Quantification of leghaemoglobin content in pea nodules based on near infrared hyperspectral imaging spectroscopy and chemometrics

Abstract

Leghaemoglobin content in nodules is closely related to the amount of nitrogen fixed by the legume–rhizobium symbiosis. It is, therefore, commonly measured in order to assess the effect of growth-promoting parameters such as fertilisation on the symbiotic nitrogen fixation efficiency of legumes. The cyanmethaemoglobin method is a reference method in leghaemoglobin content quantification, but this method is time-consuming, requires accurate and careful technical operations and uses cyanide, a toxic reagent. As a quicker, simpler and non-destructive alternative, a method based on near infrared (NIR) hyperspectral imaging was tested to quantify leghaemoglobin in dried nodules. Two approaches were evaluated: (i) the partial least squares (PLS) approach was applied to the full spectrum acquired with the hyperspectral device and (ii) the potential of multispectral imaging was also tested through the preselection of the most relevant wavelengths and the building of a multiple linear regression model. The PLS approach was tested on mean spectra acquired from samples containing several nodules and acquired separately from individual nodules. Peas (Pisum sativum L.) were cultivated in a greenhouse. The nodules were harvested on four different dates in order to obtain variations in leghaemoglobin content. The leghaemoglobin content measured with the cyanmethaemoglobin method in fresh nodules ranged between 1.4 and 4.2 mg leghaemoglobin g–1 fresh nodule. A PLS regression model was calibrated on leghaemoglobin content measured with the reference method and mean NIR spectra of dried nodules acquired with a hyperspectral imaging device. On a validation dataset, the PLS model predicted the leghaemoglobin content in nodule samples well (R2 = 0.90, root mean square error of prediction = 0.26). The multispectral approach showed similar performance. Applied to individual nodules, the PLS model highlighted a wide variability of leghaemoglobin content in nodules harvested from the same plant. These results show that NIR hyperspectral imaging could be used as a rapid and safe method to quantify leghaemoglobin in pea nodules.

Influence of Pseudomonas and Biofertisol as Foliar Spray on Nodulation Attributes and Yield of Vegetable Pea under STCR Approach

“…The next group of was due to T3, T5 and T2 for the leghaemoglobin content of 1.545, 1.493 and 1.430 mg plant -1 representing 17.2, 13.3 and an 8.5% response. The finding was in support to that of Eylenbosch et al [33] that higher leghaemoglobin content was due to better root and nodule development. Dutta and Bandyopadhyay [34] reported that phosphorus and biofertilizer application increased significantly the nodulation, leghaemoglobin content, and nitrogenase activity.…”

Effect of Microbial Inoculants on Leghemoglobin Content in Nodules and Microbial Populations in Rhizosphere of Soybean in a Vertisols

“…The higher leghemoglobin contents in this treatments due to the better root and nodules development [9,10].…”

Plant Growth Promoting Rhizobacteria (PGPR) Akar Bambu, Kacang Hijau, dan Putri Malu untuk Meningkatkan Pertumbuhan Bintil Akar Kacang Hijau

“…Warna ini menandakan adanya leghemoglobin yang berfungsi untuk mengikat oksigen (Eylenbosch et al, 2018). Oksigen tersebut dibutuhkan oleh bakteri rhizobium untuk melakukan penambatan nitrogen yang berperan dalam proses pertumbuhan (Zulkarnain, 2014).…”