An inexpensive, accurate method for measuring leaf area and defoliation through digital image analysis.

We report a protocol using a common desk-top scanner and public domain software for measuring existing leaf area and leaf area removed as a result of herbivory. We compared the accuracy and precision of this method to that of a standard leaf area meter. Both methods were used to measure metal disks of a known area, the area of soybean (Glycine max L.) leaves, and the area removed by simulating leaf feeding with a hole-punch. We varied the amount of injury across a low, medium, and high degree of simulated feeding. The mean area of 10 cm2 and 50 cm2 metal disks was more accurately estimated with the leaf area meter than the desk-top scanner. Leaf area estimates from both methods were highly correlated. The desk-top scanner accurately estimated the leaf area removed from the low, medium, or high degree of simulated leaf feeding. However, the leaf area meter overestimated low levels of simulated feeding injury. Though measuring a leaf's surface area with a desk-top scanner requires two steps (creating a digital image file and calculating the area represented by that image), the overall time required to measure leaf injury is shorter than with a leaf area meter. This relatively simple and inexpensive method of estimating leaf area and feeding damage has advantages in certain experimental situations where a prefeeding measurement of the leaf is impossible or undesirable, or when small amounts of feeding occur.