OsLIC1基因参与水稻株型调控的功能研究

CCCH型锌指蛋白是进化上比较保守的一类锌指蛋白家族，其典型的氨基酸的基序为C-X7-8-C-X5-C-X3-H，其中X为任意氨基酸，这类锌指基序一般以重复的双拷贝形式存在。本论文克隆并鉴定了一个全新的、只含有一个CCCH型锌指基序的基因，利用反义RNA策略研究该基因功能，结果发现该基因的反义转基因植株表现出叶夹角增大的表型，因此我们将该基因命名为OsLIC1（Oryza sativa Lamina Increased Leaf Angle Control 1）。生物信息学分析发现该基因定位于水稻6号染色体近端粒的一端，位于BAC克隆AP004324中。OsLIC1与通常的CCCH型锌指蛋白含有多个重复的CCCH锌指基序不同，它只含有一个CCCH型锌指基序。除了CCCH锌指结构域以外，该蛋白在靠近C-端的位置还有一段丝氨酸（Ser）富集的区域，在此区域之前，还有一个在真核生物中相对保守的，以EELR为核心基序的结构域。采用基因枪将含OsLIC1-GFP融合构建的瞬时表达载体轰击入洋葱内表皮细胞，激光共聚焦显微镜观察发现OsLIC1-GFP可以定位到细胞核中。利用酵母转录激活系统发现以EELR为核心基序的结构域具有转录激活的功能。体外核酸结合活性分析显示OsLIC1蛋白可以结合双链DNA，这些结果证明OsLIC1是一个转录因子，这也是在植物中首次发现CCCH型锌指蛋白可以作为转录因子的方式调节基因的表达。 用玉米泛素启动子（Maize Ubiquitin promoter）驱动OsLIC1基因的反义表达载体转化水稻，获得的内源OsLIC1基因表达量下降的转基因植株表现出三个明显的表型：转基因植株的叶夹角增大;转基因的株高低于对照以及转基因植株的穗粒数减少。扫描电镜观察发现转基因植株叶夹角增大是由于近轴面细胞排列发生了改变以及维管束发育受阻引起的。转基因植株的Southern Blot和RT-PCR分析，结合Western Blot分析证明了转基因植株的表型与转基因事件之间的直接联系，并证明了转基因植株中内源OsLIC1在蛋白水平的确受到了抑制。采用RT-PCR技术、Promoter：：GUS和RNA in situ杂交三种方法相结合研究OsLIC1基因的表达模式，结果表明OsLIC1基因主要在叶颈、节以及分蘖原基中表达，这与转基因植株的表型相吻合，进一步证明了转基因植株的表型与基因功能之间的关系。Affymetrix 水稻全基因组芯片分析结果显示许多受油菜素内酯诱导表达的基因在转基因植株的叶颈材料中表达量上调，RT-PCR进一步验证了这一结果。由于在转基因植株中出现的叶夹角增大的表型和水稻油菜素内酯的作用相似，而基因芯片的结果又从分子水平提供了证据和线索。进一步采用RT-PCR和Promoter：：GUS相结合的方法研究OsLIC1基因对油菜素内酯的响应，结果发现OsLIC1基因可以被油菜素内酯诱导表达。而且，OsLIC1基因的反义转基因植株与野生型相比，表现出对油菜素内酯信号更敏感的响应。根据以上结果，推测OsLIC1可能是水稻油菜素内酯信号转导途径的负调控因子。水稻油菜素内酯合成和信号转导的突变体d2-1和d61-1具有直立的叶片的特征。用反义OsLIC1转基因植株以及野生型水稻与d2-1和d61-1突变体分别进行遗传杂交，结果发现在反义OsLIC1转基因植株与d2-1和d61-1突变体的杂交F1代中，都表现出叶夹角增大的表型。但是，在F2代中，在d2-1和d61-1纯合背景下，分别表现出叶夹角增大和叶片直立的表型，说明OsLIC1上位于d2-1，而d61-1则上位于OsLIC1。这一结果进一步证明了OsLIC1是通过参与水稻油菜素内酯信号调节而发挥对水稻叶夹角的调控作用。

Refractive index and thickness analysis of natural silicon dioxide film growing on silicon with variable-angle spectroscopic ellipsometry

The refractive index and thickness of SiO2 thin films naturally grown on Si substrates were determined simultaneously within the wavelength range of 220-1100 nm with variable-angle spectroscopic ellipsometry. Different angles of incidence and wavelength ranges were chosen to enhance the analysis sensitivity for more accurate results. Several optical models describing the practical SiO2-Si system were investigated, and best results were obtained with the optical model, including an interface layer between SiO2 and Si, which proved the existence of the interface layer in this work as described in other publications.

Electrowetting On A Lotus Leaf

Electrowetting on dielectrics has been widely used to manipulate and control microliter or nanoliter liquids in micro-total-analysis systems and laboratory on a chip. We carried out experiments on electrowetting on a lotus leaf, which is quite different from the equipotential plate used in conventional electrowetting. This has not been reported in the past. The lotus leaf is superhydrophobic and a weak conductor, so the droplet can be easily actuated on it through electrical potential gradient. The capillary motion of the droplet was recorded by a high-speed camera. The droplet moved toward the counterelectrode to fulfill the actuation. The actuation speed could be of the order of 10 mm/s. The actuation time is of the order of 10 ms.

Electrowetting on a lotus leaf

Electrowetting on dielectrics has been widely used to manipulate and control microliter or nanoliter liquids in micro-total-analysis systems and laboratory on a chip. We carried out experiments on electrowetting on a lotus leaf which is quite different from the equipotential plate used in conventional electrowetting. This has not been reported in the past. The lotus leaf is superhydrophobic and a weak conductor so the droplet can be easily actuated on it through electrical potential gradient. The capillary motion of the droplet was recorded by a high-speed camera. The droplet moved toward the counterelectrode to fulfill the actuation. The actuation speed could be of the order of 10 mm/s. The actuation time is of the order of 10 ms.

Direct numerical simulation of hypersonic boundary layer transition over a blunt cone with a small angle of attack

The direct numerical simulation of boundary layer transition over a 5° half-cone-angle blunt cone is performed. The free-stream Mach number is 6 and the angle of attack is 1°. Random wall blow-and-suction perturbations are used to trigger the transition. Different from the authors’ previous work [Li et al., AIAA J. 46, 2899(2008)], the whole boundary layer flow over the cone is simulated (while in the author’s previous work, only two 45° regions around the leeward and the windward sections are simulated). The transition location on the cone surface is determined through the rapid increase in skin fraction coefficient (Cf). The transition line on the cone surface shows a nonmonotonic curve and the transition is delayed in the range of 0° ≤ θ ≤ 30° (θ = 0° is the leeward section). The mechanism of the delayed transition is studied by using joint frequency spectrum analysis and linear stability theory (LST). It is shown that the growth rates of unstable waves of the second mode are suppressed in the range of 20° ≤ θ ≤ 30°, which leads to the delayed transition location. Very low frequency waves VLFWs� are found in the time series recorded just before the transition location, and the periodic times of VLFWs are about one order larger than those of ordinary Mack second mode waves. Band-pass filter is used to analyze the low frequency waves, and they are deemed as the effect of large scale nonlinear perturbations triggered by LST waves when they are strong enough.The direct numerical simulation of boundary layer transition over a 5° half-cone-angle blunt cone is performed. The free-stream Mach number is 6 and the angle of attack is 1°. Random wall blow-and-suction perturbations are used to trigger the transition. Different from the authors’ previous work [ Li et al., AIAA J. 46, 2899 (2008) ], the whole boundary layer flow over the cone is simulated (while in the author’s previous work, only two 45° regions around the leeward and the windward sections are simulated). The transition location on the cone surface is determined through the rapid increase in skin fraction coefficient (Cf). The transition line on the cone surface shows a nonmonotonic curve and the transition is delayed in the range of 20° ≤ θ ≤ 30° (θ = 0° is the leeward section). The mechanism of the delayed transition is studied by using joint frequency spectrum analysis and linear stability theory (LST). It is shown that the growth rates of unstable waves of the second mode are suppressed in the range of 20° ≤ θ ≤ 30°, which leads to the delayed transition location. Very low frequency waves (VLFWs) are found in the time series recorded just before the transition location, and the periodic times of VLFWs are about one order larger than those of ordinary Mack second mode waves. Band-pass filter is used to analyze the low frequency waves, and they are deemed as the effect of large scale nonlinear perturbations triggered by LST waves when they are strong enough.

Numerical Study on Critical Wedge Angle of Cellular Detonation Reflections

The critical wedge angle (CWA) for the transition from regular reflection (RR) to Mach reflection (MR) of a cellular detonation wave is studied numerically by an improved space-time conservation element and solution element method together with a two-step chemical reaction model. The accuracy of that numerical way is verified by simulating cellular detonation reflections at a 19.3∘ wedge. The planar and cellular detonation reflections over 45∘–55∘ wedges are also simulated. When the cellular detonation wave is over a 50∘ wedge, numerical results show a new phenomenon that RR and MR occur alternately. The transition process between RR and MR is investigated with the local pressure contours. Numerical analysis shows that the cellular structure is the essential reason for the new phenomenon and the CWA of detonation reflection is not a certain angle but an angle range.

Small angle X-ray scattering of the colloidal crystal

The monodisperse polystyrene spheres are assembled into the colloidal crystal on the glass substrate by vertical deposition method, which is aimed at the so-called photonic crystal applications. The structural information of the bulk colloidal crystal is crucial for understanding the crystal growth mechanism and developing the various applications of colloidal crystal. Small-angle X-ray scattering (SAXS) technique was used to obtain the bulk structure of the colloidal crystal at Beamline 1W2A of BSRF. It is found that the SAXS pattern is sensitive to the relative orientation between the colloidal sample and the incident X-ray direction. The crystal lattice was well distinguished and determined by the SAXS data.

Influence of Contact Angle and Tube Size on Capillary-Driven Flow Under Microgravity

The influence of contact angle and tube radius on the capillary-driven flow for circular cylindrical tubes is studied systematically by microgravity experiments using the drop tower. Experimental results show that the velocity of the capillary flow decreases monotonically with an increase in the contact angle. However, the time-evolution of the velocity of the capillary flow is different for different sized tubes. At the beginning of the microgravity period, the capillary flow in a thinner tube moves faster than that in a thicker tube, and then the latter overtakes the former. Therefore, there is an intersection between the curves of meniscus velocity vs microgravity time for two differently sized tubes. In addition, for two given sized tubes this intersection is delayed when the contact angle increases. The experimental results are analyzed theoretically and also supported by numerical computations.

Sessile Drop in Microgravity: Creation, Contact Angle and Interface

We present in this paper the results obtained from a parabolic flight campaign regarding the contact angle and the drop interface behavior of sessile drops created under terrestrial gravity (1g) or in microgravity (mu g). This is a preliminary study before further investigations on sessile drops evaporation under microgravity. In this study, drops are created by the mean of a syringe pump by injection through the substrate. The created drops are recorded using a video camera to extract the drops contact angles. Three fluids have been used in this study : de-ionized water, HFE-7100 and FC-72 and two heating surfaces: aluminum and PTFE. The results obtained evidence the feasibility of sessile drop creation in microgravity even for low surface tension liquids (below 15 mN m (-aEuro parts per thousand 1)) such as FC-72 and HFE-7100. We also evidence the contact angle behavior depending of the drop diameter and the gravity level. A second objective of this study is to analyze the drop interface shape in microgravity. The goal of the these experiments is to obtain reference data on the sessile drop behavior in microgravity for future experiments to be performed in an French-Chinese scientific instrument (IMPACHT).

Distortion of beam shape by a rotating double-prism wide-angle laser beam scanner

We describe the rigorous results of a wide-angle laser beam scanner, obtained with the help of the vector refraction theory. Using the rigorous results, the distortion of the beam shape is discussed. The distortion to the beam varies with the different relative angles of double prisms. The scanner expands the beam in some directions while it contracts the beam in other directions. According to the conservation of energy, the distribution of the laser intensity is changed as well. (c) 2006 Society of Photo-Optical Instrumentation Engineers.

Small-dynamic-angle measurement using tangent relationship and phase-modulating technique

A method using two prisms for measurement of small dynamic angles is proposed in which the measurement is based on a simple tangent equation and a phase-modulating interferometer with a laser diode to measure dynamic optical path differences with higher accuracy. Owing to the simple tangent equation, the symmetry requirement on the two prisms in the optical configuration is eliminated, and easy measurement of the separations between two parallel beams with a position-sensitive detector is achieved. Small-dynamic-angle measurements are experimentally demonstrated with high accuracy. (C) 2007 Society of Photo-Optical Instrumentation Engineers.

The effect of the rotating double-prism wide-angle laser beam scanner on the beam shape

In this paper, we describe a wide-angle laser beam scanner and the rigorous result of the wide-angle laser beam scanner was obtained with the help of the vector refraction theory. Using the rigorous results, the distortion of the beam shape was discussed. The distortion of the beam shape is varying with the different relative angles of the double prisms. According to the conservation of the energy, the distribution of the laser intensity is changed too. (c) 2005 Elsevier GmbH. All rights reserved.

Distortion of beam shape by a rotating double-prism wide-angle laser beam scanner

We describe the rigorous results of a wide-angle laser beam scanner, obtained with the help of the vector refraction theory. Using the rigorous results, the distortion of the beam shape is discussed. The distortion to the beam varies with the different relative angles of double prisms. The scanner expands the beam in some directions while it contracts the beam in other directions. According to the conservation of energy, the distribution of the laser intensity is changed as well. (c) 2006 Society of Photo-Optical Instrumentation Engineers.

Small-dynamic-angle measurement using tangent relationship and phase-modulating technique

A method using two prisms for measurement of small dynamic angles is proposed in which the measurement is based on a simple tangent equation and a phase-modulating interferometer with a laser diode to measure dynamic optical path differences with higher accuracy. Owing to the simple tangent equation, the symmetry requirement on the two prisms in the optical configuration is eliminated, and easy measurement of the separations between two parallel beams with a position-sensitive detector is achieved. Small-dynamic-angle measurements are experimentally demonstrated with high accuracy. (C) 2007 Society of Photo-Optical Instrumentation Engineers.