Net photosynthetic carbon dioxide assimilation rate (ACO2) and transpiration (E) were determined just before the two harvests whit a portable gas exchange analyzer (LCA-4; ADC BioScientific Ltd., Hoddesdon, UK) equipped with a broadleaf chamber (cuvette window area: 6.25 cm2). The physiological measurements were carried out between 11.00 and 13.00 h on nine replicate plants per treatment. The climatic conditions (e.g., the photosynthetically active radiation, relative humidity, and carbon dioxide concentrations) were set at ambient temperature and the flow air rate was 400 mL s−1. Intrinsic water use efficiency was calculated as the ratio between ACO2 and E.
Lca 4
Manufactured by ADC BioScientific
Sourced in United Kingdom
The LCA-4 is a laboratory equipment designed for conducting leaf area measurements. It provides accurate and precise measurements of leaf area for various plant species. The LCA-4 uses advanced optical sensors and algorithms to capture and analyze leaf samples, delivering reliable data for research and agricultural applications.
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13 protocols using lca 4
1
Photosynthesis and Transpiration Measurements
2
Vine Stem Water Potential and Gas Exchange
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During the season, beginning the day after the first application, the solar noon stem water potential (Ψstem) was measured fortnightly by pressure chamber following the procedure described in [36 (link)]. Measurements were taken at sun zenith on eight primary leaves per treatment, placed inside plastic bags and sampled from eight random vines. Single leaf gas exchange measures were taken in the morning hours (8:30–10:30) on eight primary leaves, in the same day and on same vines of the Ψstem measurements, using a portable infra-red gas analyzer (LCA4, ADC BioScientific Ltd., Herts, UK) featuring a broad leaf chamber (6.25 cm2). Eight primary leaves per treatment were measured among those inserted at nodes 4–6 above the distal bunch on a main shoot. Assimilation rate (An, μmol CO2 m-2 s-1) and stomatal conductance (gs, mol H2O m-2 s-1) were obtained by measurement of inlet and outlet CO2 and H2O relative concentration. Intrinsic water use efficiency, WUEi, was instead derived as the ratio between An and gs (and then expressed in μmol CO2 mol-1 H2O). Leaf temperature was obtained by the infrared thermometer (accuracy: ± 0.5°C at 25°C) of the same instrument, on the same eight primary leaves and at the same time of photosynthesis and PPFD measurements.
3
Measuring Leaf Gas Exchange and Water Use Efficiency
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A portable gas exchange analyzer (LCA-4; ADC BioScientific Ltd., UK) was used to measure the net CO2 assimilation rate (ACO2), stomatal resistance (rs) and transpiration rate (E) just before harvesting. Based on Carillo et al.’s [69 (link)] method, ACO2 was divided by E in order to calculate the Intrinsic Water Use Efficiency (WUEi). Fully expanded leaves were chosen to carry the measurements of the leaf gas exchange, and eighteen measurements were taken.
4
Photosynthetic Efficiency of Basil
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Before harvest, a portable gas exchange analyzer (LCA-4; ADC BioScientific Ltd., Hoddesdon, UK) was used to determine the net assimilation rate of CO2, the stomatal resistance and the transpiration rate of basil plants (ACO2, rs and E, respectively). Three physiological measurements were determined per replicate. For the maximum quantum use efficiency of the Photosystem II (Fv/Fm), measurements were performed with a portable fluorometer (Plant Stress Kit, Opti-Sciences, Hudson, NH, USA), where four measurements per replicate were performed.
5
Measuring Photosynthesis and Water Use Efficiency
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Measurements were carried out inside the growth chamber in the above-described environmental conditions. Net photosynthetic CO2 assimilation rate (ACO2), stomatal resistance (rs), and leaf transpiration rate (E) were determined before the harvest on fully expanded leaves (third leaf from the top) whit a portable gas exchange analyzer (LCA-4; ADC BioScientific Ltd., Hoddesdon, UK) fitted with a broad leaf chamber head (window area: 6.25 cm2). Photosynthetic photon flux density (PPFD), RH, and carbon dioxide (CO2) concentrations were set at ambient values (420 ± 15 μmol m−2 s−1, RH 66 ± 2% and 385 ± 5 ppm, respectively), and the flow rate of air was 400 ml s−1. The intrinsic water-use efficiency (WUEi) was computed as the ratio between ACO2 and E.
6
Tomato Leaf Physiology Measurements
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On June 6 (36 DAT), leaf water potential (Ψl) measurements were performed on three replicates per treatment, using a dew-point psychrometer (WP4; Decagon Devices, Pullman, WA). The Relative Water Content (RWC) of basal and apical tomato leaves was calculated following the formula described by Jones and Turner (1978 (link)) (RWC = [FW−DW]/[TW−DW] × 100); where FW, DW and TW corresponded to fresh, dry and turgid weight, respectively.
At 58 DAT, the net CO2 assimilation rate (ACO2), stomatal resistance (rs) and transpiration rate (E) were measured with a portable gas exchange analyzer (LCA-4; ADC BioScientific Ltd., Hoddesdon, UK) equipped with a broadleaf chamber (cuvette window area, 6.25 cm2). This measurement was carried out within 2 h across solar noon (i.e., between 11.00 and 13.00) on the youngest fully expanded leaves, using six replicates for each treatment. Photosynthetically active radiation (PAR), Relative humidity (RH) and CO2 concentration (593 ± 8 μmol m−2 s−1, RH 50 ± 0.6% and 377 ± 0.6 mg kg−1, respectively) were set at ambient value and the flow rate of air was 400 mL s−1. The Water Use Efficiency (WUE) was calculated as ACO2/E.
At 58 DAT, the net CO2 assimilation rate (ACO2), stomatal resistance (rs) and transpiration rate (E) were measured with a portable gas exchange analyzer (LCA-4; ADC BioScientific Ltd., Hoddesdon, UK) equipped with a broadleaf chamber (cuvette window area, 6.25 cm2). This measurement was carried out within 2 h across solar noon (i.e., between 11.00 and 13.00) on the youngest fully expanded leaves, using six replicates for each treatment. Photosynthetically active radiation (PAR), Relative humidity (RH) and CO2 concentration (593 ± 8 μmol m−2 s−1, RH 50 ± 0.6% and 377 ± 0.6 mg kg−1, respectively) were set at ambient value and the flow rate of air was 400 mL s−1. The Water Use Efficiency (WUE) was calculated as ACO2/E.
7
Photosynthetic Rate and Leaf Traits
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Determination of net rate of photosynthesis, stomatal conductance and transpiration was done at anthesis from the top of an expanded third leaf, utilizing a system that was open LCA-4 transportable infrared gas analyzer (ADC BioScientific Ltd., Hoddesdon, UK). The optimum time to record these observations was 09:00 a.m. to 12:00 p.m. with specifications as stated; leaf chamber volume gas flow rate (v) 391 mL min-1, surface area of leaf 12.25 cm2, ambient pressure (P) 98.89 kPa, leaf chamber molar gas flow rate (U) 249 μ mol S-1, leaf chamber’s temperature (Tch) varied from 40.2 to 44.6°C, grinder flow of air per unit area of leaf (Us) 232.16 mol m-2 S-1, PAR (Q leaf) at surface of leaf was highest up to 921 μ mol m-2.
8
Measuring Leaf Gas Exchange Parameters
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Stomatal conductance (Gs) and net assimilation of CO2 (ACO2) were measured using a portable photosynthesis system (model LCA-4, ADC Bioscientific Ltd., Hoddesdon, UK) and a PLC-4N leaf chamber (11.35 cm2), configured to an open system. The third fully-expanded leaf was chosen for the analyses after the different treatments. Leaves were first equilibrated at a photon density flux of 400 μmol m−2 s−1 for at least 20 min. Gs and ACO2 were determined during the necessary seconds to stabilise the reading, once stabilised measurements were reordered every 10 s during 1 min. The measurements were made in the middle of the photoperiod in order to obtain the highest values. During all the measurements the cuvette temperature was 25 ± 2 °C, the CO2 concentration of the cuvette was 350 ± 5 ul l−1 and the leaf-to-air vapour pressure difference was 2.2 ± 0.3 kPa within the cuvette.
9
Measuring Physiological Responses in Lettuce
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In both experiments, 17 days after transplanting, the following physiological measurements were determined with a leaf gas exchange analyzer (model LCA-4, ADC BioScientific Ltd., Hoddesdon, UK) equipped with a 6.25 cm2 broadleaf chamber: net carbon dioxide assimilation rate (ACO2), stomatal resistance (rs), and transpiration (E). The leaf gas exchange parameters were quantified on 10 red and green pigmented lettuce plants per treatment by choosing one young fully expanded leaf per plant. Intrinsic water use efficiency (WUEi) was obtained as a ratio between net photosynthetic CO2 rate and transpiration. The physiological data in both experiments were analyzed by the two-way analysis of variance followed by post hoc analysis (Duncan multiple range; p < 0.05) or the independent t-test according to the level of fixed factors, namely cultivar (two levels) and NS concentration (three levels) in experiment 1 and cultivar (two levels) and light intensity (two levels) for experiment 2. Statistical analysis was carried out using the SPSS software version 21 (IBM).
10
Leaf Gas Exchange Measurements
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Leaf gas exchange measurements were conducted between 11:00 h and 13:00 h using a portable infrared gas analyzer (LCA-4; ADC BioScientific Ltd., Hoddesdon, UK) equipped with a 6.25 cm 2 leaf chamber, on nine replicate plants per treatment (three replicates per experimental unit). Net photosynthetic carbon dioxide assimilation rate (An) (µmol m -2 s -1 ) and transpiration (E) (mmol m -2 s -1 ) were determined just before each harvest on fully expanded intermediate (non-apical) leaves. The climatic conditions (e.g. the photosynthetically active radiation, relative humidity, and carbon dioxide concentrations) were set at ambient temperature and the flow air rate was 400 ml s -1 . The instantaneous Water Use Efficiency (WUEi) (µmol CO 2 assimilated per mmol of transpired water) was calculated as the ratio between An and E.
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