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Abstract
Alfalfa (Medicago sativa L.) genotypes transformed with Lc-bHLH and Lc transcription genes were developed with the intention of stimulating proanthocyanidin synthesis in the aerial parts of the plant. To our knowledge, there are no studies on the effect of single-gene and two-gene transformation on chemical functional groups and molecular structure changes in these plants. The objective of this study was to use advanced molecular spectroscopy with multivariate chemometrics to determine chemical functional group intensity and molecular structure changes in alfalfa plants when co-expressing Lc-bHLH and C1-MYB transcriptive flavanoid regulatory genes in comparison with non-transgenic (NT) and AC Grazeland (ACGL) genotypes. The results showed that compared to NT genotype, the presence of double genes (Lc and C1) increased ratios of both the area and peak height of protein structural Amide I/II and the height ratio of α-helix to β-sheet. In carbohydrate-related spectral analysis, the double gene-transformed alfalfa genotypes exhibited lower peak heights at 1370, 1240, 1153, and 1020 cm−1 compared to the NT genotype. Furthermore, the effect of double gene transformation on carbohydrate molecular structure was clearly revealed in the principal component analysis of the spectra. In conclusion, single or double transformation of Lc and C1 genes resulted in changing functional groups and molecular structure related to proteins and carbohydrates compared to the NT alfalfa genotype. The current study provided molecular structural information on the transgenic alfalfa plants and provided an insight into the impact of transgenes on protein and carbohydrate properties and their molecular structure’s changes.
Highlights
The alfalfa plant is the “queen” of foraging due to its high nutritive value, its high level of protein
How can we solve these issues? It has been found that the soluble protein in alfalfa plants can bind with proanthocyanidin (PA) to form a complex
There was a significantly higher Amide-I/II ratio in double gene alfalfa compared to NT alfalfa
Summary
The alfalfa plant is the “queen” of foraging due to its high nutritive value, its high level of protein. Extremely high soluble protein content in alfalfa plants causes major issues when ruminant livestock graze on alfalfa pasture, which include bloating causing animal death, N-to-energy unsynchronization, and nutrients being under-utilized [1,2]. It has been found that the soluble protein in alfalfa plants can bind with proanthocyanidin (PA) to form a complex. This complex can prevent protein from being degraded in the rumen but shifts protein from the rumen to small intestine. In this way, the three major issues can be solved.
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