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Variation within laminae: Semi-automated methods for quantifying leaf venation using phenoVein.

Eastyn L Newsome1, Grace L Brock1, Jared Lutz1

  • 1Department of Biology Miami University 700 E High Street Oxford Ohio 45056 USA.

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Leaf venation patterns vary within plant laminae. Accelerated methods using phenoVein software quantify leaf venation quickly and affordably, enabling crop breeding research.

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Brassica rapaareole arealeaf venationphenoVeinvenation density, venation variation

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Area of Science:

  • Plant anatomy
  • Botanical research
  • Agricultural science

Background:

  • Leaf physiological processes can differ within laminae.
  • Leaf venation heterogeneity is rarely studied due to time-consuming quantification.
  • Understanding leaf venation is crucial for crop improvement.

Purpose of the Study:

  • To introduce accelerated protocols for quantifying leaf venation.
  • To assess variation in leaf venation among *Brassica rapa* crop types and subspecies.
  • To determine if leaf venation quantification can be made more efficient.

Main Methods:

  • Utilized formaldehyde, acetic acid, and ethanol (FAA)-fixed samples stored in ethanol.
  • Employed phenoVein software for venation pattern estimation without clearing or staining.
  • Developed an R script for data analysis and used linear mixed-effects models.

Main Results:

  • Staining and clearing are unnecessary for leaf venation estimation with phenoVein and stereomicroscope imaging.
  • Significant variation in leaf venation was observed across different locations within the laminae.
  • All venation estimates required some degree of manual adjustment.

Conclusions:

  • Accelerated protocols using phenoVein enable rapid and cost-effective leaf venation data acquisition.
  • Leaf venation is a potential target for crop breeding, especially if intralaminar variation links to physiological processes.
  • Further research is needed to explore the correlation between intralaminar venation variation and physiological processes.