Beyond average protein secondary structure content prediction using FTIR spectroscopy

This paper demonstrates that secondary structure information beyond purely protein secondary structure content can be predicted from FTIR (Fourier transform infrared spectroscopy) spectra of proteins with a high degree of accuracy. Both neural networks and adaptive neuro-fuzzy inference systems (ANFISs) were employed to predict helix/sheet segment information. The best results were achieved using ANFISs with fuzzy subtractive clustering based on normalised, compressed amide I data with an average SEP (standard error of prediction, root mean of squared errors) of 1.51. Predictions for average helix/sheet length based merely on the amide I band maximum position in combination with the full-width at half-height resulted in a comparable average SEP of 1.62. This suggests the importance of information on the position and width of the amide I band maximum for the prediction of helix/sheet segment information. Finally, the most promising pattern recognition approaches found in this study were applied to a protein with an as yet unknown x-ray structure: native a1-antichymotrypsin (a1-ACT).     

Iron,silicate, and light co-limitation of three Southern Ocean diatom species

The effect of combined iron, silicate, and light co-limitation was investigated in the three diatom species Actinocyclus sp. Ehrenberg, Chaetoceros dichaeta Ehrenberg, and Chaetoceros debilis Cleve, isolated from the Southern Ocean (SO). Growth of all species was co-limited by iron and silicate, reflected in a significant increase in the number of cell divisions compared to the control. Lowest relative Si uptake and drastic frustule malformation was found under iron and silicate co-limitation in C. dichaeta, while Si limitation in general caused cell elongation in both Chaetoceros species. Higher light intensities similar to SO surface conditions showed a negative impact on growth of C. dichaeta and Actinocyclus sp. and no effect on C. debilis. This is in contrast to the assumed light limitation of SO diatoms due to deep wind driven mixing. Our results suggest that growth and species composition of Southern Ocean diatoms is influenced by a sensitive interaction of the abiotic factors, iron, silicate, and light.     

History and evolution of primary productivity studies of the Southern Ocean

The main objective of the article is to give an overview of the history and evolution of phytoplankton research in the Southern Ocean during the past century and a half. It traces the evolution of phytoplankton investigations as it went through several phases commencing with intensive collecting and cataloging and leading to the Discovery investigations with its extensive and detailed studies of Southern Ocean phytoplankton. Following, in the footsteps of the Discovery was the USNS Eltanin with its specialized cruises that centered around the study of the ecology of the primary producers and the dynamics of the lower trophic levels of the food chain. Spurred by the findings of the Eltanin cruises and with the growing concern over the impending exploitation of the Antarctic marine living resources, in particular krill, the BIOMASS program was initiated. The program was the first international collaborative effort to study the dynamic functioning of the Antarctic marine ecosystem. The success of the program has rekindled great international interest in the Southern Ocean which resulted in an explosion of programs such as SO-JGOFS, SO-GLOBEC, EPOS, and several others. In recent years, there has been a major shift in phytoplankton research in the Southern Ocean. This was in response to worldwide concern over the effects of global warming and stratospheric ozone depletion on species diversity, primary production and ecosystem function. This has led to process-oriented programs to study phenomena and processes of global significance in which Antarctica and its surrounding seas play key roles.In an effort to avoid any confusion in the strict usage of terminology, the author has opted to use the Antarctic Ocean and the Southern Ocean interchangeably in this review.     

Kinetics of dissolution of Antarctic diatom frustules and the biogeochemical cycle of silicon in the Southern Ocean

Summary In order to simulate the fate of biogenic silica generated in the surface waters of the Southern Ocean, the dissolution of silica frustules was studied for seven natural assemblages of diatoms, collected during summer 1984 in the Indian sector, and two typical Antarctic diatoms (Nitzschia cylindrus and Chaetoceros deflandrei), following the procedure of Kamatani and Riley (1979). For mean summer conditions in the surface waters of the Southern Ocean (2-3d^-1 for the natural assemblages. The silica frustules trapped by fecal pellets and by gelatinous aggregates, and rapidly transported through the cold waters of the Circumpolar Current, reach the sea bottom of either the continental shelves of the abysses without loosing much of the initial amount of silica (less than 10%). A model based on Stokes' law, modified to take in account of non ideal conditions and of the upwelling rate, is used in order to simulate the fate of silica of unaggregated particles settling down in the cold waters of the Antarctic Divergence. It supports the ideas that 1-the cycle of siliceous particles which radii are <2 m (i.e., of a part of the nanoplankton) is completely achieved in the surface layer, 2-although the biogenic silica of large unaggregated particles (radii over 25 m) may reach the seabottom (within one month to a few years) without complete dissolution, the main explanation for the accumulation of biogenic silica on Antarctic abysses remains transport by fecal pellets and gelatinous aggregates.     

Prediction of moisture, calorific value, ash and carbon content of two dedicated bioenergy crops using near-infrared spectroscopy

The potential of near infrared spectroscopy in conjunction with partial least squares regression to predict Miscanthus xgiganteus and short rotation coppice willow quality indices was examined. Moisture, calorific value, ash and carbon content were predicted with a root mean square error of cross validation of 0.90% (R² = 0.99), 0.13 MJ/kg (R² = 0.99), 0.42% (R² = 0.58), and 0.57% (R² = 0.88), respectively. The moisture and calorific value prediction models had excellent accuracy while the carbon and ash models were fair and poor, respectively. The results indicate that near infrared spectroscopy has the potential to predict quality indices of dedicated energy crops, however the models must be further validated on a wider range of samples prior to implementation. The utilization of such models would assist in the optimal use of the feedstock based on its biomass properties.     

Miocene diatom biostratigraphy at ODP Sites 689, 690, 1088, 1092 (Atlantic sector of the Southern Ocean)

Four ODP sites located between 64°S and 41°S in the eastern Atlantic sector of the Southern Ocean were investigated to refine the Miocene diatom biostratigraphic zonation tied to the geomagnetic chronology. The Miocene diatom stratigraphy from two sites located on Maud Rise (ODP Leg 113) is revised considering the progress in diatom biostratigraphic research, diatom taxonomy and magnetostratigraphic age assignment during the past 10 years. A new diatom zonation was erected for Site 1092 (ODP Leg 177) located on Meteor Rise integrating a magnetostratigraphic interpretation of the shipboard data. This zonation was also applied to Site 1088 (ODP Leg 177) located on Astrid Ridge. The study is focused to Middle and Upper Miocene sequences. It reveals latitudinal differentiations in stratigraphic species ranges and species occurrence pattern that are related to latitudinal differences in surface water masses reflecting the climatic development of the Antarctic cryosphere. Considering the latitudinal differences two stratigraphic zonations are proposed that are applicable to the northern and southern zone of the Southern Ocean, respectively. The southern Southern Ocean Miocene diatom biostratigraphic zonation consists of 16 zones in which 11 represent new or modified zones. The northern biostratigraphic zonation contains 10 diatom zones allowing a stratigraphic resolution in the range of 0.2–2 Myr. This paper also includes the taxonomic transfer of seven Miocene diatom taxa from genus Nitzschia Hassal to Fragilariopsis Hustedt.     

Colonization and community dynamics of class Flavobacteria on diatom detritus in experimental mesocosms based on Southern Ocean seawater

In order to better understand the ecology of microorganisms responsible for secondary production in the Southern Ocean the activity of Flavobacteria communities on diatom detritus in seawater mesocosms was investigated. Seawater was collected from different parts of the Southern Ocean including the Polar Front Zone (PFZ), ice-edge area of the Antarctic Zone (AZ), and a site in the AZ ice pack. Detritus from the cosmopolitan marine diatom Nitzschia closterium Ehrenberg was resuspended in mesocosms containing seawater filtered to remove particulate organic matter, including particle-associated bacteria and most eukaryotes, but retaining native planktonic bacterial assemblages. Mesocosms were incubated at 2 degrees C and samples analysed for changes in community composition using denaturing gradient gel electrophoresis (DGGE), real-time PCR and fluorescent in-situ hybridization (FISH). DGGE banding patterns and FISH images demonstrated rapid bacterial colonization of the detritus, dominated by members of class gamma-Proteobacteria, alpha-Proteobacteria and Flavobacteria. Real-time PCR data indicated members of class Flavobacteria were involved in initial colonization of detrital aggregate, however relative abundance stayed at similar levels found for the original native particle-associated populations. 16S rRNA gene DGGE banding patterns and sequence analysis demonstrated significant variation in Flavobacteria community structure occurred in the first 20 days of the experiment before community stabilization occurred. The community structures between the three mesocosms also markedly differed and major colonizers were primarily derived from detectable members of the initial particle-associated Flavobacteria community, however the abundant uncultured Flavobacteria agg58 clone-related and DE cluster 2 clades, previously identified in Southern Ocean seawater were not observed to colonize the detritus.     

Elevated carbon dioxide and ozone alter productivity and ecosystem carbon content in northern temperate forests

Three young northern temperate forest communities in the north‐central United States were exposed to factorial combinations of elevated carbon dioxide (CO₂) and tropospheric ozone (O₃) for 11 years. Here, we report results from an extensive sampling of plant biomass and soil conducted at the conclusion of the experiment that enabled us to estimate ecosystem carbon (C) content and cumulative net primary productivity (NPP). Elevated CO₂ enhanced ecosystem C content by 11%, whereas elevated O₃ decreased ecosystem C content by 9%. There was little variation in treatment effects on C content across communities and no meaningful interactions between CO₂ and O₃. Treatment effects on ecosystem C content resulted primarily from changes in the near‐surface mineral soil and tree C, particularly differences in woody tissues. Excluding the mineral soil, cumulative NPP was a strong predictor of ecosystem C content (r² = 0.96). Elevated CO₂ enhanced cumulative NPP by 39%, a consequence of a 28% increase in canopy nitrogen (N) content (g N m^?2) and a 28% increase in N productivity (NPP/canopy N). In contrast, elevated O₃ lowered NPP by 10% because of a 21% decrease in canopy N, but did not impact N productivity. Consequently, as the marginal impact of canopy N on NPP (?NPP/?N) decreased through time with further canopy development, the O₃ effect on NPP dissipated. Within the mineral soil, there was less C in the top 0.1 m of soil under elevated O₃ and less soil C from 0.1 to 0.2 m in depth under elevated CO₂. Overall, these results suggest that elevated CO₂ may create a sustained increase in NPP, whereas the long‐term effect of elevated O₃ on NPP will be smaller than expected. However, changes in soil C are not well‐understood and limit our ability to predict changes in ecosystem C content.     

Measuring carbon dynamics in field soils using soil spectral reflectance: prediction of maize root density, soil organic carbon and nitrogen content

This paper reports the development of a proximal sensing technique used to predict maize root density, soil carbon (C) and nitrogen (N) content from the visible and near-infrared (Vis-NIR) spectral reflectance of soil cores. Eighteen soil cores (0?C60?cm depth with a 4.6?cm diameter) were collected from two sites within a field of 90-day-old maize silage; Kairanga silt loam and Kairanga fine sandy loam (Gley Soils). At each site, three replicate soil cores were taken at 0, 15 and 30?cm distance from the row of maize plants (rows were 60?cm apart). Each soil core was sectioned at 5 depths (7.5, 15, 30, 45, and 60?cm) and soil reflectance spectra were acquired from the freshly cut surface at each depth. A 1.5?cm soil slice was taken at each surface to obtain root mass and total soil C and N reference (measured) data. Root densities decreased with depth and distance from plant and were lower in the silt loam, which had the higher total C and N contents. Calibration models, developed using partial least squares regression (PLSR) between the first derivative of soil reflectance and the reference data, were able to predict with moderate accuracy the soil profile root density (r ²?=?0.75; ratio of prediction to deviation [RPD]?=?2.03; root mean square error of cross-validation [RMSECV]?=?1.68?mg/cm³), soil% C (r ²?=?0.86; RPD?=?2.66; RMSECV?=?0.48%) and soil% N (r ²?=?0.81; RPD?=?2.32; RMSECV?=?0.05%) distribution patterns. The important wavelengths chosen by the PLSR model to predict root density were different to those chosen to predict soil C or N. In addition, predicted root densities were not strongly autocorrelated to soil C (r?=?0.60) or N (r?=?0.53) values, indicating that root density can be predicted independently from soil C. This research has identified a potential method for assessing root densities in field soils enabling study of their role in soil organic matter synthesis.     

Conformational studies of alanine-rich peptide using CD and FTIR spectroscopy.

The circular dichroism (CD) and Fourier transform infrared (FTIR) methods were applied to the conformational studies of alanine-rich peptide Ac-K-[A]11-KGGY-NH2 (where K is lysine, A is alanine, G is glycine and Y is tyrozyne) in water, methanol (MeOH) and trifluoroethanol (TFE). The analysis of CD-spectra of the peptide in water at different concentrations revealed that the secondary structure content depends on the peptide concentration and pH of the solution. The increase of the peptide concentration causes a decrease of alpha-helix content and, simultaneously, an increase of beta-sheet structure, while the unordered structure is the predominant one. Additional elements are discovered in MeOH and TFE but alpha-helix and beta-turns predominate. Moreover, in these solutions the percentage content of the secondary structure does not depend on the temperature. FTIR measurements, carried out at higher peptide concentration (about one order of magnitude) than these CD measurements mentioned above, revealed that in water solution the solid state beta-sheet, and aggregated structures, dominate. However, in TFE the most abundant are alpha-helix and beta-turns structures. The thioflavine T assay showed the tendency of the studied peptide for aggregate.     

Climate forcing of diatom productivity in a lowland,eutrophic lake: White Lough revisited

1. In cultural landscapes, lake response to climate can be masked by land‐use change and nutrient loss from their catchments. Palaeolimnological methods were used to reconstruct the ecological response of diatoms in a eutrophic lowland lake (White Lough, Co. Tyrone, Northern Ireland) to altered nutrient P loading and precipitation variability over c. 100 years. 2. ²¹⁰Pb‐dated sediment cores were analysed to determine diatom assemblage variability, biogenic silica concentration, geochemical phosphorus concentration and accumulation rate. Manure P and agricultural N surplus data were collated from documentary sources. Long‐term trends in annual temperature and precipitation were derived from the Armagh Observatory. 3. Diatom community turnover from 1890 until c. 1960 was limited, and assemblages were dominated by Aulacoseira subarctica; after this date, changes primarily reflected a eutrophication sequence owing to increased diffuse nutrient inputs associated with intensification of land use (external P loading increased by a factor of three). 4. Diatom and biogenic Si profiles were compared with North Atlantic Oscillation (NAO) records, an index of regional weather patterns. Biogenic Si exhibited a c. 7‐year cycle, which tracked a cycle of similar timescale in the Armagh climate record for dry summers. In turn, this cycle was related to the variation in the NAO. 5. Monitoring data from 1971 to 2007 of nitrate exports from the Blackwater River showed that these too followed a roughly 7‐year cycle at least up to 2000, in which dry summers were followed by sharp increases in nitrate export. It is argued that diatom production in White Lough reflects the cyclic behaviour in nitrate loading and the constraints that nitrogen availability places on the spring diatom bloom in a lake that is dominated by cyanobacteria.     

Viral lysis modifies seasonal phytoplankton dynamics and carbon flow in the Southern Ocean

Phytoplankton form the base of marine food webs and are a primary means for carbon export in the Southern Ocean, a key area for global pCO₂ drawdown. Viral lysis and grazing have very different effects on microbial community dynamics and carbon export, yet, very little is known about the relative magnitude and ecological impact of viral lysis on natural phytoplankton communities, especially in Antarctic waters. Here, we report on the temporal dynamics and relative importance of viral lysis rates, in comparison to grazing, for Antarctic nano- and pico-sized phytoplankton of varied taxonomy and size over a full productive season. Our results show that viral lysis was a major loss factor throughout the season, responsible for roughly half (58%) of seasonal phytoplankton carbon losses. Viral lysis appeared critically important for explaining temporal dynamics and for obtaining a complete seasonal mass balance of Antarctic phytoplankton. Group-specific responses indicated a negative correlation between grazing and viral losses in Phaeocystis and picoeukaryotes, while for other phytoplankton groups losses were more evenly spread throughout the season. Cryptophyte mortality was dominated by viral lysis, whereas small diatoms were mostly grazed. Larger diatoms dominated algal carbon flow and a single ‘lysis event’ directed >100% of daily carbon production away from higher trophic levels. This study highlights the need to consider viral lysis of key Antarctic phytoplankton for a better understanding of microbial community interactions and more accurate predictions of organic matter flux in this climate-sensitive region.Subject terms: Microbial ecology, Virus-host interactions     

FTIR spectroscopy of the complex between pharaonis phoborhodopsin and its transducer protein

pharaonis phoborhodopsin (ppR; also called pharaonis sensory rhodopsin II, psRII) is a photoreceptor for negative phototaxis in Natronobacterium pharaonis. ppR activates the cognate transducer protein, pHtrII, upon absorption of light. ppR and pHtrII form a tight 2:2 complex in the unphotolyzed state, and the interaction is somehow altered during the photocycle of ppR. In this paper, we studied the influence of pHtrII on the structural changes occurring upon retinal photoisomerization in ppR by means of low-temperature FTIR spectroscopy. We trapped the K intermediate at 77 K and compared the ppR(K) minus ppR spectra in the absence and presence of pHtrII. There are no differences in the X-D stretching vibrations (2700-1900 cm(-1)) caused by presence of pHtrII. This result indicates that the hydrogen-bonding network in the Schiff base region is not altered by interaction with pHtrII, which is consistent with the same absorption spectrum of ppR with or without pHtrII. In contrast, the ppR(K) minus ppR infrared difference spectra are clearly influenced by the presence of pHtrII in amide-I (1680-1640 cm(-1)) and amide-A (3350-3250 cm(-1)) vibrations. The identical spectra for the complex of the unlabeled ppR and (13)C- or (15)N-labeled pHtrII indicate that the observed structural changes for the peptide backbone originate from ppR only and are altered by retinal photoisomerization. The changes do not come from pHtrII, implying that the light signal is not transmitted to pHtrII in ppR(K). In addition, we observed D(2)O-insensitive bands at 3479 (-)/3369 (+) cm(-1) only in the presence of pHtrII, which presumably originate from an X-H stretch of an amino acid side chain inside the protein.     

Kinetics of intracellular carbon allocation in a marine diatom

To test models of intracellular carbon flow we measured the labelling kinetics (from ¹⁴CO₂) of major classes of cell polymers (carbohydrate, protein, lipid) and of dissolved organic carbon produced by the marine diatom Thalassiosira pseudonana Hustedt, grown at rates of 0.2 to 2.0·day^?1 under nitrogen or light limitation. Compartmental analysis indicated that tracer carbon quickly entered respiratory and excretory streams, accumulating in the cells at the rate of net production after only 25–50% of cell generation (doubling) time. Respiration rates were low (≤ 0.1 · day^?1) and suggested that illuminated cells in steady-state growth made only minor use of oxidative respiration to support cell synthesis. The tracer was quick to enter all polymers; compartmental analysis indicated that polymer labelling rates were close to the rates of mass synthesis after several hours of incubation with ¹⁴C. Polymer labelling also showed a reallocation of photosynthate from protein to carbohydrate within a few hours of perturbation (shift-down) of nutrient supply in a N-limited chemostat. In steady-state growth, the protein: carbohydrate ratio increased directly with N-limited growth rate but attained its maximum under extreme light-limitation. Carbon flow into the metabolic processes of respiration, excretion and polymer synthesis appeared to be mediated by a small and rapidly cycled pool of substrates under all steady-state growth conditions.     

pH prediction and control in bioprocesses using mid-infrared spectroscopy

An on-line pH monitoring method based on mid-infrared spectroscopy relevant to bioprocesses is presented. This approach is non-invasive and does not require the addition of indicators or dyes, since it relies on the analysis of species of common buffers used in culture media, such as phosphate buffer. Starting with titrations of phosphoric and acetic acid solutions over almost the entire pH range (2-12), it was shown that the infrared spectra of all samples can be expressed as a linear combination of the molar absorbance of the acids and their deprotonated forms. In other words, pH had no direct influence on the molar infrared spectra themselves, but only on deprotonation equilibria. Accurate prediction (standard error of prediction for pH < 0.15 pH units) was achieved by taking into account the non-ideal behavior of the solutions, using the Debye-Hückel theory to estimate the activity coefficients. Batch cultures of E. coli were chosen as a case study to show how this approach can be applied to bioprocess monitoring. The discrepancy between the spectroscopic prediction and the conventional electrochemical probe never exceeded 0.12 pH units, and the technique was fast enough to implement a feedback controller to maintain the pH constant during cultivation.     

The impact of phytoplankton on spectral water transparency in the Southern Ocean: implications for primary productivity

Spectral water transparency in the Northern Weddell Sea was studied during Austral spring. The depth of the 1-% surface irradiance level (euphotic depth) varied between 35 and 109 m and was strongly influenced by phytoplankton biomass. Secchi depths were non-linearly related to euphotic depth. In phytoplankton-poor water, the most penetrating spectral region was restricted to a relatively narrow waveband in the blue (488 nm), but the range was broader, between 488 and 525 nm when phytoplankton were abundant. Water transparency in the red spectral range was always low and only to a small extent affected by phytoplankton. Two independent procedures were used to quantify the impact of phytoplankton on spectral water transparency: (1) Regression analysis of spectral in situ vertical light attenuation coefficients in the sea, against coincident chlorophyll concentrations. This method gave chlorophyll-specific light attenuation coefficients; the y-intercept could be interpreted as a measure of light attenuation by pure water plus non-algal material. (2) Spectra of in vivo light absorption derived by spectroscopy, using phytoplankton enriched to varying degrees onto filters. Thus chlorophyll-specific absorption cross-sections were determined. Estimates obtained by both procedures were in close agreement. By integrating over the spectrum of underwater irradiance, in situ chlorophyll-specific absorption cross sections of phytoplankton suspensions, related to all photosynthetically active radiation, were calculated. Light absorption by phytoplankton for photosynthesis is accomplished mainly in the blue spectral range. Also dissolved and particulate organic matter contributed to the attenuation of blue light. Because in water poor in phytoplankton, underwater irradiance was progressively restricted to blue light, chlorophyll-specific absorption cross-sections of phytoplankton, averaged over the spectrum of photosynthetically active irradiance, increased with water depth. In water with elevated phytoplankton biomass, overall light attenuation was generally enhanced. However, because the spectral composition of underwater light changed relatively little with depth, except immediately below the water surface, light absorption cross-sections of phytoplankton changed little below 10 m depth. Vertical differences in the proportions of underwater light absorbed by the phytoplankton community here were mainly dependent on biomass variations. Because of the comparatively small attenuation of blue light by non-algal matter, the efficiency of light harvesting by phytoplankton at any given concentration of chlorophyll in Antractic waters is greater than in other marine regions. At the highest phytoplankton biomass observed by us, as much as 70% of underwater light was available for phytoplankton photosynthesis. When phytoplankton were scarce, <10% of underwater light was harvested by phytoplankton.Contribution within the European Polarstern Study (EPOS), supported by the Deutsche Forschungsgemeinschaft, Grant Ti 115/16-1 to MMT, the European Science Foundation, and by the Alfred Wegener Institut für Polar-und Meeresforschung, Bremerhaven     

Ocean acidification stimulates particulate organic carbon accumulation in two Antarctic diatom species under moderate and high natural solar radiation

Impacts of rising atmospheric CO₂ concentrations and increased daily irradiances from enhanced surface water stratification on phytoplankton physiology in the coastal Southern Ocean remain still unclear. Therefore, in the two Antarctic diatoms Fragilariopsis curta and Odontella weissflogii, the effects of moderate and high natural solar radiation combined with either ambient or future pCO₂ on cellular particulate organic carbon (POC) contents and photophysiology were investigated. Results showed that increasing CO₂ concentrations had greater impacts on diatom physiology than exposure to increasing solar radiation. Irrespective of the applied solar radiation regime, cellular POC quotas increased with future pCO₂ in both diatoms. Lowered maximum quantum yields of photochemistry in PSII (F_v/F_m) indicated a higher photosensitivity under these conditions, being counteracted by increased cellular concentrations of functional photosynthetic reaction centers. Overall, our results suggest that both bloom‐forming Antarctic coastal diatoms might increase carbon contents under future pCO₂ conditions despite reduced physiological fitness. This indicates a higher potential for primary productivity by the two diatom species with important implications for the CO₂ sequestration potential of diatom communities in the future coastal Southern Ocean.     

Multivariate FTIR analysis of substrates for protein, polysaccharide, lipid and microbe content: potential for solid-state fermentations

Components of fermentation processes such as protein, polysaccharide and lipid, as well as microbes, such as fungi grown on solid substrates, are difficult to measure in situ. The potential of Fourier Transform Infrared (FTIR) analysis of solid-state fermentations from mid-infrared absorption spectra has been investigated. The problem under consideration was to build a calibration model containing no irrelevant information to enable a multivariate mathematical approach for prediction of component concentrations. Methods for solid sample preparation and preprocessing of FTIR data were developed to assure Beer-Lambert law compliance and produce a well-conditioned multivariate system. The model was tested using composite samples of zein protein, corn starch and azolectin lipid, and corn samples containing known levels of fungal contamination. Preliminary concentration estimates were remarkably close to the correct values, with less than 5% standard error of prediction for all components measured.     

Using near-infrared reflectance spectroscopy to predict carbon, nitrogen and phosphorus content in heterogeneous plant material

The aim of this study was to produce calibration equations between near-infrared reflectance (NIR) spectra and the concentrations of carbon, nitrogen, and phosphorus in heterogeneous material: from living needles to litter in Pinus halepensis stands subjected to prescribed burnings. The aim was to determine whether calibrations should be conducted within each stage in the transformation of needles (local calibrations), giving relationships that were accurate but valid only for each particular stage, or whether it was possible to integrate the various forms of variation in needles (global calibrations) while retaining an acceptable accuracy. A principal component analysis calculated from the sample spectral data was used to distinguish three different sets, each sharing spectral characteristics and corresponding to three categories of needle: needles collected on the pines (N), falling needles (F), and litter (L), and each containing samples collected from the burnt sites and a control site. Samples representative of all the forms of variation in spectral properties were selected from within each category and their carbon, nitrogen, and phosphorus concentrations were measured using standard wet chemistry methods; these constituted the calibration sets n, f, and l. Calibrations were produced between the nutrient concentrations and the NIR spectra of the calibration sets n, f, and l and the grouped sets (n+f, f+l, n+f+l). The results of local calibrations made from each individual category showed that the carbon, nitrogen, and phosphorus concentrations were accurately predictable by NIR spectra. The global calibrations made by lumping together several categories were valid for a wider range of concentrations and for spectrally heterogeneous materials and in most cases were just as accurate as the local calibrations produced from each individual category. Received: 2 March 1998 / Accepted: 19 November 1998