Digital image analysis and chlorophyll metering for phenotyping the effects of nodulation in soybean

Leaf photosynthesis and rhizobial nitrogen fixation are the two metabolic processes of utmost importance to legume growth and development. As these processes are closely related to each other, measuring of leaf chlorophyll content can provide information on the nodulation and nitrogen fixation status of crop plants. In the present investigation, a number of soybean breeding lines consisting of near-isogenic families which are genetically segregating for the nodulation trait were utilized in field experiments carried out across three growing seasons at Vienna, Austria. For phenotyping leaf chlorophyll content, the Minolta SPAD spectrometer was applied in parallel to a simple leaf digital image analysis procedure based on a commercial digital still camera. The main objectives of the research included the comparison of SPAD metering and image analysis for determination of chlorophyll content, phenotyping of the soybean nodulation vs. non-nodulation characteristic with respect to leaf, agronomic and seed traits, and relating both chlorophyll and image analysis data to seed quality characteristics. Nodulating and non-nodulating soybean lines significantly differed in chlorophyll content from the V5 (five leaves fully developed) soybean developmental stage onwards. Apart from chlorophyll content, leaf size, plant height, number of pods per plant, 1000-seed weight, and seed protein and oil content were also affected by nodulation type. The chlorophyll content of soybean leaves as determined by SPAD metering was significantly correlated (r=−0.937) to the green color value (RGB color model) of leaf image analysis at the R3 (beginning of pod growth) soybean developmental stage. Both chlorophyll content and leaf image analysis parameters were correlated to 1000-seed weight, seed protein and seed oil content. Thus, it appears that these leaf parameters related to photosynthesis and nitrogen fixation could be utilized to determine the nitrogen status of a soybean crop and subsequently in forecasting seed quality parameters of the harvest product.