902 resultados para configuration of microscope
Resumo:
Hydrogenated nanocrystalline silicon (nc-Si:H) n-layers have been used to prepare heterojunction solar cells on flat p-type crystalline silicon (c-Si) wafers. The nc-Si:H n-layers were deposited by radio-frequency (RF) plasma enhanced chemical vapor deposition (PECVD), and characterized using Raman spectroscopy, optical transmittance and activation energy of dark-conductivity. The nc-Si:H n-layers obtained comprise fine grained nanocrystallites embedded in amorphous matrix, which have a wider bandgap and a smaller activation energy. Heterojunction solar cells incorporated with the nc-Si n-layer were fabricated using configuration of Ag (100 nm)/1T0 (80 nm)/n-nc-Si:H (15 nm)/buffer a-Si:H/p-c-Si (300 mu m)/Al (200 nm), where a very thin intrinsic a-Si:H buffer layer was used to passivate the p-c-Si surface, followed by a hydrogen plasma treatment prior to the deposition of the thin nanocrystalline layer. The results show that heterojunction solar cells subjected to these surface treatments exhibit a remarkable increase in the efficiency, up to 14.1% on an area of 2.43 cm(2). (c) 2006 Elsevier B.V. All rights reserved.
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A novel analog-computation system using a quantum-dot cell network is proposed to solve complex problems. Analog computation is a promising method for solving a mathematical problem by using a physical system analogous to the problem. We designed a novel quantum-dot cell consisting of three-stacked. quantum dots and constructed a cell network utilizing the nearest-neighbor interactions between the cells. We then mapped a graph 3-colorability problem onto the network so that the single-electron configuration of the network in the ground state corresponded to one of the solutions. We calculated the ground state of the cell network and found solutions to the problems. The results demonstrate that analog computation is a promising approach for solving complex problems.
Resumo:
Hydrogenated nanocrystalline silicon (nc-Si:H) n-layers have been used to prepare heterojunction solar cells on flat p-type crystalline silicon (c-Si) wafers. The nc-Si:H n-layers were deposited by radio-frequency (RF) plasma enhanced chemical vapor deposition (PECVD), and characterized using Raman spectroscopy, optical transmittance and activation energy of dark-conductivity. The nc-Si:H n-layers obtained comprise fine grained nanocrystallites embedded in amorphous matrix, which have a wider bandgap and a smaller activation energy. Heterojunction solar cells incorporated with the nc-Si n-layer were fabricated using configuration of Ag (100 nm)/1T0 (80 nm)/n-nc-Si:H (15 nm)/buffer a-Si:H/p-c-Si (300 mu m)/Al (200 nm), where a very thin intrinsic a-Si:H buffer layer was used to passivate the p-c-Si surface, followed by a hydrogen plasma treatment prior to the deposition of the thin nanocrystalline layer. The results show that heterojunction solar cells subjected to these surface treatments exhibit a remarkable increase in the efficiency, up to 14.1% on an area of 2.43 cm(2). (c) 2006 Elsevier B.V. All rights reserved.
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Radiant heat conversion performance dominated by the active layer of Ga0.84In0.16As0.14Sb0.86 diode has been systematically investigated based on an analytic absorption spectrum, which is suggested here by numerically fitting the limited experimental data. For the concerned diode configuration, our calculation demonstrates that the optimal base doping is 3-4 x 10(17) cm(-3), which is less sensitive to the variation of the external radiation spectrum. Given the scarcity of the alloy elements, an economical device configuration of the 0.2-0.6 mu m emitter and the 4-6 mu m base would be particularly acceptable because the corresponding conversion efficiency cannot exhibit discouraging degradation in comparison to the one for the optimal structure, the thickness of which may be up to 10 mu m. More importantly, the method we suggested here to calculate alloy absorption can be easily transferred to other composition, thus bringing great convenience for design or optimization of the optoelectronic device formed by these alloys.
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Most traditional satellite constellation design methods are associated with a simple zonal or global, continuous or discontinuous coverage connected with a visibility of points on the Earth's surface. A new geometric approach for more complex coverage of a geographic region is proposed. Full and partial coverage of regions is considered. It implies that, at any time, the region is completely or partially within the instantaneous access area of a satellite of the constellation. The key idea of the method is a two-dimensional space application for maps of the satellite constellation and coverage requirements. The space dimensions are right ascension of ascending node and argument of latitude. Visibility requirements of each region can be presented as a polygon and satellite constellation as a uniform moving grid. At any time, at least one grid vertex must belong to the polygon. The optimal configuration of the satellite constellation corresponds to the maximum sparse grid. The method is suitable for continuous and discontinuous coverage. In the last case, a vertex belonging to the polygon should be examined with a revisit time. Examples of continuous coverage for a space communication network and of the United States are considered. Examples of discontinuous coverage are also presented.
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本论文系统地研究了稀土离子的4fN-1n'l'高激发组态能级问题。利用稀土光谱理论,推导了自由离子状态下的高激发组态4fN-1n'l'的能级表达式(包括电子的库仑和旋轨作用),编写了计算机程序,首次得到了660项4fN-1n'l'(n'l'=5d,6s,6p)组态主要较低能级的详细表达式,大大扩展了以往的计算结果。同时,具备了计算4fN-1n'l'组态的全部能级表达式的能力。利用复杂晶体的化学键介电理论,研究了基质中fN-15d组态能级移动和劈裂等问题,获得了如下创新性的成果:研究了晶体中稀土离子的4fN-15d组态的禁戒跃迁能级与允许跃迁能级之间的能级差变化现象。发现fd电子间库仑作用的交换积分项是能级差的主要作用并找到了在不同基质中影响能级差发生变化的因子:he=[Σfc(i)a(i)Q(i)2]1/2,可用来分析、确定和预测Dy3+,Tb3+在不同基质中的禁戒跃迁峰的能级位置,对其它稀土离子也具有一定的指导意义。通过对自由离子能级差问题的分析,发现对不同稀土离子,能级差随f电子的增加而减小的规律,这样,无论是从横向还是纵向都可以对稀土离子的能级差进行比较,相互确定。为分析光谱中的禁戒跃迁峰提供了理论依据。研究了晶体中Ce3+、Eu2+的4fN-15d组态能级中心下移现象,发现影响其发生变化的因素与能级差的相同,但两者具有不同的变化形式,前者与玩呈一级指数关系,后者与he呈线性关系。从指数关系式中推导得到的自由离子状态下能级中心位置与实验值吻合较好。当he趋向于极大值时,得到的Ce3+、Ey2+的4fN-15d组态能级中心极限值相应于离子所含电子动能的大小。研究了晶体中Ce3+、Eu2+的4fN-15d组态能级劈裂问题,结合实验结果,发现立方场下的能级劈裂与化学键的同极化作用能,中心离子的配位数,配体离子的有效电荷以及所成键的离子性相关,并具此得到一个劈裂因子参数:Fc=EhQfi/NFc与10Dq值呈现很好的线性关系。研究结果表明,无论是4fN-15d组态的能级劈裂还是能级中心下降问题,Ce3+、Eu2+两离子都可以表达成统一的形式,显示了环境因子he与劈裂因子Fc所具有的普适性,对其它稀土离子的4fN-1n'l,高激发组态能级同样也具有理论指导意义。
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The energetics, lattice relaxation, and the defect-induced states of st single O vacancy in alpha-Al2O3 are studied by means of supercell total-energy calculations using a first-principles method based on density-functional theory. The supercell model with 120 atoms in a hexagonal lattice is sufficiently large to give realistic results for an isolated single vacancy (square). Self-consistent calculations are performed for each assumed configuration of lattice relaxation involving the nearest-neighbor Al atoms and the next-nearest-neighbor O atoms of the vacancy site. Total-energy data thus accumulated are used to construct an energy hypersurface. A theoretical zero-temperature vacancy formation energy of 5.83 eV is obtained. Our results show a large relaxation of Al (O) atoms away from the vacancy site by about 16% (8%) of the original Al-square (O-square) distances. The relaxation of the neighboring Al atoms has a much weaker energy dependence than the O atoms. The O vacancy introduces a deep and doubly occupied defect level, or an F center in the gap, and three unoccupied defect levels near the conduction band edge, the positions of the latter are sensitive to the degree of relaxation. The defect state wave functions are found to be not so localized, but extend up to the boundary of the supercell. Defect-induced levels are also found in the valence-band region below the O 2s and the O 2p bands. Also investigated is the case of a singly occupied defect level (an F+ center). This is done by reducing both the total number of electrons in the supercell and the background positive charge by one electron in the self-consistent electronic structure calculations. The optical transitions between the occupied and excited states of the: F and F+ centers are also investigated and found to be anisotropic in agreement with optical data.
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Two kinds of quantum computation systems using artificial molecules: quantum computer and quantum analog computer are described. The artificial molecule consists of two or three coupled quantum dots stacked along z direction and one single electron, In quantum computer, one-qubit and two-qubit gates are constructed by one molecule and two molecules, respectively. The coupling between two qubits in a quantum gate can be controlled by thin film electrodes. We also constructed a quantum analog computer by designing a three-dot molecule network and mapping a graph 3-colorability problem onto the network. The ground-state configuration of the single electrons in the network corresponds to one of the problem solutions, We numerically study the operations of the two kinds of the quantum computers and demonstrate that they quantum gates can perform the quantum computation and solve complex problems.
Resumo:
A novel analog-computation system using a quantum-dot cell network is proposed to solve complex problems. Analog computation is a promising method for solving a mathematical problem by using a physical system analogous to the problem. We designed a novel quantum-dot cell consisting of three-stacked. quantum dots and constructed a cell network utilizing the nearest-neighbor interactions between the cells. We then mapped a graph 3-colorability problem onto the network so that the single-electron configuration of the network in the ground state corresponded to one of the solutions. We calculated the ground state of the cell network and found solutions to the problems. The results demonstrate that analog computation is a promising approach for solving complex problems.
Resumo:
细胞生物学研究的一个重要方向是动态地控制细胞在基底上的黏附。最近,随着表面化学的研究深入,尤其是对烷基硫醇在金基底上形成自组装单层膜(self-assembled monolayers, SAMs)这一体系的研究,使得人们能在分子水平的表面上控制细胞黏附。精氨酸-甘氨酸-天冬氨酸(arginine-glycine-aspartate, RGD)序列首先是从细胞外基质蛋白中分离出来的,能够识别并非共价结合细胞膜表面的整合素受体,从而促进细胞黏附。以前的一些工作已经证实,将含有RGD的肽链连接到SAMs表面之后,能够生物特异性地黏附动物细胞。已有的手段比如光照、电压、加热、微电极、微流控以及表面纳米形貌的梯度变化,都不能真正实现可逆地控制细胞黏附,原因是这些方法所用的化学有限;这些方法也不能得到完全抗拒细胞黏附的表面,原因是这些方法产生的表面缺陷等不完整。用两种不同波长的光(紫外光和可见光)照射偶氮苯,偶氮苯会发生可逆的光致异构变化,因此,偶氮苯的光致异构性质可以用来可逆地控制细胞在表面黏附。运用含有偶氮苯的混合SAMs,偶氮苯的末端连接GRGDS肽,混合SAMs中是以末端为六聚乙二醇的硫醇为背景,该SAMs修饰而成的表面能够黏附或者抗拒细胞黏附,其表面黏附性质取决于SAMs中偶氮苯的构象。该方法提供了一种在分子水平的表面上我们所了解到的唯一能可逆控制细胞黏附的方法,该方法需要用到的光源来自于标准荧光显微镜所配置的汞灯。 为了实现在金基底表面可逆的控制细胞黏附,我们合成了如下三个化合物: 由于化合物1的溶解性很差,几乎在所有溶剂里都不溶,所以不能直接用化合物1制备SAMs;化合物2能高效地抗拒细胞的黏附;化合物3的偶氮苯末端是活化酯,能够连接GRGDS肽,从而控制细胞黏附。 将化合物2和化合物3以一定的比例均匀混合在金基底表面形成SAMs,然后将GRGDS肽连接到偶氮苯(反式)的末端(通过GRGDS肽的甘氨酸上的伯胺基与偶氮苯末端的活化酯反应),从而得到细胞黏附的表面。用紫外光照射该细胞黏附表面5-10小时,随着偶氮苯的构象由反式变为顺式,偶氮苯末端的GRGDS肽淹没在化合物2的六聚乙二醇中,得到抗拒细胞黏附的惰性表面。再用可见光照射该惰性表面1个小时,随着偶氮苯的构象由顺式变为反式,原先埋没在六聚乙二醇中的GRGDS肽伸展至单层膜的末端,又得到了细胞黏附的表面。因此,该表面能完全可逆地控制细胞在金表面黏附。 An important area in cell biology is the dynamic control of cell adhesion on substrates. Recent advancements in surface chemistry, in particular, self-assembled monolayers (SAMs) of alkanethiols on gold substrates, have permitted unprecedented control of cell adhesion via molecularly defined surfaces. The tri-peptide sequence arginine-glycine-aspartate (RGD), initially isolated from the extracellular matrix (ECM) proteins, can recognize and non-covalently bind with integrin receptors on cell membranes to promote cell adhesion. Some previous work has demonstrated that RGD peptide grafted on SAMs can allow bio-specific adhesion of mammalian cells that mimic natural adhesion. Existing technologies such as light, voltage, heat, microelectrodes, microfluidic systems and surface gradient of nanotopography, either cannot realize fully reversible control of cell adhesion, due to the limitation in the chemistry used, or cannot yield a surface completely resistant against cell adhesion, due to the imperfection of surfaces. Azobenzenes undergo reversible photo-induced isomerization rapidly at two different wavelengths of light (UV and visible light), it therefore potentially allows the reversible control of cell adhesion on a surface. By using a mixed SAMs presenting azobenzene groups terminated in GRGDS peptides in a background of hexa(ethylene glycol) groups, the surface can either accommodate or resist cell adhesion depending on the conformation of the azobenzene embedded in SAMs. This method provides the only means we know to control cell adhesion reversibly on a molecularly well-defined surface by using light generated by a mercury lamp equipped on standard fluorescence microscopes. To realize the reversible control of cell adhesion on gold surface, we synthesized three kinds of compounds as following, We found that it was difficult to obtain SAMs directly from compound 1 because of its poor solubility in almost all kinds of solvents; compound 2 can resist cell adhesion efficiently; compound 3 presents an azobenzene terminated with NHS-activated ester, which can couple with a GRGDS peptide to control cell adhesion. After coating a gold surface with compound 2 and 3 in appropriate ratios to form a SAM followed by coupling the GRGDS peptides with NHS-activated esters at the end of azobenzene (E configuration) resulted in a cell-adhesive SAM. Irradiating this cell-adhesive SAM with UV light for 5-10 h converted the E configuration of azobenzene into the Z form, the GRGDS peptides becoming masked in the PEG, resulting in a cell-resistant surface. These SAM could again support cell adhesion as a result of the conformational switch of azobenzene from Z to E with the irradiation of visible light for 1 h. This surface, therefore, allows completely reversible control of cell adhesion on a gold surface.
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本论文由三部分共4 章组成。第一部分阐述了大戟科大戟属传统中药千金子(Euphorbia lathyris L.)化学成分、生物学活性以及千金子化学成分的HPLC、UPLC-MS、GC-MS 分析成果。第二部分介绍了民族药材暖地大叶藓(Rhodobryum giganteum (Schwaegr.) Par.)的化学成分研究和结构鉴定。第三部分概述了大戟属 植物中大环二萜酯的研究进展。 第一章包括1-3 节。在第1, 2 节中报道了千金子(Euphorbia lathyris L.)95% 乙醇提取物的化学成分分离鉴定。我们采用正、反相硅胶柱层析、重结晶等各种分离方法,凭借MS、IR、NMR、X-ray 等现代仪器手段,从中共分离鉴定22 个化合物。其中8 个是高活性化合物前体-续随子烷型大环二萜及3 个巨大戟烷型二萜,还有香豆素、生物碱、甾体等类型,其中完成对5 个大环二萜酯构型的确认,对2 个二萜酯构型进行了修正。第3 节中介绍对千金子化学成分的细胞毒性、α-葡萄糖苷酶抑制活性、P-gp 表达抑制活性的模型筛选结果。 第二章包括3 节,第1 节报道不同产地千金子高效液相色谱定量分析结果。第2 节介绍了各大环二萜酯的HPLC-MS/MS 的分析结果,并且对其质谱裂解规律、UPLC-MS 快速鉴定方法做了进一步讨论。第3 节介绍了千金子挥发油成分分析。采用传统水蒸气蒸馏方法提取千金子中的挥发油,并经气相色谱-质谱联用(GC-MS)技术共分离鉴定出 49 个化合物,占挥发油总量的90.48%。 第三章包括1, 2 两节,第1 节报道了暖地大叶藓化学成分。采用正、反相硅胶,凝胶柱层析等各种分离方法和MS、IR、NMR 等解析手段,共分离鉴定10个化合物,其中一个环肽化合物为新化合物。第2 节介绍了暖地大叶藓挥发油成分分析,共分离鉴定出 52 个化合物,占其挥发油总量的85.67%。 第四章概述了大戟科大戟属植物中大环二萜酯的研究进展。 This dissertation consists of three parts. In the first part, it is elaborated that the phytochemical investigation from the traditional Chinese medicine: seeds of Euphorbia lathyris L.. Biological activity and constituents analysis by HPLC、UPLC-MS、GC-MS were reported. In the second part, it is discussed that the chemical constituents were isolated and identificated from minority nationalitical herb-Rhodobryum giganteum (Schwaegr.) Par.. The third part is a review about the progress of studies on macrocyclic diterpenes from Euphorbia. The first part is composed of 1-3 sections. The section 1and 2 is focused on the isolation and identification of chemical constituents from seeds of E. lathyris. 22 compounds were isolated from the seeds of E. lathyris. by isolation methods of column chromatography (silica gel, including reversed phase) and recrystallisation on the basis of spectroscopic methods including IR, MS, NMR and X-ray. In 8 macrocyclic and 3 ingenane diterpenes, the relative configuration of 5 macrocyclic diterpenes were confirmed, in which 2 were amended. In the third section, cell cytotoxic activity, restraining activity of α-Glucosidase and multidrug resistance (MDR) reversing activity about P-gp were tested. 5 potential revsering reagents were found. The second part is composed of 1-3 sections. In first section it is described that the quality of the chemical constituents of E. lathyris from 5 sources , which were analyzed by high-performance liquid chromatography. In addition, the fractionation rules of some macrocyclic diterpenes were discussed and Ultra Performance Liquid Chromatography/ electrospray ionization mass spectrometry (UPLC-MS) was applied for quick determination of compounds in the second section. In the third section, chemical analysis of the essential oil from seeds of E. lathyris by GC-MS were reported. The essential oil from the seeds of E. lathyris L. in Sichuan was extracted by steam distillation and 49 compounds were isolated and identified from the essential oil by gas chromatography-mass spectrometer (GC-MS). These compounds are accounted for 90.46% of the total essential oil. The second part, including section 4 and 5, is about the phytochemical investigation of R. giganteum. In the former section, ten compounds were isolated and identified. Among them, a new peptide was characterized by spectroscopic analysis including IR, MS and NMR. In the other section, 52 compounds were isolated and identified from the essential oil by gas chromatography-mass spectrometer (GC-MS). These compounds are accounted for 85.67% of the total essential oil. The third part is a review about the progress of studies on macrocyclic diterpenes from Euphorbia.
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本论文由三章组成。第一章介绍了中药蜘蛛香的化学成分的研究成果,第二章为羧甲基魔芋葡苷聚糖-壳聚糖为细胞膜的天冬酰胺酶人工细胞的研究,第三章综述了人工细胞在生物医学领域的应用。 第一章报道了中药蜘蛛香(Valeriana wallichii)根部乙醇提取物的化学成分,采用正、反相硅胶层析等分离方法和MS、NMR等多种波谱手段,从中共分离鉴定出17个化合物,分别为缬草素(valtrate,1),valechlorine(2),homobadrinal(3),baldrinal(4),乙酰缬草素(acevaltrate 5),valeriotetrate C(6),valeriotetrate B(7),对羟基苯乙酮(4'-hydroxy-acetophenone 8),7-hydroxy valtrate(9),8-methylvalepotriate(10),1,5-dihydroxy-3,8-epoxyvalechlorine A(11),二氢缬草素(didrovaltrate 12),胡萝卜苷(13),橙皮苷 (hesperidin 14),prinsepiol-4-O-β-D-glucopyranoside(15),longiflorone(16),乙基糖苷(17)。其中化合物6、7、10、和11为新化合物,化合物9、15、16为首次从该植物中得到。新化合物11为含有氯原子的刚性骨架环烯醚萜,并且确定了其绝对构型。 第二章报道了以羧甲基魔芋葡苷聚糖(CKGM)和壳聚糖(CS)为膜的固定化L-天冬酰胺酶人工细胞研究成果。利用羧甲基魔芋葡苷聚糖和壳聚糖两种生物相容性很好的天然多糖之间的静电吸引力,在非常温和的条件下制备成具有半透过性膜的人工细胞,将治疗儿童急性成淋巴细胞性白血病(ALL)的药物L-天冬酰胺酶包裹在内。通过考察温度和pH对人工细胞的影响,结果表明以CKGM- CS为膜的L-天冬酰胺酶人工细胞对温度和pH的稳定性和耐受性均高于自由酶,说明CKGM-CS对酶具有保护作用,而且小分子底物和产物可以自由进出膜内外,而包裹在膜内的生物大分子则不能泄露出来。 第三章综述了微囊化人工细胞的研究进展。 This dissertation consists of three parts. In the first part, the chemical constituents from the root of Valeriana wallichii were reported. In the second part, preparation and characteristics of L-Asparaginase Artificial cell were reported. The third part is a review on progress of microcapsule artificial cell. The first chapter is about the isolation and identification of the chemical constituents from the root of V. wallichii. Seventeen compounds were isolated from the ethanol extract of roots of V. wallichii through repeated column chromatography on normal and reversed phase silica gel. By the spectroscopic and chemical evidence, their structures were elucidated as valtrate (1), valechlorine (2), homobadrinal (3), baldrinal (4), acevaltrate (5), valeriotetrate C (6), valeriotetrate B (7), 4'-hydroxy-acetophenone (8), 7-hydroxy valtrate (9), 8-methylvalepotriate (10), 1,5-dihydroxy-3,8-epoxyvalechlorine A (11), didrovaltrate (12), daucosterol (13), hesperidin (14), prinsepiol-4-O-β-D-glucopyranoside (15), longiflorone (16), and ethyl glucoside (17). Among them, 6, 7, 10, and 11 are new compounds. 15, 16 and 9 were isolated from this plant for the first time. The absolute configuration of compound 11, an unusual iridoid bearing a C-10 chlor-group and an oxo-bridge connecting C-3 and C-8 resulting in a rigid skeleton, was confirmed. The second chapter is about the semi-permeable microcapsule of carboxymethyl konjac glucomannan-chitosan for L-asparaginase immobilization. Carboxymethyl konjac glucomannan-chitosan (CKGM-CS) microcapsules, which have good biocompatibility, prepared under very mild conditions via polyelectrostatic complexation, were used for immobilize L-asparaginase-a kind of drug for acute lymphoblastic leukemia (ALL). The activity and stability under different temperature and pH of the enzyme loaded-microcapsules were studied. The results indicated the immobilized enzyme has better stability and activity contrasting to the native enzyme. The study illustrates that the L-asparaginase could be protected in CKGM-CS microcapsules, the substrate and product could pass through the system freely.
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作物的抗旱性是一个多基因控制的、极为复杂的数量性状,植物对干旱在分子水平上的差异反应通过植物组织生理和细胞生物学水平,最终表现为植物抗旱性的不同。在我国,旱地农业超过耕地面积的50%,但水资源短缺,因此培育和选育抗旱高产作物是发展节水型农业最有效的途径。 青藏高原气候恶劣、年均降雨量少,也是世界大麦初生起源中心,因而蕴藏了十分丰富的与抗逆相关的种质资源材料,从这些特殊的资源材料克隆抗旱基因,不仅对培育抗旱、优质、高产大麦新品种具有重要理论意义和经济价值,而且对整个作物抗旱基础和育种应用研究都具重大促进作用。 为了筛选青稞(裸大麦,Hordeum vulgare ssp. vulgare)抗旱性材料,本研究选用来自青藏高原不同地区的84份青稞为材料,在叶片失水率(water loss rate, WLR)检测分析的基础上,选择失水率值差异显著的12个品种,通过相对含水量(relative water content, RWC)和反复干旱法评价其抗旱性,并通过植株对干旱胁迫下的丙二醛(MDA)含量和游离脯氨酸(free-proline)含量变化,了解不同抗旱性材料的生理反应特性。选择抗旱性强弱不同的品种各两份进行LEA2蛋白基因(Dhn6基因)、LEA3蛋白基因(HVA1基因)的克隆,比较LEA蛋白结构差异与作物抗旱性之间的关系。同时,对抗旱性不同的青稞品种受到干旱时间不同的失水变化率(dynamics water loss rate, DWLR)进行了检测;对抗旱性不同的青稞对照材料进行2 h、4 h、8 h和12 h的快速干旱处理,通过SYBR Green实时荧光定量RT-PCR技术对Dhn6基因、Dhn11基因、Dhn13基因和HVA1基因在不同抗旱性材料受到不同干旱时间处理后的相对表达水平进行了检测。本研究对LEA蛋白基因在抗旱性不同的青稞材料中的干旱胁迫分子水平上的差异反应进行了研究,也对植物的抗旱机理进行了初步探讨。主要研究结果如下: 1. 青稞苗期进行离体叶片失水率测定结果表明,来自青藏高原的84份青稞材料的WLR在0.086~0.205gh-1g-1DW之间。选择WLR低于0.1gh-1g-1DW和WLR高于0.18gh-1g-1DW的品种各6份,并对苗期分别进行未干旱及干旱12小时的处理。相对含水量检测结果表明,低失水率青稞材料干旱后的具有更高的相对含水量,盆栽缺水试验也显示叶片失水率低的材料耐旱能力强于失水率高的材料。通过水合茚三酮法测定离体叶片游离脯氨酸的含量,结果表明,所有品种未干旱处理时,游离脯氨酸含量差异不大(17.10~25.74 µgg-1FW);干旱12小时后,低失水率的品种游离脯氨酸含量明显增高(32.99~53.45µgg-1FW),高失水率品种的游离脯氨酸含量与干旱前变化不明显(P<0.05)。硫代巴比妥酸法测定离体叶片丙二醛(MDA)含量,结果显示,12份所选对照品种中,丙二醛的含量在0.97~2.74nmolg-1FW,干旱12小时后丙二醛的含量显著上升(1.46~4.74nmolg-1FW),高失水率的6个品种的丙二醛含量在未干旱和干旱处理时都明显高于低WLR品种。本研究结果表明青稞的低失水率、低丙二醛含量、高相对含水量和高脯氨酸含量具相关性(P<0.05)。综上研究,我们认为作物失水率的测定可以作为快速检测作物抗旱性的指标之一,因此,强抗旱品种喜玛拉10号(TR1)、品比14号(TR2)和弱抗旱品种冬青8号(TS1)、QB24 (TS2)被选作抗旱基因克隆和表达分析的研究材料。 2. 高等植物胚胎发育晚期丰富蛋白(late embryogenesis abundant proteins, LEA proteins)与植物耐脱水性密切相关,为了探讨青稞LEA蛋白结构差异性与植物抗旱性的关系,本研究以强抗旱品种(喜玛拉10号、品比14号)和弱抗旱品种(冬青8号、QB24)为材料,利用同源克隆法,通过RT-PCR,分别克隆了与抗旱性密切相关的Dhn6基因和HVA1基因。Dhn6基因序列分析结果表明,强抗旱品种品比14号和弱抗旱品种冬青8号Dhn6基因所克隆到的序列为1026bp,它们之间只有5个碱基的差异;喜玛拉10号和QB24克隆到的序列长963bp。在强弱不同的抗旱品种中有22个核苷酸易突变位点,相应的脱水素氨基酸序列推导结果表明,22个核苷酸突变位点中,仅有8个位点导致相应的氨基酸残基的改变,其余的位点系同义突变,另外,21个富含甘氨酸序列的缺失并没有联系作物抗旱性特征。推测这些同义突变位点的氨基酸残基对维持青稞DHN6蛋白的正常结构和功能起着非常重要的作用,也可能DHN6蛋白对青稞长期适应逆境胁迫和遗传进化的结果。对HVA1基因的序列分析结果表明,冬青8号、QB24、品比14号和喜玛拉10号的目的基因核苷酸序列全长分别为661bp、697bp、694bp和691bp,它们都包含1个完整的开放阅读框。相应的LEA3蛋白氨基酸序列结果表明,11个高度保守的氨基酸残基组成基元重复序列的拷贝数与青稞抗旱性之间没有必然关系,在强抗旱品种(喜玛拉10号、品比14号)中三个共同的氨基酸突变位点Gln32、Arg33和Ala195可能对抗旱蛋白的结构和功能有影响;另外,强抗旱青稞品种LEA3蛋白质中11-氨基酸保守基元序列拷贝数和极性氨基酸占蛋白的比例更高,推测LEA3蛋白中基元序列拷贝数和极性氨基酸占蛋白的比例对该蛋白的结构和功能影响更大。 3. LEA蛋白基因的表达水平的上调与植物的耐脱水性密切相关,我们对强抗旱性材料(喜玛拉10号、品比14号)和弱抗旱材料(冬青8号、QB24)进行干旱处理2 h、4 h、6 h、8 h和10 h的失水变化率进行测定,结果表明弱抗旱品种在2~4小时之间失水率变化最明显,而四个对照品种的失水率在8小时后和24小时的失水率值变化不大。进一步提取青稞苗期进行2 h、4 h、8 h和12 h的干旱处理后的总RNA,通过SYBR Green实时荧光定量RT-PCR技术对青稞脱水素基因(Dhn6、Dhn11和Dhn13)和LEA3蛋白基因(HVA1)的相对表达水平受干旱时间和作物抗旱性的影响进行了检测。研究发现,抗旱性不同的青稞品种随干旱处理的时间延长,Dhn6、Dhn11、Dhn13和HVA1基因的相对表达水平不同。 Dhn6基因的相对表达水平在强抗旱青稞品种干旱8小时后快速上升,但在弱抗旱青稞品种干旱处理12小时后检测到更高表达量;Dhn11基因在对照青稞抗旱品种的表达累积水平随干旱时间的延长持续下降;整个干旱过程中,Dhn13基因的相对表达水平在弱抗旱品种持续上升,在强抗旱品种中干旱处理8小时快速上升并达到最高,干旱12小时后降低。与脱水素基因相比较,强抗旱青稞品种在干旱2小时后HVA1基因的相对表达水平显著升高,相对表达量随干旱处理的时间持续上升,在干旱12小时后达到最高;与之相比较,在整个干旱过程中,弱抗旱品种的相对表达水平显著低于强抗旱品种,在干旱8小时之前弱抗旱品种的相对表达水平变化不明显;在干旱8~12小时后却显著上升。上述结果表明,不同的LEA蛋白在植物耐脱水过程中的干旱表达累积水平不同;干旱不是诱导高等植物Dhn11基因表达的主要因素;植物的抗旱性不同,不同LEA蛋白基因对干旱的反应有差异。推测某些LEA蛋白基因的干旱胁迫早期表达累积程度与植物的抗旱性直接相关;其中,Dhn11基因和Dhn12基因不同的表达模式可能与干旱调控表达顺式作用成分(dehydration responsive element, DRE)的有无或结构上的差异有关。 本研究结果认为,(1)失水率和相对含水量可作为植物抗旱性检测的指标之一;(2) DHN6同义突变位点的氨基酸残基对维持该蛋白的正常结构和功能起着重要作用;(3) 11-氨基酸保守基元序列拷贝数和极性氨基酸的比例对LEA3蛋白结构和功能有重要影响;(4)LEA蛋白表达随着干旱胁迫程度而增加,但Dhn11基因并不受干旱诱导表达;(5)作物的抗旱性不同,LEA蛋白对干旱的累积反应并不相同,干旱早期LEA蛋白的累积程度可能会影响植物的抗旱性。 Drought resistance was a complex trait which involved multiple physiological and biochemical mechanisms and regulation of numerous genes. Because its complex traits, it is difficult to understand the mechanisms of drought resistance in plants. Plants respond to water stress through multiple physiological mechanisms at the cellular, tissue, and whole-plant levels. Tibetan hulless barley, a pure line, is a selfing annual plant that has predominantly penetrated into the Qinghai-Tibetan Plateau and remains stable populations there. The wide ecological range of Tibetan hulless barley differs in water availability, temperature, soil type and vegetation, which makes it possess a high potential of adaptive diversity to abiotic stresses. This adaptive genetic diversity indicates that the potential of Tibetan hulless barley serves as a good source for drought resistance alleles for breeding purposes. 12 contrasting drought-tolerant genotypes were selected to measure relative water content (RWC), maldondialdehyde (MDA) and proline content, based on values of water loss rate (WLR) and repeated drought methods from Tibetan populations of cultivated hulless barley. As a result of the screening, sensitive and tolerant genotypes were identified to clarify relationships between characteristics of LEA2/LEA3 genes sequences and expression and drought-tolerant genotypes, associated with resistance to water deficit. In addition, dynamics water loss rate (DWLR) was measured to observe the changes on diffrential drought-tolerant genotypes. Real-time quantitative RT-PCR was applied to detect relative expression levels of Dhn6, Dhn11, Dhn13 and HVA1 genes in sensitive and tolerant genotypes with 2 h, 4 h, 8h and 12 h of dehydration. In the present study, differential sequences and expression of LEA2/LEA3 genes were explored in Tibetan hulless barley, associated with phenotypically diverse drought-tolerant genotypes. 1. The assessments of WLR and RWC were considered as an alternative measure of plant water statues reflecting the metabolic activity in plants, and the parameters of MDA and proline contents were usually consistent with the resistance to water stress. The values of detached leaf WLR of the tested genotypes were highly variable among 84 genotypes, ranging from 0.086 to 0.205 g/h.g DW. The 12 most contrasting genotypes (6 genotypes with the lowest values of WLR and 6 genotypes with the highest values of WLR) were further validated by measuring RWC, MDA and free-proline contents, which were well watered and dehydrated for 12 h. Results of RWC indicated that the values of 12 contrasting genotypes RWC ranged from 89.94% to 93.38% under condition of well water, without significant differences, but 6 genotypes with lower WLR had higher RWC suffered from 12 h dehydration. The results indicated that lower MDA contents, lower scores of WLR and higher proline contents were associated with drought-tolerant genotypes in hulless barley. Remarkably, proline amounts were increased more notable in 6 tolerant genotypes than 6 sensitive genotypes after excised leaves were dehydrated for 12 h, with control to slight changes under condition of well water. Results of MDA contents showed that six 6 tolerant genotypes had lower MDA contents than the 6 sensitive genotypes under both stressed and non-stressed conditions. As a result of that screening, drought- resistant genotypes (Ximala 10 and Pinbi 14) and drought-sensitive genotypes (Dongqing 8 and QB 24) were chosen for comparing the differential characteristics of LEA2/LEA3 genes and their expression analysis. It was conclusion that measurements of WLR could be considered an alternative index as screening of drought-tolerant genotypes in crops. 2. Late embryogenesis abundant (LEA) proteins were thought to protect against water stress in plants. To explore the relationships between configuration of LEA proteins and phenotypically diverse drought-tolerant genotypes, sequences of LEA genes and their deduced proteins were compared in Tibetan hulless barley. Results of comparing Dhn6 gene in Ximala 10 and QB24 indicated that absence of 63bp was found, except that only 5 mutant nucleotides were found. While 22 mutant sites were taken place in Dhn6 gene between sensitive and tolerant lines, 14 synonymous mutation sites appeared in the contrasting genotypes. The additional/absent polypeptide of 21 polar amino acid residues was not consistent with phenotypically drought-tolerant genotypes in hulless barley. It was deduced that synonymous mutation sites would play important roles in holding out right configurations and functions on DHN6 protein. The sequencing analysis results indicated that each cloned HVA1 gene from four selected genotypes contained an entire open reading frame. The whole sequence of HVA1 gene from Dongqing 8, QB24, Pinbi 14 and Ximala 10 was respectively 661bp, 697bp, 694bp and 691bp. Results of DNA sequence analyses showed that the differences in nucleotides of HVA1 gene in sensitive genotypes were not consistent with that of tolerant genotypes, except for absence of 33 nucleotides from +154 to +186 (numbering from ATG) in QB24. Database searches using deduced amino acid sequences showed a high homology in LEA3 proteins in the selected genotypes. Multiple sequence alignments revealed that LEA3 protein from Dongqing 8 was composed of 8 repeats of an 11 amino acid motif, less the fourth motif than Pinbi 14, Ximala 10 and QB24. Consistent mutant amino acid residues appeared in contrasting genotypes by aligning and comparing the coding sequence region, including Gln32, Arg33 and Ala195 in tolerant genotypes as compared to Asp32, Glu33 and Thr195 (Thr184 in Dongqing 8) in sensitive lines. It was concluded that consistent appearance of Gln32, Arg33 and Ala195 would contributed to functions of LEA3 protein in crops, as well as higher proportion of 11-amino-repeating motifs and polar amino acid residues. 3. Most of the LEA genes are up-regulated by dehydration, salinity, or low temperature, are also induced by application of exogenous ABA, which increases in concentration in plants under various stress conditions and acts as a mobile stress signal. Higher levels of proteins of LEA group 3 accumulated was correlated well with high level of desiccation tolerance in severely dehydrated plant seedlings. Dehydrins (DHNs), members of LEA2 protein, are an immunologically distinct protein family, and Dhn genes expression is associated with plant response to dehydration. Dynamic water loss rate was measured between sensitive genotypes and tolerant genotypes after they were dehydrated for 2 h, 4 h, 6h and 8 h. Detailed measurements of WLR at the early stage of dehydration (2, 4, 6, and 8 h) showed that WLR was stabilizing after 8 h, and there were no significant changes between these values and WLR after 24 h. Drought stress was applied to 10-day-old seedlings by draining the solution from the container for defined dehydration periods. Leaf tissues of the selected genotypes were harvested from control plants (time 0); and after 2, 4, 8, and 12 h of dehydration. Differential expression trends of Dhn6, Dhn11, Dhn13 and HVA1 genes were detected in phenotypically diverse drought-tolerant hulless barleys, related to different time of dehydration. Results of quantitative real-time PCR indicated that relative level of HVA1 expression was always higher in tolerant genotypes, rapidly increasing at the earlier stages (after 2-4 h of dehydration). However, HVA1 expressions of sensitive genotypes had a fast increase from 8 h to 12 h of stress. Significant differences in expression trends of dehydrin genes between tolerant genotypes and sensitive lines were detected, mainly in Dhn6 and Dhn13 gene, depending on the duration of the dehydration stress. The relative expression levels of Dhn6 gene were significantly higher in tolerant genotypes after 8 h dehydration, by control with notable higher expression levels after 12 h water stress in sensitive ones. The relative expression levels of Dhn13 gene tended to ascend during exposure to dehydration in drought-sensitive genotypes. However, fluctuate trends of Dhn13 expression level were detected in drought-resistant lines, including in lower expression levels of 12 h dehydration as compared to 8 h water stress. It was conclusion that (1) diverse LEA proteins would play variable roles in resisting water stress in plants; (2) expression of Dhn11 gene was not induced by dehydrated signals because of the trends of expression descended in contrasting genotypes suffered from water deficit and (3) variable accumulations on LEA proteins would be appear in diverse drought-tolerant genotypes during dehydrations. It is deduced that higher accumulations of Dhn6 and Dhn13 expression in 8 h dehydration are related to diverse drought-tolerant lines in crops. The present results indicated that different dehydrin genes would play variable functional roles in resisting water stress when plants were suffered from water deficit. The authors suggest physiologically different reactions between resistant and sensitive genotypes may be the results of differential expression of drought-resistant genes and related signal genes in plants. In addition, contrarily induced expression of Dhn11 and Dhn12 was related to dehydration responsive element (DRE) in barleys. The present study indicated that (1) measurements of WLR and RWC could be considered as one index of drought-tolerant screenings; (2) synonymous mutation sites would play important roles in holding out right configurations and functions on DHN6 protein, (3) higher proportion of 11-amino-repeating motifs and polar amino acid residues would contribute to functions on LEA3 protein, (4) the longer drought, the more accumulation on LEA proteins, except for Dhn11 gene in crops and (5) differential responses on expression of LEA protein genes would result in physiological traits of drought tolerance in plants.
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本文对不同菌种(酵母菌和运动发酵单胞菌)快速生产燃料乙醇的条件进行了研究,实现了鲜甘薯快速转化为燃料乙醇。全文分为两部分: 第一部分:酵母菌快速生产燃料乙醇的条件研究。通过单因素试验,酵母菌快速生产燃料乙醇的条件为:发酵方式采用边糖化边发酵(SSF),蒸煮温度为85 ℃,料水比2:1(初始糖浓度 210 g/kg),糖化酶用量0.75 AGU/g 鲜甘薯,接种量10%(v/w)。在最优条件下,经过24 h发酵,乙醇浓度可达97.44 g/kg, 发酵效率为92%,发酵强度为4.06 g/kg/h。由于采用了低温蒸煮和SSF,可以大大节约能耗,从而降低乙醇生产的成本。同时,利用摇瓶优化的条件,进行了10 L,100 L,500 L发酵罐的放大试验,由于发酵罐初期可以人为通氧,使菌体能迅速积累,发酵时间缩短2 h,发酵效率在90%以上。 第二部分:运动发酵单胞菌快速生产燃料乙醇条件研究。通过单因素试验和正交试验获得了发酵的最佳参数:初始pH值6.0-7.0,硫酸铵5.0 g/kg,糖化酶量1.6 AUG/kg淀粉,初始糖浓度200 g/kg,接种量12.5%(v/w)。经过21 h发酵,乙醇浓度为95.15 g/kg,发酵效率可达94%。同时对不灭菌发酵也进行了研究,发酵效率可达92%。为鲜甘薯运动发酵单胞菌燃料乙醇的工业化生产打下基础。 对发酵结束后的残糖进行了研究。通过薄层层析和葡萄氧化酶测定证明:无论是酵母菌还是运动发酵单胞菌发酵结束后的发酵液中都不含葡萄糖。经过HPLC进一步分析残糖说明:发酵液中已没有葡萄糖成分;经糖化酶水解后仍没有葡萄糖出现;但经酸水解后又出现了葡萄糖,说明结束后的残糖是一些低聚糖结构。有关残糖的结构需要进一步研究。可以通过开发高效的低聚糖水解酶来降低发酵液的残糖,提高原料的利用率。 A new technology for rapid production fuel ethanol from fresh sweet potato by different microorganisms (Saccharomyces cerevisiae and Zymomonas mobilis) was gained in this research. The paper involved two parts: Part 1: The study on fuel ethanol rapid production from fresh sweet potato by Saccharomyces cerevisiae. The following parameters of Saccharomyces cerevisiae was investigated by a series of experiments: fermentation models, cooking temperature, initial sugar concentration and glucoamylase dosage. The results showed that SSF (simultaneous saccharification and fermentation) not only reduced the fermentation time (from 30 to 24h) but also enhanced the ethanol concentration (from 73.56 to 95.96 g/kg). With low-temperature-cooking (85 ℃) using SSF, the Saccharomyces cerevisiae was able to produce ethanol 97.44 g/kg which the fermentation yield could reach to 92% and ethanol productivity 4.06 g/kg/h from sweet potato enzymatic hydrolysis. Furthermore, the savings in energy by carrying out the cooking (85 ℃) and saccharification (30 ℃) step at low temperature had been realized. The results were also verified in 10 L, 100 L and 500 L fermentor. The fermentation yield was no less than 90%. The fermentation time of fermenter was shorter than Erlenmeyer flask. This may be that the aeration in the early fermentation period is available, which lead to the rapidly commutations of biomass. Part 2: The technology of ethanol rapid production with simultaneous saccharification and fermentation ( SSF ) by Zymomonas mobilis,using fresh sweet potato as raw material was studied. The effects of various factors on the yield of ethanol were investigated by the single factor and the orthogonal experiments. As a result, the optimal technical conditions were obtained from those experiments:initial pH value 6.0-7.0, nitride 5.0 g/kg,(NH4)2SO4, glucoamylase 1.6 AUG/kg starch, inoculums concentration 12.5% (v/w). The Zymomonas mobilis was able to produce ethanol 95.15 g/kg, with 94% of the theoretical yield, from fresh sweet potato after 24 h fermentation. The fermentation efficiency of non-sterilized was also reach to 92%. We also analyzed the final fermentation residual sugars of Saccharomyces cerevisiae and Zymomonas mobilis. When the residual sugars were analyzed by thin-layer chromatogram and glucose oxidase, there was no glucose. The analysis of reducing sugars by HPLC showed that there was no glucose existed in the fermentation liquor. However, the glucose appeared after being hydrolyzed by acid. It is indicated that the residual sugars in the final fermentation liquor were the configuration of oligosaccharide, which was linked by the special glycosidic bonds. It was feasible for reducing residual sugars to develope the enzyme that can degradation the oligosaccharide.
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The newly developed multi-quasiparticle triaxial projected shell model approach is employed to study the high-spin band structures in neutron-deficient even-even Ce- and Nd-isotopes. It is observed that gamma-bands are built on each intrinsic configuration of the triaxial mean-field deformation. Due to the fact that a triaxial configuration is a superposition of several K-states, the projection from these states results in several low-lying bands originating from the same intrinsic configuration. This generalizes the well-known concept of the surface gamma-oscillation in deformed nuclei based on the ground-state to gamma-bands built on multi-quasiparticle configurations. This new feature provides an alternative explanation on the observation of two I = 10 aligning states in Ce-134 and both exhibiting a neutron character. (C) 2009 Elsevier B.V. All rights reserved.