After exudate collection, plant roots and shoots were separated, and shoots were dried at 60°C for 48 h, weighed, and ground for C and N analysis on a Vario EL elemental analyser (Elementar, Hanau, Germany). FW measurements of roots were taken before storage in 10% ethanol until root morphological analyses were conducted. Root systems were then scanned on an Epson flatbed scanner and analysed for morphological root traits using the winrhizo ®root analysis software (Regent Instruments Inc., Quebec, QC, Canada). After analysis, roots were dried at 60°C for 48 h, weighed, and ground for analysis of root C content and N content (RNC) on an Elementar Vario EL elemental analyser. SRL was calculated by dividing the total root length by the dry biomass (cm g−1). RTD was calculated by dividing the weight of the dry biomass by the root volume (g cm−3). RDMC was calculated as DW divided by FW.
Winrhizo root analysis software
Manufactured by Regent Instruments
Sourced in Canada
WinRHIZO is a software application developed by Regent Instruments for the analysis of root systems. The software provides automated tools for the measurement and quantification of various root characteristics, such as root length, root surface area, and root volume, from digitized root images.
Automatically generated - may contain errors
7 protocols using winrhizo root analysis software
1
Root Morphological Analysis Protocol
2
Comprehensive Plant Growth Analysis
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Growth parameters were measured in accordance with an earlier study (Wang et al., 2021 (link)). Shoot height and leaf dimensions were measured using a ruler. Leaf area was calculated in accordance with the following formula:
where L represented leaf length, W was leaf width, and 0.75 was a factor used for maize seedlings that consider leaf shape (Hussain et al., 2019 (link)).
Each whole plant was subdivided into shoots and roots. Roots were cleaned with tap water to remove adherent soil and then dried with absorbent paper. The fresh weight (FW) of the shoots and roots were individually measured using an analytical balance (UQINTIX65-1CN, Sartorius, Göttingen, Germany) and were then loaded into sample bags after 2 h of drying in an oven at 105°C. After that, the shoots and roots were dried to a constant weight at 80°C. The dry weight (DW) of each shoot and root was then measured using the analytical balance.
Roots were placed in an acrylic tray (400 × 300 mm) in a 1 cm depth of water. The length, volume, diameter, and surface area of the roots were measured through scanning using a flatbed scanner (1680, Epson, Nagano, Japan) and analysis with the WinRHIZO root analysis software (Pro 2007, Regent Instruments, Québec, Canada).
where L represented leaf length, W was leaf width, and 0.75 was a factor used for maize seedlings that consider leaf shape (Hussain et al., 2019 (link)).
Each whole plant was subdivided into shoots and roots. Roots were cleaned with tap water to remove adherent soil and then dried with absorbent paper. The fresh weight (FW) of the shoots and roots were individually measured using an analytical balance (UQINTIX65-1CN, Sartorius, Göttingen, Germany) and were then loaded into sample bags after 2 h of drying in an oven at 105°C. After that, the shoots and roots were dried to a constant weight at 80°C. The dry weight (DW) of each shoot and root was then measured using the analytical balance.
Roots were placed in an acrylic tray (400 × 300 mm) in a 1 cm depth of water. The length, volume, diameter, and surface area of the roots were measured through scanning using a flatbed scanner (1680, Epson, Nagano, Japan) and analysis with the WinRHIZO root analysis software (Pro 2007, Regent Instruments, Québec, Canada).
3
Seedling Root System Morphometrics
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Twelve representative seedlings of each treatment were selected and washed with distilled water, and the root system of each seedling was cut and scanned by a desktop scanner (Epson CORP, Suwa, Nagano, Japan). The images were analyzed with WinRHIZO root analysis software (Regent Instruments, Quebec, Canada) to obtain relevant indexes such as total root length, total root surface area, total root volume, and total root tip number.
4
Evaluating Rice Seedling Responses to NaCl Stress
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At the 1st, 3rd, 5th, 7th, and 9th under NaCl stress, rice seedlings of each treatment were washed with tap water, then rinsed with distilled water, blotting paper was used to absorb surface water, above- and below-ground parts were separated for the determination of morphological indicators, and a representative population of twenty seedlings was selected for each treatment. The plant height and stem diameter of each individual were measured with a ruler and vernier. The above-ground and below-ground fresh weights were measured using an electronic balance. The rice seedlings were subsequently dried at 105 °C for 30 min and dried at 80 °C to a constant weight, and the above-ground dry weight and root dry weight were determined.
Rice seedling root systems were scanned with root system scanner (Epson Perfection V800 Photo (Epson Indonesia Inc.)), and the images were analysed and calculated using WinRHIZO root analysis software (Regent Instruments, Quebec, Canada) to obtain the total root length.
Rice seedling root systems were scanned with root system scanner (Epson Perfection V800 Photo (Epson Indonesia Inc.)), and the images were analysed and calculated using WinRHIZO root analysis software (Regent Instruments, Quebec, Canada) to obtain the total root length.
5
Root Phenotypic Analysis under P Stress
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The phenotypes of each plant root in two P-level Hoagland nutrient solutions were investigated on the 60th day after planting. WinRHIZO root analysis software (Regent instruments Inc., Quebec, Canada) was used to detect the change in root phenotype under P deficiency stress, including the total root length (cm), the total root surface area (cm2), the total root volume (cm3) and the total root tip number.
6
Comprehensive Root Morphometric Analysis
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For analysis of root morphological characteristics, roots of four replicates were washed from soil using sieves (mesh size 0.5–1.0 mm) and fresh and dry biomass were recorded. Morphological characteristics were determined from fresh root samples, stored in 60% (v/v) ethanol. For analysis, root samples submerged in a water film on transparent Perspex trays, were separated with forceps and subsequently digitized using a flat-bed scanner (Epson Expression 1000 XL, Tokyo, Japan). Root length, average root diameter and the proportion of fine roots of the digitized samples were measured by applying the WinRHIZO root analysis software (Regent Instruments, Quebec, QC, Canada). Root hair length was recorded non-destructively along the root observation plane of the minirhizotrons by a video microscope (Stemi 200-c, Zeiss, Oberkochen, Germany). The digitized video images were analyzed using the AxioVision, software, Version 3.1.2.1 (Zeiss, Oberkochen, Germany).
7
Comprehensive Root Analysis in Rice
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At tillering, booting, heading, and full-ripening stages (noted as TR, BT, HD, and FR, respectively), rice plants were removed from 3 pots and rice roots were rinsed clean to determine root length and surface area through an WinRHIZO Root Analysis Software (Regent Instruments Inc., Quebec, Canada), followed by the determination of root dry weight. The remaining roots were used to measure root oxidation activity by measuring the oxidation of alpha-naphthylamine (α-NA) as described by Ramasamy et al. (1997) (link). The root tip (0–2 cm) samples were collected and immersed in liquid nitrogen and then stored at −80°C for subsequent analysis.
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