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New methods for sorghum transformation in temperate climates.

Sara Miller1,2, Asta Rønager1,2, Rose Holm1,2

  • 1Section for Plant Biochemistry, Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksbergs, Denmark.

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|July 3, 2023
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Summary

Researchers explored two genetic transformation methods for sorghum in temperate climates. While achieving low efficiency with agroinfiltration and callus induction via particle bombardment, further optimization is needed for reliable sorghum improvement in cooler regions.

Keywords:
Agrobacteriumdhurrinasesorghumtissue culturetransformationtransient

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Area of Science:

  • Agricultural Science
  • Plant Biotechnology
  • Genetics

Background:

  • Sorghum (Sorghum bicolor) is a valuable cereal crop with high drought tolerance, increasingly important in temperate regions.
  • Genetic transformation is crucial for crop improvement, but sorghum exhibits recalcitrance, particularly in cooler climates.
  • Existing transformation protocols are often optimized for warmer environments, limiting sorghum's genetic enhancement in temperate zones.

Purpose of the Study:

  • To evaluate the efficacy of two novel genetic transformation techniques for sorghum in temperate climates.
  • To optimize transient transformation using Agrobacterium tumefaciens-mediated agroinfiltration.
  • To assess stable transformation via gold particle bombardment using leaf whorls as explants.

Main Methods:

  • Transient transformation was performed using Agrobacterium tumefaciens-mediated agroinfiltration, with optimizations including dark incubation and high-density Agrobacterium cultures (OD600 = 2.0).
  • Stable transformation was attempted using gold particle bombardment on leaf whorl explants.
  • Green fluorescence protein (GFP) expression was used to assess transformation efficiency of the endogenous sorghum gene SbDHR2.

Main Results:

  • Transient transformation achieved low efficiency for GFP-tagged SbDHR2 gene expression, indicating limitations for gene localization studies.
  • Successful induction of callus and somatic embryos from leaf whorls was achieved using particle bombardment, but genetic transformation was not accomplished.
  • Both tested methods demonstrated potential but were significantly influenced by climatic conditions, requiring further refinement for routine application.

Conclusions:

  • Current Agrobacterium-mediated agroinfiltration and particle bombardment methods show limited success for sorghum genetic transformation in temperate climates.
  • Optimization of incubation conditions and explant types is necessary to overcome climatic influences and improve transformation efficiency.
  • Further research is essential to establish robust and reproducible sorghum transformation protocols for widespread application in cooler agricultural regions.