并利用KEGG通路分析技术发掘草麻黄中参与黄酮类化合物代谢途径相关基因,为克隆草麻黄黄酮合成关键基因、黄酮类化合物的生物合成提供重要的遗传资源,同时为其抗逆机制的研究和药用价值开发提供依据。     

植物系统获得性抗性研究进展

植物系统获得性抗性(Systemic Acquired Resistance,SAR)是一种能够诱导植物持续抵御病原微生物侵害的一种防御机制。SAR需要信号分子水杨酸(Salicylic Acid,SA)参与,并与能够提高抗性的病程相关蛋白(PR)的积累有关。通过对模式植物拟南芥的研究发现,异分支酸合成酶途径是合成SA的主要途径。正调控因子NPR1与转录因子TGA相互作用,诱导防卫基因表达,进而激发SAR反应。最新研究表明,脂质分子有可能是SAR反应中的移动信号分子。这些研究结果有助于进一步了解SAR反应。     

Insulin rapidly increases diacylglycerol by activating de novo phosphatidic acid synthesis

The mechanisms whereby insulin increases diacylglycerol in BC3H-1 myocytes were examined. When [3H]arachidonate labeling of phospholipids was used as an indicator of phospholipase C activation, transient increases in [3H]diacylglycerol were observed between 0.5 and 10 minutes after the onset of insulin treatment. With [3H]glycerol labeling as an indicator of de novo phospholipid synthesis, [3H]diacylglycerol was increased maximally at 1 minute and remained elevated for 20 minutes. [3H]Glycerol-labeled diacylglycerol was largely derived directly from phosphatidic acid. Insulin increased de novo phosphatidic acid synthesis within 5 to 10 seconds; within 1 minute, this synthesis was 60 times greater than that of controls. Thus, the initial increase in diacylglycerol is due to both increased hydrolysis of phospholipids and a burst of de novo phosphatidic acid synthesis. After 5 to 10 minutes, de novo phosphatidic acid synthesis continues as a major source of diacylglycerol. Both phospholipid effects of insulin seem important for generating diacylglycerol and other phospholipid-derived intracellular signaling substances.     

Emergence of drug-resistant influenza virus: population dynamical considerations

Given the considerable challenges to the rapid development of an effective vaccine against influenza, antiviral agents will play an important role as a first-line defense if a new pandemic occurs. The large-scale use of drugs for chemoprophylaxis and treatment will impose strong selection for the evolution of drug-resistant strains. The ensuing transmission of those strains could substantially limit the effectiveness of the drugs as a first-line defense. Summarizing recent data on the rate at which the treatment of influenza infection generates resistance de novo and on the transmission fitness of resistant virus, we discuss possible implications for the epidemiological spread of drug resistance in the context of an established population dynamic model.     

The shared antibiotic resistome of soil bacteria and human pathogens

Soil microbiota represent one of the ancient evolutionary origins of antibiotic resistance and have been proposed as a reservoir of resistance genes available for exchange with clinical pathogens. Using a high-throughput functional metagenomic approach in conjunction with a pipeline for the de novo assembly of short-read sequence data from functional selections (termed PARFuMS), we provide evidence for recent exchange of antibiotic resistance genes between environmental bacteria and clinical pathogens. We describe multidrug-resistant soil bacteria containing resistance cassettes against five classes of antibiotics (β-lactams, aminoglycosides, amphenicols, sulfonamides, and tetracyclines) that have perfect nucleotide identity to genes from diverse human pathogens. This identity encompasses noncoding regions as well as multiple mobilization sequences, offering not only evidence of lateral exchange but also a mechanism by which antibiotic resistance disseminates.     

Systemic signaling and acclimation in response to excess excitation energy in Arabidopsis

Land plants are sessile and have developed sophisticated mechanisms that allow for both immediate and acclimatory responses to changing environments. Partial exposure of low light-adapted Arabidopsis plants to excess light results in a systemic acclimation to excess excitation energy and consequent photooxidative stress in unexposed leaves. Thus, plants possess a mechanism to communicate excess excitation energy systemically, allowing them to mount a defense against further episodes of such stress. Systemic redox changes in the proximity of photosystem II, hydrogen peroxide, and the induction of antioxidant defenses are key determinants of this mechanism of systemic acquired acclimation.     

铜胁迫下的西瓜食酸菌转录组分析

【目的】分析西瓜食酸菌(Acidovorax citrulli)在铜胁迫下转录水平的响应特征,研究对其抗铜系统和抗铜机制,为进一步解析西瓜食酸菌的铜耐受分子机制积累数据。【方法】以西瓜食酸菌亚群I的抗铜菌株FC440铜胁迫和无胁迫培养下的菌体为材料,提取RNA并利用RNA-Seq技术获取细菌对铜胁迫的应答数据,分析数据质量、差异表达基因的类型、KEGG富集与Pathway注释、GO富集分析等转录组水平特征,并进行qPCR验证。【结果】共获得6个转录组文库,总数据量23.6 Gb;该细菌共有4 344个基因表达,其中有466个基因差异表达(上调266个,下调200个);差异表达的基因显著富集于ABC转运系统的跨膜转运、双组分系统通路的信号转导以及氨基酸代谢等约9大通路;基因表达趋势与转录组测序数据的趋势一致。【结论】铜胁迫下谷氨酸、谷胱甘肽等氨基酸代谢在西瓜食酸菌抗铜胁迫中发挥重要作用;双组分系统参与西瓜食酸菌对铜离子胁迫的响应且受到负调控;该菌中可能存在包含ABC转运体的跨膜转运、氧化还原反应、胞内束缚沉淀等多种系统互作的抗铜机制。     

烟草的TMV侵染耐受性与谷胱甘肽代谢途径的关系

为了深入了解烟草对烟草花叶病毒(TMV)的抗性机制,以耐受TMV感染品种豫烟8号和具有相同遗传背景对花叶病敏感的品种NC89为材料,通过对高通量测序获得的转录组数据进行生物信息学分析发现,二者接种TMV后的差异表达基因富集通路中共同存在谷胱甘肽代谢途径,且耐受性品种豫烟8号中该途径有增强的趋势。采用荧光定量PCR和分光光度法检测了两品种接种TMV前后谷胱甘肽代谢途径中关键酶谷胱甘肽S转移酶(GST)基因表达水平、酶活性以及谷胱甘肽(GSH)含量的变化。结果表明,接种TMV后两品种中GST基因表达水平升高,GST酶活性增强,GSH含量升高,耐受性品种豫烟8号增幅更明显。通过硫元素的丰缺试验研究了接种TMV对GST基因及与TMV侵染耐受性相关基因表达水平的影响,结果显示,+S和-S处理中两品种接种病毒后这些基因的表达趋势一致,-S处理的表达量低于+S处理。GST、PR1-a、HSP90和Catalase-3这些抗病相关基因都呈上调表达,其中豫烟8号接种后的表达量均高于NC89;而Psb A和PhotosystemⅡ10 k Da polypeptide 2个涉及光合作用的基因均呈现下调表达,但是接种后豫烟8号的下调幅度小于NC89。以上表明,在耐受TMV感染的品种中,谷胱甘肽代谢增强对细胞环境的氧化还原平衡有一定的促进作用,使光系统Ⅱ中的关键成分D1蛋白基因Psb A的表达相对稳定,从而减轻光合系统的破坏程度。     

黄果柑果实抗坏血酸、谷胱甘肽和有机酸积累特性

为探究黄果柑果实发育过程中抗坏血酸、谷胱甘肽和有机酸的积累特性及相关性,采用高效液相色谱法测定黄果柑果实有机酸和抗坏血酸的含量。结果表明:黄果柑果实发育过程中抗坏血酸的积累主要以AsA为主,含量为21.11~33.73 mg·100g^-1;谷胱甘肽的积累以GSH为主,含量为58.99~186.37 μmol·L^-1;果实中检测出7种有机酸,其中柠檬酸含量最高,为641.10~4 192.85 mg·100g^-1,总体呈“降-升-降”趋势。T-AsA含量与AsA/DHA、GSH/GSSG呈极显著相关,说明AsA/DHA和GSH/GSSG氧化还原能力对T-AsA的积累和维持有重要作用;T-GSH含量与有机酸组分呈极显著正相关,表明有机酸和T-GSH之间呈相互促进的关系;果实中T-AsA和AsA含量对主成分的影响程度相对较大;通径分析表明,柠檬酸和草酸与T-GSH的直接通径系数最大,说明柠檬酸和草酸对T-GSH的积累作用显著。     

Resistance to enediyne antitumor antibiotics by CalC self-sacrifice

Antibiotic self-resistance mechanisms, which include drug elimination, drug modification, target modification, and drug sequestration, contribute substantially to the growing problem of antibiotic resistance among pathogenic bacteria. Enediynes are among the most potent naturally occurring antibiotics, yet the mechanism of resistance to these toxins has remained a mystery. We characterize an enediyne self-resistance protein that reveals a self-sacrificing paradigm for resistance to highly reactive antibiotics, and thus another opportunity for nonpathogenic or pathogenic bacteria to evade extremely potent small molecules.     

大蒜素抗肿瘤的研究进展

大蒜素可以降低某些肿瘤的患病率。试验证明,大蒜素不仅能抑制致癌物的致癌作用,而且对肿瘤细胞的增殖也有抑制作用。目前的研究结果显示,大蒜素的抑癌机制为:抗氧化,抑制致癌物的形成,影响肿瘤细胞的细胞周期并诱导肿瘤细胞凋亡,增强机体免疫功能,对抗肿瘤药物有增敏作用等。     

中药植物川牛膝基因组从头测序及分析

川牛膝是我国重要大宗药材,但近年来品质退化严重,品种真伪混杂。为深入研究川牛膝有效成分积累的分子基础,采用第二代测序技术利用Illumina Hi-seq 2000测序平台对川牛膝进行全基因组测序,使用AByss进行初步组装,得到一个包含大量基因组序列信息的数据集,并对其进行重复序列及编码序列注释。在该测序结果基础上,初步预测得到川牛膝体内甾体合成途径,并对鉴定川牛膝真伪的SCAR分子标记进行了初步定位,为研究川牛膝主要有效成分杯苋甾酮的合成途径,改良川牛膝品质提供了基础。     

硒对甜樱桃叶片褪黑素和谷胱甘肽氧化还原循环的影响

【目的】研究不同价态和浓度硒对甜樱桃叶片褪黑素含量以及谷胱甘肽(GSH)氧化还原循环的影响,为探究外源硒、内源褪黑素和GSH氧化还原循环间的关系提供参考。【方法】2015年在四川广元进行田间试验,以两年生甜樱桃‘红蜜’和‘布鲁克斯’为试验材料,试验包括清水处理(CK)、单施Se~(6+)(来源于Na_2SeO_4)、单施Se~(4+)(来源于Na_2SeO_3)、Se~(6+)+褪黑素和Se~(4+)+褪黑素5个处理,30 d后采取叶片,测量叶片硒含量和褪黑素的含量。2016年以重庆綦江两年生甜樱桃‘布鲁克斯’叶片为试验材料进行离体试验,将甜樱桃离体叶片放置在2 mg·L~(-1) Se~(6+)处理0、1、2、3和4 d,以清水处理为对照,每天上午10:00采取叶片;将离体叶片分别置于0、0.5、1.0、2.0、4.0 mg·L~(-1) Se~(6+)溶液24 h后采取叶片,离体试验分别测量叶片硒和褪黑素的含量以及GSH氧化还原循环的物质和酶活性。【结果】田间试验中,2.0 mg·L~(-1) Se~(4+)和Se~(6+)处理显著降低了‘红蜜’和‘布鲁克斯’甜樱桃叶片褪黑素含量,且两种甜樱桃叶片褪黑素含量在Se~(6+)+褪黑素和Se~(4+)+褪黑素处理下分别比单施Se~(6+)和单施Se~(4+)处理高。离体试验中,硒处理第1天‘布鲁克斯’离体叶片褪黑素含量高于对照,之后低于对照且随着时间延长而逐渐降低。硒处理第1天迅速降低了GSH氧化还原循环的物质和酶活性,即GSH、氧化性谷胱甘肽(GSSG)、GSH+GSSG和GSH/GSSG比值低于对照,谷胱甘肽过氧化物酶(GPX)和谷胱甘肽还原酶(GR)活性也低于对照,之后随着时间的延长,GSH氧化循环效率升高并超过对照。在不同浓度Se~(6+)处理24 h下,Se~(6+)处理增加了叶片褪黑素的含量,且随着浓度的增加而增加,GSH氧化循环效率则呈下降的趋势。【结论】外源硒处理提高了甜樱桃叶片硒含量,且六价硒的效果显著高于四价硒。外源硒处理影响甜樱桃褪黑素的含量和GSH氧化还原循环。     

The many roles of computation in drug discovery

An overview is given on the diverse uses of computational chemistry in drug discovery. Particular emphasis is placed on virtual screening, de novo design, evaluation of drug-likeness, and advanced methods for determining protein-ligand binding.     

De novo crypt formation and juvenile polyposis on BMP inhibition in mouse intestine

Little is known about the signaling mechanisms that determine the highly regular patterning of the intestinal epithelium into crypts and villi. With the use of mouse models, we show that bone morphogenetic protein (BMP)-4 expression occurs exclusively in the intravillus mesenchyme. Villus epithelial cells respond to the BMP signal. Inhibition of BMP signaling by transgenic expression of noggin results in the formation of numerous ectopic crypt units perpendicular to the crypt-villus axis. These changes phenocopy the intestinal histopathology of patients with the cancer predisposition syndrome juvenile polyposis (JP), including the frequent occurrence of intraepithelial neoplasia. Many JP cases are known to harbor mutations in BMP pathway genes. These data indicate that intestinal BMP signaling represses de novo crypt formation and polyp growth.     

烟草叶片发育过程中抗坏血酸-谷胱甘肽循环清除H2 O2 的研究

[目的]研究烟草叶片发育过程中H2O2积累以及抗坏血酸-谷胱甘肽(AsA-GSH)循环清除H2O2能力的变化。[方法]以烟草(Nicotiana tabacumL.cvNC89&JYH)为材料,研究了叶片发育过程中H2O2积累与AsA-GSH循环的关联。[结果]结果表明,抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)活性,AsA、GSH含量在叶片幼嫩及成熟时期维持较高水平,衰老时下降较快,与H2O2含量存在相关性,且AsA-GSH循环系统在运行过程中保持平衡。[结论]AsA-GSH循环系统的运行与H2O2代谢存在相关性。     

外源硒调控植物重金属胁迫机制的研究进展

硒是对植物有益的微量元素，在植物生长发育以及抗性方面起着重要的作用。越来越多的研究发现，外源添加硒可以有效缓解植物重金属胁迫，且相关研究也逐渐深入。总结归纳了近年来植物对硒的吸收、转运和积累规律，以及硒缓解重金属胁迫机制的相关研究进展，并比较了硒对不同种类重金属胁迫缓解机制的异同点，以期为提高植物抵御重金属胁迫能力提供理论基础。     

利用突变体揭示植物重金属镉抗性和积累机制的研究进展

重金属镉是土壤中毒性最强的污染物之一,不但严重影响植物生长发育,而且通过植物体内积累进入食物链危害人类健康。突变体不但是作物改良的重要资源,也是基因功能研究的基础材料。总结了利用各种突变体对植物镉抗性和积累的分子机理所做的研究和取得的丰硕成果,在此基础上提出了参考模式植物在作物上利用突变体开展作物抗镉、低积累机制及分子育种研究的策略,为作物抗镉和低积累品种的选育提供理论指导。     

Germline mutations in genes within the MAPK pathway cause cardio-facio-cutaneous syndrome

Cardio-facio-cutaneous (CFC) syndrome is a sporadic developmental disorder involving characteristic craniofacial features, cardiac defects, ectodermal abnormalities, and developmental delay. We demonstrate that heterogeneous de novo missense mutations in three genes within the mitogen-activated protein kinase (MAPK) pathway cause CFC syndrome. The majority of cases (18 out of 23) are caused by mutations in BRAF, a gene frequently mutated in cancer. Of the 11 mutations identified, two result in amino acid substitutions that occur in tumors, but most are unique and suggest previously unknown mechanisms of B-Raf activation. Furthermore, three of five individuals without BRAF mutations had missense mutations in either MEK1 or MEK2, downstream effectors of B-Raf. Our findings highlight the involvement of the MAPK pathway in human development and will provide a molecular diagnosis of CFC syndrome.