Winrhizo image analysis software

Manufactured by Regent Instruments

Sourced in Canada

WinRHIZO is an image analysis software designed to quantify and analyze root systems. It provides measurements of root length, surface area, volume, and other morphological parameters from digital images of root samples.

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Lab products found in correlation

Expression std 4800 scanner, by Epson (2 mentions) Perfection v850 pro scanner, by Epson (1 mentions)

11 protocols using winrhizo image analysis software

Root Decomposition in Forest Ecosystem

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Air-dried root branches of 1.0000 ± 0.0010 g of each order class were placed in litterbags (5 cm × 5 cm, mesh size 120 μm). Four plots (5 m × 5 m) were randomly set up in the original forest. For each root order, duplicate sets of litterbags were placed horizontally at a soil depth of 10 cm in each plot late in the growing season (mid-October) 2013. One litterbag of each branch order was randomly harvested from each plot on five occasions: early spring 2014 (mid-April), late autumn 2014 (mid-October), early spring 2015, late autumn 2015, and early spring 2016. In the laboratory, soil particles and other extraneous materials were removed. Cleaned roots were oven-dried at 65 °C to constant mass and weighed.
The root diameter of different branch orders was measured using WinRHIZO image analysis software (Regent Instruments, Quebec, QC, Canada). For each sampling date, specific litters of the same branch order were pooled for chemical analyses after the determination of the dry mass. The concentrations of total carbon (C) in the root samples were determined using the dichromate oxidation-ferrous sulfate titration method and the total nitrogen (N) and phosphorus (P) were measured using the methods of Kjeldahl and phosphomolybdenum yellow spectrophotometry after digestion with hydrogen peroxide and sulfuric acid, respectively.

Li J., You C., Zhang L., Li H., Tan B., Liu Y., Wang L., Liu S., & Li Z. (2021). Do root modules still exist after they die?. Forest Ecosystems, 8(1).

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Liquorice Root Morphometric Analysis

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The roots of cultivated liquorice were scanned with an Epson Expression/STD 4800 scanner (Seiko Epson Corporation, Nagano, Japan), and WinRHIZO image analysis software (Regent Instruments Inc., Quebec, QC, Canada) was used to derive the root length, surface area, volume, average diameter, the numbers of tips and forks, and the distribution of root lengths.

Chen M., Yang G., Sheng Y., Li P., Qiu H., Zhou X., Huang L, & Chao Z. (2017). Glomus mosseae Inoculation Improves the Root System Architecture, Photosynthetic Efficiency and Flavonoids Accumulation of Liquorice under Nutrient Stress. Frontiers in Plant Science, 8, 931.

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Root Morphological Analysis

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Before the root biomass assessment, different root orders were positioned to minimize overlap and placed on a atbed scanner. Root morphological parameters, including total root length (TRL), total root surface area (TRSA), average length of individual root (ALIR), and average diameter of individual root (ADIR) for different orders, were measured using WinRHIZO image analysis software (Regent Instruments, Quebec, QC, Canada). Then, the speci c root area (SRA) was calculated by dividing the root surface area by the dry weight of the roots used for scanning.

Chen L., Mi J., Hao L., He F., Yang H., Wan X., Zhang F., & Lin T. (2022). Effects of simulated nitrogen deposition on the ecophysiological responses of Populus beijingensis and P . cathayana under intra- and interspecific competition.

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Measuring Plant Growth and Morphology

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On the 30th day after carrying out the Mn treatment, 10 individuals were randomly selected in each treatment. A vernier caliper with an accuracy of 0.1 mm was used to measure the plant height and taproot length, respectively. Then, the plants were washed with deionized water. After the plants were dried, the root morphology was measured with an Epson scanner (expression 11,000 xl, Japan). The root surface area, root volume, root diameter, and fibrous root number were scanned and analyzed by WinRHIZO image analysis software (2013e, Regent Instruments Inc., Canada). The leaf area was measured with the same method. Finally, the plant individuals were put in the oven, scalded at 95°C for 10 min, and baked at 65°C to constant weight, and the dry weight was weighed. The specific leaf area (SLA, the ratio of leaf area and leaf dry weight) was also calculated.

Kuang X., Wang W., Hu J., Liu W, & Zeng W. (2022). Subcellular distribution and chemical forms of manganese in Daucus carota in relation to its tolerance. Frontiers in Plant Science, 13, 947882.

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Root Decomposition Dynamics across Orders

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Air-dried root branches of 1.0000 ± 0.0010 g of each order class were placed in litterbags (5 cm⋅5 cm, mesh size 120 µm). Four plots (5 m ⋅ 5 m) were randomly set up in the original forest. For each root order, duplicate sets of litterbags were placed horizontally at a soil depth of 10 cm in each plot late in the growing season (mid October) 2013. One litterbag of each branch order were randomly harvested from each plot on ve occasions: early spring 2014 (mid April), late autumn 2014 (mid October), early spring 2015, late autumn 2015, and early spring 2016. In the laboratory, soil particles and other extraneous materials were removed. Cleaned roots were oven-dried at 65 °C to constant mass and weighed.
Root diameter of different branch orders was measured using WinRHIZO image analysis software (Regent instruments, Quebec, QC, Canada). For each sampling date, speci c litters of the same branch order were pooled for chemical analyses after determination of the dry mass. The concentrations of total carbon (C) in the root samples was determined using the dichromate oxidation-ferrous sulfate titration method and the total nitrogen (N) and phosphorus (P) were measured using the methods of Kjeldahl and phosphomolybdenum yellow spectrophotometry after digestion with hydrogen peroxide and sulfuric acid, respectively.

Li J., You C., Zhang L., Li H., Tan B., Liu Y., Wang L., Liu S., & Li Z. (2021). Do Root Modules Still Exist after they Die?.

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Quantifying Root Distributions in Soil Columns

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Root distribution was measured in soil columns at 82 and 102 DAE. Each sector (tube) were carefully dug and cut down into 40 cm segments at the top of each column. The segments were immersed in water for 1 h and the roots from each soil layer were rinsed with tap water. Plant debris, weeds, and dead roots were sorted concurrently from ‘live’ roots by hand according to (Gwenzi et al., 2011 (link)). Live roots from each sector were evenly spread in a plastic tray containing deionized water and scanned using a flatbed scanner (300 dpi). Root images were analyzed using WinRhizo image analysis software (Regent Instruments, Quebec, Canada). The software was configured to measure RLD and RSD. After scanning, the roots were oven-dried at 80°C for 48 h and root dry mass was weighed to calculate RMD; the RLD, the RSD and the RMD were expressed as cm, cm², and mg per unit volume (cm³) of soil, respectively.

Chen Z., Tao X., Khan A., Tan D.K, & Luo H. (2018). Biomass Accumulation, Photosynthetic Traits and Root Development of Cotton as Affected by Irrigation and Nitrogen-Fertilization. Frontiers in Plant Science, 9, 173.

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Root Morphology Analysis of S. miltiorrhiza

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Thirty days after pathogen inoculation, the roots of S. miltiorrhiza were scanned with an Epson Expression/STD 4800 scanner (Seiko Epson Corporation, Nagano, Japan), and the root length, root projArea, and root surfArea were derived with WinRHIZO image analysis software (Regent Instruments Inc., Quebec, QC, Canada).

Pu C., Ge Y., Yang G., Zheng H., Guan W., Chao Z., Shen Y., Liu S., Chen M, & Huang L. (2022). Arbuscular mycorrhizal fungi enhance disease resistance of Salvia miltiorrhiza to Fusarium wilt. Frontiers in Plant Science, 13, 975558.

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Quantifying Fine Root Morphology

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For fine roots samples, the volume of each soil sample square was divided into two uniform ones according to the depth of every 10 cm. Therefore, for two altitudes and three provenances, a total of 60 soil blocks were obtained. The soil blocks were placed into plastic bags and then washed gently by running water through sieve nets to get them clean. Live roots were distinguished from dead roots according to the difference in root shape, elasticity and color (Freschet and Roumet, 2017 (link); Wang et al., 2019 (link)). All live fine roots were transported to the laboratory, stored at – 20°C, and then processed for morphological trait analysis.
The fine root samples were arranged on a transparent plate and scanned using an Epson Perfection V850 Pro scanner at a resolution of 400 dpi. Scanned images were analyzed for root length, root surface area, root projected area, and root volume using the WinRHIZO image analysis software (Regent Instruments Inc., Quebec, Canada). Fine root length density (FRLD, m m^{− 3)} and fine root surface area (FRSA, m² m^{− 3}) were calculated as root mass, root length, and root surface area per soil block volume, respectively. Specific root length (SRL, m g^-1) was calculated as root length per unit root dry mass. Fine root averaged diameter (FRAD, mm) was the ratio of the total projected area to the total root length.

Li D., Li G., Xi B., Gan J., Wen D., Cao F., Suo F., Li J., Ma B, & Guo B. (2023). Response mechanism of growth and gypenosides content for Gynostemma longipes cultivated at two altitude habitats to fine root morphological characteristics. Frontiers in Plant Science, 14, 1143745.

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Measurement of Effective Root Length

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The root distribution was measured in the soil columns at 39, 54, 69, 84 and 99 DAE. Three tubes per treatment were carefully dug from the ground and cut into 20 cm (2015) or 10 cm (2016) segments (beginning at the top of each column). The segments were immersed in water for 1 h. Then, the roots from each soil layer were placed in a 0.5 mm sieve and rinsed with running water. Simultaneously, debris, weeds, and dead roots were sorted from the living roots by hand during washing based on the procedure described by Gwenzi et al.^{49 (link)}. The living roots were placed in deionized water and stored in a refrigerator prior to analysis. Living roots from three of the columns were evenly spread in a plastic tray filled with deionized water and scanned using a flatbed scanner (300 dpi). The root images were analyzed using the WinRhizo image analysis software (Regent Instruments, Quebec, Canada), which was configured to measure the root length. After scanning, the roots were oven-dried at 80 °C for 48 h and weighed. The effective root length was the length of the root with a diameter less than 0.5 mm, which represented the primary part with a role in nutrient absorption^{50 (link)}.

Chen Z., Ma H., Xia J., Hou F., Shi X., Hao X., Hafeez A., Han H, & Luo H. (2017). Optimal pre-plant irrigation and fertilization can improve biomass accumulation by maintaining the root and leaf productive capacity of cotton crop. Scientific Reports, 7, 17168.

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Detailed Root Growth Analysis

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Root growth and distribution within each soil columns was determined at 70 and 140 DAP. Six tubes from each treatment were carefully dug out at ground level on each sampling date. The soil columns were cut into 20 cm slices starting from the top of each column. The slices were dipped in water at approximately 60 min and rinsed with tap water. Roots were collected in a 0.5 mm sieve using a water jet. Debris, weeds, and dead roots were separated from ‘live’ roots by hand according to a method used by Gwenzi et al. (2011) (link). There were few dead roots and the live roots were stored in deionized water for further analysis.
Live roots from three of the columns were spread out on a plastic tray contained deionized water and scanned using a flatbed scanner (300 dpi). Root images were analyzed using WinRhizo image analysis software (Regent Instruments, Quebec, QC, Canada). The software was configured to measure root length and root volume. After scanning, the roots were oven-dried at 60°C for 48 h and weighed. The root vigor was measured using the triphenyltetrazolium chloride (TTC) method (Luo et al., 2014 (link)).

Zhang H., Khan A., Tan D.K, & Luo H. (2017). Rational Water and Nitrogen Management Improves Root Growth, Increases Yield and Maintains Water Use Efficiency of Cotton under Mulch Drip Irrigation. Frontiers in Plant Science, 8, 912.

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