The structural basis of ribosome activity in peptide bond synthesis

Using the atomic structures of the large ribosomal subunit from Haloarcula marismortui and its complexes with two substrate analogs, we establish that the ribosome is a ribozyme and address the catalytic properties of its all-RNA active site. Both substrate analogs are contacted exclusively by conserved ribosomal RNA (rRNA) residues from domain V of 23S rRNA; there are no protein side-chain atoms closer than about 18 angstroms to the peptide bond being synthesized. The mechanism of peptide bond synthesis appears to resemble the reverse of the acylation step in serine proteases, with the base of A2486 (A2451 in Escherichia coli) playing the same general base role as histidine-57 in chymotrypsin. The unusual pK(a) (where K(a) is the acid dissociation constant) required for A2486 to perform this function may derive in part from its hydrogen bonding to G2482 (G2447 in E. coli), which also interacts with a buried phosphate that could stabilize unusual tautomers of these two bases. The polypeptide exit tunnel is largely formed by RNA but has significant contributions from proteins L4, L22, and L39e, and its exit is encircled by proteins L19, L22, L23, L24, L29, and L31e.     

砂石地基质栽培番茄日光温室CO_2浓度的变化规律研究

以越冬茬番茄为试验作物,研究了番茄砂石地基质栽培日光温室内CO2浓度在不同生育进程中的变化规律.结果表明:生育前期,室内CO2浓度日变化呈不规则的"U"型,生育后期,为不规则的"W"型.不同季节通风对室内CO2浓度的影响不同.在温度较高的生育前期,通风前温室CO2浓度高于大气浓度,通风降低了温室CO2浓度;在温度较低的生育后期,通风前温室CO2浓度低于大气浓度,通风可提高室内CO2浓度.但为了保持温室温度,生育后期通风时间应较短,风口也应较小,室内CO2浓度一直处于较低水平,不能满足作物光合作用的需要.番茄苗期以外各生育阶段的CO2浓度均低于800μL/L,且每天低于外界大气CO2浓度的时间达6.0～8.3h.因此,砂石地日光温室番茄基质栽培需要进行CO2施肥.     

GPCR engineering yields high-resolution structural insights into beta2-adrenergic receptor function

The beta2-adrenergic receptor (beta2AR) is a well-studied prototype for heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptors (GPCRs) that respond to diffusible hormones and neurotransmitters. To overcome the structural flexibility of the beta2AR and to facilitate its crystallization, we engineered a beta2AR fusion protein in which T4 lysozyme (T4L) replaces most of the third intracellular loop of the GPCR ("beta2AR-T4L") and showed that this protein retains near-native pharmacologic properties. Analysis of adrenergic receptor ligand-binding mutants within the context of the reported high-resolution structure of beta2AR-T4L provides insights into inverse-agonist binding and the structural changes required to accommodate catecholamine agonists. Amino acids known to regulate receptor function are linked through packing interactions and a network of hydrogen bonds, suggesting a conformational pathway from the ligand-binding pocket to regions that interact with G proteins.     

开放式空气中CO2浓度增高（FACE）对常规粳稻蛋白质和氨基酸含量的影响

为了明确未来大气CO2浓度升高对水稻蛋白质营养品质的影响,2009年利用稻田开放式空气CO2浓度增高(FACE,FreeAirCO2 Enrichment)系统,以武运粳21、扬辐粳8号和武粳15为供试品种,研究大田生长期CO2浓度升高200μmol.mol-1对常规粳稻蛋白质营养品质的影响。结果表明:大气CO2浓度增加使所有供试品种精米蛋白质含量平均下降5.6%,使氨基酸、必需和非必需氨基酸总量平均分别下降7.6%、6.7%和7.9%,均达极显著水平。大气CO2浓度增加使供试品种精米必需氨基酸占氨基酸总量百分比显著增加,使非必需氨基酸占氨基酸总量百分比显著下降,但对精米中必需和非必需氨基酸的相对含量无显著影响。从氨基酸组分看,大气CO2浓度增加使供试品种精米中7种必需氨基酸和8种非必需氨基酸的含量均显著或极显著下降。CO2处理与品种对精米蛋白质含量、氨基酸总量、必需和非必需氨基酸总量以及部分氨基酸组分有一定的互作效应,武运粳21上述参数对高浓度CO2的响应大于扬辐粳8号或武粳15对应参数的响应。以上结果说明本世纪中叶大气中的CO2浓度将使粳稻蛋白质营养品质下降,不同品种下降幅度存在一定差异。     

1-MCP处理对圆黄梨MA包装果实冷藏效果研究

研究不同MAP袋和1-MCP浓度对圆黄梨冷藏特性影响。结果表明：0.02 mm PE袋维持果实可滴定酸含量、色泽,抑制黑心,效果好于0.04 mm PVC袋。1-MCP处理有利于保持MAP贮藏果实货架期品质、果柄保鲜和抑制黑心,对0.02mm PE袋果实生理的调节作用大于0.04 mm PVC袋;0.5μL/L1-MCP处理可抑制0.04 mm PVC袋果实黑心,但贮藏后期（180 d）风味变淡,甚至出现异味。MAP袋冷藏圆黄梨可忍受4.7%CO2,无黑皮病、果肉褐变等伤害,但果实品质下降,CO2浓度应以〈3%为宜。圆黄梨最佳贮藏条件：0.02 mm PE袋＋1-MCP处理,180 d货架期品质良好,其余处理可短期贮藏（≤90 d）。此外,试验发现果实SSC与黑心指数显著负相关。     

辣椒矮秆突变体的表型及其对外源激素的响应

于EMS诱变辣椒自交系6421种子所构建的突变体库获得1份矮秆突变体(命名为E29),观察E29的表型和细胞学特征,研究E29突变性状的遗传规律和赤霉素(GA3)、24-表油菜素内酯(24-e BR)及生长素(IAA)对E29种子萌发和幼苗生长的影响。结果表明:与自交系6421相比,E29表现出植株矮化,叶片变大、变厚,叶色变深等特征;显微观察结果显示,E29变矮是由于其茎秆伸长区细胞长度缩短,叶片变大、变厚是由于其叶肉细胞变大及细胞数增加;遗传分析结果表明,突变性状由单隐性基因控制;外施不同质量浓度的激素,发现4.0 mg/L GA3处理显著促进E29的根与下胚轴伸长,1.0~4.0 mg/L的IAA处理显著抑制E29的根与下胚轴伸长,0.5~2.0 mg/L的24-e BR处理对E29的根和下胚轴伸长影响不明显,各激素处理均不能使之恢复至野生型6421的根与下胚轴长度,表明E29为BR不敏感型突变体,其突变性状与GA3、IAA调控无关。     

Studies on Single Cell Culture in vitro in Wheat--The variation of grain protein content and its fractions from regenerated plants

On the basis of previous studies dealing with the variation of major agronomic and yield characteristics of regenerated plants derived from single cell culture in vitro of common wheat (Triticum aestivum L.Cultivar NE 7742),the grain protein content and its fractions from regenerated plants with stable agronomic characteristics were studied from 1992 to 1995.The results showed that the variation of grain protein content and its fractions in somaclones from single cell culture in vitro were very significant and the range was very wide (11.53-17.70%).Several types of variation were found in the studies,espercially the type with higher protein content than that of cultivar NE 7742(non-culture parent).Among them.-20.69% of lines the grain protein content was significantly higher than that of NE 7742 and combined with high yielding potential.The tendency of variation of the four protein fractions showed that the variation of albumin was not obvious and maintained the same level as NE7742,the content of gliadin increased in some somaclones and decreased in others.However,the percentages both globulin and glutenin tended to increase.The variation of total amount of structural protein and the ratio between globulin and glutenin tended to increase.the variation of total amount of structural protein and the ratio between globulin and albumm was mainly influenced by globulin under the condition of culture in vitro.The variation of total amount of storage protein and the ratio between gliadin and glutenin was mainly by glutenin.The results mentioned above bemonstrated that the induction and screening of somaclonal variation could be an effective way in wheat improvement in combining high protein content with high yield.     

不同氮源及配比对钝顶螺旋藻的影响研究

本文就不同氮肥及其添加量对钝顶螺旋藻的生长量和蛋白质含量等方面进行了研究和探讨.结果表明:量添加尿素和碳酸氢铵都有助于钝顶螺旋藻生长量和蛋白质含量的增加,尿素的最佳添加量为0.02g/L·d,碳酸铵的最佳添加量为0.03g/L ·d.     

从芸薹属作物同工酶和蛋白质表型差异探讨杂种优势机理

以芸薹种(Brassica campestrisL.,syn.B.rapaL.)的3个亚种,即白菜(B.campestrisL.ssp.chinensisM ak ino)、大白菜(B.campestrisssp.pekinensisO lsson)和芜菁(B.campestrisssp.rapifera(Matzg.)S insk)等芸薹作物为研究材料,对其杂种优势形成进行同工酶和蛋白质分析,结果表明:不同品种在同一生育期或同一品种在不同生育期酶谱和蛋白质表型有差异,但把各品种不同生育期的谱带叠加,所得到的总谱带是相同的;品种间杂交其F1的同工酶和蛋白质谱带不存在“杂种带”或“互补带”;酶谱和蛋白质表型上的差异,可能是芸薹属作物杂种优势产生的生理基础之一。     

普通小麦TaZIP29-7B基因的克隆及表达研究

金属离子在小麦生长过程中起重要作用,但土壤中金属离子含量过高,会对小麦生长造成损害。锌铁转运蛋白ZIP家族(ZRT,IRT-like protein,ZIP)广泛存在于植物中,参与多种金属离子的转运,也能提高植物的耐盐性。利用同源克隆技术获得 AtZTP29在小麦中的同源基因TaZIP29-7B,并获得其启动子序列,利用生物信息学技术对所获得的基因以及启动子序列进行初步分析,预测启动子顺式作用元件,通过实时荧光定量PCR(Quantitative real-time PCR,qRT-PCR)技术分析小麦 TaZIP29-7B基因在根和叶中在不同金属离子溶液处理下的表达情况,以研究该基因在小麦体内中对重金属转运的作用。结果表明:序列分析显示 TaZIP29-7B基因(登录号MK203894)编码277个氨基酸,TaZIP29-7B蛋白为疏水性蛋白,定位在细胞膜上,存在信号肽,属于分泌蛋白,第89～263位置间的结构功能域ZIP,属于典型的ZIP超级家族结构域,具有8个跨膜区;进化分析显示TaZIP29-7B与单子叶植物山羊草(Aegilops tauschii)ZIP29蛋白同源性最高;启动子中存在多种响应胁迫的顺式作用元件;qRT-PCR结果显示,小麦 TaZIP29-7B基因在外界重金属盐的变化下表达量有所变化。说明研究结果为进一步改良小麦抗盐性提供了参考。     

根际CO_2浓度对网纹甜瓜生长和根系氮代谢的影响

【目的】探讨根际CO2浓度对网纹甜瓜生育的影响。【方法】采用雾培植株根际通CO2处理方式,在开花结果期对网纹甜瓜进行不同浓度的根际CO2处理,研究根际CO2浓度对网纹甜瓜植株生长和根系氮代谢的影响。【结果】在网纹甜瓜果实发育期间,根际2500μL·L-1和5000μL·L-1CO2浓度处理同350μL·L-1(对照)处理相比,植株生长受到明显抑制。根系硝态氮(NO3--N)含量、氨态氮(NH4+-N)含量、ATP酶活性、硝酸还原酶(NR)活性和谷氨酸脱氢酶(GDH)活性均表现出先增后降的变化趋势,且5000μL·L-1处理变化的幅度大于2500μL·L-1处理。根系中可溶性蛋白质含量、谷氨酰胺合成酶(GS)活性、谷草转氨酶(GOT)活性和谷丙转氨酶(GPT)活性在整个处理期间,均随着根际CO2浓度的升高而降低。根际高浓度CO2使根系伤流液中氨基酸的总量和大部分氨基酸的含量下降。【结论】在网纹甜瓜果实发育期间根际CO2浓度长期达到2500μL·L-1以上时,植株生长和果实发育受到抑制,根系对氮的吸收、代谢能力下降,植株氮循环水平降低。     

Na2CO3胁迫对马蔺幼苗生长量及生理特性的影响

为了探讨马蔺(Irjs lacteal Pall.var.chinensis(Fisch.)Koidz)在胁迫条件下的适应性反应,揭示其抗盐碱能力,设置了9个不同的Na2CO3浓度处理马蔺幼苗,并测定了逆境胁迫下马蔺幼苗的株高、相对电导率和MDA质量摩尔浓度以及游离脯氨酸质量分数的变化。试验结果表明:马蔺对浓度为0～125 mmol/L范围Na2CO3的胁迫作用有抗性(或耐性);Na2CO3浓度高于125 mmol/L,对马蔺幼苗有不可恢复的伤害作用;相对透性和膜脂过氧化作用的影响可以用来反映马蔺所受盐害的程度,游离脯氨酸质量分数大小可作为评价马蔺耐盐碱性的指标。     

外置式秸秆反应堆对日光温室内部环境及番茄光合特性的影响

【目的】研究外置式秸秆反应堆在日光温室越冬茬番茄的生产过程中,对日光温室环境因子及番茄光合特性的影响。【方法】以无CO₂发生装置的日光温室为对照温室,以利用外置式秸秆反应堆补充CO₂的日光温室为处理温室,观测在番茄不同生育阶段（苗期、结果期、转色期）温室内CO₂含量、气温、空气相对湿度等环境因子的日变化以及对番茄净光合速率、蒸腾速率、水分利用率和产量的影响。【结果】日光温室内CO₂含量的日变化曲线呈不规则的“N”形,空气湿度昼夜变化曲线呈“V”形。与对照温室相比,在番茄不同生育期,处理温室10:00-11:30 CO₂含量提高了230 μL/L,13:30-16:00提高了84～150 μL/L;夜间平均气温提高了1.5～2.0 ℃,空气相对湿度平均提高了4%～5%;并可显著提高不同生育阶段番茄的净光合速率和水分利用率;极显著降低苗期蒸腾速率,显著提高结果期和转色期番茄的蒸腾速率,产量和经济效益分别提高13 707 kg/hm²和59 310元/hm²。【结论】外置式秸秆反应堆能有效改善越冬茬番茄栽培过程中日光温室内的环境条件,缓解晴天日光温室内CO₂的亏缺,增加番茄的净光合速率和水分利用率,最终提高产量和经济效益。     

Arctic lakes and streams as gas conduits to the atmosphere: implications for tundra carbon budgets

Arctic tundra has large amounts of stored carbon and is thought to be a sink for atmospheric carbon dioxide (CO(2)) (0.1 to 0.3 petagram of carbon per year) (1 petagram = 10(15) grams). But this estimate of carbon balance is only for terrestrial ecosystems. Measurements of the partial pressure of CO(2) in 29 aquatic ecosystems across arctic Alaska showed that in most cases (27 of 29) CO(2) was released to the atmosphere. This CO(2) probably originates in terrestrial environments; erosion of particulate carbon plus ground-water transport of dissolved carbon from tundra contribute to the CO(2) flux from surface waters to the atmosphere. If this mechanism is typical of that of other tundra areas, then current estimates of the arctic terrestrial sink for atmospheric CO(2) may be 20 percent too high.     

Molecular cloning and functional characterization of apple U-box E3 ubiquitin ligase gene MdPUB29 reveals its involvement in salt tolerance

An E3 ubiquitin ligase gene(Genbank accession no.: MD01 G1010900) was cloned from the Royal Gala apple genome(Malus×domestica Borkh.). Sequence analysis showed that the length of the MdPUB29 gene was 1 275 bp, encoding 424 amino acids. Phylogenetic tree analysis indicated that the apple E3 ubiquitin ligase exhibited the greatest sequence similarity to Pyrus×bretschneideri. The predicted protein structural domain of MdPUB29 showed that it contained a U-box domain. qRT-PCR analysis showed that Md PUB29 was expressed widely in different tissues of the Royal Gala apple species, and was highly expressed in the root, while the expression of MdPUB29 was significantly inhibited by exogenous NaCl. Immunoblotting assays revealed that MdPUB29 protein abundance in tissue cultures of the Royal Gala apple accumulated under NaC l stress conditions. Three-dimensional protein structure prediction indicated that MdPUB29 was highly homologous with AtPUB29. The growing potential of MdPUB29-expressing apple calli and Arabidopsis were much stronger than that of the control under salt stress conditions, suggesting that MdPUB29 may positively regulate salt tolerance.     

水稻黄矮病毒结构蛋白的分离及其小鼠腹水抗体的制备

从人工接种感染黄矮病毒(Rice Yellow Stunt Virus,RYSV)的水稻茎杆中提取得到了部分纯化的RYSV制品,病毒制品经Tricine-SDS-PAGE得到分子量分别为84 kD,60 kD,31 kD和29.5 kD的4条主要蛋白带和分子量为170kD和43kD的两条次要蛋白带。根据弹状病毒结构蛋白命名法,按分子量大小依次命名为L,G,N,NS,M_1和M_2蛋白。从电泳凝胶上切下G和N蛋白带,制成抗原免疫BALB/C小鼠;制得G和N蛋白小鼠腹水抗体。经Dot-blot试验测得N蛋白抗体效价为1:1000,G蛋白抗体效价为1:400。     

氮肥对小麦籽粒营养品质的影响

本试验主要研究了氮肥施用量和施用时期对两个小麦品种(鲁麦8号和泰山1号)氮素营养、籽粒产量和营养品质的影响。试验表明,在本试验条件下,每亩纯氮施用量从10公斤增至20公斤,对两个品种的籽粒产量、籽粒蛋白质含量的影响不显著(5%水平),但却显著增加其蛋白质产量;在各施氮期中,拔节期处理的籽粒产量最高,揚花肥提高籽粒蛋白质含量和蛋白质产量的效应最大,并且各施肥处理的籽粒产量、籽粒蛋白质含量和蛋白质产量均比对照高。试验还表明,合理运筹施氮时期和施氮量可成功地打破籽粒产量和蛋白质含量间的负相关关系,使籽粒产量和蛋白质含量得到同步提高,并有效地增加单位籽粒中量重的赖氨酸含量。     

小麦WPBF与高分子量谷蛋白基因上游Prolamin-Like box的特异结合

【目的】克隆普通小麦胚乳特异表达转录因子基因WPBF,对其原核表达条件进行优化,并验证其编码蛋白和小麦高分子量谷蛋白亚基基因（HMW-GS）上游Prolamin-Like box的结合特性,为小麦HMW-GS表达调控机制的阐明奠定基础。【方法】利用RT-PCR扩增得到WPBF编码区,将其连接到pET-21a-MBP载体中,并分别在E.coli T7 Express和Origami B（DE3）菌株中诱导表达。依次利用Amylose和Ni-NTA亲和层析柱纯化重组蛋白,最后通过凝胶迁移阻滞试验（EMSA）对重组蛋白的DNA结合特性进行分析。【结果】在33℃的Origami B（DE3）菌株中表达含MBP和His双标签的重组WPBF大部分以可溶性形式存在,其表达量占细菌总蛋白的48.0%。重组WPBF和含HMW-GS上游Prolamin-Like box的两类DNA探针能发生特异结合,但重组WPBF和含“TGCAAAG”基序DNA探针的结合强度高于其和含“TGCAAG”基序DNA探针的结合。【结论】建立了优化的WPBF原核表达系统,并获得具有生物活性的重组蛋白;重组WPBF能与HMW-GS启动子来源的两种Prolamin-Like box特异性结合, 但结合能力存在差异。     

羊草几丁质酶ClassⅡ基因的克隆、生物信息学分析及原核表达

根据已报道的羊草几丁质酶基因的EST序列,利用RACE技术,从100 mmol/L Na2CO3胁迫的羊草叶片中克隆得到羊草几丁质酶基因cDNA序列全长(GenBank登录号为GQ397277),命名为LcChi2基因。经序列测定和生物信息学分析,结果表明该序列开放阅读框为771 bp,编码256个氨基酸,为Ⅱ类几丁质酶,属于19家族,分子量约为27.4 kDa,预测等电点为8.67,与小麦(Triticum aestivum)、大麦(Hordeum vulgare)、黑麦(Secale cereale)和水稻(Oryza sativa)等植物的几丁质酶具有高度的同源性,序列相似性分别为96.5%、965%、95.2%和80.5%,推测LcChi2蛋白与其他几丁质酶执行相似的功能。在大肠杆菌中进行了原核表达,为后期植物转化的验证准备多克隆抗体。本研究所获得的信息为今后对羊草几丁质酶基因功能的进一步研究奠定了基础。