Chlorophyll content, net photosynthesis, transpiration rate, and leave angle of six shoots of each genotype were measured on the fifth to tenth terminal of mature leaves from the base of the shoot at 30 d after infestation. Chlorophyll content (ChlSPAD value) was estimated using a portable chlorophyll meter (CCM-200, Opti-Sciences, Tyngsboro, MA, USA). Light-saturated net photosynthesis was measured with a portable infrared gas analyzer (LI-6200, LI-COR, Lincoln, NE, USA). Transpiration rate was measured with a portable porometer (LI-1600, LI-COR, Lincoln, NE, USA). Leaf angle was measured using an Accupar LP-80 ceptometer (Decagon Devices, NE Hopkins Court, Pullman, WA, USA).
Ccm 200
Manufactured by Opti-Sciences
Sourced in United States
The CCM-200 is a compact and versatile chlorophyll content meter that provides a non-destructive way to measure the chlorophyll content in plant leaves. It utilizes dual-wavelength optical absorbance technology to determine the chlorophyll content index (CCI) of the leaf sample.
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Lab products found in correlation
Li 6200, by LI COR (2 mentions)
Accupar lp 80 ceptometer, by METER Group (2 mentions)
Fms 2, by Hansatech (2 mentions)
Chlorophyll content meter, by Opti-Sciences (2 mentions)
Licor 6200, by Opti-Sciences (2 mentions)
Handy pea, by Hansatech (2 mentions)
Li 1600, by LI COR (1 mentions)
Lci sd system, by ADC BioScientific (1 mentions)
Ap4 system, by Delta-T Devices (1 mentions)
Lcpro, by ADC BioScientific (1 mentions)
Flash 2000, by Thermo Fisher Scientific (1 mentions)
28 protocols using ccm 200
1
Leaf Physiological Responses to Infestation
2
Photosynthetic Capacity Evaluation in Plants
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Foliar chlorophyll content index [CCI; the ratio of % transmission 569 at 931 nm and % transmission at 653 nm (Parry et al., 2014 (link))] was measured in upper and lower canopy leaves in week 3. Measurements of CCI were taken from the center leaflet of the three youngest fully-expanded fan leaves and from three fan leaves at the bottom of each plant using a chlorophyll meter (CCM-200; Opti-Sciences, Hudson, NH, United States). The CCI measurements were averaged, for the upper and lower canopy leaves, respectively, on a per plant basis.
The ratio of variable to maximum fluorescence (Fv/Fm) emitted from photosystem II in dark-acclimated leaves exposed to a light-saturating pulse is an indicator of maximum quantum yield of photosystem II photochemistry (Murchie and Lawson, 2013 (link)). In week 5, during the first 8.5 h of the PAR photoperiod (i.e., before daily UV exposure), the middle leaflet of the youngest fully-expanded fan leaf from each plant was dark acclimated for 15 min and then Fv/Fm measurements were taken with a fluorometer (FluorPen FP 100; Drasov, Czech Republic).
The ratio of variable to maximum fluorescence (Fv/Fm) emitted from photosystem II in dark-acclimated leaves exposed to a light-saturating pulse is an indicator of maximum quantum yield of photosystem II photochemistry (Murchie and Lawson, 2013 (link)). In week 5, during the first 8.5 h of the PAR photoperiod (i.e., before daily UV exposure), the middle leaflet of the youngest fully-expanded fan leaf from each plant was dark acclimated for 15 min and then Fv/Fm measurements were taken with a fluorometer (FluorPen FP 100; Drasov, Czech Republic).
3
Evaluating Leaf Photosynthesis Capacity
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Eight shoots of each treatment were selected to measure leaf chlorophyll content and net photosynthesis. The measurement was taken on the fifth to tenth terminal mature leaves from the base of the shoot 20 d after infestation. A portable chlorophyll meter (CCM-200, Opti-Sciences, Tyngsboro, MA, USA) was used to measure leaf chlorophyll content. Light-saturated net photosynthesis was also measured with a portable infrared gas analyzer (LI-6200, LI-COR, Lincoln, NE, USA).
4
Measuring Leaf Physiology Traits
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Ten intact leaves were randomly selected from each treatment and control to measure the stomatal conductance (SC), and chlorophyll content index (CCI). For CCI, portable and non-destructive device (CCM-200; Opti-Sciences Inc., Hudson, NH, USA) was used. Fully expanded leaves were taken for chlorophyll content measurements.
Decagon leaf Porometer (Model SC-1) was used to measure the stomatal conductance. The leaves were placed in between the sensor head of the portable device. The stomatal conductance (mmol m−2 s−1) and temperature were displayed on the device screen.
Decagon leaf Porometer (Model SC-1) was used to measure the stomatal conductance. The leaves were placed in between the sensor head of the portable device. The stomatal conductance (mmol m−2 s−1) and temperature were displayed on the device screen.
5
Leaf Greenness Measurement using Chlorophyll Meter
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A portable chlorophyll meter CCM 200 (Opti-Sciences, Tyngsboro, MA, USA) was used to measure leaf greenness of the plants. Chlorophyll concentration was calculated using the formulae from Strain and Svec (1966) .
6
Physiological Measurements of Plant Responses
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Physiological measurements (chlorophyll fluorescence, chlorophyll content, and gas exchange) were performed on the fifth youngest expanded leaf of three plants per variety and treatment, after 8 weeks of treatment. Chlorophyll fluorescence was monitored using a fluorescence monitoring system (FMS II; Hansatech Instruments, Norfolk, United Kingdom) according to Aubert et al. [44 (link)]. The quantified parameters were photosystem II efficiency (ϕPSII) and nonphotochemical quenching (NPQ) [71 (link)]. The chlorophyll content index (CCI) was measured using a chlorophyllometer (Opti-Sciences, CCM-200), and the measurement was taken three times on the same leaf. Gas exchange was measured using an infrared gas analyzer (IRGA, ADC BioScientific LCI-SD system, Hoddesdon, United Kingdom). The temperature and relative humidity in the cuvette were set at 21 °C or 28 °C according to the growth chamber. The quantified parameters were instantaneous net photosynthesis rate (Ai), instantaneous net transpiration rate (E), and stomatal conductance (gs).
7
Portable Chlorophyll Content Indexing
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A portable, nondestructive, and lightweight instrument (CCM-200; Opti-Sciences Inc., Hudson, NH, USA) was used to estimate CCI for early and late rice, and it provided instantaneous, in situ information. The CCM-200 was adopted to take CCI values from the first fully expanded functional leaf on each plant. Total of 10 plants were measured randomly in each plot, and three CCI readings per leaf, including one reading around the midpoint of leaf blade and two readings three cm apart from midpoint. These values were averaged for the mean CCI reading of each leaf (Peng et al., 1993 (link)). CCI in our research was standardized with the following equation:
where x was the observed CCI values, max and min were the sample maximum and minimum of CCI in each growing stage.
where x was the observed CCI values, max and min were the sample maximum and minimum of CCI in each growing stage.
8
Measuring Plant Physiological Responses
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The 5th‐node leaves of 10 plants per treatment were measured at the beginning of the experiment and 2 weeks after inducing stress. The chlorophyll content index (CCI) was measured using a chlorophyllometer (Opti‐Sciences, CCM‐200), and three measurements were taken per leaf. An automatic porometer (AP4 System, Delta‐T Devices) was used to measure the stomatal conductance. Gas exchange was measured using an infrared gas analyzer (IRGA ADC BioScientific LCI‐SD system, serial No. 33413). The instantaneous water use efficiency (WUEi) was calculated as WUEi = Ai/Ei.
9
Measuring Plant Growth and Physiology
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Plant height was measured for each treatment at the sampling time. After ARA, the nodule number and fresh nodule weight were counted for each plant. Then, all the plant samples were dried at 60°C for 48 h to determine the dry weights of the aboveground part and the root. Chlorophyll content of the last but one ternate compound leaf was measured just before the plant was sampled by the portable chlorophyll detector (CCM-200, OPTI-Sciences, United States). All the data were obtained in triplicate for the subsequent statistic analysis.
10
Tracking Leaf Senescence and Root Dynamics
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Photographic documentation of leaves was collected along with chlorophyll measurements to better illustrate the relationship between the two parameters during the senescence process. Chlorophyll levels were measured several times during the growth season using a CCM-200 plus Chlorophyll Content Meter (Opti-Sciences). Changes in the morphology of fine roots were examined several times during the growth season. This was done by removing the rhizotrons from the chamber and taking photos of the root systems, and immediately returning them back into the chamber. The same 30 plants were analyzed each time.
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