Chlorophyll measurement

The UOR tows of August 1979 (Figure 4), July 1981, and July 1982 (Figures 5 and 6) showed chlorophyll concentrations of 10-20 mg/m in the stratified waters (usually highest adjacent to the tidal front), and in June 1980 chlorophyll concentrations close to 10 mg/m were measured at the front. The increased abundance of zooplankton, coincident with the high concentrations of chlorophyll measured at the front, is shown in Figure 4c (abundance of Calanus helgolandicus Claus, the major herbivorous cope-pod in the plankton samples). 

Larsson, U., Norling, L., Carlberg, S., L66f, S., Tolstoy, A., v.Brockl, K., Elizarjeva, V., Kaiser, W, Lassig, J., Makinen, L, Melvasalo, T., 1978. Intercalibration of methods for chlorophyll measurements in the Baltic Sea. Merentutkimuslaitoksen Julkaisu/Havsforskningsinstitutets, Skrift243, 63-76. 

Phytoplankton provides a good indication of lake trophic state, measurable for example as chlorophyll a concentration, and responds quickly and predictably to changes in nutrient status. Relatively standard methods exist for evaluation of functional and non-taxonomic structural (biomass, chlorophyll measurements) characteristics of algal communities. 

Chlorophyll measurements were performed as described by Lichten-thaler and Wellburn (5). 

Napolitano, G. E. (1994) The relationship of lipids with light and chlorophyll measurements in freshwater algae and periphyton. Journal of Phycology, 30, 943-50. 

Beddard, G.S., Fleming, G.R., Porter, G., Searle, G.F.W., and Synowiec, J.A. (1979). The fluorescence decay kinetics of in vivo chlorophyll measured using low intensity excitation, Biochim. Biophys. Acta, 545, 165-174. 

A method of detecting herbicides is proposed the photosynthetic herbicides act by binding to Photosystem II (PS II), a multiunit chlorophyll-protein complex which plays a vital role in photosynthesis. The inhibition of PS II causes a reduced photoinduced production of hydrogen peroxide, which can be measured by a chemiluminescence reaction with luminol and the enzyme horseradish peroxidase (HRP). The sensing device proposed combines the production and detection of hydrogen peroxide in a single flow assay by combining all the individual steps in a compact, portable device that utilises micro-fluidic components. 

In the bacterial reaction center the photons are absorbed by the special pair of chlorophyll molecules on the periplasmic side of the membrane (see Figure 12.14). Spectroscopic measurements have shown that when a photon is absorbed by the special pair of chlorophylls, an electron is moved from the special pair to one of the pheophytin molecules. The close association and the parallel orientation of the chlorophyll ring systems in the special pair facilitates the excitation of an electron so that it is easily released. This process is very fast it occurs within 2 picoseconds. From the pheophytin the electron moves to a molecule of quinone, Qa, in a slower process that takes about 200 picoseconds. The electron then passes through the protein, to the second quinone molecule, Qb. This is a comparatively slow process, taking about 100 microseconds. 

One apparent discrepancy between the spectroscopic data and the crystal structure is that no spectroscopic signal has been measured for participation of the accessory chlorophyll molecule Ba in the electron transfer process. However, as seen in Figure 12.15, this chlorophyll molecule is between the special pair and the pheophytin molecule and provides an obvious link for electron transfer in two steps from the special pair through Ba to the pheophytin. This discrepancy has prompted recent, very rapid measurements of the electron transfer steps, still without any signal from Ba- This means either... 

Abridged Spectrophotometry. It is not always necessary to obtain complete spectrophotometric curves in order to measure physical characteristics related to color. The procedure can often be considerably simplified by some abridged form of spectrophotometry. Measurements may be made only at critical wave lengths or wave-length bands, as has been done to determine chlorophyll degradation (1, 8). In such instances the real problem that faces the investigator is to establish the critical wave lengths. 

Measured end points are photosynthesis as the incorporation of radiolabelled H C03 ( C) and bacterial activity as the incorporation of radiolabelled thymidine (thym) fluorometric measurements basal fluorescence (Fo) and photon yield (Y) chlorophyll-a concentration (chl-a) species composition (spp) and the biovolume of algae obtained after algal counting (biovolume)... 

Fig. 4 Predicted versus observed summer Anoxic Factor (AF) in (a, b) Foix Reservoir (Spain), (c, d) San Reservoir (Spain), (e, f) Brownlee Reservoir (USA), and (g, h) Pueblo Reservoir (USA). The results have been arranged to place the systems along a gradient of relative human impact (Foix Reservoir at the top, Pueblo Reservoir at the bottom). Predictions are based on linear models using different independent variables (in brackets) Inflow = streamflow entering the reservoir during the period DOCjjiflow = mean summer river DOC concentration measured upstream the reservoir CljjjAow = mean summer river CU concentration measured upstream the reservoir and Chlepi = mean summer chlorophyll-a concentration measured in the epilimnion of the reservoir. The symbol after a variable denotes a nonsignificant effect at the 95% level. Solid lines represent the perfect fit, and were added for reference. Modified from Marce et al. [48]...

The fluorescent lifetime of chlorophyll in vivo was first measured in 1957, independently by Brody and Rabinowitch (62) using pulse methods, and by Dmitrievskyand co-workers (63) using phase modulation methods. Because the measured quantum yield was lower than that predicted from the measured lifetime, it was concluded that much of the chlorophyll molecule was non-fluorescent, suggesting that energy transfer mechanisms were the means of moving absorbed energy to reactive parts of the molecule. 

Havaux, M. Lannoye, R. (1985). Drought resistance of hard wheat cultivars measured by a rapid chlorophyll fluorescence test. Journal of Agricultural Science, Cambridge, 104, 501-4. 

Because of the role these algae play in the oceans biological productivity and their impacts on climate due to the removal of carbon dioxide, satellite sensors have been employed to measure the chlorophyll a contents in oceans, lakes, and seas to indicate the distribution and abundance of biomass production in these water bodies. Detection is set at the specific reflectance and absorption wavelengths of the light from the upper layer of the ocean where photosynthesis occurs. 

In a similar way, microalgal biomass on the sediment surface can be estimated by measuring the chlorophyll contents in benthic microalgae, which are single-celled microscopic plants that inhabit the top 0 to 3 cm of a sediment surface and are sometimes referred to as microphytobenthos. These organisms are the primary food resources of benthic grazers such as shellfish and numerous finfish species. 

Because a chlorophyll molecule contains a closed circuit of ten conjugated double bounds to absorb light, spectrophotometric (UV-Vis) and fluorometric measurements are satisfactory to identify and estimate amounts of chlorophyll a and chlorophyll b, usually the only ones present in fresh plant extracts. The basis of numerous spectrophotometric determinations reported in literature is that chlorophylls strongly absorb at 500 to 700 nm in the visible region and show a large typical band around 400 nm. 

All the analytical methods mentioned to separate, identify, and quantify chlorophylls and derivatives consume time, money, and samples. As alternatives, industries have been employing non-destructive methods for surface color measurements that are not only indirectly related to chlorophyll content, but may also estimate the pigments directly in tissues, leaving the sample intact and enabling serial analyses in a relatively short time. Eood color affects consumer acceptance and is an important criterion for quality control. Color vision is a complex phenomenon that depends on both the total content and number of pigments and also on absorption, reflectance and emission spectra of each compound present. 

Additional research with Lemna species has indicated that parameters other than direct growth measurements may be more sensitive In this bioassay. Table VII compares growth and chlorophyll content of L. minor as affected by catechln. Final frond number was inhibited by 1000 pM catechln and stimulated at lower concentrations of 50 and 100 pM. Chlorophyll content on a per-frond basis, however, was consistently Inhibited by catechln and was concentration dependent to 100 pM. 

---