RS was conducted weekly for examining plants starting 33 DAI and continued until 75 DAI. Raman spectra were taken from plants (top three fully expanded leaves in the areas between the leaf veins) with a handheld Resolve Agilent spectrometer equipped with 830 nm laser source. Over the course of the whole experiment, spectra were taken in the same location and at approximately the same time on the day. Locations for spectral acquisition were randomly taken across plant leaves; 2–3 spectra were taken per one plant leaf. In total, 50 spectra were collected per sample (Control, FOC and Drought groups) per day. The following experimental parameters were used for all collected spectra: 1 s acquisition time, 495 mW power, and baseline spectral subtraction by device software (Agilent, Santa Clara, CA, United States). Fifty spectra were collected from each group of plants at the given timepoint; no distinctions were made for individual plants within each group.
Resolve agilent spectrometer
Manufactured by Agilent Technologies
Sourced in United States
The Resolve Agilent spectrometer is a versatile analytical instrument designed for spectroscopic analysis. It provides high-resolution data for a wide range of applications. The core function of the Resolve Agilent spectrometer is to perform precise spectroscopic measurements and data collection.
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4 protocols using resolve agilent spectrometer
1
Raman Spectroscopy Analysis of Plant Responses
2
Raman and FT-IR Analysis of Extracts
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Raman spectra of the extracts were collected using a hand-held Resolve Agilent spectrometer equipped with 831 nm laser source. The following experimental parameters were used for all collected spectra: 1s acquisition time, 495 mW power. Spectral background was corrected using iterative polynomial smoothing method. FT-IR spectra were acquired on Perkin Elmer 100 spectrometer equipped with attenuated total reflectance (ATR) module. For each reported spectrum, 15 spectra were recorded with a resolution of 4 cm-1 in the range of 4000–560 cm cm-1. A background spectrum was acquired immediately before the measurement. Both IR and Rama n spectra shown in the manuscript are raw spectra without any smoothing or pre-processing.
3
Rice and Corn Spectral Analysis
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Spectra from intact seeds of 15 different rice varieties (Figure 4 ) were collected using a hand-held Resolve Agilent spectrometer (Agilent Technologies, Santa Clara, CA, USA) equipped with 830 nm laser. The following experimental parameters were used for all collected spectra: 1 s acquisition time, 1 accumulation, 495 mW power, surface scanning mode, and baseline spectral subtraction by device software. Spectra from the corn cob were acquired with the same settings as described for rice except that the instrument was in through-barrier mode, the offset distance was changed from 0–5 mm, and 10 accumulations were used instead of 1.
4
Raman Spectroscopy for Herbicide Resistance
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Raman spectra were determined with a handheld Resolve Agilent spectrometer (Agilent, Santa Clara, CA, United States) equipped with an 830-nm laser source. The spectra were collected with 1-s acquisition time and 495-mW power. Four spectra were collected from each leaf from four quadrants on the adaxial side of the leaf of each resistant and susceptible plant before herbicide treatment (D0) and after 1 day (D1) and 2 days (D2) of herbicide application. For each of the treatment groups (resistant and susceptible) at each time point, 30 spectra were collected, resulting in a total of 1584 spectra. Following this, the spectra were baselined using the handheld instrument software. Initially, spectra were collected from one herbicide-resistant (TX15-14-1) and one susceptible (TX15-13-2) population only for standardization of the instrument. Later, about 600 Raman spectra from leaves of three herbicide-resistant (TX15-14-1, TX15-10, and TX15-12-2) and four herbicide-susceptible (TX16-10, TX15-2, TX15-29, and TX-13-2) populations (Figure 2 ) were collected and normalized to the 1438-cm–1 band, representing CH2 and CH3 vibration (Table 1 ; Farber et al., 2019b (link)). This chemical group is present in nearly all biological molecules, which makes the normalization unbiased to any specific chemical component of the weed leaf.
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