Gene-Transformation-Induced Changes in Chemical Functional Group Features and Molecular Structure Conformation in Alfalfa Plants Co-Expressing Lc-bHLH and C1-MYB Transcriptive Flavanoid Regulatory Genes: Effects of Single-Gene and Two-Gene Insertion.
Alfalfa(Medicago sativaL.) genotypes transformed withLc-bHLHandLctranscription 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 inalfalfaplants when co-expressingLc-bHLHandC1-MYBtranscriptive 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 (LcandC1) 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-transformedalfalfagenotypes exhibited lower peak heights at 1370, 1240, 1153, and 1020 cm-1compared 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 ofLcandC1genes resulted in changing functional groups and molecular structure related to proteins and carbohydrates compared to the NTalfalfagenotype. The current study provided molecular structural information on the transgenicalfalfaplants and provided an insight into the impact of transgenes on protein and carbohydrate properties and their molecular structure's changes.
Department of Animal and Poultry Science, College of Agriculture and Bioresources, University of Saskatchewan, Saskatoon, SK S7N5A8, Canada. rgv863@mail.usask.ca.
Ravindra G
RG
Heendeniya
Department of Animal and Poultry Science, College of Agriculture and Bioresources, University of Saskatchewan, Saskatoon, SK S7N5A8, Canada. peiqiang.yu@usask.ca.
Peiqiang
P
Yu
D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000737:chemistry; D016000:Cluster Analysis; D005419:Flavonoids / Q000737:chemistry / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D000455:Medicago sativa / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008968:Molecular Conformation; D016254:Mutagenesis, Insertional; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D013057:Spectrum Analysis; D013329:Structure-Activity Relationship; D014158:Transcription, Genetic
D002241:Carbohydrates; D005419:Flavonoids; D010940:Plant Proteins
alfalfa plant; function groups; gene transformation; molecular spectroscopy; molecular structure; nutrition and structure interaction