title: Non-destructive determination of floral staging in cereals using X-ray micro computed tomography (µCT).
abstract: BACKGROUND
  Accurate floral staging is required to aid research into pollen and flower development, in particular male development. Pollen development is highly sensitive to stress and is critical for crop yields. Research into male development under environmental change is important to help target increased yields. This is hindered in monocots as the flower develops internally in the pseudostem. Floral staging studies therefore typically rely on destructive analysis, such as removal from the plant, fixation, staining and sectioning. This time-consuming analysis therefore prevents follow up studies and analysis past the point of the floral staging.
  
  
  RESULTS
  This study focuses on using X-ray µCT scanning to allow quick and detailed non-destructive internal 3D phenotypic information to allow accurate staging ofArabidopsis thalianaL. and Barley (Hordeum vulgareL.) flowers. X-ray µCT has previously relied on fixation methods for above ground tissue, therefore two contrast agents (Lugol's iodine and Bismuth) were observed in Arabidopsis and Barleyin plantato circumvent this step. 3D models and 2D slices were generated from the X-ray µCT images providing insightful information normally only available through destructive time-consuming processes such as sectioning and microscopy. Barley growth and development was also monitored over three weeks by X-ray µCT to observe flower development in situ. By measuring spike size in the developing tillers accurate non-destructive staging at the flower and anther stages could be performed; this staging was confirmed using traditional destructive microscopic analysis.
  
  
  CONCLUSIONS
  The use of X-ray micro computed tomography (µCT) scanning of living plant tissue offers immense benefits for plant phenotyping, for successive developmental measurements and for accurate developmental timing for scientific measurements. Nevertheless, X-ray µCT remains underused in plant sciences, especially in above-ground organs, despite its unique potential in delivering detailed non-destructive internal 3D phenotypic information. This work represents a novel application of X-ray µCT that could enhance research undertaken in monocot species to enable effective non-destructive staging and developmental analysis for molecular genetic studies and to determine effects of stresses at particular growth stages.
authors.affiliation: School of Agriculture and Food Science, University College Dublin, Dublin, Ireland.|Agricultural and Environmental Sciences, University of Nottingham, Sutton Bonington Campus, Loughborough, LE12 5RD UK.
authors.forename: Saoirse R
authors.initials: SR
authors.lastname: Tracy
authors.affiliation: Plant and Crop Science, School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, LE12 5RD UK.
authors.forename: José Fernández
authors.initials: JF
authors.lastname: Gómez
authors.affiliation: Agricultural and Environmental Sciences, University of Nottingham, Sutton Bonington Campus, Loughborough, LE12 5RD UK.|Centre for Plant Integrative Biology (CPIB), University of Nottingham, Sutton Bonington Campus, Loughborough, LE12 5RD UK.
authors.forename: Craig J
authors.initials: CJ
authors.lastname: Sturrock
authors.affiliation: Plant and Crop Science, School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, LE12 5RD UK.|Centre for Plant Integrative Biology (CPIB), University of Nottingham, Sutton Bonington Campus, Loughborough, LE12 5RD UK.
authors.forename: Zoe A
authors.initials: ZA
authors.lastname: Wilson
authors.affiliation: Plant and Crop Science, School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, LE12 5RD UK.
authors.forename: Alison C
authors.initials: AC
authors.lastname: Ferguson
keywords: Arabidopsis thaliana; Barley; Floral staging; Flower development; Hordeum vulgare L.; Image analysis; X-ray computed tomography