Photoacoustic Spectroscopy and its Application in Plant Science

The principles of photoacoustic spectroscopy are outlined. This new and still developing technique enables one to detect non-radiative de-excitation processes. It is possible to distinguish between signals emenating from different depths of the sample. In plant science photoacoustic spectroscopy has mainly been applied in photosynthesis research. Here it allows the detection of gross oxygen evolution and gives information about the energy balance of the primary photosynthetic reaction.     

The Optical Absorption Coefficient of Maize Seeds Investigated by Photoacoustic Spectroscopy

The knowledge about a seed??s optical parameters is of great relevance in the seed technology practice. Such parameters provide information about its absorption and reflectance, which in turn is related to its color, quality, and health condition. The objective of the present study was to determine the optical absorption coefficient ?? for maize seeds (Zea mays L.) by means of the photoacoustic spectroscopy (PAS). Untreated seeds (I) and seeds dyed with methyl red (II) were used in this investigation. In addition, conventional reflectance measurements (obtained with the integrating sphere) were performed to validate PAS absorption measurements. The results show that the absorption spectra and reflection data of the seed samples are complementary. When used with thermally thick and optically opaque seeds, PAS may be considered as a potential diagnostic tool for the characterization of the seeds.     

广西不同产地番石榴叶总黄酮含量测定

目的对广西不同产地番石榴叶中总黄酮含量进行测定。方法应用紫外分光光度法,以芦丁为对照品,于357nm下测定吸光度,计算番石榴叶中总黄酮的含量。结果在1.7~14.3范围内总黄酮含量与吸光度A值线性关系良好,平均回收率=99.28%,RSD=3.69%,R2=0.9993。结论该方法简便可靠、快速、重现性好,适用于番石榴叶总黄酮含量测定。采收地区不同,番石榴叶中总黄酮的含量也不同。     

油菜种子类胡萝卜素总量测定方法的研究

类胡萝卜素是植物中最重要的一类色素,具有抗氧化、预防心血管疾病和癌症、延缓衰老等生物学功能,与人类健康关系密切.培育高类胡萝卜素油菜品种能有效地提升菜籽油的营养保健价值和稳定性,是今后油菜品质育种的一个新方向.研究建立快速、准确和简便的油菜种子类胡萝卜素总量测定方法,是发掘高类胡萝卜素油菜种质资源和开展高类胡萝卜素油菜育种的前提条件.本实验研究了不同提取溶剂、时间、料液比等因素对油菜种子中类胡萝卜素的提取效果和测定结果的影响,确定了最佳的提取和测定条件.建立了快速、准确和简便的油菜种子类胡萝卜素总量测定方法,并测定了30份不同类型油菜种质资源种子中类胡萝卜素总量,为进一步发掘高类胡萝卜素油菜种质资源和开展高类胡萝卜素油菜育种创造了条件.     

植物叶片的相位解析光声光谱和光声相位谱

用相位解析法获得的未损伤植物叶片叶肉组织的光声光谱,与叶片的光声相位谱进行了比较．发现植物叶片的光声相位谱与叶绿体光吸收带之间呈现互补关系．叶片表皮的光声相位谱存在持有的波谷结构．不同调制频率的光声相位谱有较大的差异．这些现象表明光声相位谱与光声光谱一样,也可对未损伤的层状生物样品作深度剖面分析．     

On-Line Monitoring of Nitrogenase Activity in Cyanobacteria by Sensitive Laser Photoacoustic Detection of Ethylene

A new and extremely sensitive method for measuring nitrogenase activity through acetylene reduction is presented. Ethylene produced by nitrogenase-mediated reduction of acetylene is detected by using laser photoacoustics (LPA). This method possesses a detection limit making it 3 orders of magnitude more sensitive than traditional gas chromatographic analysis. Photoacoustic detection is based on the strong and unique absorption pattern of ethylene in the CO(inf2) laser wavelength region (9 to 11 (mu)m). The high sensitivity allowed on-line monitoring of nitrogenase activity in a culture of the heterocystous cyanobacterium Nodularia spumigena, which was isolated from a water bloom in the Baltic Sea. This setup makes it unnecessary to take subsamples from the culture and avoids long incubations in sealed vials. The fast response of the LPA technique allows measurement of real-time dynamic changes of nitrogenase activity. The method was used to analyze in vivo saturation of nitrogenase by acetylene in N. spumigena. It is demonstrated that 20% acetylene does not saturate nitrogenase and that the degree of saturation depends on light intensity. With concentrations of acetylene as low as 2.5% it is possible to assess the degree of saturation and to extrapolate to total nitrogenase activity. In N. spumigena nitrogenase activity becomes independent of light intensity above 20 to 80 (mu)mol of photons m(sup-2) s(sup-1) at 20% O(inf2).     

Increased Tolerance to Photoinhibitory Light in Paraquat-Resistant Conyza bonariensis Measured by Photoacoustic Spectroscopy and CO(2)-Fixation

Tolerance to photoinhibition was compared between a paraquat-resistant and a sensitive biotype of Conyza bonariensis (L.). Cronq. Photoinhibitory damage was measured as a decrease in oxygen evolution or energy storage using photoacoustic spectroscopy, or as a decrease of ¹⁴CO₂-fixation. Prior to exposure to high fluence rates, both biotypes had similar quantum yields of oxygen evolution and energy storage. After exposure to high intensity light, the resistant biotype continued to evolve oxygen and to store energy with a high quantum yield while both energy storage and oxygen evolution were severely reduced in the sensitive biotype. CO₂-fixation was less rapidly inhibited in the resistant biotype compared to the sensitive one. The data show that the paraquat resistant biotype with its high constitutive levels of the chloroplast localized enzymes of the oxygen detoxification pathway, is also partially protected from photoinhibition. This supports the theory that an enhanced radical scavenging system can give temporary protection against photooxidative damage from a variety of sources.     

P-NMR Spectroscopy of Roots of Intact Corn Seedlings

The application of ³¹P nuclear magnetic resonance spectroscopy to the study of metabolism in roots of intact corn seedlings is described. ³¹P-NMR spectra of developmentally distinct parts of primary roots of whole seedlings are presented. The spectra are of quality comparable to those of excised pieces of plant tissue.     

Methanogenic activity tests by Infrared Tunable Diode Laser Absorption Spectroscopy

Methanogenic activity (MA) tests are commonly carried out to estimate the capability of anaerobic biomass to treat effluents, to evaluate anaerobic activity in bioreactors or natural ecosystems, or to quantify inhibitory effects on methanogenic activity. These activity tests are usually based on the measurement of the volume of biogas produced by volumetric, pressure increase or gas chromatography (GC) methods. In this study, we present an alternative method for non-invasive measurement of methane produced during activity tests in closed vials, based on Infrared Tunable Diode Laser Absorption Spectroscopy (MA-TDLAS). This new method was tested during model acetoclastic and hydrogenotrophic methanogenic activity tests and was compared to a more traditional method based on gas chromatography. From the results obtained, the CH(4) detection limit of the method was estimated to 60ppm and the minimum measurable methane production rate was estimated to 1.09(.)10(-3)mgl(-1)h(-1), which is below CH(4) production rate usually reported in both anaerobic reactors and natural ecosystems. Additionally to sensitivity, the method has several potential interests compared to more traditional methods among which short measurements time allowing the measurement of a large number of MA test vials, non-invasive measurements avoiding leakage or external interferences and similar cost to GC based methods. It is concluded that MA-TDLAS is a promising method that could be of interest not only in the field of anaerobic digestion but also, in the field of environmental ecology where CH(4) production rates are usually very low.     

SO(2) Effect on Photosynthetic Activities of Intact Sugar Maple Leaves as Detected by Photoacoustic Spectroscopy

Short-term (4 hours) effect of different concentrations of SO₂ fumigation on in vivo photochemical activities of sugar maple (Acer saccharum Marsh.) leaves was investigated using photoacoustic spectroscopy. The relative quantum yield of O₂ evolution (ratio of O₂ signal to the photothermal signal) and photochemical energy storage are increased by 0.05 microliter per liter of SO₂. This increase is more pronounced in 5 to 7 year old saplings than in 3 month old seedlings. Both oxygen-relative quantum yield and energy storage of seedlings are inhibited by increased concentrations of SO₂ and the inhibition is concentration dependent. The inhibition is greater in seedlings than in saplings at 2 microliters per liter of SO₂, indicating the more susceptible nature of seedlings. The present study indicates a concentration dependent differential effect of SO₂ on photochemical activities of sugar maple leaves.     

Selective Excitation on Tip-Enhanced Raman Spectroscopy by Pulse Shaping Femtosecond Laser

In this paper, we propose a scheme for achieving ultrafast coherent control of the selective excitation among three excited states in stimulated Raman scattering process on a tip-enhanced Raman spectroscopy (TERS). The center frequencies of the pump and Stokes laser pulses are 14,000 cm⁻¹ and 12,500 cm⁻¹, and their spectral bandwidths are both 700 cm⁻¹. By properly modulating the spectral phase distribution and cutting the frequency components, the stimulated Raman transition probabilities of two excited state keep maximal, while the other one can be suppressed to zero. The shaped pump and Stokes pulse irradiate obliquely into the TERS nanostructure containing a single layer molecule. The impulse response in temporal and frequency domain is calculated by using finite-difference time-domain (FDTD) simulation followed by Fourier transform. The frequency components and the relative phase are same with the corresponding input pulses, but the intensities are enhanced by more than 10 times. Compared with the case without the TERS nanostructure, the probability of selective excited Raman transition increases by more than 4 orders of magnitude, and the selective depressed Raman peak keeps at 0.

     

近红外漫反射光谱法快速测定药用植物淫羊藿总黄酮含量

利用一种基于近红外漫反射光谱技术的淫羊藿总黄酮快速定量分析方法, 测定了淫羊藿属(Epimedium)7个种中总黄酮的含量。研究结果表明, 近红外光谱经二阶导数处理、主成分分析聚类及加权多元离散校正处理后, 采用改进最小二乘法回归得到的定标模型的预测效果最佳; 定标样品集预测方程的定标决定系数、交互验证标准差及交互验证相关系数分别为0.985 5、0.023 0和0.044 3; 检验样品集方程的定标决定系数、检验标准差及偏差分别为0.965 1、0.018和0.006, 达到了快速测定淫羊藿总黄酮含量的要求。该方法为淫羊藿总黄酮的快速定量分析提供了新的依据。     

Molecular Weights of Vesicular Stomatitis Virus and Its Defective Particles by Laser Light-Scattering Spectroscopy

Laser light-scattering spectroscopy has been used to determine the diffusion coefficients of vesicular stomatitis virus and three of its defective particles in order to calculate their molecular weights.     

Scavenging of Benzylperoxyl Radicals by Carotenoids

Carotenoids scavenge simple lipid-like alkylperoxyl radicals. However, the rate constant is too low to be determined directly and the mechanism is likewise not known with certainty [Mortensen, A. and Skibsted, L.H. (1998) FEBS Lett . 426 , 392-396]. It is demonstrated that carotenoids react with peroxyl radicals only slightly more reactive than lipidperoxyl radicals neither by electron transfer nor by hydrogen atom donation, but by adduct formation. Benzylperoxyl radicals are scavenged by the carotenoids &#103 -carotene and canthaxanthin with a second-order rate constant of at least 1 &#117 &#50 &#117 10 6 &#117 M &#109 1 &#117 s &#109 1 by formation of an adduct which decays in a first-order reaction.     

小球藻中类胡萝卜素类化合物的高效液相色谱分离条件的优化

采用超声破碎法和甲醇:二氯乙烷=2:1(v/v)溶剂来萃取小球藻中类胡萝卜素类化合物.分别采用了3种不同的色谱分离方法,最后确定小球藻中类胡萝卜素类化合物的最优色谱分离条件为甲醇(A):乙氰(B)=90:10(v/v),柱温为室温,流速1ml/min.用ZOBAX SB-C18色谱柱和ZOBAX Eclipse plus C18色谱柱进行分离试验时发现,ZOBAX Eclipse plus C18色谱柱的分离效果良好.最后确定小球藻中叶黄素含量为2.312mg/g.     

甘薯抗旱初步鉴定及渗透胁迫对抗氧化生理指标的影响

选用抗旱性有差异的6份甘薯栽培品种为试验材料,在田间鉴定抗旱性的基础上,通过室内PEG-6000模拟水分胁迫,测定胁迫下与抗旱性相关的9个生理生化指标,采用抗旱指数和隶属函数相结合的方法筛选与抗旱性密切相关的生理生化指标,对其抗旱性进行评价。结果表明:经田间抗旱鉴定,广薯87和徐薯22的抗旱性强,日紫0602和宁紫薯1号的抗旱性差;在PEG-6000模拟水分胁迫下,甘薯叶片NOS、CAT、T-AOC和T-SOD活性随着PEG浓度增高,其活性增强;Pro、MDA、Prot和T-AA含量也增加;尤其在25%PEG-6000胁迫处理24 h,甘薯叶片NOS、T-AOC、Pro的相对值与抗旱指数呈显著正相关,RWC相对值与抗旱指数呈极显著正相关,这4个指标的加权隶属函数值D与抗旱指数呈显著正相关(r=0.8944);因此,可用这4个指标的D值对甘薯抗旱性进行初步鉴定。     

Rapid Staining and Enumeration of Small Numbers of Total Bacteria in Water by Solid-Phase Laser Cytometry

The nucleic acid stain SYBR Green I was evaluated for use with solid-phase laser cytometry to obtain total bacterial cell counts from several water sources with small bacterial numbers. Results were obtained within 30 min and exceeded or equaled counts on R2A agar plates incubated for 14 days at room temperature.