B vitamin quantification: LR-93 (NAM-2) seed tissues using UPLC/MRM MS performed in 2022

Overview
Objectives

  • Characterize the B vitamin profile of the LR-93 recombinant imbred lines using UPLC/MRM MS.

  • Utilize existing genotypic datasets to perform QTL analysis and look for trait-associated genetic markers for use in the breeding program for genetic improvement of B vitamin levels in lentil.

Parent Research Study
B vitamin quantification in wild and cultivated lentil seed tissues using ultra-performance liquid chromatography-selected reaction monitoring mass spectrometry
Experiment Summary

LR-93 (NAM-2) recombinant imbred lines were evaluated to assess the variation on B vitamin concentration cross genotypes and seed tissue types (seed coat, cotyledon and whole seed). Seeds will be grown in 3 site-years during 2022 and 2023. Specifically, it will assess B1 (thiamine), B2 (riboflavin and riboflavin-5-monophosphate), B3 (nicotinamide and nicotinic acid), B5 (pantothenic acid), B6 (pyridoxine, pyridoxamine and pyridoxal), B7 (biotin), and B9 (folate). It will also map QTL associated with B vitamin concentrations to identify regions in the genome responsible for enhanced vitamins profiles.

Germplasm
Germplasm Genus
Lens
Germplasm Scientific Name
Lens culinaris
Germplasm Collection
LR-93 (NAM-2; CDC Redberry x 3339-3)
Phenomics
Key Measurements
  • Per B-vitamin: Concentration in Seed Coat (ug/g)
  • Per B-vitamin: Concentration in Cotyledon (ug/g)
  • Per B-vitamin: Concentration in Whole Seed (ug/g)
Experimental Design
Experimental Site Locations
Aberdeen, Saskatchewan, Canada
Timepoints
  • 2022
Biochemical Technique
ultra-performance liquid chromatography-selected reaction monitoring mass spectrometry (UPLC/MRM MS)
Experimental Design

  • Whole seeds for each genotype were air dried at 45°C until a moisture <13% was reached. Seeds from each plot were then divided into two groups 1) whole seeds, 2) cotyledons. Seeds from the second group were separated into cotyledons and seed coats using a Satake huller TM05C(2) and then seed coats were removed.

  • Seed samples were ground to a fine powder using an Udy grinder. Equal amounts powdered sample per plot for a single genotype were combined for each of 1) whole seeds, 2) cotyledons. These samples were stored at -80°C until analysis.

  • B vitamins were extracted from these samples and then measured using ultra-performance liquid chromatography-selected reaction monitoring mass spectrometry (UPLC-SRM MS) analysis with resulting values being expressed in micrograms (ug) of B vitamin per gram (g) dry weight of initial powdered sample.

Attribution
Researchers
Data Collector
Jeremy Marshall
Data Custodian
Data Curator
Ruobin Liu
Associated Datasets

  • LR-93 (NAM-2) B Vitamin Concentration Dataset (XLSX, TSV)

    This data consists of the concentration (ug B vitamin per gram dry weight of initial powdered sample) for seven B vitamins. Samples consist of 1 site-year, 98 recombinant inbred lines and either whole seed or cotyledons. The field plots per genotype were pooled into a single composite sample. The B vitamins were measured using ultra-performance liquid chromatography-selected reaction monitoring mass spectrometry (UPLC-SRM MS).

  • LR-93 (NAM-2) Field Phenology Dataset (XLSX, TSV)

    This data consists of phenology measurements for the plots eventually harvested for B vitamin quantification. Specifically, emergence, flowering time and maturity dates/days are recorded. Additionally, opportunistic root rot measurements were taken.

Funding
Research Organization
University of Saskatchewan
Funding Grant
EVOLVES: Enhancing the Value of Lentil Variation for Ecosystem Survival
  • Kirstin E Bett
  • Albert Vandenberg
University of Saskatchewan

2019-2023