skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Molecular Structural Changes in Alfalfa Detected by ATR-FTIR Spectroscopy in Response to Silencing of TT8 and HB12 Genes

Journal Article · · International Journal of Molecular Sciences (Online)
DOI:https://doi.org/10.3390/ijms19041046· OSTI ID:1628379
ORCiD logo [1];  [2];  [1];  [1];  [1];  [1]
  1. Univ. of Saskatchewan, Saskatoon, Canada. Dept. of Animal and Poultry Science, College of Agriculture and Bioresources
  2. London Research and Development Centre, Agriculture and Argi-Food Canada, London, ON, Canada
+ Show Author Affiliations

This study investigated the spectral changes in alfalfa molecular structures induced by silencing of Transparent Testa 8 (TT8) and Homeobox 12 (HB12) genes with univariate and multivariate analyses. TT8-silenced (TT8i), HB12-silenced (HB12i) and wild type (WT) alfalfa were grown in a greenhouse under normal conditions and were harvested at early-to-mid vegetative stage. Samples were free-dried and grounded through 0.02 mm sieve for spectra collections with attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy. Afterwards, both univariate and multivariate analyses were conducted on amide, carbohydrate and lipid regions. Univariate results showed that silencing of TT8 and HB12 genes affected peak heights of most total carbohydrate (TC) and structural carbohydrate (STC), and structural carbohydrate area (STCA) in carbohydrate regions; and β-sheet height, amide areas, and ratios of amide I/II and α-helix/β-sheet in amide region; and symmetric CH2 (SyCH2), asymmetric CH2 (AsCH2) and (a)symmetric CH2 and CH3 area (ASCCA) in the lipid region. Multivariate analysis showed that both hierarchy cluster analysis (HCA) and principal component analysis (PCA) clearly separated WT from transgenic plants in all carbohydrate regions and (a)symmetric CH2 and CH3 (ASCC) lipid region. In the amide region, PCA separated WT, TT8i and HB12i into different groups, while HCA clustered WT into a separate group. In conclusion, silencing of TT8 and HB12 affected intrinsic molecular structures of both amide and carbohydrate profiles in alfalfa, and multivariate analyses successfully distinguished gene-silenced alfalfa from its parental WT control.

Research Organization:
Brookhaven National Lab (BNL), Upton, NY (United States); Lawrence Berkeley National Lab (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
USDOE Office of Science (SC); Natural Sciences and Engineering Research Council of Canada (NSERC-Individual Discovery Grant and NSERC-CRD Grant); Saskatchewan Agriculture Strategic Research Program Fund; Agricultural Development Fund (ADF); SaskMilk; Saskatchewan Forage Network (SNK); Western Grain Research Foundation (WGRF); SaskPulse Growers; SaskCanola; POGA; China Scholarship Council (CSC)
Grant/Contract Number:
SC0012704; AC02-05CH11231
OSTI ID:
1628379
Journal Information:
International Journal of Molecular Sciences (Online), Vol. 19, Issue 4; ISSN 1422-0067
Publisher:
MDPICopyright Statement
Country of Publication:
United States
Language:
English

References (28)

The Use of Gene Modification and Advanced Molecular Structure Analyses towards Improving Alfalfa Forage journal January 2017
Analysis of ATR-FTIR Absorption-Reflection Data from 13 Polymeric Fabric Materials Using Chemometrics journal January 2015
Effects of heat processing methods on protein subfractions and protein degradation kinetics in dairy cattle in relation to protein molecular structure of barley grain using advanced molecular spectroscopy journal March 2018
Fourier Transform Infrared Spectroscopy (FTIR) and Multivariate Analysis for Identification of Different Vegetable Oils Used in Biodiesel Production journal March 2013
Measurement and characterization of external oil in the fried waxy maize starch granules using ATR-FTIR and XRD journal March 2018
Serum-based diagnostic prediction of oral submucous fibrosis using FTIR spectrometry journal January 2018
Predicting the cell-wall compositions of Pinus radiata (radiata pine) wood using ATR and transmission FTIR spectroscopies journal October 2017
Using Fourier transform IR spectroscopy to analyze biological materials journal July 2014
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 journal March 2017
NMR Characterization of C3H and HCT Down-Regulated Alfalfa Lignin journal October 2009
Co-downregulation of the hydroxycinnamoyl-CoA:shikimate hydroxycinnamoyl transferase and coumarate 3-hydroxylase significantly increases cellulose content in transgenic alfalfa (Medicago sativa L.) journal October 2015
Transformation with TT8 and HB12 RNAi Constructs in Model Forage ( Medicago sativa , Alfalfa) Affects Carbohydrate Structure and Metabolic Characteristics in Ruminant Livestock Systems journal October 2015
Gene-Silencing-Induced Changes in Carbohydrate Conformation in Relation to Bioenergy Value and Carbohydrate Subfractions in Modeled Plant (Medicago sativa) with Down-Regulation of HB12 and TT8 Transcription Factors journal May 2016
Structural Makeup, Biopolymer Conformation, and Biodegradation Characteristics of a Newly Developed Super Genotype of Oats (CDC SO-I versus Conventional Varieties): A Novel Approach journal February 2010
Characterizing the molecular structure features of newly developed hulless barley cultivars with altered carbohydrate traits (Hordeum vulgare L.) by globar-sourced infrared spectroscopy in relation to nutrient utilization and availability journal July 2014
Investigating Molecular Structures of Bio-Fuel and Bio-Oil Seeds as Predictors To Estimate Protein Bioavailability for Ruminants by Advanced Nondestructive Vibrational Molecular Spectroscopy journal October 2017
Physiochemical Characteristics and Molecular Structures for Digestible Carbohydrates of Silages journal October 2017
Molecular basis of protein structure in proanthocyanidin and anthocyanin-enhanced Lc-transgenic alfalfa in relation to nutritive value using synchrotron-radiation FTIR microspectroscopy: A novel approach journal September 2009
Carbohydrate and lipid spectroscopic molecular structures of different alfalfa hay and their relationship with nutrient availability in ruminants journal November 2017
Microwave Irradiation Induced Changes in Protein Molecular Structures of Barley Grains: Relationship to Changes in Protein Chemical Profile, Protein Subfractions, and Digestion in Dairy Cows journal July 2014
Relationship of feeds protein structural makeup in common Prairie feeds with protein solubility, in situ ruminal degradation and intestinal digestibility journal August 2014
Impact of ethanol bioprocessing on association of protein structures at a molecular level to protein nutrient utilization and availability of different co-products from cereal grains as energy feedstocks journal October 2014
Univariate and multivariate molecular spectral analyses of lipid related molecular structural components in relation to nutrient profile in feed and food mixtures journal February 2013
Detect changes in lipid-related structure of brown- and yellow-seeded Brassica Carinata seed during rumen fermentation in relation to basic chemical profile using ATR-FT/IR molecular spectroscopy with chemometrics journal December 2014
Mid-Infrared Spectral Characteristics of Lipid Molecular Structures in Brassica carinata Seeds: Relationship to Oil Content, Fatty Acid and Glucosinolate Profiles, Polyphenols, and Condensed Tannins journal July 2014
MicroRNA156 as a promising tool for alfalfa improvement journal December 2014
Protein secondary structures (α-helix and β-sheet) at a cellular level and protein fractions in relation to rumen degradation behaviours of protein: a new approach journal November 2005
Alfalfa-most important perennial forage legume in animal husbandry journal January 2009

Cited By (2)

Effects of silencing TT8 and HB12 on in vitro nutrients degradation and VFA production in relation to molecular structures of alfalfa ( Medicago sativa ) journal September 2019
Physicochemical Properties of African Catfish Mucus and Its Effect on the Stability of Soya Milk Emulsions journal January 2020

Similar Records

Molecular Basis of Protein Structure in Proanthocyanidin and Anthocyanin-Enhanced Lc-transgenic Alfalfa in Relation to Nutritive Value Using Synchrotron-Radiation FTIR Microspectroscopy: A Novel Approach
Journal Article · Thu Jan 01 00:00:00 EST 2009 · Spectrochimica Acta, Part A · OSTI ID:1628379
Resolution of coi-dominant phytoplankton species in a eutrophiclake using synchrotron-based Fourier transform infraredspectroscopy
Journal Article · Mon Oct 09 00:00:00 EDT 2006 · Phycologia · OSTI ID:1628379
Principal Component Analysis-Based Pattern Analysis of Dose-Volume Histograms and Influence on Rectal Toxicity
Journal Article · Sat Sep 01 00:00:00 EDT 2007 · International Journal of Radiation Oncology, Biology and Physics · OSTI ID:1628379

Related Subjects

Biochemistry & Molecular Biology
Chemistry