基于叶绿素荧光参数的小麦叶片叶绿素相对含量估算方法

叶绿素含量是植物学和农业相关研究领域常用的生理指标.叶绿素含量和叶片光合功能密切相关,但是现有的叶绿素含量的测定方法无法实现叶绿素含量和光合功能的同步测定和关联分析.为解决该问题,本研究通过测定35个小麦品种旗叶的SPAD值和叶绿素荧光诱导动力学曲线,分别使用不同时间的快速叶绿素荧光动力学曲线的荧光值,以及33个常用荧...     

Vertical distribution and composition of phytoplankton under the influence of an upper mixed layer

The vertical distribution of phytoplankton is of fundamental importance for the dynamics and structure of aquatic communities. Here, using an advection-reaction-diffusion model, we investigate the distribution and competition of phytoplankton species in a water column, in which inverse resource gradients of light and a nutrient can limit growth of the biomass. This problem poses a challenge for ecologists, as the location of a production layer is not fixed, but rather depends on many internal parameters and environmental factors. In particular, we study the influence of an upper mixed layer (UML) in this system and show that it leads to a variety of dynamic effects: (i) Our model predicts alternative density profiles with a maximum of biomass either within or below the UML, thereby the system may be bistable or the relaxation from an unstable state may require a long-lasting transition. (ii) Reduced mixing in the deep layer can induce oscillations of the biomass; we show that a UML can sustain these oscillations even if the diffusivity is less than the critical mixing for a sinking phytoplankton population. (iii) A UML can strongly modify the outcome of competition between different phytoplankton species, yielding bistability both in the spatial distribution and in the species composition. (iv) A light limited species can obtain a competitive advantage if the diffusivity in the deep layers is reduced below a critical value. This yields a subtle competitive exclusion effect, where the oscillatory states in the deep layers are displaced by steady solutions in the UML. Finally, we present a novel graphical approach for deducing the competition outcome and for the analysis of the role of a UML in aquatic systems.     

浮游植物中叶绿素a提取方法的比较与改进

对水质监测中叶绿素a的提取方法进行改进,采用反复冻融-浸提方法提取叶绿素a,即将过滤水样的滤膜于-20℃冰箱内和室温下反复冻融3～5次后,放进90%丙酮溶液中浸提20h。与标准方法中的研磨法对比,该方法具有人为误差小、稳定性好、结果准确及操作简单安全等优点,适合运用于常规的水质监测。采用该方法时过滤水量对湖水水样的测定结果有影响(P<0.01),对于纯微囊藻液影响不明显(P>0.05)。因此,在对不同叶绿素水平或营养水平的水体采样时,应注意水样的水量。通过方差分析表明该方法中采用醋酸纤维滤膜、微孔滤膜和玻璃纤维滤膜对结果没有显著影响,对这3种滤膜可以进行自由选择。     

Extraction of chlorophyll a from freshwater phytoplankton for spectrophotometric analysis

The efficiency of pigment extraction forms the crux of the spectrophotometric analysis of chlorophyll a. The alcoholic solvents, methanol and ethanol, proved to be superior to acetone and acetone with DMSO. Homogenisation and sonication did not improve the extraction in the alcoholic solvents. Boiling at 100°C had an adverse effect whereas complete extraction of the pigments was obtained at the solvents boiling point and allowing the samples to stand for 24 h in the dark.     

基于因子分析的苜蓿叶片叶绿素高光谱反演研究

因子分析是一种能够将具有错综复杂关系的变量归结为少数几个综合因子的多变量统计分析方法,在降低数据维数的同时又可以保存足够的信息,这为处理信息量丰富但冗余较大的高光谱数据提供了一种有效方法。本文利用2010年9月23日采集的16个样点的苜蓿叶片反射率及叶绿素含量数据,采用因子分析方法,分别提取苜蓿叶片反射率光谱400～900nm,以及可见光400nm～760nm和近红外760～900nm光谱区的公共因子,分析因子载荷分布、载荷总量对公共因子与叶绿素含量相关性的影响。利用逐步回归法建立基于公共因子的叶片叶绿素反演模型,并将反演模型与光谱指数建立的模型进行对比。研究表明,1）公共因子与叶片叶绿素的相关性,在更大程度上是与该因子在各个波段上载荷分布有关,而不是总载荷量;2）对波谱进行分区建立的反演模型略优于全区因子分析建立的反演模型;3）与常用于叶片叶绿素含量反演的光谱指数CARI、MCARI、mND680、mND705、mSR705、TVI、DmSR、BGI、BRI相比,因子分析建立的叶绿素反演模型精度更高。     

2006年夏季南黄海浮游植物叶绿素a分布特征及其环境调控机制

根据2006年夏季南黄海生态调查资料,分析了浮游植物现存量(叶绿素a)的空间分布特征及其主要环境调控机制.结果表明:南黄海浮游植物叶绿素a浓度范围为0.07～12.17 mg/m3,平均值为1.42 mg/m3;浮游植物叶绿素a平面分布呈近岸混合区高于外海层化区的特征,总体上随水体盐度和营养盐梯度的变化而变化,其中浅海陆架锋区对浮游植物有明显的聚集作用,垂直结构中层化区叶绿素次表层最大值现象明显,且最大值的深度和量值具有明显的区域差异;叶绿素a浓度与主要环境因子的相关性分析表明,在较大空间尺度上营养盐是浮游植物的主要调控因子,温度、光照和浮游动物摄食等也在一定空间和程度上对浮游植物的生长产生影响.     

Spectrofluorimetric method for the determination of large chlorophyll a/b ratios

A practical spectrofluorimetric calibration method at room temperature is described for determining large chlorophyll a/b ratios on direct extracts from plant material of pigments in 80% aqueous acetone, dimethyl sulfoxide, and N,N-dimethylformamide. The method is based on the work of Boardman and Thorne [Boardman NK and Thorne SW (1971) Biochim Biophys Acta 253: 222–231] who used diethyl ether as solvent. We repeated the calibration in diethyl ether and find significantly different parameters for the calibration curve. The range of standards in this work included solutions with chlorophyll a/b ratios of 10–125 in dimethyl sulfoxide, and of 10–220 in the other solvents. Fluorescence emission spectra were found to be a highly sensitive method for assessing chlorophyll purity. We determined the limits of sensitivity for each solvent from the calibration data. The empirically determined slope of the calibration curve was shown to be related to intrinsic properties of the chlorophylls in solution; this allows predictions of the performance of the method in other solvents.     

The influence of external perturbations on the functional composition of phytoplankton in a Mediterranean reservoir

The Salton Sea is a large, shallow, endorheic, polymictic, saline lake in Southern California sustained by runoff from local agricultural and municipal wastewater. As a closed-basin lake in an arid region with declining water supply, it is becoming increasingly saline. Due to its eutrophic status and episodic wind-driven mixing, basin-wide anoxia events are becoming more common during summer. Sequencing of 16S rRNA gene clones obtained from the water column and sediments provided the first molecular assessment of bacterial community structure in the Salton Sea. Similar to other saline habitats, the water column community was dominated by Gamma- and Alphaproteobacteria, and Bacteroidetes, although there was significant seasonal variability in diversity, richness, and the specific genera recovered. Sediments showed high diversity and richness and were dominated by sequences from cultured and uncultured phyla typical of marine sediments, especially the Deltaproteobacteria. This study has provided baseline data about the bacterial communities found in this important California water body and may serve as important indicators of change as the lake becomes increasingly saline over time or as remediation efforts are enacted. Handling editor: Darren Bade     

The influence of fluctuating light intensities on species composition and diversity of natural phytoplankton communities

The influence of fluctuating light intensities on phytoplankton composition and diversity was investigated for 49 days under semi-continuous culture conditions with sufficient nutrient supply, using phytoplankton assemblages from Lake Biwa, Japan. Light conditions were either periodically changed from high intensity (100 µmol photons m^-2 s^-1) to low intensity (20 µmol photons m^-2 s^-1) at intervals of 1, 3, 6 and 12 days, or fixed to constant intensities (permanent high and low light levels). All treatments additionally experienced a day:night cycle of 16:8 h. Phytoplankton abundance increased and reached a saturation level on day 19 of the treatment with permanent high light, but increased continuously until the end of the experiment (day 49) in the treatment with permanent low light intensity. In treatments with periodically changing light intensities, the phytoplankton abundance reached saturation levels between these dates. Under phytoplankton abundance saturation, chlorophytes predominated in the treatment with permanent high light, while either cyanophytes or diatoms were abundant under permanent low light intensity. Treatments with changing light supply had chlorophyte- and cyanobacteria-dominated replicates as well as replicates with balanced proportions of both. Furthermore, species diversity, measured by the Shannon index, was low in cultures under permanent light intensity, while slow fluctuating light at the scale of 3 -12 days resulted in an increased diversity index. These results indicate that species composition and diversity of the phytoplankton were affected by the periodically changing light regime in the order of days, and suggest that temporal changes in weather conditions are a major impediment to competitive exclusion of phytoplankton species in nature.     

The influence of chlorophyll fluorescence on the light gradients and the phytochrome state in a green model leaf under natural conditions

Abstract. The effect of chlorophyll fluoresence on the spectral light gradients within a model green leaf was examined under different light qualities (day-light, sunset, canopy) and different quantum efficiencies. Light fluxes within the leaf tissue are nearly doubled in the emission domain of fluorescence but the effect on the phytochrome photoequilibrium is very small.     

The effect of cerium (III) on the chlorophyll formation in spinach

The effect of Ce³⁺ on the chlorophyll (chl) of spinach was studied in pot culture experiments. The results showed that Ce³⁺ could obviously stimulate the growth of spinach and increase its chlorophyll contents and photosynthetic rate. It could also improve the PSII formation and enhance its electron transport rate of PSII as well. By inductively coupled plasma-mass spectroscopy and atom absorption spectroscopy methods, it was revealed that the rare-earth-element (REE) distribution pattern in the Ce³⁺-treated spinach was leaf>root>shoot in Ce³⁺ contents. The spinach leaves easily absorbed REEs. The Ce³⁺ contents of chloroplast and chlorophyll of the Ce³⁺-treated spinach were higher than that of any other rare earth and were much higher than that of the control; it was also suggested that Ce³⁺ could enter the chloroplast and bind easily to chlorophyll and might replace magnesium to form Ce-chlorophyll. By ultraviolet-visible, Fourier transform infrared, and extended X-ray absorption fine structure (EXAFS) methods, Ce³⁺-coordinated nitrogen of porphyrin rings with eight coordination numbers and average length of the Ce-N bond of 0.251 nm.     

In situ microcosm experiments on the influence of nitrate and light on phytoplankton community composition

Phytoplankton blooms are fundamental features of coastal ecosystems, but the processes that select for blooms of certain species are not well understood. The aim of this work was to investigate experimentally the interaction of light and nutrients (nitrate) in structuring phytoplankton community composition in Pelorus Sound, New Zealand. Microcosm experiments were conducted in situ nine times throughout the year, providing controls and treatments for increased nutrients and decreased light. Nitrate availability was found to be limiting to phytoplankton growth during spring and summer. Small- to medium-sized, chain-forming diatom taxa such as Chaetoceros sp., Skeletonema sp., Pseudonitzschia sp. and Thalassiosira sp. responded most rapidly to nitrate enrichment, increasing their biovolume up to 32-fold during the 5-day experiments. A long-term phytoplankton monitoring database showed that these taxa have historically dominated the phytoplankton assemblage, suggesting that intense competition for nitrate is a key component in structuring the phytoplankton community. Many of the taxa that were able to withstand light reduction in the shaded treatments were rare historically in Pelorus Sound, suggesting that light is secondary to nitrate availability in structuring the phytoplankton community composition in this coastal embayment.     

UV radiation and phosphorus interact to influence the biochemical composition of phytoplankton

1. Numerous laboratory studies have shown that food quality is suboptimal for zooplankton growth. However, little is known about how food quality is affected by the interaction of potential global change factors in natural conditions. Using field enclosures in a high altitude Spanish lake, seston was exposed to increasing phosphorus (P) concentrations in the absence and presence of UV radiation (UVR) to test the hypothesis that interactions between these factors affected the biochemical and stoichiometric composition of seston in ways not easily predicted from studies of single factors. 2. Phosphorus enrichment increased the content of total fatty acids (TFA), ω3‐polyunsaturated fatty acids (ω3‐PUFA) and chlorophyll‐a : carbon (Chl‐a : C) and C : N ratios in seston. The pronounced increase in ω3‐PUFA was largely explained by the enhancement of 18:3n‐3 (α‐linolenic acid). In contrast, P‐enrichment lowered the content of highly unsaturated fatty acids (HUFA), the HUFA : PUFA ratio and, at high P loads, seston C : P ratio. Although phytoplankton assemblages dominated by Chlorophytes were not rich in HUFA, seston in the control had substantially higher 20:4n‐6 (arachidonic acid, ARA) content (79% of HUFA) than did P‐enriched enclosures. 3. The UVR increased the content of ω3‐PUFA and TFA in seston at the two ends of the trophic gradient generated at ambient and high concentrations of P, but decreased seston C : P and HUFA at all points on this gradient. ARA was not detected in the presence of UVR. 4. The interaction between P and UVR was significant for seston HUFA and C : P ratios, indicating that the effect of UVR in reducing HUFA (decreased food quality) and C : P ratios (enhanced food quality) was most pronounced at the low nutrient concentrations characteristic of oligotrophic conditions and disappeared as P increased. Therefore, any future increase in UVR fluxes will probably affect most strongly the food quality of algae inhabiting oligotrophic pristine waters although, at least in the Mediterranean region, these effects could be offset by greater deposition of P from the atmosphere.     

The influence of ultraviolet-B radiation on the growth, pigment production and chlorophyll fluorescence of Norway spruce seedlings

Norway spruce (Picea abies (L.)Karst.) from seven seed sources was grown in a greenhouse with 8.3 and 14.7 kJ·m⁻²·d⁻¹ m UV-B_BE (biologically effective UV-B: 280–320 nm) irradiation, and with no supplemental irradiation as control. The seedlings total biomass (dry weight) and shoot growth decreased with high UV-B treatment but spruce from low elevation seed sources were more affected. The seedlings grown at the highest UV-B irradiance (14.7 kJ·m⁻²·d⁻¹) showed from 5 to 38% inhibition of total biomass and 15 to 70 % shoot growth inhibition. Norway spruce populations from higher altitude seed sources manifested greater tolerance to UV-B radiation compared to plants from low altitudes. Changes in phospholipids and protective pigments were also determined. The plants grown at the lower UV-B irradiance (8.3 kJ·m⁻²·d⁻¹) showed greater ability to concentrations UV-B-absorbing pigments then control plants. Chlorophyll a fluorescence parameter R_fd, (R_fd=(F_m-F_s)/F_s) showed a significant decrease in needles of UV-B treated plants and this correlated with the altitude of seed source. Exposure to UV-B affect levels of the ratio of variable to maximum fluorescence (F_v/F_m). Results from this study suggest that the response to increased levels of UV-B radiation is depended upon the ecotypic differentiation of Norway spruce and involved changes in metabolites in plant tissues.     

The influence of copepod and krill grazing on the species composition of phytoplankton communities from the Scotia Weddell sea

The influence of copepods (mainly Oithona sim-ilis) and krill (Euphausia superba) grazing on the species composition of plankton communities in ship board con tainers was investigated during the spring and post spring period in the Scotia Weddell Sea in the Antarctic ocean. Numbers of grazers were experimentally manipulated in containers with natural phytoplankton assemblages. With ratural levels of copepods but no krill a high (700–950 g C·l¹, ca 30 g chl a·.l¹) phytoplankton biomass developed. In these cultures large diatoms, e.g. Corethron criophilum and chains of Thalassiosira sp., made up 80% of total phytoplankton cell carbon at the end of the experiment. In cultures with elevated numbers of copepods (5X or 10X the natural level) phytoplankton biomass was somewhat reduced (ca 23 g chl a · l¹) compared to cultures with natural copepod abundance, but still high. Phytoplankton species composition was on the other hand greatly influenced. Instead of large diatoms these cultures were dominated by Phaeocystis pouchetii (70%) together with small Nitszchia sp. and Chaetoceros neogracile (20%). In containers with krill (both juveniles and adults), but without elevated numbers of copepods, phytoplankton biomass rapidly approached zero. With 10X the in situ level of copepods, krill first preyed on these before Corethron criophilum and Thalassiosira sp. were grazed. When krill were removed a plankton community dominated by flagellates (60–90%), e.g. Pyramimonas sp. and a Cryptophycean species, grazed by an unidentified droplet-shaped heterothrophic flagellate, developed. These flagellates were the same as those which dominated the plankton community in the Weddell Sea after the spring bloom. A similar succession was observed in situ when a krill swarm grazed down a phytoplankton bloom in a few hours. Our experiments show that copepods cannot control phytoplankton biomass in shipboard cultures even at artificially elevated numbers. Krill at concentrations similar to those in natural swarms have a great impact on both phytoplankton biomass and species composition in shipboard cultures. Both copepods and krill may have an impact on phytoplankton species composition and biomass in situ since the rates of phytoplankton cell division were probably artificially increased in shipboard cultures compared to natural conditions, where lower growth rates make phytoplankton more vulnerable to grazing. A similarity between phytoplankton successions in containers and in situ, especially with respect to krill grazing, supports the conclusion that grazing may structure phytoplankton communities in the Scotia-Weddell Sea.Data presented here were collected during the European Polarstern Study (EPOS) sponsored by the European Science Foundation     

The annual cycles of phytoplankton biomass

Terrestrial plants are powerful climate sentinels because their annual cycles of growth, reproduction and senescence are finely tuned to the annual climate cycle having a period of one year. Consistency in the seasonal phasing of terrestrial plant activity provides a relatively low-noise background from which phenological shifts can be detected and attributed to climate change. Here, we ask whether phytoplankton biomass also fluctuates over a consistent annual cycle in lake, estuarine–coastal and ocean ecosystems and whether there is a characteristic phenology of phytoplankton as a consistent phase and amplitude of variability. We compiled 125 time series of phytoplankton biomass (chlorophyll a concentration) from temperate and subtropical zones and used wavelet analysis to extract their dominant periods of variability and the recurrence strength at those periods. Fewer than half (48%) of the series had a dominant 12-month period of variability, commonly expressed as the canonical spring-bloom pattern. About 20 per cent had a dominant six-month period of variability, commonly expressed as the spring and autumn or winter and summer blooms of temperate lakes and oceans. These annual patterns varied in recurrence strength across sites, and did not persist over the full series duration at some sites. About a third of the series had no component of variability at either the six- or 12-month period, reflecting a series of irregular pulses of biomass. These findings show that there is high variability of annual phytoplankton cycles across ecosystems, and that climate-driven annual cycles can be obscured by other drivers of population variability, including human disturbance, aperiodic weather events and strong trophic coupling between phytoplankton and their consumers. Regulation of phytoplankton biomass by multiple processes operating at multiple time scales adds complexity to the challenge of detecting climate-driven trends in aquatic ecosystems where the noise to signal ratio is high.     

The influence of vertical mixing on the photoinhibition of variable chlorophyll a fluorescence and its inclusion in a model of phytoplankton photosynthesis

The maximum effective quantum yield of photosystem II was estimatedfrom measurements of variable chlorophyll a fluorescence [(F'_m- F'_o)/F'_m = F'_v/F'_m] in samples of phytoplankton collectedfrom various depths in Chaffey Reservoir, Australia. Duringstratified conditions, F'_v/F'_m showed depth-dependent decreasesas irradiance increased during the morning, and increases asirradiancereduced in the afternoon. Wind-induced mixing disrupted thediel pattern, but even under well-mixed conditions a verticalgradient in F'_v/F'_m remained. Differences in F'_v/F'_m valuesbetween samples incubated at fixed depths and unconstrainedlake samples enabled identification of the phytoplankton mixingdepth. Recovery of F'_v/F'_m was modelled as a function of timeand the degree of F'_v/F'_m inhibition, while damage was considereda function of photon dose. A combined, numerical model was fittedto diel sequences of F'_v/F'_m to estimate rate constants fordamage and repair. Recovery rate constants ranged from r = 0.7x 10^-4 to 9.1 x 10^-4 s^-1, while damage rate constants rangedfrom k = 0.03 to 0.22 m² mol photon^-1. A fluorescence-basedmodel of photosynthesis was used to investigate the effectsof wind speed, euphotic depth and mixed layer depth on photoinhibition.At different mixing rates, depth-integrated photosynthesis wasenhanced by up to 16% under the conditions tested, while increasesof 9% occurred between phytoplankton with different measureddamage and repair characteristics.     

Structural and functional organization of the photosystems in spinach chloroplasts. Antenna size,relative electron-transport capacity,and chlorophyll composition

The structural and functional organization of the spinach chloroplast photosystems (PS) I, II_α and II_β was investigated. Sensitive absorbance difference spectrophotometry in the ultraviolet (?A₃₂₀) and red (?A₇₀₀) regions of the spectrum provided information on the relative concentration of PS II and PS I reaction centers. The kinetic analysis of PS II and PS I photochemistry under continuous weak excitation provided information on the number (N) of chlorophyll (Chl) molecules transferring excitation energy to PS II_α, PS II_β and PS I. Spinach chloroplasts contained almost twice as many PS II reaction centers compared to PS I reaction centers. The number N_α of chlorophyll (Chl) molecules associated with PS II_α was 234, while N_β = 100 and N_{PS I} = 210. Thus, the functional photosynthetic unit size of PS II reaction centers was different from that of PS I reaction centers. The relative electron-transport capacity of PS II was significantly greater than that of PS I. Hence, under light-limiting green excitation when both Chl a and Chl b molecules are excited equally, the limiting factor in the overall electron-transfer reaction was the turnover of PS I. The Chl composition of PS I, PS II_α and PS II_β was analyzed on the basis of a core Chl a reaction center complex component and a Chl $\text{a}\text{b-}\text{LHC}$ component. There is a dissimilar Chl $\text{a}\text{b-}\text{LHC}$ composition in the three photosystems with 77% of total Chl b associated with PS II_α only. The results indicate that PS II_α, located in the membrane of the grana partition region, is poised to receive excitation from a wider spectral window than PS II_β and PS I.     

Evidence for localisation of accumulated chlorophyll cation on the D1-accessory chlorophyll in the reaction centre of Photosystem II

Absorption and circular dichroism spectra of Photosystem II (PS II) reaction centres (RC) were studied and compared with spectra calculated on the basis of point-dipole approximation. Chlorophyll cation was accumulated during a light treatment of PS II RC in the presence of artificial electron acceptor silicomolybdate. Light-induced difference spectra and their calculated counterparts revealed the location of accumulated cation at the accessory chlorophyll of the D1 protein subunit.     

Using DCMU-fluorescence method for the identification of dominant phytoplankton groups

For the identification of ecologically significant dominant groups of phytoplanktonic algae a polychromatic DCMU-induced fluorescence method is recommended. A special fluorometer equipped with a system of replaceable filters is used to differentiate three regions of the spectrum (410 ± 20, 510 ± 20 and 540 ± 10 nm) that can excite the basic light-harvesting pigments. Total and differential (for every algal taxon studied) chlorophyll a calculated from the fluorescence signals is in good agreement with biomass estimates from direct cell counts for several different trophic types of aquatic systems. This is made possible by the vizualization of the ratios of fluorescence signal values in their own coordinates: first, to decide whether it is necessary to correct linear equations in order to eliminate negative solutions; second, to determine the possibility of nulling the negative solution if a point is situated close to a side of the triangle; and third, to reduce the number of linear algebraic equations to two if the points are situated along one of the triangle sides or to one if the points are gathered at the apex. The polychromatic DCMU-induced fluorescence method can be used for monitoring natural phytoplankton populations to detect changes in their taxonomic structure.