Accelerating leaf area measurement using a volumetric approach

Plant Methods. 2022 May 9;18(1):61. doi: 10.1186/s13007-022-00896-w.

Abstract

Background: Despite the advances in the techniques of indirect estimation of leaf area, the destructive measurement approaches have still remained as the reference and the most accurate methods. However, even utilizing the modern sensors and applications usually requires the laborious and time-consuming practice of unfolding and analyzing the single leaves, separately. In the present study, a volumetric approach was tested to determine the pile leaf area based on the ratio of leaf volume divided by thickness. For this purpose, the suspension technique was used for volumetry, which is based on the simple practice and calculations of the Archimedes' principle.

Results: Wheat volumetric leaf area (VLA), had a high agreement and approximately 1:1 correlation with the conventionally measured optical leaf area (OLA). Exclusion of the midrib volume from calculations, did not affect the estimation error (NRMSE < 2.61%); however, improved the slope of the linear model by about 6%, and also reduced the bias between the methods. The error of sampling for determining mean leaf thickness of the pile, was also less than 2% throughout the season. Besides, a more practical and facilitated version of pile volumetry was tested using Specific Gravity Bench (SGB), which is currently available as a laboratory equipment. As an important observation, which was also expectable according to the leaf 3D expansion (i.e., in a given 2D plane), it was evidenced that the variations in the OLA exactly follows the pattern of the changes in the leaf volume. Accordingly, it was suggested that the relative leaf areas of various experimental treatments might be compared directly based on volume, and independently of leaf thickness. Furthermore, no considerable difference was observed among the OLAs measured using various image resolutions (NRMSE < 0.212%); which indicates that even the superfast scanners with low resolutions as 200 dpi may be used for a precision optical measurement of leaf area.

Conclusions: It is expected that utilizing the reliable and simple concept of volumetric leaf area, based on which the measurement time might be independent of sample size, facilitate the laborious practice of leaf area measurement; and consequently, improve the precision of field experiments.

Keywords: Image resolution; LAI; Optical leaf area; Phenotyping; Volumetric leaf area.