Macf-Ij, automated method for measuring color and leaf area through digital images [Macf-Ij, método automatizado para medir color y área foliar mediante imágenes digitales]
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Leaf color and area are important characteristics in plant physiology and genetic studies, but data collection is laborious and requires optical integrators and colorimeters. Nowadays, data collection could be automated through digital image analysis (DIA). Free software for DIA to measure leaf area or color is available, but user intervention is required to record dimensions or leaf color. This study shows the Macf-IJ routine for DIA to measure leaf area and color. Results from DIA of seven geometric figures were compared to physical measurements with a LI-COR 3000® Leaf Area Integrator. Additionally, leaf area was measured in 13 herbaceous plants, and color and relative chlorophyll content (RCC) was recorded for Rhynchosia minima (L.) DC with a SPAD 502®. DIA was performed with Digitizer 4.3.5® and Black Spot Leaf. Duration of each measurement was recorded for each case. The area measured with the integrator for any figure varied less than 8.96 %25 with Macf-IJ and less than 1.82 %25 with photographs. R2 between methods was 0.999 (p≤0.001). With exception of five grasses, leaf area measured with the integrator correlated well with the Macf-IJ (r > 0.989, p≤0.001); Macf-IJ, Black Spot Leaf and Digimizer® had an R2 greater than 0.99 (p≤0.001). Duration was not significantly different between methods, but Macf-IJ registered additional data such as length (cm), width (cm), circularity (0-1), and red (R), green (G), and Blue (B) colors. There was correlation between RCC and green (r=-0.946, p≤0.01), red (r=-0.941) and blue (r=-0.881) colors and between RGB obtained the Digimizer® and CRC (r=-0.939, -0.942 and -0.811for red, green and blue colors, respectively). Macf-IJ took 4.43 s to measure leaves, while RCC took 38.37 s (p≤0.001). Macf-IJ is a quick and precise method that measures area, dimensions and leaf color through scanned images and digital photography.
