保护酶POD在木薯种质抗螨中的功能初探

为了从木薯和害螨互作的角度初步阐明保护酶POD在木薯种质抗螨中的功能。本研究通过荧光定量PCR和酶活性分析手段测定了朱砂叶螨取食抗、感螨木薯种质后,POD的表达量和酶活性分别在木薯和朱砂叶螨中的变化情况。结果表明,朱砂叶螨取食1d和8d后,感螨木薯种质BRA900体内MePOD的表达量和POD酶活力仅分别较为害前提高1.13倍、0.97倍和1.08倍、1.10倍,而在抗螨木薯种质C1115体内则分别较为害前提高2.35倍、2.15倍和1.89倍、1.81倍,显著高于感螨木薯水平。朱砂叶螨取食感螨种质BRA9001d和8d后,保护酶TcPOD的基因表达量和POD酶活分别是为害前的0.99倍、1.16倍和1.11倍、1.17倍,而取食抗螨种质C11151d和8d后,保护酶TcPOD的基因表达量和POD酶活分别降低至为害前的0.53倍、0.52倍和0.51倍、0.55倍,显著低于取食感螨木薯水平。本研究初步证实了木薯保护酶POD受螨害后活性显著增强能够有效减轻氧化损伤,朱砂叶螨POD显著抑制阻碍其为害木薯,从而形成木薯的抗螨性。上述结果为将POD作为潜在基因资源应用于抗螨木薯分子育种提供了理论基础。     

siRNA介导的PRR11表达抑制导致肺癌细胞系基因表达谱变化的分析

Proline rich 11(PRR11)是本课题组鉴定的一个新的肿瘤相关基因。为研究PRR11介导肺癌发生发展相关的分子机制,本研究分析了PRR11表达被抑制后人肺癌细胞系H1299的全基因组基因表达谱的变化。首先,采用siRNA抑制H1299细胞中PRR11的表达,提取总RNA,采用基因芯片分析全基因组基因表达谱的变化。然后,对呈现差异表达的基因进行GO和Pathway富集分析,并对部分重要的候选基因进行定量RT-PCR验证。基因芯片结果表明,采用siRNA有效抑制H1299细胞中PRR11表达后,共有550个基因的mRNA水平出现明显变化,其中139个基因表达上调,411个基因表达下调。生物信息学分析结果表明,上述差异表达的基因显著富集于细胞周期和MAPK通路。定量RT-PCR验证分析结果表明,PRR11表达抑制后确实可导致多个与细胞周期和肿瘤发生发展密切相关的基因(包括DHRS2、EPB41L3、CCNA1、MAP4K4、RRM1、NFIB)呈现显著的表达变化。这些结果提示,PRR11可能通过上述通路和/或基因的表达变化参与肺癌的发生发展过程。     

薏苡种子形态性状多样性评价

薏苡是优质的药食兼用作物。本研究对我国36份薏苡种质的种子形态性状进行了主成分分析和聚类分析。结果表明,36份薏苡种质具有较高的形态性状多样性。不同薏苡种质的4个种子质量性状间存在差异。单因素方差分析显示11个种子数量性状在不同薏苡种质间呈极显著差异。数量性状的变异系数在8.43%~68.01%之间,种壳与外种皮重量的变异系数最大而种仁长度的变异系数最小。主成分分析将36份薏苡种质分为中部及南部栽培种质、北部栽培种质和野生种质3大类,3类种质的形态特征存在明显差异。聚类分析将36份薏苡种质分为5大类群,即中部及南部栽培种质、北部栽培种质和3个类群野生种质。初步筛选出6份可能具有较高抗肿瘤活性的薏苡种质FJNJ、TJ-BK-1、TJ-BK-2、MY-BK-1、MY-BK-2和JYSD。     

几种薏苡的过氧化物酶和多酚氧化酶同工酶分析

用聚丙烯酰胺凝胶电泳（PAGE）的方法,对薏苡,野生薏苡和水生薏苡的几个不同种质分别进行了过氧化物酶（POD）和多酚氧化酶（PPO）的同工酶酶谱分析,结果表明：POD和PPO含量丰富,活性强,酶谱复杂,酶带清晰,稳定,种间差异明显。结果可以作为薏苡研究种质亲缘关系的基础。     

泾阳茯砖茶中“金花菌”的分离、鉴定及抗氧化酶活性

目的分离鉴定泾阳茯砖茶中的金花菌,探讨其发酵产物的抗氧化酶活性。方法采用涂布平板法分离纯化泾阳茯砖茶中的"金花菌",通过其ITS序列分析进行分子鉴定。利用抗氧化酶试剂盒测定不同时期"金花菌"发酵液的抗氧化酶活性。结果从茶叶中分离出1株"金花菌"J17,通过ITS序列分析初步确定其为赤散囊菌(Eurotium rubrum)。在J17菌株发酵后第1、7、10、14和21天,其发酵液中过氧化物酶(POD)活性(U/mL)分别为0.22±0.11、1.22±0.13、2.46±1.64、7.35±2.77和45.19±1.54;过氧化氢酶(CAT)活性(U/mL)分别为0.06±0.00、0.45±0.00、1.52±0.20、2.35±0.20和2.75±0.37;总超氧化物歧化酶(T-SOD)活性(U/mL)分别为25.55±10.12、50.90±3.72、76.77±6.75、89.07±3.33和36.34±12.32。发酵14d左右,3种抗氧化酶活性均较高,与茶砖在发花车间的发花时间一致。茯砖茶沸水冲泡液和沸水冲洗液中POD和CAT活性差异无统计学意义,而沸水冲泡液的T-SOD活性显著高于沸水冲洗液。结论泾阳茯砖茶的抗氧化酶活性与茶砖发花期产生的赤散囊菌有关,泾阳茯砖茶适用于热水冲泡或煮沸后饮用。     

梨树腐烂病抗性种质筛选及相关生理生化特性研究

以46份不同类型梨树种质资源的休眠期枝条为试材,连续2年分析枝条的腐烂病抗病性,并测定枝条韧皮部组织POD、SOD活性,以及总酚含量和组织疏松度;采用酶联免疫分析法(ELISA)测定抗病品种‘秋白’和感病品种‘茌梨’枝干中内源水杨酸(SA)和茉莉酸(JA)的含量;并依据转录组文库中SA和JA合成相关基因(PAL、ICS、AOS和AOC)片段的序列信息,通过实时荧光定量(qRT-PCR)技术分析基因的相对表达量。结果表明:(1)不同类型的梨树种质资源之间腐烂病发病程度表现出较大的差异和多样性,并以秋子梨种类抗性最强,西洋梨抗性最弱。(2)梨树种质资源种类腐烂病发病程度与其枝条韧皮部POD、SOD活性和组织疏松度无显著相关性,与枝条韧皮部总酚含量呈显著正相关关系。(3)在病菌接种情况下,抗病品种‘秋白’枝干中SA和JA含量显著上升,同时其相关合成酶基因PAL和AOS表达量也显著升高,而感病品种‘茌梨’枝干中SA和JA含量变化不明显,相关合成酶基因PAL、ICS、AOS和AOC的表达量则显著下降。研究发现,梨树不同类型种质的腐烂病抗性有明显差异,其发病程度与枝条韧皮部总酚含量呈显著正相关关系;内源SA和JA可能参与抗病梨树种质资源对腐烂病的抵御过程。     

油松雌性不育系的POD同工酶和蛋白质多肽分析

用聚丙烯酰胺凝胶电泳技术分析了油松雌性不育系和可育系雌配子体发育关键时期的POD和蛋白质多肽的变化。结果表明不育系POD同工酶在雌配子体处于几十个游离核时期活性较高,在雌配子体游离核停止分裂期活性降低,雌配子明显败育的后期,该酶的活性又有所增强。不育系的POD同工酶谱与可育系存在差异,出现了Rf值分别为0.39、0.77两条特异谱带,Rf为0.77的谱带不仅稳定而且表达量很高,Rf值为0.56的谱带表达量明显低于可育株。蛋白质多肽的分析结果表明不育系中存在分子量为18．7kD的特异蛋白质多肽,分子量为38．3kD的蛋白质多肽表达量明显低于可育系。     

干旱胁迫对烟草两品种幼苗生理生化特性及NtDEGP5基因表达的影响

以烟草Nicotianatabacum抗旱品种NC82和干旱敏感品种云烟87幼苗为材料,利用PEG-6000对幼苗进行干旱胁迫处理,研究干旱胁迫对烟草不同品种Fv/Fm、SOD活性、POD活性、MDA含量等生理生化指标及NtDEGP5基因表达的影响,为进一步开展NtDEGP5基因的功能研究提供依据。结果表明,干旱胁迫对烟苗的Fv/Fm、SOD活性、POD活性、MDA含量有影响,随着干旱的持续,Fv/Fm呈逐渐下降趋势,POD、SOD活性和MDA含量呈先升高后降低的趋势;不同品种对干旱胁迫的响应有差异,在干旱胁迫0～9 h,云烟87幼苗的Fv/Fm降幅大于NC82,POD、SOD活性、MDA含量的增幅小于NC82,胁迫12h后与对照差异不显著。干旱胁迫下不同品种、不同器官间的Nt DEGP5基因表达有差异,云烟87幼苗根、茎、叶中的表达量均较低,NC82幼苗根中的表达量较低,但干旱胁迫9～12 h后叶和茎的表达量较高。NC82幼苗叶片NtDEGP5基因表达量与Fv/Fm、POD活性、MDA含量呈显著正相关。Fv/Fm、SOD活性、POD活性、MDA含量可作为烟草苗期抗旱性评价指标,NtD...     

低温胁迫下烤烟幼苗叶片光合作用和抗氧化能力基因差异表达谱

对低温(5—7℃)胁迫下烤烟"K326"幼苗叶片光合指标、膜氧化水平及其抗氧化指标进行测定,并利用数字化基因表达谱技术进行基因差异表达分析。低温胁迫后烤烟幼苗叶绿素含量、光合能力显著下降,脯氨酸含量、丙二醛含量上升,超氧化物歧化酶活性、过氧化氢酶活性、抗坏血酸含量和谷胱甘肽含量均显著上升。低温胁迫后有2357个基因发生了显著差异表达,其中1673个基因表达上调、684个基因表达下调,其分子功能、细胞位置和主要代谢过程均涉及光系统、膜氧化系统和抗氧化系统。对涉及到的代谢过程进行分析,结果表明:光合天线蛋白调控基因表达量均显著下降、光合作用的主要调控基因表达量多数表现为显著下调、而与氧化能力相关的谷胱甘肽代谢差异表达基因大多数显著上调。基因差异表达谱分析结果和低温胁迫后叶片光合能力、抗氧化能力生理生态指标测定结果基本一致,为进一步研究低温胁迫对作物的生态影响和研究基因克隆与功能提供基础。     

铜绿假单胞杆菌QS相关基因差异表达的研究

选取100个与铜绿假单胞杆菌（Pseudomonas aeruginosa）群感效应（quorum-sensing,QS）相关的基因,克隆这些基因片段于pMD-18T载体,测序鉴定,点样制备cDNA基因芯片。制备cy3-dCTP／cy5-dCTP标记的探针,与芯片杂交。初步研究了处于不同生长期的铜绿假单胞杆菌基因的表达差异。指数中期和平台初期相比,有9个QS基因表达量最著增加,有6个基因表达量显著下降。利用芯片做针对铜绿菌假单胞杆菌药物的筛选：妥布霉素（Tobramycin）给药后细菌基因发生差异表达。证明了该cDNA芯片用于药物筛选的可行性。在国内首次研制开发了QS相关基因的cDNA芯片。应用基因芯片技术建立的铜绿假单胞杆菌QS相关基因研究平台,为找到能较好抑制铜绿假单胞杆菌正常生长的药物研究提出新的解决方法。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Physiological characteristics of various species of strains of carboxydobacteria

Seven strains of aerobic carbon monoxide-oxidizing bacteria (carboxydebacteria) when growing on CO as sole source of carbon and energy had doubling times which ranged from 12–42 h. The activity profiles obtained after discontinuous sucrose density gradient centrifugation indicated that the CO-oxidizing enzymes are soluble and the hydrogenases are membrane-bound in all strains examined. The CO-oxidizing enzymes of Pseudomonas carboxydohydrogena, Pseudomonas carboxydoflava, Comamonas compransoris, and the so far unidentified strains OM2, OM3, and OM4 had a molecular weight of 230,000; that of Achromobacter carboxydus amounted to 170,000. The molecular weights of the CO-oxidizing and H₂-oxidizing enzymes turned out to be identical. The cell sonicates were shown to catalyze the oxidation of both CO and H₂ with methylene blue, thionine, phenazine methosulfate, toluylene blue, dichlorophenolindophenol, cytochrome c or ferricyanide as electron acceptors. Methyl viologen, benzyl viologen, FAD⁺, FMN⁺, and NAD(P)⁺ were not reduced. The spectrum of electron acceptors was identical for all strains tested. Neither free formate, hydrogen nor oxygen gas were involved in the CO-oxidation reaction. Methylene blue was reduced by CO at a 1:1 molar ratio. The results indicate that CO-oxidation by carboxydobacteria is catalyzed by identical or similar enzymes and that the reaction obeys the equation CO+H₂OCO₂+2H⁺+2e^- as previously shown for Pseudomonas carboxydovorans.Dedicated to Otto Kandler remembering almost three decades of enjoyable cooperation     

Modulation of allostery of pyruvate kinase by shifting of an ensemble of microstates

Since the introduction of the concepts of allostery about four decades ago, much advancement has been made in elucidating the structure-function correlation in allostery. However, there are still a number of issues that remain unresolved. In this review we used mammalian pyruvate kinase (PK) as a model system to understand the role of protein dynamics in modulating cooperativity. PK has a triosephosphate isomerase (TIM)(α/β)₈ barrel structural motif. PK is an ideal system to address basic questions regarding regulatory mechanisms about this common (α/β)₈ structural motif. The simplest model accounting for all of the solution thermodynamic and kinetic data on ligand-enzyme interactions involves two conformational states, inactive E^T and active E^R. These conformational states are represented by domain movements. Further studies provide the first evidence for a differential effect of ligand binding on the dynamics of the structural elements, not major secondary structural changes. These data are consistent with our model that allosteric regulation of PK is the consequence of perturbation of the distribution of an ensemble of states in which the inactive E^T and active E^R represent the two extreme end states. Sequence differences and ligands can modulate the distribution of states leading to alterations of functions. The future work includes: defining the network of functionally connected residues; elucidating the chemical principles governing the sequence differences which affect functions; and probing the nature of mutations on the stability of the secondary structural elements, which in turn modulate allostery.     

（鱼句）亚科花（鱼骨）型鱼类骨骼系统的比较

对我国花型Hemibarbuspattern鱼类作了骨骼系统比较,结果表明,此类型鱼类脑颅较长,副蝶骨平直或稍弯曲,眶蝶骨腹纵嵴发达(铜鱼Coreius septentrionalis例外),下颞窝和咽突中等大,基枕骨后突发达;脑颅中的上筛骨的后突、侧筛骨的外筛突,蝶耳骨的外突、上耳骨的后突、围眶骨和后颞窝等均有明显的差异;咽颅中的舌颌骨、尾舌骨、鳃盖骨和下咽齿的列数等又有显著的区别;附肢骨骼中的腰带骨、脊椎骨中的复合神经骨和第4椎骨腹侧的悬器等也有不同之处。据此,这些差异和区别可作为属间或种间的分类依据。