Summary
• Over the last decade, technological developments have made it possible to quickly and nondestructively assess, in situ, the chlorophyll (Chl) status of plants. We evaluated the performance of these optical methods, which are based on the absorbance or reflectance of certain wavelengths of light by intact leaves.
• As our benchmark, we used standard extraction techniques to measure Chla, Chlb, and total Chl content of paper birch (Betula papyrifera) leaves. These values were compared with the nominal Chl index values obtained with two hand-held Chl absorbance meters and several reflectance indices correlated with foliar Chl.
• The noninvasive optical methods all provided reliable estimates of relative leaf Chl. However, across the range of Chl contents studied (0.0004–0.0455 mg cm−2), some reflectance indices consistently out-performed the hand-held meters. Most importantly, the reflectance indices that performed best 

were not those most commonly used in the literature.
• We report equations to convert from index values to actual Chl content, but caution that differences in leaf structure may necessitate species-specific calibration equations.

https://nph.onlinelibrary.wiley.com/doi/epdf/10.1046/j.0028-646X.2001.00289.x

An evaluation of noninvasive methods to estimate foliar chlorophyll content

Use of leaf meters to provide an instantaneous assessment of leaf chlorophyll has become common, but calibration of meter output into direct units of leaf chlorophyll concentration has been difficult and an understanding of the relationship between these two parameters has remained elusive. We examined the correlation of soybean (Glycine max) and maize (Zea mays L.) leaf chlorophyll concentration, as measured by organic extraction and spectrophotometric analysis, with output (M) of the Minolta SPAD-502 leaf chlorophyll meter. The relationship is non-linear and can be described by the equation chlorophyll (μmol m−2)=10(M0.265), r2=0.94. Use of such an exponential equation is theoretically justified and forces a more appropriate fit to a limited data set than polynomial equations. The exact relationship will vary from meter to meter, but will be similar and can be readily determined by empirical methods. The ability to rapidly determine leaf chlorophyll concentrations by use of the calibration method reported herein should be useful in studies on photosynthesis and crop physiology.

/pdf/calibration-of-the-minolta-spad-502-leaf-chlorophyll-meter-19qjo8vtml.pdf

Calibration of the Minolta SPAD-502 leaf chlorophyll meter.

Some red edge parameters in the first derivative reflectance curve (wavelength, amplitude and area of the red edge peak) were studied to evaluate plant chlorophyll content, biomassand RelativeWater Content (RWC).Plants of Capsicum annuum and Phaseolus vulgaris under different nitrogen and water availabilities, and plants of Gerbera jamesonii with different hydric status were studied. A high correlation was found between chlorophyll content and the wavelength of the red edge peak (λre ), and between LAI (leaf area index)and the amplitude of the red edge peak (drr e ), but the area of the red edge peak (σ680–780 nm) was the best estimator of LAI. Thus, red edge was found valuable for assessment of plant chlorophyll concentration and LAI, and therefore nutritional status. Water stress also affected drre, but only when the stress was well developed.

The red edge position and shape as indicators of plant chlorophyll content, biomass and hydric status.

Relationships between chlorophyll concentration ([chl]) and SPAD values were determined for birch, wheat, and potato. For all three species, the relationships were non-linear with an increasing slope with increasing SPAD. The relationships for birch and wheat were strong (r
 2 ∼ 0.9), while the potato relationship was comparatively weak (r
 2 ∼ 0.5). Birch and wheat had very similar relationships when the chlorophyll concentration was expressed per unit leaf area, but diverged when it was expressed per unit fresh weight. Furthermore, wheat showed similar SPAD–[chl] relationships for two different cultivars and during two different growing seasons. The curvilinear shape of the SPAD–[chl] relationships agreed well with the simulated effects of non-uniform chlorophyll distribution across the leaf surface and multiple scattering, causing deviations from linearity in the high and low SPAD range, respectively. The effect of non-uniformly distributed chlorophyll is likely to be more important in explaining the non-linearity in the empirical relationships, since the effect of scattering was predicted to be comparatively weak. The simulations were based on the algorithm for the calculation of SPAD-502 output values. We suggest that SPAD calibration curves should generally be parameterised as non-linear equations, and we hope that the relationships between [chl] and SPAD and the simulations of the present study can facilitate the interpretation of chlorophyll meter calibrations in relation to optical properties of leaves in future studies.

Evaluating the relationship between leaf chlorophyll concentration and SPAD-502 chlorophyll meter readings

Summary
• Reproductive traits are key characteristics for predicting the response of communities and ecosystems to global change.
• We used meta-analysis to integrate data on eight reproductive traits from 159 CO 2 enrichment papers that provided information on 79 species.
• Across all species, CO 2 enrichment (500–800 µl l −1 ) resulted in more flowers (+19%), more fruits (+18%), more seeds (+16%), greater individual seed mass (+4%), greater total seed mass (+25%), and lower seed nitrogen concentration, (N) (−14%). Crops and undomesticated (wild) species did not differ in total mass response to elevated CO 2 (+31%), but crops allocated more mass to reproduction and produced more fruits (+28% vs +4%) and seeds (+21% vs +4%) than did wild species when grown at high CO 2 . Seed [N] was not affected by high CO 2 concentrations in legumes, but declined significantly in most nonlegumes.
• Our results provide robust estimates of average plant reproductive responses to CO 2 enrichment and demonstrate important differences among individual taxa and among functional groups. In particular, crops were more responsive to elevated CO 2 than were wild species. These differences and the substantial decline in seed [N] in many species have broad implications for the functioning of future natural and agro-ecosystems.

Plant reproduction under elevated CO2 conditions: a meta-analysis of reports on 79 crop and wild species

Two types of nondestructive chlorophyll meters were compared with a standard, destructive chlorophyll measurement technique. The nondestructive chlorophyll meters were 1) a custom built, single-wavelength meter, and 2) the recently introduced, dual-wavelengh, chlorophyll meter from Minolta (model SPAD-502). Data from both meters were closely correlated with destructive measurements of chlorophyll (r2 = 0.90 and 0.93; respectively) for leaves with chlorophyll concentrations ranging from 100 to 600 mg m-2, but both meters consistently overestimated chlorophyll outside this range. Although the dual-wavelength meter was slightly more accurate than the single-wavelength meter (higher r2), the light-scattering properties of leaf cells and the nonhomogeneous distribution of chlorophyll in leaves appear to limit the ability of all meters to estimate in vivo chlorophyll concentration.

/pdf/inherent-limitations-of-nondestructive-chlorophyll-meters-a-4g1ehwpt5n.pdf

Inherent limitations of nondestructive chlorophyll meters: a comparison of two types of meters.

Struvite formation and decomposition characteristics for ammonia and phosphorus recovery: A review of magnesium-ammonia-phosphate interactions

Farming in space: environmental and biophysical concerns

The effect of elevated [CO2] on wheat (Triticum aestivum L. Veery 10) productivity was examined by analysing radiation capture, canopy quantum yield, canopy carbon use efficiency, harvest index and daily C gain. Canopies were grown at either 330 or 1200 micromoles mol-1 [CO2] in controlled environments, where root and shoot C fluxes were monitored continuously from emergence to harvest. A rapidly circulating hydroponic solution supplied nutrients, water and root zone oxygen. At harvest, dry mass predicted from gas exchange data was 102.8 +/- 4.7% of the observed dry mass in six trials. Neither radiation capture efficiency nor carbon use efficiency were affected by elevated [CO2], but yield increased by 13% due to a sustained increase in canopy quantum yield. CO2 enrichment increased root mass, tiller number and seed mass. Harvest index and chlorophyll concentration were unchanged, but CO2 enrichment increased average life cycle net photosynthesis (13%, P < 0.05) and root respiration (24%, P < 0.05). These data indicate that plant communities adapt to CO2 enrichment through changes in C allocation. Elevated [CO2] increases sink strength in optimal environments, resulting in sustained increases in photosynthetic capacity, canopy quantum yield and daily C gain throughout the life cycle.

Adaptation to high CO2 concentration in an optimal environment: radiation capture, canopy quantum yield and carbon use efficiency

The use of higher plants as the basis for a biological life support system that regenerates the atmosphere, purifies water, and produces food has been proposed for long duration space missions. The objective of these experiments was to determine what effects microgravity (μg) had on chloroplast development, carbohydrate metabolism and gene expression in developing leaves of Triticum aestivum L. cv. USU Apogee. Gravity naive wheat plants were sampled from a series of seven 21-day experiments conducted during Increment IV of the International Space Station. These samples were fixed in either 3% glutaraldehyde or RNAlater™ or frozen at −25°C for subsequent analysis. In addition, leaf samples were collected from 24- and 14-day-old plants during the mission that were returned to Earth for analysis. Plants grown under identical light, temperature, relative humidity, photoperiod, CO2, and planting density were used as ground controls. At the morphological level, there was little difference in the development of cells of wheat under μg conditions. Leaves developed in μg have thinner cross-sectional area than the 1 g grown plants. Ultrastructurally, the chloroplasts of μg grown plants were more ovoid than those developed at 1 g, and the thylakoid membranes had a trend to greater packing density. No differences were observed in the starch, soluble sugar, or lignin content of the leaves grown in μg or 1 g conditions. Furthermore, no differences in gene expression were detected leaf samples collected at μg from 24-day-old leaves, suggesting that the spaceflight environment had minimal impact on wheat metabolism.

Oscar Monje

Papers

Inherent limitations of nondestructive chlorophyll meters: a comparison of two types of meters.

Struvite formation and decomposition characteristics for ammonia and phosphorus recovery: A review of magnesium-ammonia-phosphate interactions

Farming in space: environmental and biophysical concerns

Adaptation to high CO2 concentration in an optimal environment: radiation capture, canopy quantum yield and carbon use efficiency

Microgravity effects on leaf morphology, cell structure, carbon metabolism and mRNA expression of dwarf wheat